PACIFIC ISLANDS TREE SEED CENTRE Manual of operating procedures for seed collection, documentation, treatment and storage
PACIFIC ISLANDS TREE SEED CENTRE Manual of operating procedures for seed collection, documentation, treatment and storage
Prepared by:
Elina Young Basil Gua Bronwyn Clarke John Doran Paul Macdonell
Version 1 March 2014
©Copyright Secretariat of the Pacific Community (SPC) 2014 All rights reserved. SPC authorises partial reproduction or translation of this material for scientific, educational or research purposes, provided that SPC and the source document are properly acknowledged. Permission to reproduce the document and/or translate for commercial purposes must be requested in writing (spc@spc.int). Original SPC artwork may not be altered or separately published without permission. Original text: English Secretariat of the Pacific Community Cataloguing-in-publication data Young, Elina Pacific Islands Tree Seed Center: manual of operating procedures for seed collection, documentation, treatment and storage / prepared by Bronwyn Clarke, John Doran, Paul Macdonell, Elina Young and Basil Gua 1. Seeds – Oceania. 2. Vegetables – Seeds – Oceania. 3. Trees – Seedlings – Oceania. 4. Seeds – Testing – Oceania. 5. Seeds – Storage – Oceania.
The and tree imp to e to c
Ge abl nism PIC of P
At t Agr sou PIC Am tree incl imp
I. Young, Elina. II. Gua, Basil, III. Clarke, Bronwyn. IV. Doran, John. V. Macdonell, Paul. VI. Title. VII. Secretariat of the Pacific Community. 581.995
F
AACR2
ISBN: 978-982-00-0716-1 Produced by: Land Resources Division Secretariat of the Pacific Community Private Mail Bag Suva Fiji Website: www.spc.int/lrd For more information please contact: lrdhelpdesk@spc.int
The lish Cen see pla
As coll Aus bot of t qua Its see
The the reg abl
Design and layout: SPC Land Resources Publication Unit, Nabua, Fiji
INO Dire Sec Suv i
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
Foreword
re
t The genetic resources in native and exotic trees represent a reserve of vast economic, social and environmental importance for Pacific Islanders. Every day, they use hundreds of different tree species for various purposes – for timber, food, fuel or medicine, for cultural purposes, for improving soil fertility or for environmental protection. Certain tree species that are well adapted to extreme conditions will be vital in supporting the resilience of Pacific Island production systems to climate change and severe climatic events such as cyclones and storm surge. Germplasm supply and exchange are essential for the conservation, management and sustainable use of forest genetic resources in Pacific Island countries and territories (PICTs). Mechanisms for the efficient, effective and safe exchange of priority germplasm within and between PICTs are critical to sustaining genetic diversity within the region and enhancing the livelihoods of PICT communities. At the regional meetings of Pacific Heads of Agriculture and Forestry Services and Ministers of Agriculture and Forestry in Samoa in 2008, the Strategy and Action Plan on Forest Genetic Resource Conservation and Management in the Pacific was adopted as the framework under which PICTs would develop and implement activities and policies for the development of their resources. A major recommendation of the Regional Strategy and Action Plan was to establish a regional tree seed centre to assist island countries to collect and share germplasm of priority species, including regionally important timber, fruit, nut tree and shrub species, and species of particular importance for atolls, for resistance to cyclones, and for coastal protection. The Secretariat of the Pacific Community, in collaboration with its development partners, established and began operating a regional tree seed centre (now called the Pacific Islands Tree Seed Centre or PITSC) in 2012 as mandated under the Regional Strategy and Action Plan. The tree seed centre acts as a regional focal point for coordination and implementation of priority germplasm collection, storage, distribution and research. As part of the further development of the centre, this manual of operating procedures for seed collection, documentation, treatment and storage was developed in collaboration with CSIRO’s Australian Tree Seed Centre. The manual documents the methodology and reasoning behind both field and laboratory practices used in the collection, processing, storage and management of tree seed collections. The procedures reflect the importance of the genetic and physiological quality of seed, which is a key factor in the success or failure of the establishment of any crop. Its main aim is to standardise procedures used by staff working at the PITSC and in national tree seed centres in SPC member countries and territories. The information in this manual is also likely to be of interest to a much wider audience across the region. It should thus contribute to improved outcomes for tree planting initiatives across the region and ensure that with effective conservation and utilisation, forest genetic diversity is a valuable tool in meeting the challenges of food and nutritional security and climate change.
INOKE RATUKALOU Director, Land Resources Division Secretariat of the Pacific Community Suva, Fiji PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
ii
Acknowledgments 1
This manual is adapted from the CSIRO document “Australian Tree Seed Centre Operations Manual, Gunn 2000”
2
Version 1 of this Manual was prepared while the authors were recipients of a traineeship at CSIRO’s Australian Tree Seed Centre (ATSC), Canberra. This opportunity was made possible by funding support from Australian Aid (AusAID), Australian Centre for International Agricultural Research (ACIAR), EU - Facilitating Agricultural Commodity Trade in Pacific (FACT) and Commonwealth Scientific Industrial Organisation (CSIRO). Special thanks to Bronwyn Clarke, Paul Macdonell, Debbie Solomon and John Doran of ATSC for providing training and assisting in preparation of this Manual.
3.1
3
3.2
3.3
3.4
3.5
3.6
3.7
3.8
3.9
3.1
3.1
3.1
3.1
3.1
iii
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
tre
eewas for dity O). ran
Table of Contents
Pages
1
Background.........................................................................................................1-0
2
Supply of Seed to PITSC......................................................................................2-0
3
Seed Collection in the Pacific Island Countries..............................................3-0
3.1
Planning Seed Collections...................................................................................3-1
3.2
Determining Species Locations..........................................................................3-2
3.3
Determining Flowering and Collection Times...................................................3-2
3.4
Seed Type.............................................................................................................3-4
3.5
Purpose for Collection and Required Quantity................................................3-5
3.6
Access to Customary Lands................................................................................3-5
3.7
Field Survey and Tree Marking............................................................................3-6
3.8
Breeding Systems..................................................................................................3-7 3.8.1 The Main Types of Mating Systems............................................................3-7 3.8.2 Types of Flowers...........................................................................................3-7 3.8.3 Types of Pollinators and Flowering Levels................................................3-7
3.9
The Concept of Provenance Collection...........................................................3-8 3.9.1 What is a Provenance or Population........................................................3-9 3.9.2 Minimum Requirements of a ProvenaNce Collection...........................3-9
3.10
Selection of Provenances.................................................................................3-11
3.11
Types of Collections............................................................................................3-13 3.11.1 Provenance Collections from Natural Forests....................................3-13 3.11.2 Collections from Plantations..................................................................3-14 3.11.3 Collections from Seed Orchards..........................................................3-14
3.12
Methods of Collection.......................................................................................3-15 3.12.1 Tree Climbing using Spurs and Harness...............................................3-15 3.12.2 Catapult..................................................................................................3-17 3.12.3 3.12.4 3.12.5 3.12.6 3.12.7 3.12.8
Throwing Rope.......................................................................................3-17 Pole Saws and Hooks.............................................................................3-18 Bow and Arrow.......................................................................................3-18 Other Climbing Aids..............................................................................3-19 Collections from Felled Trees................................................................3-19 Collecting off the Ground.....................................................................3-20
3.13
Harvesting............................................................................................................3-21
3.14
Bagging and Transportation.............................................................................3-22
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
iv
3.15
Recording Field Data........................................................................................3-24 3.15.1 Botanical specimens..............................................................................3-27
3.16
Appendices to Section 3..................................................................................3-28 3.16.1 Equipment Checklist for the field.........................................................3-29 3.16.2 Flowering and Fruiting Phenological Data Form................................3-32 3.16.3 Seed Collection Data Sheet.................................................................3-33 3.16.4 Seed Collection Data Key....................................................................3-34 3.16.5 PITSC Code of Practice for Seed Collecting......................................3-35 3.16.6 List of Priority Species/Genera for attention by PITSC at CePaCT....3-36 3.16.7 A Rough Guide to Flowering and Seed Collection Times of Some Important Commercial Species............................................................3-37
4
Seed Processing..................................................................................................4-0
4.1
Processing Methods.............................................................................................4-1 4.1.1 Drying............................................................................................................4-2 4.1.2 Shaking and Beating..................................................................................4-3 4.1.3 Sieving...........................................................................................................4-3 4.1.4 Soaking.........................................................................................................4-3 4.1.5 Fermenting...................................................................................................4-4 4.1.6 Maceration..................................................................................................4-5 4.1.7 Washing.......................................................................................................4-5 4.1.8 Flotation.......................................................................................................4-5 4.1.9 Winnowing...................................................................................................4-6
5
Seed Testing.........................................................................................................5-0
5.1
Sampling................................................................................................................5-1
5.2
Purity Analysis........................................................................................................5-3 5.2.1 Physical purity..............................................................................................5-3 5.2.2 Genetic purity..............................................................................................5-4
5.3
Seed Dormancy...................................................................................................5-4 5.3.1 Procedures to break seed-coat dormancy............................................5-5 5.3.1.1
Boiling and hot water treatments.....................................................5-6
5.3.1.2
Acid scarification................................................................................5-6
5.3.1.3
Scarification or cracking of the seedcoat.......................................5-7
5.3.1.4
Manual nicking....................................................................................5-7
5.3.2 Procedures to overcome embryo dormancy.........................................5-7 5.3.3 Procedures for removing inhibitory substances......................................5-8 5.4
Germination Testing............................................................................................5-8
5.5
5.6
5.7
5.8
5.9
5.1
6
6.1
6.2
5.4.1 Test conditions............................................................................................5-9 5.4.1.1 5.4.1.2 5.4.1.3
v
Germination containers and substrate............................................5-9 Controlling fungi..................................................................................5-9 Moisture................................................................................................5-10
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
6.3
5.5 5.6
Temperature......................................................................................5-10 Light....................................................................................................5-10
5.4.1.6
Germination counts..........................................................................5-10
5.4.2 Evaluation...................................................................................................5-11 5.4.3 Re-testing....................................................................................................5-12 5.4.4 Vigour test.................................................................................................5-12 Indirect Viability Tests.........................................................................................5-13 Moisture Content................................................................................................5-14 5.6.1 Oven method............................................................................................5-15 5.6.1.1
Low constant temperature oven method.....................................5-15
5.6.1.2
High constant temperature oven method....................................5-15
5.7
Authenticity Test.................................................................................................5-15
5.8
Laboratory Hygiene...........................................................................................5-16
5.9
Laboratory Safety...............................................................................................5-16 5.9.1 Fungicides on seed..................................................................................5-16 5.9.2 Fumigated seed.......................................................................................5-17 5.9.3 Use of vermiculite......................................................................................5-17 5.9.4 Pre-treatment of seed using acid..........................................................5-17 5.9.5 Burns from boiling water and ovens.......................................................5-17 5.9.6 Repetitive movements and confined movements sitting at lab benches..............................................................................................5-17
5.10
Appendices to Section 5..................................................................................5-18 5.10.1 Germination and Storage information for PITSC priority tree species...................................................................................................5-19 5.10.2 Germination Test Sheet........................................................................5-24
6
5.10.3 Tolerance Tables...................................................................................5-25 5.10.3.1 5.10.3.2
A. Maximum tolerated range between weighed replicates.....5-25 B Maximum tolerated ranges between replicates of 25 seeds..5-26
6
Storage.................................................................................................................6-0
6.1
Principles of Storage.............................................................................................6-3 6.1.1 Moisture Content........................................................................................6-3 6.1.2 Temperature................................................................................................6-4 6.1.3 Atmosphere................................................................................................6-4
6.2
Storage procedures at PITSC for orthodox seed.............................................6-4 6.2.1 Fumigation...................................................................................................6-4 6.2.2 Seed Storage...............................................................................................6-5 6.2.2.1 Temperature.........................................................................................6-5
7
0
5.4.1.4 5.4.1.5
6.3
6.2.2.2 Control of seed moisture and atmosphere......................................6-5 Storage Procedures at PITSC for Recalcitrant Seed........................................6-6
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
vi
vii
6.4
Maintaining Seed Identity in Storage.................................................................6-7
6.5
Appendices to Section 6....................................................................................6-8 6.5.1 Draft Seed Centre Flowchart.....................................................................6-8
7
Quarantine Procedures......................................................................................7-0
8
Documentation for Seed Exchange..................................................................8-0
8.1
The Process............................................................................................................8-1
8.2
Material Transfer Agreement..............................................................................8-1
8.3
Other Requirements for Seed Movement.........................................................8-1
8.4
Appendices to Section 8....................................................................................8-2 8.4.1 Draft Consignment Note...........................................................................8-2 8.4.2 Draft Explanation of Codes Used in Seed Consignments......................8-3 8.4.3 Seed Evaluation Form for Inclusion with PITSC Seed Shipments...........8-4 8.4.4 Draft Material Transfer Agreement (MTA)...............................................8-6
9
Glossary................................................................................................................9-1
10
Bibliography.......................................................................................................10-1
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
M do
0
1
PACIFIC ISLANDS TREE SEED CENTRE Manual of operating procedures for seed collection, documentation, treatment and storage
1. Background
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
1-0
1
Th cifi ap Sa Ex ou the (PI ter
Th wh imp suc pro tre lec
Th cen me tion to Th ticu
Th me eco the list es. ge shi pro util de sci ties PIT fun an
1 Background The updated Forest Genetic Resources (FGR) Action Plan 2007–2015 for the Pacific, formally endorsed by the Pacific Heads of Agriculture and Forestry Services and approved by Ministers of Agriculture and Forestry at their regional meeting in Apia, Samoa in September 2008, recognises the pivotal role that “Germplasm Supply and Exchange” has on all other priority Forest Genetic Resources R&D themes throughout Pacific Island countries and territories (PICTs) (SPC 2009). It recommended that the Secretariat of the Pacific Community (SPC) establish a regional tree seed centre (PITSC) within its Centre for Pacific Crops and Trees (CePaCT) in Suva, Fiji as a matter of priority. The key activities proposed for the are to assist PICTs to collect, safely store and share when appropriate, and genetically improve tree and shrub germplasm of high regional importance. This may include species of importance for timber, non-timber products such as fruit, nuts and essential oils, conservation of genetic diversity, soil and coastal protection, cultural significance, and carbon sequestration. It is also to serve as a centre of excellence providing technical support, training and advice on germplasm collection, storage and propagation techniques, as well as tree improvement strategies. These objectives imply that PITSC will not be regarded as a commercial seed storage centre but focused on research quantities of valuable tree seeds sourced from SPC member nations for research and safe storage and distribution. Many of the Pacific nations have established their own seed storage facilities but these facilities are mainly to meet the seed needs for replanting programmes and research within that country. This, however, does not solve or cater for the needs of the region as a whole, and particularly other small island member nations. The initial focus will be on the Priority Species List which has been determined by the member countries. The priority list contains both species that are natural to the island ecological systems and exotics (land races) that became part of their programmes in the respective islands. As additional species are identified or excluded from the priority list by the regional member nations, the list will be updated subject to available resources. PITSC will undertake research on the species it stores, mainly focused initially on germination and storage trials. However, as funding becomes available and partnerships develop, other research activities could include nursery trials, field establishment procedures, taxonomy, genetic variation studies, tree improvement programmes, and utilisation studies among other activities that are relevant to further knowledge and development of these species. It is envisaged that this information will be published in scientific journals and technical data sheets so it is available at all levels. These activities call for both qualified staff and financial resources to enable implementation. Once PITSC becomes fully established and well equipped with appropriate resources (staff, funds, etc.), the member nations will need to be supported with relevant level training and workshops to up-skill local staff.
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
1-1
This manual outlines the operating procedures that for PITSC will use to obtain, germination test and store seed. It sets out the guidelines for member nations to follow when planning, collecting, documenting, handling, cleaning and transporting seed for PITSC. Planning seed collections in the islands may vary in some ways given the scattered islands and the mode of transport commonly used; however, the principles and methods of collecting seeds will be the same. Some of the commonly used methods of collecting seeds that may be relevant in the island nations are described with procedures and instructions outlined in this manual. The importance of this manual is to guide those who are handling seeds for their research or planting programmes in the aspects of genetic and physiological quality of seeds, which have a major bearing on the success or failure of the establishment of the crop, whether it be at research stage or for commercial application (Gunn 2001).
M do
1-2
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
ain, to ing ven rinsed bed ual s in ing rch
PACIFIC ISLANDS TREE SEED CENTRE Manual of operating procedures for seed collection, documentation, treatment and storage
2. Supply of Seeds to PITSC
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
2-0
2
PIT its It is foll ing me wit ser of t
2 Supply of Seed to PITSC PITSC hopes to have the active participation of all member nations so that it may meet its goal to assist them to conserve and share high quality tree seeds of priority species. It is imperative that the country agencies responsible for carrying out seed collections follow common protocols and practices acceptable to international standards in collecting, processing and handling of seeds that are to be supplied to the Centre. Only seeds meeting these requirements will be received in the Centre for storage and exchange with participating Pacific Islands countries and territories. Stringent control will be observed to ensure that sharing is fair and equitable and will not endanger local species of the importing country.
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
2-1
M do
PACIFIC ISLANDS TREE SEED CENTRE Manual of operating procedures for seed collection, documentation, treatment and storage
3. Seed Collection in the Pacific Island Countries
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3-0
3
Th Ag tion as On gat trie inte
Se gat son are con dlin kno mu
3.
Se com Isla and sca qui ces pla con • • •
3 Seed collection in the Pacific Island Countries The Pacific Islands Tree Seed Centre will be mostly sourcing its seeds from Forestry or Agriculture seed centres in member countries, or organising collaborative seed collection expeditions with local agencies in specific instances. The list of species identified as priority species in each country will be the target species for collection and storage. Ongoing collections may be required where seed is required for continuing research to gather valuable data that can be useful for management of the species. All the countries will have some form of agreement with the centre to address issues concerning intellectual property (IP) rights and benefit sharing. Seed collection is not a new activity in the Pacific Islands. Many Pacific Island peoples gather and collect seeds for food, ceremonies, growing plants and various other reasons associated with their prevailing lifestyles. The traditional collection techniques are generally acceptable to gather seed for these purposes. However, in the context of conservation, domestication and development of the species, the selection and handling of seeds for these purposes will differ from traditional practices. The available knowledge, technology and acceptable practices in collection, processing and storage must be adequately adhered to for best results.
3.1 Planning Seed Collections Seed collection involves good planning and decision-making to achieve the best outcomes in terms of seed quantity and quality for the intended end use. Within the Pacific Islands, planning for seed collection requires a good knowledge of species occurrence and must consider the contingencies of undertaking collections on often numerous scattered islands. This will aid planning for the mode of transport and when it is required during the collection expedition, especially if the seeds require immediate processing back at base. Collections from seed stands and plantations may be easier to plan, but again transport may be a critical factor in seed processing. Some common considerations in planning seed collection in the Pacific are: • • •
the purpose of collection and if collecting bulk or individual tree seedlots the required quantity of seed and the number of individuals, or trees in the bulk determining the distribution of the species and where collection will occur
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3-1
• • • • • • • • • • • • • •
determining similar species within the area and knowing the differences obtaining access to customary lands/obtaining permits to collect if needed determining the seed type — orthodox or recalcitrant? determining peak flowering and collection season the size of the collection team field survey and tree marking collection method how to reduce the bulk of the material collected, if at all how to transport the seed collected where seed extraction and cleaning will occur whether the nursery is ready to sow recalcitrant seed, since it has minimal storage time what training is required for seed collecting unit staff whether sufficient funds are available to allow the objectives of the collection to be met having a list of high, medium and low priority alternative species in case seed crops are poor on target species.
3.2 Determining Species Locations To collect seeds from the natural forest, a good knowledge of the sites where the species of interest occurs is essential. Locating these sites during the planning of the seed collecting expeditions is vital. Ways to identify potential collection sites include: • • • • •
literature search of published information and internet search talking to whomever has done some work on the same species talking to the local village people who have traditionally used these species having an understanding of co-occurrence with other species and vegetation types field reconnaissance (if the species is new and information is scarce).
3.3 Determining Flowering and Collection Times A critical factor in planning seed collection is for the collecting trip to be scheduled to coincide with the peak maturation of the fruit/seed crops. Seeds that are collected at peak maturity are in most cases superior to seeds that were collected early (green) or too late. Generally, they are easier to clean, store for longer periods of time and take less effort to collect. When fleshy fruit is ripe it tends to change colour from a green to a yellow, orange, red or black colour, the flesh becomes softer and some fruits start to
3-2
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
fall rat ope ano
Sp ma at h lati ripe be can sho lec pro
Mo for see see this bas the col ofte and ten for be the how exp the ogy or
Th in t ser nor is i for es lim
ion
peeed
ion
d to at or ake n to t to
fall to the ground. For dry fruits they become firmer, change to a brownish colour, may rattle in pods, and the fruits may start to dehisce. A cut test, whereby the fruit is cut open and the endosperm is observed to be firm and white and not pale and milky, is another indication of maturity. Species that occur over large areas and variable environments tend to have different maturation times. As a generalisation, populations at lower altitudes ripen sooner than at higher altitudes, populations closer to the equator (lower latitude) ripen before higher latitude populations and populations that are exposed to the sun (north facing slopes) ripen sooner than southern facing slopes. Year-to-year differences in maturity may be affected by differences in temperature and rainfall patterns. These generalisations can be used to help plan the collection. Some generalisations might also be exploited should seed not be available at the chosen location. For example, if you were to collect on a north facing slope and all the seed had fallen to the ground, then look at more protected areas that may ripen at a later time than exposed sites. Monitoring the timing of peak flowering season/s, fruit development and seed maturity for different species is essential to determine the patterns of variation. Whether good seed crops are produced each year, how often and how long the species flowers, if the seed crop is retained on the tree for a period after maturity are all valuable to know. If this information is not known then the species must be monitored on at least a monthly basis, in multiple locations for at least 24 months. A form that can be used to gather these data is provided in Appendix 3.16.2. Closely monitoring the species will allow the collecting team some accuracy in timing their collection to capture the seed crop in the often limited time it is at its peak maturity. Where possible early and late seed crops, and seed crops resulting from a low flowering event should be avoided. These crops tend to develop from a few trees and are more likely to be inbred. If the collection is for plantation development the risk of collecting a genetically inferior seed crop should be borne in mind as this could influence the success of the crop and the economics of the whole venture. There is data available for some of the species on the priority list, however it is incomplete particularly for the rainforest species. This signals the need to expend more effort in carrying out data collection on flowering and fruiting seasons of the island tree species. One way is to observe and record flowering and fruiting phenology of any of the priority species encountered whilst undertaking any seed collections or other work using the form in Appendix 3.16.2. The precise timing of seed maturation to seed shed in different species varies greatly in the tropics. One classical example is the effect of altitude in the same area on Paraserianthes falcataria in the Solomon Islands, where individuals at high altitude don’t normally synchronise with those of the coastal areas when flowering. Such information is important to know so that close monitoring is carried out before committing resources for seed collection. This difference in flowering times may indicate separate provenances as pollen flow from coastal to high altitude provenances cannot occur or is severely limited (see Section 3.9 for a discussion)
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3-3
3.4 Seed Type
3.
There are three main categories relating to the handling and storage behaviour of seed — orthodox, intermediate and recalcitrant. Orthodox seed tolerates drying to below 5% moisture content and can be stored at low temperatures (<0°C). Recalcitrant seed is the opposite, tolerating no to very little drying and is sensitive to low temperatures (tropical >20°C, temperate >0°C). The intermediate seed category falls between these two seed types and mainly contains the species that don’t conform to the orthodox or recalcitrant categories (see Table 3.1 for a comparison). The seed category must be known before collection as it influences how the seed is treated post-harvest. Excess temperatures are to be avoided for all fruit that has a high moisture content, in particular recalcitrant seed, as well as for immature fruit and extracted orthodox seed. Recalcitrant seed requires oxygen to remain viable, and is packed loosely in containers with holes. The recalcitrant fruits respire and in this process consume oxygen and produce heat. Orthodox seed maintains viability best when the moisture content is reduced quickly and temperatures are kept low following extraction. For all seed types to maintain their highest viability, their moisture content, the temperature they are exposed to and access to oxygen (recalcitrant) have critical thresholds which need to be considered and controlled as best as possible in the field. (Refer to Section 3.14 Bagging and Transportation, Section 4 Seed Processing, and Section 6.1 Principles of Storage for more information.)
Se me ed res of t ma tion spe col jus veg com
Table 3.1. Generalisations of the differences between orthodox and recalcitrant seeds (from Thomsen 2000)
3-4
Orthodox seed
Recalcitrant seed
Desiccation tolerant
Desiccation sensitive
Tolerate low temperatures
Can be sensitive to temperatures below 15°C
Has low moisture content at shedding
Has relatively high moisture content at shedding
Includes dormant and non-dormant species
Usually no dormancy
Perennial, annual, woody and herbaceous species
Mostly perennial and woody species
Found in all ecosystems
Often found in humid ecosystems (if very desiccation sensitive)
Usually small seeds
Often large, fleshy seeds
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
Figu mor coll
If th kep asp end
3.
Ap fro ces uni app rela
eed ow eed res ese or
eed has ruit and his est ing ent, cal eld. and
3.5 Purpose for Collection and Required Quantity Seeds are normally collected for scientific studies, revegetation programmes or commercial planting in national or private reforestation programmes. Based on the intended use of the seeds, collections are either small quantities kept as individual trees for research, or bulked large quantities for revegetation. Whatever the intended end use of the seed, the same rules apply as outlined in Section 3.9.2 and Section 3.11.1. The main difference is that seed for revegetation will mainly be collected as a â&#x20AC;&#x153;bulk collectionâ&#x20AC;? where approximately equal quantities of seed from different trees of the same species and from one population are mixed together. It is very important that seed is collected from many trees (relative to population size) within the population, and not just a few trees. The objective of revegetation is generally to recreate a self-sustaining vegetation community that will prevail over many generations. If the seed used only comes from a few trees, the next generation will likely be highly inbred and fail. Numbering if a bulk collection
Numbering if an individual tree collection
Figure 3.1. Bulk seed consists of approximately equal quantities of seed from at least five trees (normally 10 or more), whereas individual tree collections keep the seed from all trees separate, and are labelled with a different collectorâ&#x20AC;&#x2122;s number.
If the seed collection is for research purposes, then typically the trees sampled will be kept separately during collecting, cleaning and storage. This involves more work in all aspects compared with collecting bulked seed. It is important, therefore, to know the end use of the seed for efficiency in the field.
3.6 Access to Customary Lands Apart from established seed stands in Forestry or private companies, seeds sourced from the natural forest normally come from customary land owned by people. To access customary land is at times difficult. Therefore, team leaders of the seed collection unit must be prepared to skilfully negotiate with people whom they have never met. One approach in the islands is to contact the village chiefs of the target community and build relationships by presenting awareness talks and meetings beforehand. The practice
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3-5
of promising payments should be discouraged where possible, as this often leads to suspicion and may result in unnecessary demands of expensive compensation. Where possible tokens of appreciation can be arranged (e.g. people may accept an offer to supply them with seedlings for planting). Where fixed access fees are offered, a clear explanation of the basis of their calculation should be given. Again, how this can be effectively approached will depend on each country and needs good communication skills of the team leader of the collection unit to professionally deliver with success. PITSC has a code of practice (Appendix 3.16.5) which outlines the approved actions of staff whilst undertaking seed collections. The main aim is to protect the environment from which the seed comes and also show the land owners that staff will act responsibly within the forest.
3.7 Field Survey and Tree Marking Areas are earmarked for collecting seeds, or the species selected may be new and this might require preliminary activities prior to the actual seed collecting event to ensure a successful collection. A survey of the target site to locate and verify existence of the species there must be done before seed maturity. If funding resources permit, marking of the desired trees, using the rules to overcome possible chance of selecting related trees, must be done at this time. Collection of seeds on trees encountered with good crops can be done simultaneously during the survey provided a good proportion of the population is seen to be fruiting before doing so. A field survey must be done if there are doubts about the extent of the species’ existence in the target area. When undertaking a pre-collection survey of the site the following factors should be observed (reasons given in brackets): • the presence of any similar species at site (to ensure only the target species is collected) • the health of the plants (a. so they can survive branch removal; b. so disease susceptible individuals are not spread to new plantings) • the presence of weed species (so precautions can be taken to avoid spreading them to new areas) • the number of target plants in population (should normally be large to better ensure genetic diversity) • how much seed is available (so there is enough for the objectives of the collection without disadvantaging the plant community) • the stage of maturity (ensuring the seed is at peak maturity so it handles and stores well) • whether there have been recent collections in the area (to ensure the collection activity does not put too much stress on trees to recover).
3.
Av (tru
Th inte
Via tur • • •
Wh eve (i.e len tha Ho to d win bee fro
Ma (he or the flow rod the
Wi dis the me be
Ta mo
3-6
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
to ere to ear be ion
ons ent nsi-
his ure the ing ted ood the ere be
pe-
3.8 Breeding Systems A very brief and simplified account of the main reproductive strategies of Angiosperms (true flowering plants) follows, as the Priority Species List comprises this plant type. This section explains the varying characteristics of species and how these differences interact within the environment effecting the genetic quality of the seed available.
3.8.1 THE MAIN TYPES OF MATING SYSTEMS
Viable seed develops when the pollen tube fertilises an ovule within the flower structure. There are three main systems used by plants: • • •
fertilisation only occurs when the pollen comes from a different plant (self-infertile) fertilisation only occurs when the pollen comes from itself (self-pollinating) fertilisation is a mixture of pollen from itself and different plants (mixed mating).
When fertilisation occurs between unrelated plants, it is said to be “outcrossed”. However, fertilisation can occur from a different plant which is closely related by descent (i.e. they have originated from the same mother tree, but have different fathers (pollen)), and is “partially inbred”. Natural population structure within a forest may mean that trees near each other are closely related as seed dispersal could be minimal. However, this is less pronounced with species that have fleshy fruits, as animals tend to disperse them over larger distances, compared with seeds that are dispersed by the wind and gravity. When seed dispersal distances are minimal it is likely that pollen has been provided by a close relative and it is “partially inbred”. When fertilisation occurs from the plants own pollen it is said to be “inbred”.
3.8.2 TYPES OF FLOWERS
et-
Many tree species have flowers that are bisexual with both male and female parts (hermaphroditic). Some species have unisexual flowers where they have either male or female parts. Unisexual species either have separate male and female flowers on the same plant (monoecious), or on different plants with only male or only female flowers (dioecious). Another category is where a plant has both male and hermaphroditic inflorescences (andromonoecious). Typically, the male inflorescences are on the outside of the plant while the hermaphroditic inflorescences are within the canopy.
the
les
Wind is a very efficient pollination mechanism and results in the pollen cloud being dispersed over large areas providing an excellent opportunity for gene flow throughout the population. Insects and animals are also effective pollinators; however, environmental variables can impact on their success. Therefore, their characteristics need to be understood to decide how to conduct a provenance collection.
dis-
oid
col-
3.8.3 TYPES OF POLLINATORS AND FLOWERING LEVELS
Table 3.2 lists different groups of pollinators and the likely range they would effectively move pollen in an area. Using the effective range as a guideline gives an indication
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3-7
of the minimum population structure for effective gene flow through the population. For example, if the target species was very scattered throughout the population with an average of one tree every 500 metres, and is pollinated by flies and bees which have an effective range of 200–300 metres, then there is likely to be very little pollen exchange between the trees. For another example, if the target species has many trees within the population, but flowering was very sparse at an average of one tree every 500 metres and the flies and bees are still the primary pollinators, then there is still likely to be little pollen exchange. In both of these examples the seed that subsequently develops on the few trees that flowered is likely to be highly inbred and not suitable for collection. As there is little flowering, the pollinators are attracted to other species or different areas, and if few trees are flowering, then they generally stay within the same tree and feed, leading to inbred seed.
con new
Sometimes one to a few trees will have heavy seed crops and almost all other trees very little to none. These heavy-crop trees should either be avoided in a collection or the proportion of seed collected from them controlled to be approximately equal to the other trees. When only a few trees flower heavily the pollinator visits to these flowers is very high compared with low flowering trees, and the pollen mainly comes from itself, therefore leading to inbred seed. So beware of the “loaded” tree.
When there is good flowering throughout the population the pollinators move from tree to tree and there are high visitation rates per tree. This promotes a high rate of gene flow throughout the population and seed produced from such an event is very suitable for a provenance collection. It is representative of the population and contains a high amount of genetic diversity. Seed that comes from one mother tree could have over 100 different fathers (pollen) represented (= high genetic diversity), whereas when flowering is low, the fathers represented will also be low (= low genetic diversity). Table 3.2. Types of pollinators, the potential distance they can move pollen, whether the group is involved with dispersal of the seed, and example species Pollinator Groups Wind Flies, Bees, Moths, Wasps etc.
Effective Range 100 + km <1km
Seed Dispersal Yes Very rarely
Birds Small mammals Bats
5–100 km 1–30 km > 100 km
Yes Yes Yes
Species Examples Casuarina Pterocarpus indicus, Canarium spp., Santalum spp., Endospermum spp. Terminalia catappa Terminalia catappa
If t sim con com
Th It is tion the
Fo the ma on. can big for fer • • • • •
Th Yo can are ano flec pro As tha
3.9 The Concept of Provenance Collection A group or population of plants that exchange genes through pollen and/or seed generally define a provenance. It is this common gene flow that keeps the differences within the group smaller than between groups (other populations). When a physical or biological barrier prevents gene flow, then a separate group of trees evolve to the prevailing
3-8
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
Un for tha me sho me of t
on. with ich len any ree e is senot her hin
conditions in the area they occupy. When these differences become significant, then a new provenance is formed.
ees or the s is elf,
ree ene ble igh ver hen
ved
m p.
erhin ogiing
If the site and environmental conditions at different locations for the one species are similar, then the gene structure in the populations may be similar. However, if they are contrasting then differences are likely to emerge after a few generations as different combinations of gene pairing will be successful in the varying environments. There can be large variation within a population, which is known as genetic diversity. It is this diversity that we use in tree breeding to select traits suitable for forest plantations. Therefore, seed collections for provenance research need to capture as much of the genetic diversity within the population as possible. 3.9.1 WHAT IS A PROVENANCE OR POPULATION?
For seed collection activities “provenance” should refer to the name of the place where the seed was collected. It can also be referred to as a “population”. The provenance may be labelled by the forest name, village name, province name, region name and so on. The main consideration is that enough details on location are recorded so the site can be identified (e.g. on maps). This is so that later the site can be identified unambiguously for follow-on collections or other work. This topic can be complex; however, for seed collections only one concept needs to be understood — plants respond differently to different environmental conditions. Therefore if: • • • • •
the soil type changes the altitudinal changes are great you change aspect (e.g. from the north side of a mountain range to the south) there is a change in vegetation type you move a substantial distance signalling a potential change in environment
Then, keep the seed separate between each change and call it a different provenance. You do this because there could be differences in the genetic makeup of the trees which cannot be seen in the forest. Previous studies have shown that the points listed above are cues to a change in the gene frequencies, which determines one provenance from another. Therefore, the provenances sampled and kept separate by location should reflect the observed changes throughout the range of the species. The “common-garden” provenance trials that follow will determine if the provenances are genetically different. As a seed collector you should concentrate on collecting seed from a target population that meets the genetic qualities detailed in Section 3.9.2.
3.9.2 MINIMUM REQUIREMENTS OF A PROVENANCE COLLECTION
Unless the objectives of the collection dictate otherwise, then the population chosen for sampling needs to meet certain minimum requirements. This gives the best chance that the seed collected is of high genetic quality and is suitable for plantation establishment, revegetation, and research. Populations that do not meet these requirements should be avoided, unless for research (range wide collection, outlier populations, fragmented populations), or a deliberate decision is taken to sample an inferior population of trees for some other reason. You should consider:
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3-9
• • • • • • • • •
the species range population size type of mating system (Section 3.8.1) type of pollinators (Section 3.8.3) cross pollination between plants (Section 3.8.3) distance of seed dispersal neighbourhood inbreeding changes in environmental and geographic boundaries (Section 3.9.1) individuals with an abnormally high proportion of seed.
As a general guideline the population should contain a minimum of 200 trees of the target species. At least 100 trees should have some seed, with at least 50 of these with medium to heavy seed crops. These numbers are arbitrary as different species have different requirements, but the population should ideally be large and the more trees with seed the better (see Section 3.8.3). Medium to heavy widespread seed crops indicate that the flowering was good and gene flow between trees is likely to be high. When the population is large (>200 trees) it generally contains high levels of genetic diversity. If the flowering level is high and widespread throughout the population then the seed will likely contain high levels of genetic diversity. If flowering was low and patchy then the seed will likely have low levels of genetic diversity and not adequately represent the population. Setting a minimum distance between seed trees aims to minimise the chance of collecting from trees that are related by recent descent, (i.e. they share the same mother). As a generalisation, seed dispersal by wind and gravity is mainly within the horizontal distance of the tree height. Therefore, nearby trees have a good chance of being related, which is known as “neighbourhood inbreeding”. Fleshy fruits are generally dispersed by birds and mammals. Whether the fruit is eaten from the tree or eaten from the forest floor they have an unknown and inconsistent dispersal distance from one metre to many kilometres. Regardless of the fruit type, for tall forest trees a distance between seed trees of at least twice the average height of the trees is used, generally 100 metres. This minimises the chance of collecting related trees, as some seed from fleshy fruits will remain below the mother tree, as well as most of the seed from gravity and wind dispersed species. In summary, the minimum requirements for a provenance collection are (and also see Section 3.11.1): • • • • • • • •
3-10
large inter-breeding population medium to heavy widespread seed crop collected trees 100 metres or more apart minimum of 10 trees to capture a good representation of the genetic variation in the population for large bulk collections aim to collect a modest amount of seed from many trees, rather than a lot of seed from a few trees similar amounts of seed from each tree so the seedlot is not biased towards particular trees only collect seed resulting from synchronous flowering events document the collection properly and securely (Section 3.15).
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3.
Th will ost pro dic As wit ava the occ ing the det
If a info of p mo larg 2–4 tria spe
Th gre los Re ma con in V mo und tion wh asp • • • • •
the with ave ees ops gh. etic hen and ely
coler). oning ally om one nce ally om vity nce
n
3.10 Selection of Provenances The species distribution, end use of seed, resources available and access to land will affect the selection of provenances for collection. In the Solomon Islands Xanthostemon melanoxylon can only be sourced from two sites, either in Boeboe, Choiseul province or San George, Isabel province. Therefore the species distribution largely dictates the selection of provenance. In contrast, Terminalia catappa occurs across Asia and the Pacific, from India in the west to Polynesia in the east, and the variation within the species is quite large. In this instance the end use of the seed and resources available will dictate what provenances are collected. If breeding for large edible nuts then collections should be from the Solomon Islands where large fruited provenances occur. However, the timber of this species is frequently used for construction of buildings and canoes where taller, straighter provenances should be selected. As well as the intended end use, the resources available to sample this wide ranging species will determine what countries/regions can be collected. If a breeding programme is going to be developed, the first step is to gather as much information about the species as possible from every conceivable source. The testing of provenances collected will normally be through provenance/progeny trials on two or more target sites. This becomes quite a logistics exercise for rainforest species with large fruit sizes. To test 100 families in a replicated trial of Terminalia catappa, then 2–4 kg of fruit per tree (depending on fruit size) totalling 200–400 kg, is required for one trial. Therefore, careful selection of provenances that span the wide distribution of the species is required. There are many approaches to selecting appropriate provenances and they will vary greatly with the objectives of the programme. For example, for Endospermum medullosum (whitewood) in Vanuatu, two trials were established under the South Pacific Region Initiative on Forest Genetic Resources (SPRIG) project on Espiritu Santo. The main aim of the project was to provide improved germplasm for growers and also conserve the genes of the few remaining natural trees that had been over-harvested in Vanuatu. The species occurs in Indonesia (West Papua or Irian Jaya), PNG, Solomon and Santa Cruz Islands, and more than 12 islands in Vanuatu. Collections were undertaken on six of Vanuatu’s islands ranging from 6 to 20 individuals per population, totalling 100 individual trees, plus another three trees from PNG Oro provenance which were removed soon after establishment. The researchers had to consider many aspects including: • • • • •
the project aim — to conserve Vanuatu genetic material (i.e. only represent Vanuatu in the trials) that there had been significant harvesting of this species with most assessable trees already harvested only small populations remained, potentially leading to inbred seed superior trees may have already been harvested only leaving inferior trees to interbreed and therefore to collect seed from likely variation across the species range (is seed from all islands required?)
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3-11
• • •
this is a dioecious species (separate male and female trees), therefore, are there enough trees of each sex within the population for good outcrossing? the stands needed to be accessible by boat, vehicle or foot financial and labour resources.
The following summing up of this topic is taken from Gunn (2001). Some species are found over a wide range of environments and cover extensive areas. For instance, in the Pacific Islands species like Terminalia catappa, Cordia subcordata, Calophyllum inophyllum and Barringtonia spp. occur across Asia and the Pacific Islands. Variation within these widely distributed species may sometimes be as great as the variation from between closely related species. Other species have a more limited distribution which; however, may sometimes consist of isolated provenances adapted to specific environmental conditions. Others again, like Xanthostemon melanoxylon, may occur naturally on very limited areas but still be genetically variable and adaptable to a variety of conditions when planted. For species in which comprehensive provenance trials have already been conducted, the published results are an important source of information when determining which provenances to focus on. Sampling provenances within species can be split according to two distinct requirements: 1. Sampling methods for species introduction trials 2. Wide-ranging sampling of many provenances to represent part or whole of the distribution for use in provenance trials For the first requirement, where there is little known about the species variation, several provenance collections should be made to include: • sampling from that part of the natural range where the species appears to be growing best
3.
Th (Se wit eve cal ens lati are see due see is l
Th the abi hea tre sho
Fo of p col pro hav larg
• part of the range that most closely matches the climate for which the seed is required
Th 200
• marginal sites within the natural range
Se the To fro twi use
For the second requirement the number of sources sampled will depend on the extent of the natural distribution, the diversity of the species, ease of access, seed availability, time available, money, staff resources, and other resources available to mount a collecting expedition. Knowledge of the breeding system of the target species and its pollen and seed dispersal mechanisms will assist in determining the collection strategy.
No tre am los pro in a ma alre 3-12
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3.11 Types of Collections
as. ata, Isas ted ted on, ble
ed, ich
re-
the
evbe
d is
ent bilta its gy.
3.11.1 PROVENANCE COLLECTIONS FROM NATURAL FORESTS
There are two types of collections â&#x20AC;&#x201D; bulk collections and individual tree collections (Section 3.5). The only difference is that a bulk collection represents multiple trees within the provenance and the individual tree collection has the seed from each and every tree kept separate. The procedure to collect seed from the provenance is identical for all other aspects. The following guidelines are the current best practice, which ensures adequate sampling of trees to represent the genetic variability in the population, and that the seed is suitable for research or revegetation. Initially survey the area of collection to determine the extent of the population and how widespread the seed crop is. During this time mentally select which trees you will collect from, giving due consideration to seed availability (quantity of seed per tree) and accessibility (can seed-bearing branches be reached with the equipment available), and if the population is large enough (also see Section 3.7). There should be at least 100 inter-breeding trees within the population to ensure that there has been good cross-fertilisation and the collection represents the genetic variability within the population. This can be seen by many of the trees having medium to heavy seed crops. If seed crops are light and patchy then it is assumed that not many trees were flowering and seed is more likely to be inbred. In this case another site should be found with better seed crops. For a new provenance, collect from a minimum of 10 well-spaced trees. In the case of proven provenances showing high levels of genetic diversity, it may be desirable to collect from up to 100 or more trees as part of a base population for intensive breeding programmes. Larger numbers of trees per locality are sampled after provenance trials have shown which provenances are best and where there is a requirement to obtain large quantities of seed. The following points provide the guidelines for selecting which trees to sample (Gunn 2001): Selections should aim to sample unrelated trees that cover the genetic variability of the population. In natural forests nearby trees are likely to share the same mother. To reduce the probability of sampling trees that are siblings, seed should be collected from trees which are at least seed-fall distance apart from each other; this means about twice the average height of the trees (Eldridge et al. 1993). One hundred metres is a useful rule of thumb for tall forest trees. Normally no particular attention is given to selecting against poor and average form trees in natural forests, as environmental and competition effects are unknown. For example, the trees at a site which is prone to cyclonic winds would appear deformed from losing branches in the strong winds; however, the seed from this population grown in a protected area may perform well. When there is a mix of poor and good formed trees in a population a decision to only collect from better than average formed trees can be made. While the performance of poor formed trees is unknown, good formed trees are already known. PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3-13
When collecting seed for revegetation purposes no selection is done as the aim is to collect as much genetic diversity within the population as possible. The only exception is to avoid trees showing signs of disease or stress. Selected trees must be carrying a medium to heavy mature seed crop. It is desirable to collect approximately equal quantities of seed from each tree. If the seed type is of orthodox behaviour, then quantities in addition to immediate demand can be collected for the seed store to maintain seed stocks and to meet future demands.
3.11.2 COLLECTIONS FROM PLANTATIONS
Seed collections from plantations should only be considered where appropriate information on the origin of the seed used to establish the plantation is available. The stand must contain an adequate genetic base in terms of the species, provenance and the number of unrelated parent trees (Eldridge et al. 1993). Collections would not normally be made from plantations that have been established from seedlots comprising fewer than 20 unrelated seed trees. The plantation should have been thinned at least once to remove inferior phenotypes, and then parent trees should be selected on growth and form to obtain a genetic gain compared with the natural stand/provenance. The seed collection data sheet is still used to record the collection details except that a clear reference under “Location” is made that the collection is from a planted stand together with the name of the original source (provenance).
3.11.3 COLLECTIONS FROM SEED ORCHARDS
Well-designed and managed seed orchards are a means of obtaining large quantities of highly genetically improved seed, and seed production areas for modestly improved seed. For recalcitrant species it may be necessary to establish seed production areas or seed orchards to have ready access to adequate supplies of seed for the planting programmes. Any species selected for this strategy will be in high demand and warrant an improvement programme to be initiated. The planning, design, layout and management of seed orchards is beyond the scope of this manual; however, for seed collection it is important to know an orchard’s history and to consider these points: • origin of the material used to establish the orchard (provenance and family origin, numbers of families, and whether it is a first-generation orchard using material collected from natural stands, or whether it is an advanced-generation orchard based on material collected from plantations or a breeding programme) • field layout (if family identity has been retained) • history of the orchard — considerations include: its objective — to confirm that it suits your present purpose; selection methods and effect on tree characteristics; extent of thinning and its effects on genetic diversity; how to avoid collecting from trees that have flowered out of phase with the others in the orchards (early or late flowering), as this seed may be highly inbred. It is highly recommended not to collect seed from orchards until at least 50% of individual trees (or clones, in the case of a clonal orchard), flower and set seed to produce the crop that is being collected
3-14
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3.
PIT ees exp me dep uns job are the
Tra pod and tise is g tre nes
Fo spu acc abo bot
Tra tra Ca sup
â&#x20AC;˘ it will generally be appropriate to maintain separate individual seedlots of the best trees in the orchard with individual tree identity retained â&#x20AC;˘ when recording the seed orchard details, provide information on reference documents describing the seed orchard, its physical location, whether it is a seed production area (SPA), seedling seed orchard (SSO) or clonal seed orchard (CSO), and the original genetic material (natural provenance source). Where possible provide a reference document describing the history of the seed orchard â&#x20AC;˘ seed orchard seed is generally more valuable than seed from natural provenances, so greater care is needed during harvesting, to avoid disrupting later crops
s to ion
ble s of ted
orand the ally wer e to and
hat and
ies ved eas ing ant
ope ory
mily ing ening
firm aroid the t is % of set
3.12 Methods of Collection PITSC promotes a safe working environment where the health and safety of employees and contractors is very important, especially those working in the forests that are exposed to additional safety issues. This section outlines the various seed collection methods that are known to be available in the island nations. The method used will depend upon the conditions at the collection site, (e.g. if it is raining or windy, it may be unsafe to climb). Here experience and common sense prevails. If it is not safe to do the job then reassess the method employed and maybe return on another day when there are more favourable conditions. Where possible, minimise any climbing and remain on the ground, as it is safer.
3.12.1 TREE CLIMBING USING SPURS AND HARNESS
Traditionally Pacific Islanders have climbed trees for food gathering (fruits, beans, pods, greens, hunting) and for some ceremonial rituals. These skills still remain intact and freeform tree climbing without the aid of modern climbing equipment is widely practised. Whilst PITSC respects these traditional methods, the risk of injury to the climber is greater than when using modern equipment. It promotes, therefore, the climbing of trees for seed collection by climbers with proper training and the use of at least a harness and rope. This method is described below. Forestry seed collection and other similar activities have seen the technique of using spurs and a safety harness introduced to the Pacific Islands. It is particularly useful for accessing the crowns and collecting seed from very tall trees, especially those that are above 20 metres. It has been recognised as feasible and practically appropriate for both research quantity and for commercial seed collections. Training in this method is to be from an experienced and accredited climber and the trainee must be secured with an advanced line (as described under Section 3.12.2 Catapult) and belayed (rope is secured by attaching to another person to offer stable support) while the required skills are learnt.
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3-15
Ca can sec (e. sec The ver the g) the tor pro effe
Figure 3.2 LHS â&#x20AC;&#x201D; Using single rope technique to ascend the tree. RHS â&#x20AC;&#x201D; Selection of climbing gear used for seed collection.
The climber wears a harness with two pole straps (one each side), spurs and safety equipment (helmet, gloves, long pants, sturdy boots) and has a climbing rope generally at least twice the length of the height to be climbed to descend on without having to use the spurs. One pole strap is placed around the bole of the tree and attached to the other side of the harness. The spurs are driven into the tree and the pole strap adjusted to support the climber on a 30 to 40 degree angle from the bole. The climber walks up the tree using the spurs and continually raising and adjusting the pole strap so they are adequately supported. When a branch is reached, the other pole strap is placed over that branch and around the back of the bole and attached to the opposite side of the harness. Once pressure is taken up, the first pole strap is released and the climber proceeds above that branch. When the crown is reach, a high point is set where the climbing rope is passed around the back of a solid branch in a crutch and attached to the central anchor point of the harness. A friction hitch (prussic knot) is attached to the other side of the rope to hold the climber in position. The climber can then move about the crown and remove branches/fruit. When finished they can descend on the rope or use the spurs. Figure 3.2 gives an example of equipment that might be required for climbing a tree bole using spurs. Spurs are best suited for trees with bark that is sufficiently deep and soft, but firm enough to enable the spur to penetrate and grip securely. Keep the spurs properly sharpened and tightened during use. Always have protectors over the spurs when walking on the ground or during transport. The main disadvantage of spurs is that they may damage the tree when the spur penetrates the bark. The spurs should be cleaned between different trees, or at least different areas, to minimise the spread of pathogens. Tree climbing should only be performed by trained personnel.
3-16
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
No mo sm me see
Figu repl in tw
Th see upo des tog tak dia be the
3.12.2 CATAPULT
Catapults are common in the Pacific Islands and this could be a method local people can simply adopt utilising locally available resources. Catapults are very effective in securing a light line over tree branches which can be used to transport a heavier line (e.g. a throwing rope — Section 3.12.3) higher than 12 metres within the crown, or to secure an advance line for climbing as has been done in Fiji for Agathis macrophylla. The free standing catapult “Big Shot” is considerably larger than the normal hand-held version and is mounted on a 2.4 metre pole. The pole is held upright with one hand and the other hand stretches the rubber sling holding a weight (throwing bag, 225 to 450 g) attached to a cord or fishing line on a reel, either on the ground or hand held facing the branch by a second person, as with a hand-held catapult (Figure 3.3). The operator lines up the target before letting go the sling. It is estimated that the weight can be propelled to a vertical height in excess of 25 metres and is arguably (Gunn 2001) more effective than a bow and arrow (Section 3.12.5).
for
ety ally to the ted up hey ced e of ber the d to the out or
ree and urs urs s is uld ead
Note that the use of a catapult and rope (or rifle) to break off seed bearing limbs is often more convenient and efficient than climbing tall trees, especially when only collecting small quantities of seeds. The “Big Shot”, with a light but strong rope, is the preferred method of collecting Endospermum medullosum seed in a seed orchard in Vanuatu.
Figure 3.3 LHS – Big shot catapult equipment showing the sling, replacement rubber, fishing line on reel, weight bags and the pole in two pieces. RHS – The big shot in action.
3.12.3 THROWING ROPE
This method is simply the throwing of a rope over seed bearing branches up to 12 metres, depending upon the users’ ability. The rope is thrown over the desired branch, and the ends of the rope brought together on the ground. The rope is pulled down to completely break off the branch, taking good care as the broken branch descends. To use this method a rope of suitable diameter (4–6 mm) and length of at least twice the height of the tree branch (12 m) must be available. Consider the strength of live-branches of the species being collected and the strength of the number of people in the collecting party before deciding on the size
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3-17
of the branch to target. Larger live branches of up to 100 mm can sometimes be easily pulled down. This method is not commonly practised in the Pacific though its application is simple and could be easily adopted to use on trees with lower hanging branches (if branches are higher, then a catapult should be used). Positioning the rope towards the end of the branch gives greater flex when trying to break it. Careful consideration of the size of the branch to minimise impact on the seed tree is required, and if repeat collections are required from that tree.
3.12.4 POLE SAWS AND HOOKS
Trees that have seed-bearing branches close to the ground can be easily harvested by standing on the ground and removing seeds from them. In many forest species in the tropics, this can be engineered in a managed seed orchard where trees are kept as low as possible by pollarding. For instance, Acacia mangium can be managed in a height range of 8–10 metres. This facilitates the use of pole pruners from the ground to collect pods from mid to lower crowns. This method works best if a tarpaulin or net is spread on the ground under the crown of the seed tree. The entire branch with fruits is lopped using the pole pruner and falls onto the spread tarpaulin. With typical vigorous growth of commercial tree species in the tropics the desire to keep them at a preferred height for seed collection is not that easy to achieve due to the rapid regrowth of the trees.
3.12.5 BOW AND ARROW
The bow and arrow is a tool commonly used in the Pacific for hunting. It is not commonly used for collecting tree seeds but given the lessons learnt at the Australian Tree Seed Centre (ATSC) on its possible use for seed collecting it can be easily adopted by Pacific Island seed collectors, where it can be a useful alternative (Figure 3.4). Depending on the operator’s skill in using it, the bow and arrow can reach a distance of over 50 metres, and can do a similar job to the catapult. It can be used to shoot a fishing line or a suitable cord over a selected branch of a seed tree. As with using the catapult, a suitable sized rope is then attached to one end of the fishing line and the rope pulled over the branch. The choice to use the rope to pull and break off the branch depends on the size of the branch and manpower to do this. It could also be used in combination with a flexible saw or to haul a ladder up the tree. Where the collector needs to climb the tree, the rope can be used to ascend to the tree crown using the “single rope technique” (Stubsgaard, cited in Gunn 2001)
Figu atta
3.1
Lad abo als
Figu the t 3-18
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
sily cahes rds ion eat
Gunn (2001) describes the bow and arrow and accessories used successfully by ATSC. They use a recurve break-down longbow with a draw weight of 13.5–18 kg (30–40 lb.) and a wooden or aluminium riser. Modified fibreglass fishing arrows are attached to a 22.5 kg (50 lb.) breaking strain fishing line which is spooled on to an archery fishing reel mounted on the front of the bow. The arrow tips are weighted and covered with a rubber bung. Great care must be taken when shooting the arrow to ensure the line is not tangled or likely to catch on the bow, user or surrounding vegetation. A short length (2–4 m) of weaker breaking strain line (6.8 kg (15 lb.) breaking strain)) should be connected between the arrow and main line. The weaker line is designed to break should the line be impeded immediately after firing, thereby allowing the arrow to continue rather than jerking back and endangering the operator. A face visor should also be used.
by the ow ght ect ead ped
to e to
mree ted .4). nce ta the the nch d in tor the
Figure 3.4 Bow and arrow with fishing line attached to get a rope into the crown of the tree.
3.12.6 OTHER CLIMBING AIDS Ladders — home-made ladders using bamboo poles provide access to a height of about 12 metres. More sophisticated ladders made from aluminium (Figure 3.5) are also available. 3.12.7 COLLECTIONS FROM FELLED TREES Plantations and native forest areas that are assigned to clear felling provide an opportunity to collect large quantities of seed without the usual consideration of causing minimal damage to the trees. Collection from plantations is generally suitable when large quantities of seed are required for routine plantation establishment, as long as the original seed source is known, is a mix of at least 20 different trees, grows well, preferably has been thinned to remove inferior trees, and a better seed source is not available. If harvest plans have been done well in advance (12 months) then arrangements can be made to collect seed from multi-species natural forest as the required crop (species) is ripe, or, if a plantation, coups are harvested only when the seed is ripe. Otherwise collecting seed from felled trees may only be possible where harvesting coincides with fruit maturity. Figure 3.5 Bamboo or aluminium ladders can be used to gain access to seed crops in the tree crown with the aid of a pole hook (Gunn et al. 2004).
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3-19
Seeds can be sourced for research purposes if the forest meets the minimum requirements for the research programme or from a plantation if the species is part of an improvement programme that requires selection of superior phenotypic trees (also known as candidate plus trees [CPTs]). Selection of CPTs involves surveying the plantation and selecting the very best one or two trees per hectare based on a predetermined set of traits that are often scored for each potential CPT in a specially prepared form (e.g. Appendix 2 in Howcroft 2002). When working near tree felling operations safety is the number one concern and safety equipment must be worn, especially hard hats and high visibility clothing. Watch out for tree limbs still hanging in standing trees which could fall, plus felled trees on the ground which could move when walked on.
Fo son • • • • •
3.12.8 COLLECTING OFF THE GROUND
Collecting seed which has fallen onto the ground is a very common practice in the Pacific and a very safe method to use (Figure 3.6). It is best suited to bulk collections of large fruit or seed as in the case of a number of rainforest species. Fruit containing sound seed should be collected as soon as possible after shedding to minimise fungal, insect and animal attack and thus reduce the incidence of mortality and poor germination. In the case where seed or fruit is in the process of shedding at the time of collection, large tarpaulins can be strategically spread out on the ground or in purpose built traps (Figure 3.6A) to catch the fruit or seed from under the harvested tree.
3.
On rea Tre ma ens stic ties of c in t app cm onl aw the lea the ing con the
Fo dry of r
Figure 3.6. A – Funnel for catching seed during natural seed fall (Doran et al. 1983). B – Some fruits are gathered from the forest floor (Lauridsen et al. 1987).
3-20
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
Wh ma for the the sta
remwn ion set e.g.
ety for und
the ons ing gal, naecuilt
ered
For research purposes this practice is not encouraged for the following reasons (Thomson 1995): • • • • •
uncertainty regarding the source tree risks of contamination from morphologically similar seeds of nearby related species their possible low physiological quality, compared with those obtained direct from the crown. This may be due to collecting a higher proportion of immature, empty, unsound or insect damaged seed, and early onset of deterioration or germination greater risk of contamination of the fruit or seed surface with soil-borne pathogenic fungi impractical for the collection of fine seed.
3.13 Harvesting Once the seed crop has been removed from the tree, the fruit needs to be harvested ready for transport, temporary storage, drying (if orthodox), extraction and cleaning. Tree seed harvesting is essentially a manual task in which as much of the unnecessary material like branches, twigs and leaves are removed in order to reduce the bulk. This ensures seed cleaning is not hampered by impurities and minimises the risk of large sticks puncturing the container. The degree to which the crop should be free of impurities needs to be a balance between ease of harvesting and transport versus the ease of cleaning. It is generally easier with large fruit to pluck or cut them from the branch in the field, as it saves time back at the base (Figure 3.7). With small fruit a different approach is generally used. For example with Eucalyptus deglupta, branches 5–10 cm may be cut from the tree. They are too large to transport and the fruit is small and only located within the leaves at the end of the branch. The tips of branches are cut away to significantly reduce the bulk, only leaving small branches (<1 cm), leaves and the fruit. If all the leaves are removed it is very time consuming. If the dried fruit and leaves are handled carefully then they are easily separated from the seed; however, if the dried leaves are crushed, they may break up into a similar size as the seed, making separation difficult. A judgement call is necessary in the field on which is least time consuming. It is normal to remove at least half the leaves as thick layers of leaves slow the drying process and promote fungal attack. For dehiscent fruit (e.g. Eucalyptus) which release their seed upon drying, the fruit will dry quicker if attached to the twig. Leaving the fruit attached has the added advantage of reducing the workload by not having to pluck off individual fruit. When collecting small-sized seed, consider impurities collected in the material that may be hard to remove during the cleaning process (e.g. dust, dirt, gravel, sawdust, foreign seeds). When it is raining the leaves are wet and dirt and gravel may stick to them making it hard to gather without these impurities. Therefore, after collection allow the leaves to surface dry, shake them and transfer to a clean sheet before the seed starts to extract from the fruit.
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3-21
Figure 3.7. LHS – Reducing the bulk of whitewood seed by removing the leaves from the fruits. RHS – Large fleshy fruits are individually collected with no leaves and branches.
3.14 Bagging and Transportation During the pre-collection planning a decision will be made on how to handle the harvested material, particularly when dealing with recalcitrant seeds and/or large fruits that may weigh in excess of 100 kg. The type of seed (orthodox or recalcitrant), the type of container/bag/sheet for carrying seed, whether dehiscent or indehiscent fruit, condition of the fruit, quantity of fruit, carrying capacity of the vehicle, climatic conditions for drying, time available to clean the seed in the field, and the distance to seed processing facilities are to be considered.
Un sho rec bot ma (e. mo mo are (e. ene see see tra mid bet
Aft and has (e. tre fiel wri cla “1 o tre ber Ap
It is from this point forward that the collector’s actions can have an impact on the physiological quality of the seedlot. The temperature, moisture content, insects, fungi and the type of container can all affect the length of time that the seed will remain viable. Understanding how these affect the seed and taking appropriate actions to circumvent problems will ensure that the seed arrives in the seed store in good condition. See Section 3.4 for an explanation of the seed types and in particular how temperature, moisture content and oxygen effect their viability. Once bagged, care must be taken to ensure that the fruits are not damaged or lost during transportation. At the time of bagging check there are no holes through which seed can escape. Sheets and open containers containing fruit must be kept upright and secured effectively to minimise the risk of seed loss. The fruit must be checked at least daily for fungal or insect activity and any problems addressed accordingly. The seed should be protected from direct sun and high temperatures. If transporting by sea then ensure protection from salt water spray and heat from the sun.
3-22
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
Figu spe repr
arge
arhat e of ion rying
ysiand ble. ent See re,
ost ich and ast eed hen
Unless collecting a fruit that requires fermentation to break the pulp down, the bag used should allow gas exchange. For orthodox seed this allows the moisture to escape, for recalcitrant seed it allows gas exchange so the seed does not die. For most collections, both individual tree and bulk collections, either collection sheets measuring approximately 1.8 m X 1.8 m (made from calico or a cotton synthetic fibre mix) or calico bags (e.g. 100 cm X 50 cm depending on seed size) are used. The fabric must allow free air movement to avoid the crop from turning mouldy, particularly where the environment is moist. For fleshy fruits, plastic containers with holes or open weave bags (jute, copra) are used. In some cases cloth bags will rot as the pulp breaks down, creating holes (e.g. with Terminalia catappa fruit). Recalcitrant seed will normally have some moistened coconut husks (or similar material) in the container to retain the moisture of the seed. In all instances the seed must be loosely packed to allow air flow; for orthodox seed, to promote even drying and stop fungal attack and, for fleshy fruits and recalcitrant seeds, to prevent excess heat accumulation and an anaerobic environment in the middle and bottom of the bag where fermentation will take place. Small containers are better to minimise this effect. After the harvest is complete, the fruit must be bagged and clearly labelled both inside and out (label example shown in Figure 3.8). For labelling in the field, each collector has their own sequential numbering system starting with 1 and prefixed by their initials (e.g. Paul Macdonell — PM1, PM2, PM3, etc.). A separate number is issued to each tree collection. In the case of a bulk collection representing a provenance, then a single field number is used to identify the bulk collection. Additional information should be written on the label, (e.g. the name of the provenance, date, and species) for added clarity. When more than one bag is used for a collection number, then write on the label “1 of 2” and “2 of 2” if there are two bags so that mistakes are not made. The individual tree number then becomes a permanent identifier throughout the system with the number linked to the seed and documentation at all times (see Seed Collection Data Sheet, Appendix 3.16.3).
FRONT
Duplicate
BACK
Figure 3.8. An example label used for each bag or sheet used. The collector’s number is circled, and the date, species and provenance are included on two separate labels. PM322 will never be used again, it will only ever represent this tree.
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3-23
3.15 Recording Field Data Recording accurate and complete data for the seedlot is a very important step in a provenance collection. This information completes the collection and without it the seed will not be accepted by PITSC. When undertaking research on a species it is essential to know where the seed came from, from how many plants, and details on the seed crop. It is also beneficial to know details about the individual trees collected from, but not essential. This section goes through the data fields on the seed collection sheet by explaining the information to include in each section, then an example of the section with data. A key is provided in Appendix 3.16.4 showing the values available to use. Character
Description
Species
To be written out in full giving genus and species (and subspecies).
Latitude(Lat) and Record the latitude and longitude for the first collection tree as this is generally the Longitude(Long) easiest access point to the site, which future collectors will use. If using a GPS, ensure the coordinate system is set to “WGS84”. Other grid systems may be 200 metres to a few kilometres out. If no GPS is available then it may be possible to use Google Earth. Seedlot
This is allocated in the seed laboratory and is a unique number issued to each provenance collection.
Location
Provide sufficient detail for future collectors to return to the same site. The most appropriate information varies from site to site. Geographical features such as mountains, rivers and/or distances along roads or rivers or specific locations within forest areas are useful locators. If latitude and longitude are from a GPS then a less detailed description is required here.
Ha
Ve (ve
Sp fre
As
Slo
Se Tim
Bu
Province/Country
Record country name, province and nearest local village, (e.g. Boeboe, Choiseul province Solomon Islands) where the collection was made.
Flo
Altitude
One reading in metres above sea level representing the mean for the collection site, best taken from a GPS or Google Earth. The range of altitude can be entered under “comments” if significant variation or against individual trees.
So
pH
So
3-24
Provenance:
Use the nearest village name, forest name, region name, province name, etc.
Climate:
Used for classification of climate, based on Koppen (1923).
Association
List the most dominant/co-dominant associated species together with related
Includes:
information on their frequency and mean height.
Comments:
Additional information that is relevant for the collection.
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
Ge
Pr
Ro
na the esthe om, eet ion e.
he PS, 00 to
ch
ost as thna
eul
on ed
Habitat:
Description of the environment in which the collection is made (e.g. river, ridge top, estuary).
Veg’n Type (vegetation):
Ranges from montane rain forest, through coastal forests to forest plantations. Developed by L. Thomson (pers. comm. 2013) after Mueller-Dombois and Fosberg (1998).
Sp freq (Species An estimation of how frequently the species occurs in the population, from “Rare” frequency): to “Abundant”. Aspect:
Compass direction in which the slope of the collection site is facing. For example if you are standing on a north facing slope and facing north, you would be looking down-slope.
Slope:
The average steepness of the slope ranging from “Level” (0°) to “Cliffs” (61–80°).
Seed crop & Timing:
Size of crop ranging from light to heavy, relative to typical crops for that species and the timing of collection (early, peak or late).
Buds &Timing:
Relates to presence of flower buds ranging from light to heavy (or absent) and timing (early, peak, late).
Flowers & Timing: Relates to presence of flowers ranging from light to heavy (or absent) and timing (early, peak, late). Soil texture:
Field texture (Soil Bolus Test) is a measure of the behaviour of a small handful of soil when moistened, kneaded into a ball and then pressed out between thumb and forefinger. The resulting behaviour of the bolus is compared with the texture grades listed in the seed collection datasheet key (Appendix 3.16.4), ranges from “Sand” to “Clay”. Refer to Northcote (1979) and McDonald et al. (1998).
pH:
Tested in the field using representative soil sample from a depth of 10–15 cm. It is more reliable if two or more tests are undertaken to cover the range of sites. Avoid testing near roads or other areas where soil is disturbed as these areas may have a non-representative pH.
Soil colour:
Visual estimation of the colour of moist soil, which can indicate a soil’s history and some likely properties, (e.g. dark soils generally have a higher nutrient status than pale soils).
Geology:
Based on collector’s knowledge or reference to geological maps for the area. Often difficult to determine accurately. Draw on local knowledge.
Predation:
Level of predation of the seed crop being light, moderate or heavy, and predator (birds, insects or other).
Root sucker:
Present or absent. A root sucker is described as a shoot arising from below the ground level either from the root or a rhizome (NAS 1980).
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3-25
Cr de bra an
Cr (H
La an
No
Te
Da
Co ind
Co
Se Via
Colln No (Collection Number):
3-26
Each field worker records their collections whether they be botanical or seed collections according to a sequential field number prefaced with the collector’s initials as described earlier under Section 3.14 (Bagging and Transportation). It is important that the collector’s initials are unique to avoid any duplication with other collectors using the same system. A separate number is allocated to each tree for identification purposes. Apart from being recorded on the data sheet, the same number is used on the seed label, botanical label, and any other collection item which is linked to the tree. It is good practice to enter field numbers into a field botanical book.
Bot Spec (Botanical Specimen):
Indicate whether a botanical specimen was taken.
Photo No:
If a photo was taken record the identifying number.
Ht (m) (Height in metres):
Height of tree in metres, either measured or an educated estimation.
Age:
The age of wild trees is unknown; however, the tree can usually be categorised into an age class – Young (Y), Maturing (Mg), Mature (M), or Over Mature (OM).
Bole - dbh (cm):
Diameter at breast height (DBH) is measured on the upper side of a slope at 1.3 metres above ground level by measuring the circumference of the tree with a DBH tape, (a DBH tape’s markings convert the circumference measurement to a diameter).
Bole–Form:
The straightness of the bole is assessed as poor (three or more defects) to excellent (no defects) or multi-stemmed.
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
It is Th fles com • • • •
Wh as gen is t usi 3.9 and an be
ed r’s . It ith ch he on oa
ed M).
1.3 ha to
el-
2 Crown– This is a comparison between trees of the same species within a stand. Three d e n s i t y ( D e n ) , options are given for each character. branching (Brn) and width (Wdt): Crown–height (Ht%):
% Is estimated as the bole length from the overall tree height given as a percentage. For example a 20 m tree with a 10 m bole = 50%; a 30 m tree with 10 m bole = 33%.
Latitude, Longitude If a GPS is available, record the location of every individual tree collected. and Altitude: Notes:
Record observations related to this tree, if necessary.
Team:
Names of people involved in undertaking the collection.
Date:
The date the seed was collected.
Collected as – individuals
Number of individual trees collected from.
Collected as – bulk If a bulk collection is done, the number of trees represented in the bulk seedlot. Seed mass (g) & Viab/10g:
Recorded after the seed has been cleaned, weighed and tested for germination in the laboratory.
3.15.1 BOTANICAL SPECIMENS
It is not common practice in the Pacific Island nations to collect botanical specimens. This is mainly due to the difficulty and inconvenience of drying large-leaved and large fleshy fruited species, that otherwise are easily identified from other species. Some common reasons for obtaining a specimen include: • • • •
to confirm the identity of the species to assist with botanical studies when a species is found in a new area to act as a reference library for the collector or organisation.
When collecting botanical specimens it is ideal to have as many identifying features as possible which includes flowers, fruit, seed, bark, and mature leaves. However, it is generally not possible to collect flowers and fruit at the same time as the seed collector is there to collect mature fruit. Therefore, collect the best possible specimen, label it using the collector’s number, and press it between newspaper in a plant press (Figure 3.9). Careful monitoring is required in the first few days to ensure it does not go mouldy, and the newspaper may require changing a number of times as it becomes moist. As an alternative to collecting a botanical specimen, or in addition to, a digital camera can be used to take close-up photos of identifying features.
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3-27
Forestry, Fiji.
3.
3.1
Au Co Mo -T -T
Of Figure 3.9. LHS – A plant press made from split bamboo. RHS – A pressed botanical specimens labelled with the collector’s number.
Bo
Se
Fie
Ma
Re
Ru
Co
Ba
Ba
Big
Bin
Bo
3-28
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
with
3.16 Appendices to Section 3
3.16.1 EQUIPMENT CHECKLIST FOR THE FIELD Authorities/Landowners Collection permits or agreements Movement approval - Travel request - Travel plan Office equipment Booking board/file Seed collection data sheets Field note books Maps Reference material on flora etc. Rulers, pens, pencils Collection equipment
Bow - string - arrows - reel with line - face shield Bow saw with blades Camera Chainsaw - spares to include bars, chain, sprocket, plug, diaphragm, starter rope, sharpening equipment - fuel (2 stroke) - oil for bar - sharpening equipment - protective clothing
Bags collecting - large - medium
Compass
Bags seed - calico - envelopes
Flexible saw
Big Shot catapult
Geologistâ&#x20AC;&#x2122;s pick
Binoculars
Global positioning system (GPS)
Botanical press - paper - plastic bags - jewellers tags - straps - boxes for dried specimens -specimen book with record of next field collection number
Height measuring instrument
Secateurs
- screw jars containing alcohol
Diameter tape
Gaffa (Gaffer) tape for repairs (e.g. sheets & equipment)
Helmets for all party members Needles pH kit Pruning saw-long handle and attachment
Seed identification labels
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3-29
1 1
Sheets for collecting
Tree climbing harness
Sieves - large - fine
Vehicle items
Em me or
Brake fluid
Ex
String
Extra fuel
Tarpaulins
Electrical wire
Life vic me
Throwing rope – 25 m (4-6 mm diameter)
Hydraulic jacks with levers
Wool bales Tree climbing equipment Big Shot head and 2.4 m pole Big Shot fine line (45 m) Big Shot sling replacement
Levers for tyre repairs
Wa pro rai
Puncture repair kit
Sa
Spare inner tyre tubes
Em be
Oil
Climbing rope (50 m static x 12 mm diam)
Spare parts to include: - air pump - bolts - fan belt - filters - fuses - jumper leads - radiator hoses
Spur guard
Spare tyres
Harness carry bag
Tool kit to include spanners, screwdrivers, pliers, shifter, wheelbrace, hammer, grease
Big Shot throw bag (450 g) Carabiners – 2 steel and 2 aluminium
Helmet Kit bag Pole pruner with extendable handle Pole straps Prusik rope (2 m x 8 mm diam) Pulleys
Tow rope Winch operating switch Wire Boat items
Rope bag
Anchor (of appropriate size to stop boat drift)
Sheathed saw (24 cm)
Bailers
Climbing spikes
Buoyant heaving line at least 15 m long
Throwing rope – 25 m x 5 mm diam. Plus weight
3-30
Insulating tape, clips
Se vic
Distress equipment such as watertight flashlight, distress flares
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
Fir
Ha EH
Mo
Su
Mi
Ax
Ca loa
Ro
Ta
Ta
Wa of
We
w-
p
First aid k - stan - rem Emergency Communication equipment such as GPS & mobile phone or Marine VHF radio, EPIRB
Hard hat
Extra fuel
Sunscree
Life jackets or Personal Flotation Device of appropriate size for all party members
Miscella
Set of oars (manual propelling device)
Cargo ne
Mobile p
Axes
Rope
Waterproof containers or bags to protect seed from sea water and/or rain on boat
Tape me
Tape – m
Safety items
Waterpro
Emergency position indicating radio beacon (EPIRB)
Wet wea
First aid kit - standard kit - remote area kit Hard hat, earmuffs (heavy duty EH12 32DB) Mobile phones Sunscreen 15+ Miscellaneous Axes Cargo nets, straps etc. for securing loads Rope Tape measuring – 30 m Tape – masking Waterproof torch for each member of the party Wet weather gear
-
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3-31
3-32
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
Slope Facing: Altitude:
Location of observation: GPS co-ordinates (latitude/longitude):
3.16.3
/
/
Female Buds
Female Flowers
/ /
Male Flowers
Female Buds
Female Flowers
/
Male Flowers
Female Buds
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/ / Fruit / / / / Male buds / / SEED COLLECTION Male Flowers / /
Female Flowers
/ /
Male buds
Fruit
/ /
Male buds
/
/
Male Flowers
Fruit
/ /
Male buds
/
/
Female Flowers
/
/
Male Flowers
Female Buds
Fruit
/
Male buds
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
Apr/Date / /
DATA SHEET
/
/
/
/
/
Female Flowers
/
/ /
Female Buds
Fruit
/
/
Male Flowers /
/
/ /
Mar/Date / /
Feb/Date / /
Tree No Phenology of ---Jan/Date Date --------------------> / / Male buds /
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
May/Date / /
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
Jun/Date / /
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
Jul/Date / /
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
Aug/Date / /
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
Sep/Date / /
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
Oct/Date / /
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
Nov/Date / /
Fruit: I - initiated; E - early; F - fully developed; ------------ R - rare; C - common; A - abundant; ------------- G - green; C - changing from green; R - ready for collection; S - shedding.
Flowers: I - initiated; E - early; F - fully developed; ------------ R - rare; C - common; A - abundant
Flower Buds: I - initiated; E - early; F - fully developed; ------------ R - rare; C - common; A - abundant
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
Dec/Date / /
Observations to be made as close as possible to the same date each month. When observing trees use the following symbols to record the development relating to flower bud, flowering and fruiting development.
Species:
from Gunn et al. 2004
Observers Name:
FLOWERING AND FRUITING PHENOLOGICAL DATA FORM
Flowering and Fruiting Phenological Data Form - Individual Trees
3.16.2
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3-33
/
/
/
Geology:
Predation status:
Root sucker:
Coppice: Age
Slope:
Seed crop:
Bud:
Flowers:
Colln No
Team:
/
Bole dbh Form (cm)
Soil colour:
Aspect:
Ht (m)
pH:
Sp freq:
Film No
Soil texture:
Bot sp
/
/ / /
/
/
/
/
/
/
/
/
/
/
/
/
/
Date:
Den
Crown Brn W dt
Map name: Ht (%)
Association includes:
/
/ /
/
/ /
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/ /
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
/
Longitude
Bulk: Individuals:
Collected as
Freq:
Lat:
Altitude
Ht (m)
Comments:
Koeppen Climate Class
State:
Long:
/ / DATA SHEET / SEED COLLECTION / / / / / / /
/
/
Latitude
Description/ notes:
Provenance name for Database:
/ / / / / Fruit / / / / / / / / / / / Male buds / / / / / SEED COLLECTION DATA SHEET Male Flowers / / / / / Female Buds / / P A C IF IC IS/L A N D S T R E/ E S E E D C E/N T R E Female Flowers / / / / / FORESTS AND TREES PROGRAMME Fruit S ecr etar iat/of the/P acific C ommunity / / - C eP aC / T B /uilding, N ar / er e C/ omplex, N/ ar er e,/ S uva, F ij/i
/
Veg'n type:
Habitat:
Location:
Species:
3.16.3
/
Female Buds
Female Flowers
/ /
Male Flowers
/
Male buds
Fruit
/
/
/ /
/
/
/
/
/
/
/
/
/
/
Total:
Alt (m):
Seedlot:
/
/
/
/
/
Seed weight (g)
/
/
/
/
/
/
/
/
/
Viab/ 10g
3-34 R = Rare
Mangroves growing in intertidal zone Trees growing in agricultural setting To be reserved for more highly disturbed forests - usually after clear felling or shifting cultivation (not after cyclone or sustainable, low intensity logging)
> 1000 m elevation < 5 m elevation & within 1 km of coastline
500-1000 m elevation
< 500 m elevation
Forest plantations = FP SPECIES FREQUENCY A = Abundant C = Common O = Occasional UC = Uncommon INDIVIDUAL TREE CHARACTERISTICS Age class Bole form Crown Density (Den) Young = Y Poor (3 or more defects) = P Sparse Maturing = Mg Fair (2 defects) = F Medium mature = M Good (1 defect) = G Heavy Overmature = O Excellent (no defects) = E Coppice = C Multi-stemmed = M Not applicable = NA Branching (Brn) Crown width (Wdt) Crown height (Ht) as a proportion Light = L Narrow = N of the tree height given as a Medium = M Spreading = S percentage Heavy = H Broad spreading = BS SOIL TEXTURE Behaviour of soil bolus Sand (S) little or no coherence, cannot be moulded Loamy sand (LS) slight coherence minimal ribbons of 5 mm Sandy loam (SL) bolus just coherent but very sandy to touch, will form short ribons to 2 cm Loamy sand (L) bolus coherent and rather spongy, no obvious sandiness but may be somewhat greasy to touch, if much organic material present will ribbon to 2.5 cm Sandy clay loam (SCL) sandy to touch, sand visible, ribon 2.5-4 cm Clay loam (CL) bolus coherent, plastic and smooth to manipulate, ribbons to 4.5 cm Clay (C) smooth plastic bolus, can be moulded into rods without fracture, ribbons > 7.5 cm
= LTF = FSW = UTF = MRF = MCF = CF = CBF = CSF = SDF = SDT = SDM = MGF = AGF = SF
SEED COLLECTION DATA KEY
VEGETATION TYPE Variant Main Lowland tropical forest - Freshwater swamp forest Upland tropical forest Montane rainforest - Cloud forest Coastal forest - Beach - Strand Forest Seasonally dry forest, woodlands and scrub - Talasiga in Fiji - Maquisa Scrub in New Caledonia Mangrove forests Agroforests Secondary forests
3.16.4 Level (0°) Undulating Gently inclined (1-3°) Mod. Inclined (4-10°) Steep (11-23°) Very steep (24-37°) Precipitous (38-60°) Cliffs (61-80°)
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
Yes No Undetermined
= Y = No = U
COPPICE ABILITY ROOT SUCKERING ABILITY
Avian (A) / Heavy (H) Insect (I) / Medium (M) Other (O) / Light (L)
PREDATION STATUS OF SEED CROP
e.g. River, creek, drainage line, floodplain, plain, rocky outcrop, undulating hills, rocky slope, plateau, swamp, disturbed area, salt lake, sand dune, estuary, escarpment, etc.
HABITAT
Seed, bud, flower crop Light (L) / Early (E) Medium (M) / Peak (P) Heavy (H) / Late (L)
PHENOLOGY
L U G M S VS P C
SLOPE
3.1
= U
Undetermined
PITSC CODE OF PRACTICE FOR SEED COLLECTING
1. All collections are to be made in a manner sympathetic to maintaining conservation of the species and integrity of the population. 2. Fruit removal: i. Where possible individual fruits should be removed rather than cutting branches. ii. When cutting branches, no more than one quarter of the crown should be removed. This typically amounts to about four branches. Under no circumstance should the tops be removed from trees. 3. Trees for collection must be a minimum distance apart of at least two tree heights. 4. All branches must be removed from the road and must not obstruct traffic or road maintenance. Large branches should be cut into sections. 5. Landowners must be consulted to negotiate terms and conditions of access before collections. 6. Every effort is to be made to advise landowner of your presence before the collection and a courtesy call must be made on completion of the collection. 7. Due care and attention are to be taken when using vehicles to minimise road damage especially under wet conditions. Gates must be left as found. Any damage to property to be reported immediately to the landowner or manager. 8. Staff must conduct themselves in such a manner that they will be welcomed back. 9. The trip must be followed up with a letter of thanks and, where appropriate, indication of the outcome of collections.
Sandy clay loam Clay loam Clay
(SCL) (CL) (C)
sandy to touch, sand visible, ribon 2.5-4 cm bolus coherent, plastic and smooth to manipulate, ribbons to 4.5 cm smooth plastic bolus, can be moulded into rods without fracture, ribbons > 7.5 cm
3.16.5
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3-35
3-36
Species
Occurrence
Artocarpus altilis (breadfruit)
Primary Uses
Seed Type
Notes
Indonesia, Papua New Guin- Fruit, multi-purpose ea, W Melanesia, Micronesia
Recalcitrant
Candidate for vegetative propagation
Bamboo species
Widely dist. in Pacific, also elsewhere
Furniture, flooring
Recalcitrant
Commonly propagated vegetatively
Barringtonia species (cut nut)
Widely dist. in Pacific, also elsewhere
Species specific â&#x20AC;&#x201C; nut, multi-purpose, coastal stabilisation
Recalcitrant
Candidate for vegetative propagation
Canarium indicum (nagai Widely dist. in Pacific, also nut) elsewhere
Edible nut, veg. oil, timber, firewood
Recalcitrant
Targeted for commercial development in Papua New Guinea, Solomon Islands and Vanuatu
Casuarina equisetifolia (coast she-oak)
Widely dist. in Pacific, also elsewhere
Multi-purpose, coastal stabilisation
Orthodox
Cocus nucifera (coconut)
Widely dist. in Pacific, also elsewhere
Food, coastal stabilisation, wood etc.
Recalcitrant
Cordia subcordata (island walnut)
Widely dist. in Pacific, also elsewhere
Wood for crafts, coastal stabilisation etc.
Orthodox
Endospermum spp. esp. E. medullosum (whitewood)
Widely dist. in Pacific, also elsewhere
Timber, multi-purpose
Variable depending on species
Flueggea flexuosa (poumuli)
Philippines to Vanuatu, exTimber, multi-purpose otic in Samoa and elsewhere
Orthodox
Inocarpus fagifer (Tahitian chestnut)
Widely dist. in Pacific, also elsewhere
Edible nut, wood, coastal stabilisation etc.
Recalcitrant
Candidate for vegetative propagation
Intsia bijuga (kwila)
Widely dist. in Pacific, also elsewhere
Timber, multi-purpose
Orthodox
Candidate for vegetative propagation
Azadirachta indica (neem)
NE India, naturalised in Fiji, as well as India, Ghana, Kenya & Puerto Rico, exotic in other Pacific countries and elsewhere
Medicinal, insecticide
Recalcitrant
Paraserianthes falcataria Philippines to Solomon (syn. Albizia falcataria) Islands (falcata)
Timber, soil improvement
Orthodox
Superior provenance sources from Solomon Islands.
Pandandus tectorius (thatch screw pine)
Widely dist. in Pacific, also elsewhere
Food, multi-purpose
Recalcitrant
Selected clones propagated by branch cuttings
Pometia pinnata (tuan)
Widely dist. in Pacific, also elsewhere
Timber
Recalcitrant
Candidate for vegetative propagation.
Pterocarpus indicus (rosewood)
SE Asia, Papua New Guinea, Solomon Islands, Vanuatu
Timber, multi-purpose
Orthodox
Candidate for vegetative propagation
Santalum spp. (sandalwood)
Fiji, Tonga, Vanuatu, New Caledonia, also elsewhere
Essential oil
Orthodox
Candidate for vegetative propagation
Tectona grandis (teak)
India, Myanmar and Thailand
Timber
Orthodox
Improved material available from Solomon Islands
Terminalia spp. especially T. catappa sea almond) and T. richii (malili)
Widely distributed in tropics and subtropics
Timber, edible nuts, multiple uses, cyclone resistance
Variable, species of interest most likely orthodox
Candidate for vegetative propagation
Spondias dulcis (Wi)
Widely distributed in Pacific Islands, often an ancient introduction
Fruit
Recalcitrant
Candidate for vegetative propagation
Swietenia macrophylla (mahogany)
Introduced in many Pacific countries
Timber
Short-lived orthodox
Candidate for vegetative propagation
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
A ROUGH GUIDE TO FLOWERING AND SEED COLLECTION TIMES OF SOME IMPORTANT COMMERCIAL
LIST OF PRIORITY SPECIES/GENERA FOR ATTENTION BY PITSC
3.16.7
3.16.6
al
ve
ve
ve
gs
ve
ve
ve
n
ve
ve
KEY: # = all year; F = Fiji; V = Vanuatu, S = Samoa; SI = Solomon Islands; PNG = Papua New Guinea; T = Tonga; H - Hawaii; P = Philippines Flowers Fruits Reference Jan F Feb–Apr SI; Feb–March & June V; Jan–March F #1 Dec-Feb in F Apr-Oct in F #11 Feb-Apr SI & V #6 Aug PNG Sep-Dec PNG #4 #F #7 Dec-Feb F Apr-Oct F #11 # F; Jan-March V; Aug-Sep V; Sep-# S; #1 Jan-March Aug-Sept #2 #F #F #7 Nov-Jan & Apr-Jun S Jun-Jan S #8 Mar-May F Jun-Oct F #11 Mar-Apr & Oct-Nov PNG Dec-Jan & Jun-Sep PNG #4 May-Jun & Oct-Nov SI Sept-Mar V #1 Nov-Dec PNG Jan PNG #4 Sep-Dec for SI Jan-Feb for SI #5 Jul-Jan F Jan-Jun F # 11 Dec-Jan (Jun-Aug) PNG (Jan-Mar) Aug-Sep PNG #4 May-Jul PNG; Aug-Oct SI; Oct-Jan & peak in Canarium indicum Jul-Aug V Nov V #1 Canarium indicum Jul-Sep PNG, SI, V, F, S Sep-Jan PNG, SI, V, F, S #6 Canarium indicum Jan-Apr SI Aug-Oct SI #10 Casuarina equisetifolia # # #1 Casuarina equisetifolia Feb-Mar & May-Jun F May-Aug F #11 Cocos nucifera # # #1 Cordia subcordata # # #1 Endospermum medullosum Dec-Oct PNG Feb-May (Sep-Oct) PNG #4 Endospermum medullosum Jan-March SI; Feb-May & Aug-Oct V #1 Eucalyptus deglupta Nov-Mar for SI Jan-Dec PNG; Mar-May for SI #4, #5
Species Agathis species Agathis species Agathis species Agathis alba Agathis macrophylla Agathis macrophylla Alphitonia zizyphoides Alphitonia zizyphoides Alphitonia zizyphoides Alphitonia zizyphoides Alphitonia zizyphoides Artocarpus altilis Barringtonia procera Calophyllum inophyllum Calophyllum inophyllum Calophyllum inophyllum Canarium indicum
A ROUGH GUIDE TO FLOWERING AND SEED COLLECTION TIMES OF SOME IMPORTANT COMMERCIAL SPECIES
ve
3.16.7
ve
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3-37
3-38
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
Pometia pinnata Pometia pinnata Pometia pinnata Pometia pinnata Pterocarpus indicus
Paraserianthes falcataria Paraserianthes falcataria Paraserianthes falcataria Paraserianthes falcataria
Intsia bijuga Intsia bijuga Intsia bijuga Intsia bijuga Pandanus tectorius
Inocarpus fagifer Inocarpus fagifer Intsia bijuga
Species Flueggea flexuosa Flueggea flexuosa Flueggea flexuosa Hibiscus tiliaceus Inocarpus fagifer
May-Jun SI May-Jul Nov-Jan in SI; Jan-Apr in Fiji; Feb-Mar in T; Mar/Aug/Nov in S Mar-May & Oct-Dec PNG Mar-May F Mar F & S Dec - Apr PNG; July - Jan & March V; March– Apr F; Jan-Feb SI
KEY: # = all year; F = Fiji; V = Vanuatu, S = Samoa; SI = Solomon Islands; PNG = Papua New Guinea; T = Tonga; H - Hawaii; P = Philippines
Epi Tanna and Aneityum - Dec–Jan (–Apr)
Vanuatu - Torres and Banks Is.(June–) October–December (–Mar.) Santo, Malo, Maewo - (July–) August–October (–Nov.) Malekula, Ambrym - Oct–Dec(–April);
Solomon Is- Western Province (March–) June–July (–August) Malaita Province (May–) Oct.–Nov. (–Dec.); Makira Province (July–) Oct.–Dec. (–Jan.); Choiseul/Santa Isobel June- July; Santa Cruz Group Sept–Dec
Oct PNG Mar-May SI Feb-May & Oct-Dec S Sep-Oct Dec-Mar in F; Feb-March/Jun-Jul/Oct-Nov in S; # & peak in Jan-Apr in P Mar-Apr & Aug-Dec PNG Dec-Mar F # sporadically May–Nov PNG;
#1
#4 #7 #8
#1
#4 #5 #8 #6
#4
KEY: # = all year; F = Fiji; V = Vanuatu, S = Samoa; SI = Solomon Islands; PNG = Papua New Guinea; T = Tonga; H - Hawaii; P = Philippines Flowers Fruits Reference Jan, May-Nov # S; # V; Jan, May-June P; Mar, May-Aug # S; # V; #1 Jun-Jul SI Aug-Sep & Jan-Feb SI #5 #S #S #8 # # #1 Jan - April V; Nov - Feb PNG & SI; Jan–March #1 Nov–Dec for PNG, SI & V & May–July F #F #F #7 Jan-Mar & Jul S # max Nov-May S #8 # & peak in Aug PNG; May-Jun V; Oct-May &Apr-Nov S # & peak in Feb PNG; Dec-Jan V; Apr-Oct F; #1 Oct-Feb & Jun-Jul S Jan – Dec PNG Jan-Dec (Feb) #4 Jan-May & Oct-Dec F; May-Jun V; Apr-May, Jul-Aug & Nov S Dec-Feb V; Oct-Feb & Jun-Jul S #6 Oct-May F April-Oct F #7 #S #8 March–May F February–April F
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
3-39
# & Nov–Jan V; Jan–Mar & Jun–Aug F Jan-Dec PNG #F May-Oct for SI May-Sep PNG May-Jan for SI Jun-Nov in F Dec-Feb in PNG; May-Jun in V; Jun-Jul & FebMar in S; Sept-Dec & Feb-May in T Dec-Apr PNG Feb-Mar SI; Dec-May S # sporadic; Jun-Jul & Feb-Mar S; Nov-Mar PNG Mar-Jul in F Nov-Mar S
Fruits
#9 #11 #8
#1 #4 #5 #8
#4 #7 #5 #4 #5 #11
#1
Reference
10. Evans, B. 1999. Nangai (Canarium spp.) seed collection, Vanuatu. SPRIG, CSIRO, Canberra.
8. Whistler, AW. 2004. Rainforest Trees of Samoa. Isle Botanica, Hawaii USA. 9. Thomson, L. & Uwamariya, A. 1999. Terminalia catappa Linn. Full Data Sheet. CABI Electronic Forestry Compendium.
7. Keppel, G. & Ghazanfar SA. 2006. Trees of Fiji: a guide to 100 rainforest trees. SPC, Suva, Fiji.
6. SPRIG Species Database. 2005. SPRIG, CSIRO, Canberra.
3. Gunn, BV. 2001. ATSC Operations Manual 4. Gunn, BV., Agiwa, A., Bosimbi, D., Brammal, B., Jarua, L., Uwamariya, A. 2004. Seed Handling & Propagation of PNG’s Tree Species 4. Bena, G., 2012. Solomon Islands seed collection work plan.
1. Elevitch, CR. (ed.). 2006. Traditional Trees of Pacific Islands. Permanent Agriculture Resources, Hawaii. 2. Orwa, C., Mutua, A., Kindt, R., Jamnadass, R., Simons, A. 2009. Agroforestree Database: a tree reference and selection guide version 4.0 (http://www.worldagroforstry.org/af/treedb/)
# sporadic H, F, V; Oct-Jan V; Jan in F Sep-Jan S
Terminalia catappa Terminalia catappa Terminalia richii
References:
# in T; Oct-Jan in V Nov-Mar PNG Jan-Feb SI; #S
Oct–Nov & Feb F; Jun–Sept & Nov-Dec T; Jan - Apr & July & Oct V Aug-Oct PNG Aug-Dec F Oct-May for SI Mar-Apr PNG Feb-May & Sep-Oct for SI Jan-May in F
Flowers
Terminalia catappa Terminalia catappa Terminalia catappa Terminalia catappa
Spondias dulcis Spondias dulcis Swietenia macrophylla Tectona grandis Tectona grandis Tectona grandis
Santalum spp
Species
KEY: # = all year; F = Fiji; V = Vanuatu, S = Samoa; SI = Solomon Islands; PNG = Papua New Guinea; T = Tonga; H - Hawaii; P = Philippines
Epi Tanna and Aneityum - Dec–Jan (–Apr)
M do
PACIFIC ISLANDS TREE SEED CENTRE Manual of operating procedures for seed collection, documentation, treatment and storage
4. Seed Processing
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
4-0
3
4
Th the the ties ery Se fur • • • • • •
Th car see dep
No me are dam am usi cal tem is s con wh col
4.
Th inc dire and Sc
4 Seed Processing The area available for seed storage in most seed centres is limited, and more so when the seed requires refrigeration or freezing to maximise its storage life. It is necessary, therefore, to reduce the bulk by removing the seed from the pulp, fruit and other impurities. The main exception is when the seed is going to be immediately sown in the nursery, and the pulp does not interfere with sowing the seed or the germination process. Seed processing may involve drying, depulping, extraction of the seed from the fruit, further cleaning and/or fumigation to: • • • • • •
remove gross impurities to facilitate further cleaning, eg. leaves, twigs, dirt, etc. dry dehiscent fruit to accelerate separation of seed from fruit remove the pulp from fleshy fruit to reduce bulk, minimise fungal problems and reduce the risk of viability loss clean the seed to achieve maximum purity and viability; reduce the moisture content of orthodox seeds fumigate the seed to kill insects contained in or on the seed.
These processes should be carried out as soon as possible following collection and care must be taken to avoid damage to the seed and maintain the identity of each seedlot (individual-tree or bulked). Methods for processing are many and varied and depend very much on the type of fruit, seed and equipment available. Normally small quantities of seed can be cleaned by hand without the use of equipment. However, when dealing with large quantities different techniques and equipment are used to speed up the process. When a “new” method is being trialled which might damage the seed, or for recalcitrant seed, it is good practice to hand clean a small amount and immediately conduct a germination test. After cleaning the remaining seed using the “new” method, take a sample and conduct another germination test. Physical damage to the seed coat is generally seen. However, the damage to moisture and temperature sensitive recalcitrant seeds cannot be seen. To check if the “new” method is successful, the results from both germination tests should be similar. Probert (2003) considers the effect of field drying on seed viability and discusses ways of determining what is optimal for a particular species in relation to the ambient conditions during seed collection.
4.1 Processing Methods There are many texts available describing processing methods for seed of forest trees including Gunn (2001), Schmidt (2007) and Willan (1985) and interested parties are directed to these for more information. Seed of desiccation sensitive seed (recalcitrant and intermediate seed) will require particular care, texts that discuss these include Schmidt (2007) and Berjak & Pammenter (2003). PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
4-1
Most of the methods available, however, involve manual handling the seed through a number of processes until it is clean enough for storage. Sometimes mechanical methods are available for species which over the years have proven to be difficult to clean, and specialised equipment has been made. Generally seed collectors have to be inventive and adapt whatever equipment is available to complete the task. Depending upon the circumstances most of these techniques can be undertaken in the field or back at base. It is important to maintain the labelling and separation of different individuals and bulked seedlots during processing. Equipment needs to be cleaned between every individual or bulked seedlot to maintain the integrity of the collection. Commonly used processing methods available in the Pacific include:
4.1.1 DRYING
Fruit of dehiscent species require drying so they open and allow the seed to eject. The material harvested is laid out on sheets either in the full sun, under a tree in the shade or under a covered shed with airflow from at least two opposite sides. It should be spread out as a thin layer so that air can pass around and through the material to allow it to dry evenly and quickly. Initially it should be turned over at least twice daily to assist with even drying. If possible the material can be suspended above the ground to assist with air flow using temporary constructed or purpose built racks (Figure 4.1 – LHS).
On out gro the reg det ext
Oft of s see
Th Th see sm is u wo
The method used will depend on the type of seed, if the material is green (not yet fully mature), and where you are. Some seeds can be dried in the full sun (Figure 4.1 – RHS), others will lose viability quickly and require protected drying. For recalcitrant seed, immature or green fruit, moist fruit or species with sensitive seed (e.g. Toona, Grevillea robusta), the material should be aerated in the shade to avoid excessive rapid drying and high temperatures which may have an adverse effect on the viability of the seed.
Fig wire
Figure 4.1. LHS – a seedlot elevated on wooden frame and shade cloth. RHS – field drying orthodox seed
4-2
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
Fle to sof rele crit
ugh cal to to ndd or
ked ual ing
The ade be ow sist sist .
4.1.2 SHAKING AND BEATING
Once dehiscent fruits have opened some force is generally required to get all the seed out. It may require a gentle tap to dislodge the seed or maybe forceful hits on the ground. When using this method carefully monitor for damage to the seed coat. As the seed ejects from the fruit it will fall to the bottom of the bag or the sheet, therefore regularly remove it to avoid unnecessary damage. Check in the bottom of the fruit to determine that all seed has ejected. More drying may be required if the seed is not extracting. Often there will be leaves and sticks mixed in with the material harvested. The process of shaking and beating may break this material into smaller pieces the same size as the seed. If this happens then cleaning will be more difficult and time consuming.
4.1.3 SIEVING
There are many aperture sized sieves available to remove impurities from the seed. The best sized sieve is chosen for the task which may be larger or smaller than the seed, depending upon the task. If dust and small particles are being removed then one smaller than the seed is used, if larger particles are being removed then a larger size is used. They can either be purchased, or made by drilling holes in buckets or making wooden frames with various sized wire bottoms (Figure 4.2).
ully 1– ant na, pid the
Figure 4.2. Handmade sieves. LHS – holes drilled in buckets. RHS – wooden frames and different wire mesh sizes
4.1.4 SOAKING
Fleshy fruits normally require soaking in water for a period of time (often 1 to 7 days) to soften the pulp and allow easier removal. The amount of time varies for thin pulp, soft fleshy pulp, fibrous pulp and felty pulp. As the flesh breaks down it ferments and releases heat into the water. This must be monitored to ensure it stays below the critical threshold for the species. The water is normally changed at least twice daily.
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
4-3
Some species loose viability quickly if left immersed in water for any length of time. Therefore they are spread on shade cloth in the shade and regularly wet with water to break the pulp down till soft enough for removal (Figure 4.3 â&#x20AC;&#x201C; LHS).
Figure 4.3. LHS- Keeping fruits moist in the shade to soften the pulp. RHS â&#x20AC;&#x201C; Equipment used to soak, remove and clean fleshy fruits
Fig
4.1.5 FERMENTING
Some orthodox species with a tough pulp require fermentation to break it down (Figure 4.4). These species are deliberately left soaking in the same water and allowed to ferment. For some species it does not affect the viability of the seedlot.
Aft ing bet 4.5
Ge allo rub (Fi
Flo mo as to den pro age goi so Figure 4.4. Lilypilly (Acmena smithii) is left in the same water and fermented to soften the pulp before cleaning
4-4
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
Wh atta see cie (Fi
4.1.6 MACERATION
me. r to
d to
Figure 4.5. Using hands to macerate the flesh from the seed
wn wed
After the pulp of the fruit has become soft it can be removed from the seed by macerating the pulp/seed/water mixture with mortar and pestle of some form. It may be rubbed between your hands, rubbed against a wire sieve or some other rough surface (Figure 4.5). The aim of this process is to remove as much pulp from the seed as possible.
4.1.7 WASHING
Generally, on washing, the seed will sink to the bottom of the pulp/seed/water mixture allowing the pulp to be scooped from the mixture. Final separation can be done by rubbing the seed by hand on a correct sized sieve with water flowing over the sieve (Figure 4.3 LHS).
4.1.8 FLOTATION
Flotation is the process where the almost cleaned seedlot is immersed in water to remove insect attacked seed, underdeveloped seed, and foreign material the same size as the seed. These are generally less dense than the sound seed and therefore float to the surface where they are scooped up and removed. It is possible to change the density of the water by adding sugar to increase the density (less floats) or petroleum products to decrease the density (more floats). However petroleum products may damage the seed and they are not recommended. Before application ensure the seed is not going to imbibe water and start the germination process and only apply in the morning so the seed can be immediately dried and not left wet overnight. Whitewood (Endospermum medullosum) fruit is prone to heavy insect (wasp larvae) attack and if not promptly removed, the larvae from the infected fruit will move to sound seed and consume the whole seedlot. Therefore, the first step in processing this species is the flotation method to remove the fruits that float. These have live insects (Figure 4.6).
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
4-5
Figure 4.6. Hollow and shrivelled seed which floated to the top when immersed in water
4.1.9 WINNOWING
Winnowing is the process of using air movement to separate different size, mass or surface area objects in the seedlot. At its simplest level the seed is tipped from one container to another container in a gentle breeze. The light material blows away and the heavier seed falls to the container below. Repeat as necessary tipping from different heights and strengths of wind to adjust the effect. It is a good idea to do this on a clean sheet so if the seed blows away it can be retrieved. The same principle can be used with an electrical fan (Figure 4.7). However it mainly works well when the material being removed is substantially lighter than the seed, and it is slowly fed past the fan.
Fig ma Per
Figure 4.7. Using an electrical fan to separate different sizes in the seedlot
A more controlled method of winnowing can be constructed by making a rectangular box with internal divisions, a clear front that can be removed for cleaning, and a strong vacuum cleaner (Figure 4.8). The seedlot is trickle fed down from the top and the different air pressures as set by the user separate it into three different classes, the heaviest on the left and the lightest on the right. All the material is retained so if the seed is lighter than the foreign material, it will be captured in one of the right hand compartments. The vacuum is attached to the two holes in the back of the unit shown as nine small circles. These holes are plugged or left open to vary the amount of airflow. The more holes left unplugged, the greater the air pressure, and the larger the size of the material that will move to the right. This allows greater control over the separation of the seed, whereas the container to container method will only separate fragments that are much different to the seed and are lighter.
4-6
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
d to
ent ea he her rial he ng do ple he ast
Figure 4.8. LHS - a commercially produced winnowing machine in action. RHS - the winnowing machine can be easily copied by making a timber box with internal divisions, a removable clear Perspex front, and attaching a strong vacuum cleaner to the outlets.
lar ng difavid is artne he he of hat
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
4-7
M do
PACIFIC ISLANDS TREE SEED CENTRE Manual of operating procedures for seed collection, documentation, treatment and storage
5. Seed Testing
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
5-0
5
Go tion for Th the ity
Kn spe por det ent mo nat rele run ods tria
Ide to P gra to c pen Wh be the cle tes pro bes lot. tes com
5.
Th hav is u
5 Seed Testing Good seed testing requires reliable (optimum or near optimum conditions of germination for the species under test) and repeatable standard methods to ensure that uniform and reproducible results are obtained for every individual seed test conducted. The procedures described in this section are based on current available information for the Centreâ&#x20AC;&#x2122;s target priority species and uses the principles of accuracy and repeatability described by the International Seed Testing Association (ISTA) (2011). Knowledge of optimum conditions of germination for many Pacific Island tree and shrub species is limited, as research to date has mainly been confined to commercially important agricultural and horticultural crops across all regions. The Centre will focus on determining the initial germination of each bulked seedlot and individual tree lot that is entered into the PITSC system and to monitor the seed during storage. It also aims to modify and build upon the information presented here for breaking dormancy, germination conditions, viability tests, vigour, purity and moisture content determinations, as relevant to its target priority tree species. In addition it will aim to work with countries to run joint trials to further develop these protocols and information. The Centreâ&#x20AC;&#x2122;s methods will be updated over time, based on experience and/or results from these research trials. Ideally, in-country testing of seed should be carried out for all seedlots before dispatch to PITSC and should also be done routinely for seed used for in-country planting programs to provide guidance to end users. Standard seed testing requires the capacity to control all parameters; in particular standard germination tests require relatively expensive equipment such as germination cabinets with temperature and light control. While facilities are being developed, some in-country forestry seed centres may not be able to adhere to the strict ISTA guidelines. However, as long as seed to be sent to the PITSC for storage and/or exchange has demonstrated acceptable viability and is clean and free of pests and foreign seeds, the PITSC can perform the full germination test when the seed arrives at the Centre. If good records are kept of equipment and procedures used by in-country staff, their test results can still inform end users about best handling methods to obtain a good nursery and planting outcome from that seedlot. The accumulation of results and experience from both formal and informal seed tests gradually establishes a reference frame against which following results can be compared (Schmidt 2007).
5.1 Sampling The comparatively small size of some of the seedlots that will come into PITSC will have implications for the number of seeds used per test and whether a particular test is undertaken.
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
5-1
Seed testing is carried out on a sample which is a small ‘representative’ part of the seedlot. Sampling is the first step for every seed test and it is essential to obtain a representative sample of the right size so that the result is indicative of the whole seedlot. Obtaining a representative sample determines the validity of test results, in particular for a large seedlot. There are two techniques used depending upon the quantity of seed, and the size and number of containers within which it is stored. Prior to testing the following procedures are to be followed – for more information see ISTA (2011), Schmidt (2007), Poulsen et al. (1998), Bonner et al. (1994) and Willan (1985). • When practical (<10kg up to 20kg) the whole seedlot should be thoroughly mixed to randomly distribute its contents throughout the container. Settling may occur during transport or during storage and if the sample is taken from the top with no mixing, the test result may not reflect the whole seedlot. After thoroughly mixing, three random samples containing approximately 100 seeds are taken from the container and all mixed together to form one “composite” sample. • For seedlots that are stored in multiple containers and, in particular, larger containers (20kg – 60 kg), mixing the whole seedlot as part of the sampling procedure is impractical. Samples are instead taken from three levels within the container, generally using a “Seed Trier” (Figure 5.1-LHS) and mixed with samples taken from each container to form a ‘composite’ mix. When there are more than five containers of a seedlot refer to Table 5.1 for the number of containers to sample to obtain a ‘composite sample’.
Fig sam qua
5.
Table 5.1: Guidelines for the number of containers to obtain samples from to form the “composite sample” for representative testing
No. of containers
No. of containers to sample
Up to 5
every container
6–30
1 in 3 containers (minimum of 5)
Over 30
10 containers or at least 1 in every 5
The composite sample is then further reduced until a working sample which is approximately twice the amount of seed required for the test is obtained. There are a number of methods used for mixing and sub-sampling. The simplest method is to spread the composite sample on a clean flat surface (lab bench), divide into 4 to 8 equal portions and alternate portions rejected leaving sufficient seed for the tests (Figure 5.1-RHS). Other methods range from a Boerner gravity-fed divider for large seedlots (>10 kg), electrically-driven Gamet divider for smaller weights (<10 kg) and gravity-fed soil dividers.
5-2
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
Tre see sub nen the and pre tru see und res ing if th cie rem
the eplot. ular of
see lan
be er. om ghom
ger ure erach fa site
te
oxiber the ons S). g), vid-
Figure 5.1 LHS: A seed Trier which has holes evenly spaced along its length to obtain small samples within a container of seed. RHS: Dividing the composite sample to reduce the seed to the quantity required
5.2 Purity Analysis
5.2.1 PHYSICAL PURITY
Tree seeds may contain impurities such as twigs, leaf matter, fruit particles, soil, foreign seed and other material. A purity analysis is often the first test to be carried out since subsequent tests (except moisture content) are done only on the pure seed component. As defined by ISTA (2011), the objective of the purity analysis is to determine the composition by weight of the pure seed as a percentage of seed of other species and inert matter (impurities). Seeds of other species and the types of other matter present in the batch should be identified as far as is possible. The distinction between true seeds of the species under investigation and trash can be unclear for some tree seeds, especially those that are de-winged (Bonner et al. 1994). Pure seed refers to the undamaged, undersized, shrivelled, immature or germinated seed and pieces of seed resulting from breakage that are more than half their original size identified as belonging to the species under consideration (ISTA 2011). The purity test will be more difficult if there are impurities the same sizes as the seed, particularly for small seeds. For species that are not normally cleaned to pure seed (eg. teak), the impurity that normally remains is classed as part of the pure seed.
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
5-3
PITSC’s normal procedure is to make a visual observation to determine if the seed is clean enough for phytosanitary purposes to place directly into storage and is free of damage by insects or other injuries. In cases where it is generally easy to clean the seed using rapid cleaning methods (e.g. separating fine seeds from impurities using sieves) then it is expected that the seed will be almost free of impurities. However, where mechanical methods are not effective for the separation of seed from particles then it may be necessary to accept some level of impurities in the seedlot. This situation is best avoided by ensuring the fruit are sufficiently free of impurities at the time of harvesting as discussed in sections 3.12 Methods of Collection and 3.13 Harvesting. PITSC will not routinely undertake purity tests beyond visual observation for cleanliness of > 95%, unless requested or required for a particular purpose (i.e. export of seed). To avoid receiving seedlots that contain excessive impurities, the Centre advises suppliers to send only clean seedlots. When required, the physical purity will be determined by following the principal rules under ISTA (2011). However for comparatively small size seedlots that do not have enough seeds, the sample size can be reduced. Once purity is determined the seed is returned to the seedlot. Purity test results are recorded on the Germination Test Sheet. The following procedure is used: •
•
The total mass of the sample is weighed then the pure seed is separated from the impurities and weighed separately.
•
The percentage of pure seed is calculated as follows:
Sample mass to contain at least 700 seed units. For fine seed use the mean germination/10g figure contained in the Germination Standards for the species in question. (e.g. Casuarina equisetifolia has a mean germination of 2637/10g which when converted to 700 seeds = 2.7g).
5.2.2 GENETIC PURITY
Collection of seeds from indigenous trees and shrubs growing in the wild does not warrant such systems where certified seeds are produced under strict controls as is done for agricultural crops. Genetic variations within individual species and geographic areas of occurrence are not well documented for many of the PITSC priority species.
5.3 Seed Dormancy ‘Seed dormancy’ refers to a condition where despite being provided with optimum germination conditions i.e. temperature, moisture, light and oxygen, a viable seed is prevented from germinating. To a large degree, dormancy is genetically controlled (Bonner et al. 1994) and this has enabled agriculturalists to breed out dormancy in crops.
5-4
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
Ho art atio of c
Do bry •
•
Ma bef son ban occ the an
So the it is tion this see tre sem ful
To com usu do tec app info be
d is of the ing er, les uae of ng. ess To ers
les ave d is eet.
es 0g
m
not s is hic s.
erreonps.
However, in woody species and particularly those from wild populations, no such artificial selections have been made, making seed dormancy an important consideration for many species. The environmental conditions during seed maturation and time of collection can influence the degree of dormancy. Dormancy can be grouped into two broad categories relating to the seed coat, the embryo, or both. â&#x20AC;˘
Seed coat dormancy relates to a physical, chemical or mechanical barrier provided by the seed coat. In acacias, for example, the seed coat does not allow moisture to penetrate to begin germination. In Paraserianthes falcataria the seed coat contains chemicals that prevent germination while in Tectona grandis moisture can penetrate the seed coat but germination is inhibited because the embryo cannot swell because of the mechanical barrier of the strong seed coat.
â&#x20AC;˘
Embryo dormancy is when inhibiting substances usually within the embryo or surrounding tissue prevent germination as is the case in some tree species.
5.3.1 PROCEDURES TO BREAK SEED-COAT DORMANCY
Many natural occurring Pacific Island tree species will not require any pre-treatment before sowing as they have evolved in an environment that does not have defined seasons. Typically, dormancy occurs within a species that needs to remain in the seed soil bank for a period of time until conditions are favourable for germination. Dormancy may occur in rainforest species that are dispersed by animals and need to pass through their stomach, using the acid to remove the pulp, wear through the seed coat or remove an inhibiting substance. Some trees species especially those with hard coated seed and/or waxy cuticle around the seed are immune to water and gaseous exchange. To aid germination and ensure it is rapid and uniform some form of pre-sowing treatment is necessary. The germination testing will determine which treatment is the most appropriate for the seedlot, and this can vary from season to season and different provenances. Fresh or immature seed (green and slightly shrunken in appearance) may not require as severe a pretreatment as that prescribed in the standards and, for some tree species with soft or semi-permeable seedcoats, a pre-treatment is not required and, in fact, may be harmful to the seeds. To determine the best method to overcome dormancy, the method trialled must be compared against the manual nicking technique. Manual nicking of the seed coat will usually give the highest germination rate for the seedlot, however, it is not practical to do this for large amounts of seeds sown in the nursery. Therefore, the results from the techniques listed below are compared against manual nicking to determine the most appropriate pre-sowing treatment for that seedlot. The germination standards provide information on historic pre-treatment requirements by species which will generally still be the best method. The recommended methods are:
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
5-5
5.3.1.1 Boiling and hot water treatments
5.3
Boiling water, pour and soak: Seeds are placed in glass beakers (100 ml) and precisely 10 times the volume of boiling water to seed is added to it. Seeds are then left to soak for approximately 24 hours at room temperature before sowing. The soaking process provides the opportunity for the seed to imbibe/absorb water and speed up germination.
Sc out wh So the Wit sca me est cau as see flow
Boiling water, immersion for 1, 2 or 5 minutes: Water is first brought to the boil then the seed (placed in a perforated steel tea infuser or tied in a piece of cloth) is immersed in the boiling water for the designated time. It is then removed from the heat source and placed directly into a germination dish or in water at room temperature and allowed to soak for approximately 24 hours before sowing. Hot water treatments: Although seeds of some tree species require some form of boiling water pre-treatment in order to aid germination, there are a number of species or specific seedlots which respond better to a hot water treatment (90°C for 1 minute). Heat treatment: Species like Tectona grandis (teak) require this treatment to aid germination. Seeds are placed in an oven for 24 hours at 80°C then removed and placed straight into a germination dish with moist vermiculite. In the nursery, seeds are sown on the germination beds, covered with a thin layer of soil then dry grass or coconut leaves is spread on top then burnt. 5.3.1.2 Acid scarification Acid scarification is seldom used with preference given to alternative methods that are safer and easier to apply. This method is, however, recommended as an alternative treatment for seeds of species with very thick seed coats, e.g. Acacia stenophylla, and is commonly used in Africa for the treatment of indigenous acacias. Seeds are soaked in concentrated sulphuric acid (95%, 36N) at room temperature for a designated time (30-120 min) (Bonner 1974) depending on the species then removed from the acid and rinsed under running water for at least 10 minutes. This can be done by placing the seeds contained in a perforated steel tea infuser in a 1 litre glass beaker and allowing the water to run through the beaker. CAUTION: Extreme care is necessary when handling concentrated acid – only trained staff should carry out this procedure which must be conducted in a fume cupboard. Never pour water into undiluted acid; rather, in disposing of the acid, pour a small quantity of acid into running water and be mindful of the gases given off by this procedure. Laboratory coats, glasses, face masks and chemical resistant gloves must be worn. A concentrated solution of potassium or sodium bicarbonate may be used as an antidote against accidental splashes (Laurie 1974). In addition wash the affected area in running water or use an eye wash bottle and seek medical attention if required.
5-6
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
5.3
Ma wh coa teu nic sm Sin the can and
In soa Th typ
Wh som •
•
releft ing up
hen sed and d to
of ies e).
erced wn nut
are ive and
or a ved one ker
ned
mall robe an rea
5.3.1.3 Scarification or cracking of the seedcoat Scarification wears down the seed coat allowing water absorption and may be carried out by hand especially for laboratory purposes or by mechanically operated scarifiers which rotate the seed contained in a drum against a rough surface like sand paper. Some factors to take into consideration when using this method are the roughness of the surface, duration of the scarification plus the amount and thickness of the seedcoat. With the ease and success of boiling water treatments, seeds are seldom mechanically scarified. Poulsen and Stubsgaard (1995) provide information on three methods for mechanical scarification of hard coated seed which was developed by the Danida Forest Seed Centre and includes (i) the ‘seedgun’ which propels seed against a hard wall causing the seed coat to crack on impact (ii) a hot wire ‘glow burner’ which is similar to a soldering iron for use in manually treating individual seeds and (iii) for treating larger seedlots a ‘mechanical burner’ which uses a hot glowing thread and continuous seed flow. 5.3.1.4 Manual nicking Manual nicking is often used to decide optimum germination of a seedlot especially where boiling water treatments have not been successful. A small section of the seed coat at the cotyledon (distal) end of the seed is removed with the use of either secateurs, nail clippers or a scalpel blade. Due to the time this operation takes, manual nicking is not suitable for a large number of seeds but is useful as a research tool for small numbers of seeds or to check the results of other pre-treatment techniques. Since it overcomes the problem of seed-coat variation, manual nicking usually gives the most reliable results. To split thick seed coated species (e.g. Macadamia) a vice can be used. However, Marunda (1990) reported that nicked seeds are less vigorous and more susceptible to fungal attack.
5.3.2 PROCEDURES TO OVERCOME EMBRYO DORMANCY
In the Pacific Islands, chemicals are hardly ever used in overcoming dormancy and soaking in water (either hot or cold) plus nicking are the pre-treatments normally used. This is mainly due to financial constraints and the recalcitrant behaviour of many seed types. While chemical treatments are not routinely used to overcome physiological dormancy, some problem species respond to the following: • Gibberellic acid –. Santalum album is routinely bubbled in a 0.01-0.05% gibberellic acid (GA) to water solution for 16 - 24 hours to speed up and create more uniform germination. ISTA (2011) also refers to the GA3 method for breaking physiological dormancy in seed. • Hydrogen peroxide - Seedcoats are cut to expose the radicle and incubated in a 1% hydrogen peroxide (H2O2) solution for 48 hours in the dark with alternating temperatures of 20 and 30oC. Radicle growth is measured after 3 to 4 days (Bonner et al. 1994). This method is not practical for very small seeds and may take 7 to 8 days to get a result.
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
5-7
• Citric acid – seeds are soaked in a 1% citric acid solution for 48 hours or combined with stratification (Bonner et al. 1994). • Potassium nitrate (KNO3) ~0.2% KNO3 solution, prepared by dissolving 2g KNO3 in 1 litre of water, is used to saturate the germination substrate at the beginning of the test (ISTA 2011). This method is used for a number of agricultural and vegetable seeds as indicated in the ISTA rules. For additional information on the types and methods for breaking dormancy see: Baskin & Baskin (2003), Boland et al. (1980), Bonner et al. (1994), Doran et al. (1983), Langkamp (1987), Schmidt (2007), Willan (1985).
5.3.3 PROCEDURES FOR REMOVING INHIBITORY SUBSTANCES
Seeds of some tree species contain inhibitors in the seed coat that prevent or delay germination and in such instances the inhibitor is leached out by placing the seed under running water for either several hours or days or soaking the seed in a large volume of water that is changed at frequent intervals (every 6-12 hours). Other leaching methods that have been used include alkaline solutions. Soaking in water to remove inhibitory substances from the seed coat is the most common method used in the Pacific with differences in length of time (most common is 1224 hours), and temperature of the water depending on the species. In the Solomon Islands, Tectona grandis is soaked for 24 hours then laid out in the sun for another 12 hours until the seeds are dry before they are sown. Flueggea flexuosa seed is soaked in cold water for 12 to 24 hours and directly sown with no drying, whilst Paraserianthes falcataria seed is soaked in cold water or hot water (60-70°C) before sowing.
5.4 Germination Testing For seed users it is important to have an accurate estimate of the number of healthy, vigorous plants suitable for field planting that can be produced from a given quantity of seed from a particular seedlot. A low germination percentage can indicate the need for changes in collection and/or storage practices if the seed is found to have poor physiological quality. For these reasons all seed requires an initial germination test, and likely followed by further tests on the remaining seed in storage after certain periods of time have lapsed. Testing of the highly recalcitrant species on the PITSC priority species list should still be carried out (e.g. for research purposes and as a check on nursery practices with that species), but may need to be run in parallel with sowing the rest of the seedlot in the nursery. If there is some excess seed to immediate needs then experiments on moisture content and storage temperature can be conducted. In this way information can be gathered for that species and may assist with planning of future collections.
5-8
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
Ge see see cul for see ma
Th in a bef see (Se not bet
Tab slig
5
The Lar sm wit hus ting wh ger can up as def
5
Fun imm Fun pre Se
ned
in 3 the ble
kin ng-
lay unme eth-
m12-
sun osa ilst ore
hy, y of for ysiand s of
still hat the oisbe
5.4.1 TEST CONDITIONS
Germination tests should, where possible, be carried out using a known number of seeds per replicate. The standard procedure is to separate out 25 randomly selected seeds for each replication (see Section 5.1) which are then weighed in order to calculate germination/10g prior to being placed on the germination substrate. However, for fine seed it is not practical and in many cases not possible to count the number of seeds. Tests therefore, are carried out on a known mass basis containing approximately 50 germinants per replicate. The number of replicates to obtain an average estimate of the germination percentage in a seedlot varies with the total mass of the seedlot as shown in Table 5.2. However, before averaging the test results to obtain an overall germination percentage for the seedlot, they must be examined to ensure they do not vary greatly between each other (See Tolerance Tables in Appendix 5.10.3). If this occurs then either the seedlot was not mixed properly prior to sampling, or the germination test conditions were not even between replicates. Table 5.2. The number of replicates required to test a bulked seedlot for germination, which varies slightly depending upon if a known number of seed is used per test, or the seed is weighed. Seed Mass (Kg)
Replicates â&#x20AC;&#x201C; known number of seeds
Replicates â&#x20AC;&#x201C; known mass of seed
up to 8
3
4
8 to 12
12
12
12 +
16
16
5.4.1.1 Germination containers and substrate The size of germination container used will vary with the size of the seed being tested. Large plastic containers (with or without lids) may be used for large sized seed, and smaller containers like 9 cm diameter petri dishes for smaller sized seed. The substrate within the container may vary from vermiculite, agar gel, sterilised river sand, coconut husks, cotton wool, etc. The seed must not be immersed in water, but rather placed sitting upon a moist substrate. A piece of filter paper is only placed on top of the substrate when the seed is small enough to fall between the substrate, making counting the germinants difficult. If a suitable sized piece of filter paper is not available then these can be cut by hand from paper towel or any other absorbent paper that does not break up when wet. When the seed has leachates within the seed, filter paper is not used as leachates may concentrate on the paper and can cause the radicles to become deformed. 5.4.1.2 Controlling fungi Fungal problems are generally associated with poor quality seed as in the examples of immature, damaged or old seeds which has lost considerable germination and vigour. Fungal development is also associated with seeds subject to insect attack or where the pre-treatment has been too severe. Established hygienic practices discussed under Section 5.8 will provide effective preventative measures in the control of fungi. Other
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
5-9
laboratory practices include preventing seeds from touching each other, adequate aeration, removal of decayed seed, avoiding pre-treatments that cause injury to the seed and keeping the substrate moist with no signs of free water. Where chemical controls are required the seeds are soaked for 10 minutes in a 1% solution of sodium hypochlorite followed by a rinse and surface drying before sowing to treat for possible external infections. A 10% sodium hypochlorite solution or a 30% solution of hydrogen peroxide (H2O2) for 20 minutes is recommended by Bonner et al. (1994). Studies by Yuan et al. (1990) on the presence of fungi on germinating seedlings of some Australia tree species with and without sterilisation found only a weak correlation between the frequency of seed germination and the level of fungal infection (contamination above a level of 60% did not result in further depression in germination frequency). 5.4.1.3 Moisture With the use of vermiculite, distilled water should be added to the substrate (28 ml for 30 ml of vermiculite). The substrate or filter paper should be wet but not saturated. Excessive moisture that can lead to fungal and bacterial problems should be avoided. Ensure that the container lid fits firmly and check the moisture level regularly, particularly in the cabinets set at 30 to 35°C. 5.4.1.4 Temperature Temperatures for germination of most species are in the range of 15 to 35oC. Seed is tested under constant temperatures with the exception of some tree species for which there is empirical evidence that an alternating temperature of 30oC daytime and 20oC at night may be advantageous. Seed of tropical species tend to have optimum temperatures of 25-35°C. 5.4.1.5 Light Light is required for germination in many tree seeds (Willan, 1985). Fluorescent light is as effective as natural daylight and is preferred in testing, because the wavelength and intensity can be standardised, within limits. Cool white fluorescent lamps are recommended because of their light quality and low heat emission. Light should be evenly distributed over the tests, with intensity within the range of 750 – 1250 lux. Seeds should be subjected to light for only part of the test period, 8 – 12 hours in every 24 hours is usual but a longer or shorter period may be beneficial to seeds of some species. 5.4.1.6 Germination counts All tests are recorded on a ‘Germination Test Sheet’ (Appendix 5.10.2). The sheets record details of the seedlot, method of test, replication weight, date of germination count and number of germinants. Counts should be carried out at regular intervals. The number of counts per week depends on the rate of germination and ranges from one to two times per week. The test period can vary from 10 days to over a month and should not be concluded if it is obvious that germination is likely to continue. With old seeds particularly those that have been stored in the cool room or deep freeze, germination often takes longer.
5-10
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
Th are at pla inc ger
On ed non pot use atta big of w see the see
Ste
Th tion see the cal
Th to a an exp ger see
Hig ess mig or find bot into
In PIT to t is a
ereed ols hlonal ide al. pency of
for ed. ed. cu-
d is ich C at ra-
t is and mnly eds 24 pe-
ets ion The e to uld eds ion
5.4.2 EVALUATION
The time at which a germinant is counted as normal varies. In some species, counts are made once the seed coat has been shed while for others the radicle must be at least three times the length of the seed. Once counted, the germinants are either planted out in the greenhouse or discarded. Germinants that are considered abnormal include albinos and those with abnormal cotyledons, radicles, and hypocotyls. Mouldy germinants should also be recorded as indicated on the Germination Test Sheet. On completion of the germination test, a squash test which is a count of non-germinated seed is made. For species with small seeds, a pair of tweezers is used to squash non-germinated seed and any seed found to have a firm white embryo is considered potentially viable. For species with bigger seeds or hard seed coats, forceps can be used for soft seed otherwise the seed is subject to a cut test. Seeds that have been attacked by insects should also be recorded. The count of viable seed for species with bigger seed should be recorded according to hard or soft seed. This is an indication of whether the pre-treatment was insufficient (i.e. high % of hard seed) or whether the seed coat was soft which indicates that the pre-treatment had been effective but that the germination conditions were not right. A record should also be made for mouldy seed as this reflects injured or dead seed. Steps taken when assessing results The number of normal seedlings produced is calculated and converted to a germination figure per 10g. This is a more conservative and realistic figure than referring to seed viability where it includes sound non-germinated seed in the final figure. Where the number of seeds is known, a figure for average germination percentage is also calculated. The germination results are compared with the ISTA tolerance tables (Appendix 5.10.3) to assess whether the variation between replicates is within acceptable tolerances. If an inexperienced seed tester has run the tests, the results should be shown to an experienced staff member to determine whether the test should be accepted. If the germination figures between replicates are beyond the accepted tolerances, then the seedlot must be retested. High squash test figures (>25% of total germination), are not acceptable making it necessary for the tester to seek an explanation. Some reasons that the tester will explore might be that the seedlot contained too many dead or damaged seed, pre-treatment or germination conditions were sub-optimal or there was operator error. Based on the findings, the tester must decide whether the seeds need to be re-cleaned, retested, both of which may require a cut test, or whether to accept the test and enter the results into the system. In the case of germination tests on bulk seed, the results are used for updating the PITSC germination standards. The results of the test are transferred from the test sheet to the provenance sheet, seed record card and seed database. The pre-treatment code is also included on the seed database.
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
5-11
5.4.3 RE-TESTING
Re-tests are carried out on seed where the initial test gave unsatisfactory results (see above), where there have been changes in the composition of the seedlot (e.g. recleaning, new bulk or seed added to current bulk) or after each 2 year period in storage (5 years for long lived orthodox seed). For initial retests and where the composition of the seedlot has been changed, the retest is comprised of four replications. Re-test figures are recorded on the card and seed database. It has been found that following a period in storage, some tree species like acacia seeds often require a more severe pre-treatment compared with the initial test (ATSC unpublished results). It may therefore be more effective when testing acacia seeds after 5 years in storage to use the standard pre-treatment method plus a more severe method. For seeds stored for long periods the following guidelines should be used in determining what action to take following a drop in germination over the previous test results: For seedlots of orthodox species stored in air-conditioned rooms (18-200C) with MC < 8%, if the average annual germination capacity for a species drops more than 6% (compare germination retest figures with original figures across the range of seedlots for a given species) then serious consideration should be given to recommending that the species be routinely stored in the cool room (3-50C). Once a seedlot has dropped its viability by 35% over the original test figure, then an assessment must be made on whether to replace it if an alternative seedlot is not already in the seed store. Once germination for a seedlot drops below 50% of the original figure, a decision must be made on whether to discard the seedlot from the system. In determining whether to discard the seed, consideration must be given to the value of the seed i.e. whether it is the only seedlot represented in the system, amount of seed involved and can it be replaced.
5.4.4 VIGOUR TEST
To determine the potential for rapid, uniform emergence and development of normal seedlings under a wide range of field conditions, a vigour test is used. According to Bonner et al. 1994, seed vigour declines more rapidly than the ability to germinate. Specific vigour tests are not routinely carried out, however, it is important that a vigour assessment based on germination data is made when conducting 2 (5) -year retests particularly where the seed is more than 10 years old. For most tree seed, the rate of germination is the most practical expression of vigour and this assessment is determined by the time taken in days for 50% germination to be achieved. Alternatively, compare the time taken for germination to be completed with the standards or, if available, the previous test for the specific seedlot. If the time taken for germination
5-12
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
to b fina it s can stu she cou see
5.
Cu len via firm ple and to e it is cle me Th old
Sq usi cut
Th see min
Se
Th twe
Fo sor pre wh
Co Gu
see reage
re-
cia SC eds ere
mins: MC 6% ots hat
asady
ust her her be
mal to ate. viear the t is naor, ion
to be completed (when 90% of the seed has germinated) is one third greater than the final count day recommended for a given species (refer germination standards), then it should be considered that there is an unacceptable lack of vigour. An assessment can also be made on the development of individual germinants as to whether they are stunted, have growth abnormalities and whether or not they have sufficient vigour to shed the seed coat. When using germination data to determine vigour, more frequent counts are required i.e. every one or two days. For additional information on this test see Bonner (1984) and Willan (1985).
5.5 Indirect Viability Tests Cutting test: This is a simple indicative viability test in which the seed is cut open lengthwise and the endosperm inspected to determine whether the seed is potentially viable or not. This method is not suited to fine seed - seeds deemed to be good are firm, white to ivory, sometimes green in colour with the endosperm taking up the complete space inside the seed coat. Non viable seeds are discoloured (grey), shrunken, and/or damaged which includes insect attack. The PITSC uses this method as a tool to estimate the quality of seed at the time of collection in the field, determine whether it is mature enough to collect and is also used for some species to assess whether the cleaning process is sufficiently rigorous. At the completion of a germination test the method is used to determine the condition of those seeds which have not germinated. The method is fairly reliable for healthy, fully mature fresh seed but less reliable for older seeds and seeds that were collected slightly immature. Squash test: The method is similar to a cut test except that the seed is squashed often using a pair of tweezers. It is more applicable to fine seed with a soft seed coat where cutting in half is not a practical option. The main application for this method at the PITSC is to determine which remaining seeds are viable following a germination test. This method can also be used to determine the viability of a seedlot as follows: Seeds are first soaked in water for 1 to 4 days. The water is then drained off and individual seeds are squeezed gently using a pair of tweezers and visually inspected to assess the number of viable seeds. For fine soft oily or moist seed, spread the seed samples between two pieces of absorbent paper (brown) then roll a glass bottle or rolling pin over the seeds with enough pressure to crush them against the paper. Viable seeds will leave a stain on the paper whereas dead seed and chaff will not stain. Count the stains to determine the number of viable seeds per unit weight (Quayle and Gunn 1998).
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
5-13
Excised embryo test: Seeds are soaked for 1 â&#x20AC;&#x201C; 4 days before the embryos are excised and placed on moist filter paper in a petri dish (Willan 1985). Embryos are germinated under constant light for 10 to 14 days at the temperature nominated in the germination standards. The method is slow and suitable for larger seeds. The technique could have application for determining whether seed dormancy can be attributed to the seed coat or the embryo by germination of seed with the seed coat attached compared to germination of the excised embryo.
Th Eq
Tetrazolium chloride: This method uses (TZ) (2, 3, 5-triphenyl tetrazolium chloride) to differentiate living from dead tissue by staining live tissue red. The concentration normally used should be 1.0%. For specific instruction on the procedure to follow refer to the ISTA Rules (ISTA 2011). Hydrogen peroxide: Bonner et al. (1994) provide the following technique where the seedcoats are cut to expose the radicle and incubated in 1% hydrogen peroxide (H2O2) in the dark with alternating temperatures of 20 and 30oC. Radicle growth is measured after 3 to 4 days and then the seeds are placed in fresh hydrogen peroxide. Radicle growth is measured again at 7 and 8 days. This test method was developed on barley and is used on many North American conifers. Evaluation is based on radicle growth: 5 mm growth or more is good; less than 5 mm growth is classed as uncertain; no growth is non viable. The method is not practical for very small seed, and has only been tested on conifers amongst tree seeds. X-ray: This method offers a quick estimate of seed viability and is non destructive but can only be applied to seed with a diameter over about 5 mm. PITSC does not have access to the equipment to apply this method.
5.6 Moisture Content The two most important factors affecting seed longevity in storage are moisture content (MC) of the seed along with storage temperature. It is important, therefore, to be able to determine seed MC accurately for processed seed, when drying seed as part of reducing the moisture content prior to storage or assessing the effects of storage conditions on seed moisture. Determining the moisture content of orthodox species is generally not required as the normal drying process will reduce this to an acceptable level. Recalcitrant seed is another story whereby the seed of some species (e.g. mangroves) do not tolerate any desiccation, some a little drying and others a bit more. It is therefore important to know the moisture content of this seed when undertaking a germination test to increase knowledge on the species and to explain the results. There are a number of methods used to determine moisture content of seed and the oven drying methods prescribed by ISTA (2011) are routinely used by PITSC. Other methods designed for more rapid results include electric meters and infrared driers. Probert et al. (2003) discuss non-destructive measurement of seed moisture.
5-14
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
5
Pri to 1 the of aga 30 anc wit tha
5
The ture tem res
Wh
5.
Th es spe the of h
exmierque the red
de) ion fer
the O2) red cle ley h: 5 wth ted
but ave
ent e to ucons ally Redo ore ion
the her ers.
5.6.1 OVEN METHOD
The oven method follows the procedures prescribed in the ISTA Rules (ISTA 2011). Equipment requirements and other factors to consider • • • • • • • • • •
Fan forced oven Aluminium containers with numbered base and lid Desiccator and silica gel Balance, accurate to 0.01g Moisture content test sheet Tongs, gloves Ensure oven has reached desired temperature before use Two representative seed samples with a weight in excess of 4 g Any seed >10 mm in diameter should be ground up to facilitate drying or sliced into 5 mm thick sections. Refer to ISTA Rules (ISTA 2011) for allowable tolerance between replicates.
5.6.1.1 Low constant temperature oven method Prior to drying, seed is weighed in a lidded aluminium container and the oven is heated to 103oC before drying the seed for 17 hr ± 1 hr. Time starts when the oven returns to the nominated temperature following the placement of the seed in the oven and closure of the door. The container tops are removed when placed in the oven and replaced again after completion in the oven prior to being cooled in a desiccator with silica gel for 30 to 45 minutes. The seed and container are re-weighed after cooling. Check the balance is tared between weighing. This method is used for most species especially those with high moisture contents or oily seed. Under ISTA (2011) rules, Table 9B specifies that all tree seed should be tested using the low constant temperature oven method. 5.6.1.2 High constant temperature oven method The procedures are the same as above except that the seed is subject to a temperature of 130 to 133°C for 1 hour. This method has been compared with the low constant temperature oven method for a range of eucalypt species and found to give similar results. When the seeds are large, they may be cut into pieces to aid drying.
5.7 Authenticity Test The assessment of seedling characteristics may help the identification process in cases where species have similar adult botanical characteristics or where it is possible that species may have hybridised. This requires raising seedlings in order to authenticate the species or to assist in decisions where seedlots are suspected of being mixed or of hybrid origin.
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
5-15
5.8 Laboratory Hygiene On the completion of any test or counting procedures, the laboratory seed testing area should be cleaned. All used petri dishes should be soaked overnight in disinfectant (1% Ammonia) and washed thoroughly in hot water. Disinfectant should be used to wipe down laboratory benches. Tweezers, used for seed counting, are soaked in 70% ethanol solution with distilled water between germination counts on each dish to avoid fungal contamination between replicates. All equipment should be cleaned between seedlots to avoid contamination.
5.9 Laboratory Safety PITSC staff should exercise on behalf of their organisation, the required “duty of care”. While responsibility for health and safety is a prime function of all levels of line management, staff are responsible for complying with all occupational health and safety instructions and taking action to avoid, eliminate or minimise risks to themselves and others. Staff must promptly report every new identified hazard, incident or accident in the workplace and contribute to developing safe working instructions for equipment and activities with health and safety risks. Whilst PITSC activities associated with the lab should be considered as ‘low risk’ there are however, a number of specific activities or materials for which there is a potential risk. Material Data Sheets (MDS) for all chemicals and materials used should be kept in an accessible and well publicised location in the laboratory. Safe working instructions should be written and displayed near equipment such as CO2 fumigation, oven etc.
5.9.1 FUNGICIDES ON SEED
PITSC discourages treating seed with fungicides. When ordering seed particularly from overseas countries, it should be requested that the seed not be dusted with any fungicide. Where a fungicide is applied, information on the fungicide should be provided with the seed shipment. Seed which has been treated with a fungicide should on arrival at the PITSC be handled with care. Staff should use gloves, and a face mask. Where considered appropriate, the seed should be washed using the laminar flow facilities available at CePaCT. For seed required by the phytosanitary regulations of the recipient country to be treated with a fungicide prior to dispatch or for other purposes, the officer should take the same precautions as described above.
5-16
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
PIT risk by circ the fum
Un an res
Ple Cu use kno
Us ing hea
Ta out sta sta
rea ant to 0% oid een
eâ&#x20AC;?. anety and t in ent
ere tial
t in ons
5.9.2 FUMIGATED SEED
PITSC routinely fumigates seed with carbon dioxide, which does not put the user at risk. However, there are occasions where the seed is fumigated with methyl bromide by quarantine on arrival in the country or by the dispatching organisation. Under these circumstances, it is important to allow for adequate aeration of the seed by spreading the seed out in a well ventilated area away from people for sufficient time to allow the fumes to be dispersed.
5.9.3 USE OF VERMICULITE
Under the Material Safety Data Sheet (MSDS Ref. AP91R3), vermiculite is regarded as an irritant if inhaled. For personal protection against respiratory problems, wear a filter respirator suitable for dust and minimise dust generation during handling.
5.9.4 PRE-TREATMENT OF SEED USING ACID
Please refer to the text under Section 5.3.1.2 Current MSDS sheets for all chemicals and other hazardous substances stored and used in the laboratory should be kept in a clearly marked folder and all staff should know where to find it.
5.9.5 BURNS FROM BOILING WATER AND OVENS
Use heat proof gloves when handling hot equipment. Wear safety glasses when working with boiling water. Make sure you are wearing enclosed shoes. Wear an apron of heavy material that can be removed quickly to avoid a burn from spilled hot liquid. 5.9.6 REPETITIVE MOVEMENTS AND CONFINED MOVEMENTS SITTING AT LAB BENCHES Take regular breaks when doing activities that involve repetitive movements and carry out stretches of key muscles involved e.g. neck, arms, and wrists. Have your workstation set up adequately with ergonomic design and equipment. Use rubber mats for standing tasks to reduce fatigue in lower limbs.
om unded val ere ies
ted me
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
5-17
SEED CENTRE Manual of operating procedures for seed collection, documentation, treatment and storage
5.10 Appendices to Section 5 5.10.1 Germination & storage information for PITSC priority species 5.10.2 Germination test sheet 5.10.3 Tolerance tables 5.10.3.1 Maximum tolerated range between weighed replicates
5.10.3.2 Maximum tolerated range between replicates of 25 seeds
5.10.1
GERMINATION & STORAGE INFORMATION FOR PITSC PRIORITY TREE SPECIES
PACIFIC ISLANDS TREE
5-18
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
8900
depends R on the fruit size (fruit weight range could be 0.25-6kg)
40004500
10-30
2
1-3
Artocarpus altilis
Azadirachta indica
Barringtonia procera
R
I
O
I
Alphitonia zizyphoides
30,00035,000
Agathis species
seed type
avge. fruit per no. kg seeds per fruit
200 #4
recomm. temp for germination deg C
replicate weight
viable seed per kg
7
7-21
14
7
2-7
60-90
28
16
14
avge time avge time start germiend gernation (days) mination (days)
no pre-treatment
no pre-treatment is required
no pre-treatment
water soak 12-24 hours for seed >10 months old
no pre-treatment is required
pre-treatments
river sand, forest soil or coir (fibre of coconut husk)
deep, permeable sandy soils
depends on variety - deep, fertile well drained soils while some prefer light & medium soils (sand, sandy loams, loams and sandy clay loams)
cover with thin layer (2-3mm) sand, loamy soil or potting medium
standard potting medium with good drainage
substrates
dried seeds stored in shady, cool (19-25°C [66-77°F) and low humidity (ca 50%) conditions
hermetically sealed containers and kept at 3-5°C (38–41°F).
seed has no dormant period, germinates immediately and is unable to withstand desiccation. Seed loses viability within a few weeks and can’t be stored in a fridge. Fresh seed moisture content 60%. Storage temperature 15°C (59°F)
hermetically sealed containers in cool, dry conditions
seed should be dried to around 9–13% moisture content and kept as cold as possible, preferably at –13°C (9°F)
storage
GERMINATION & STORAGE INFORMATION FOR PITSC PRIORITY TREE SPECIES
Species
5.10.1
n,
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
5-19
reference
#1
can also grow in a wide variety of soil types including difficult sites where most species do not perform well.
#1
#1 & #5
seeds are rarely grown #1 & #4 because they do not grow up or develop true to form. Seeds are normally cooked in the fruits and consumed throughout the Pacific (rarely in Polynesia) due to them being high in protein, low in fat and good source of vitamins and minerals. Expected germination of fresh seed 90-95%.
some seedlots take several months to germinate
to avoid reduction in #1 viability during storage, it is recommended that dewinged seeds be sown immediately following collection and processing
comments
5-20
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
90009600
2 million
1
1-3
1-4
1
1-4
1
avge.
Calophyllum inophyllum
Canarium indicum
Casuarina equisetifolia
Cocos nucifera
Cordia subcordata
Endospermum medullosum
Eucalyptus deglupta
Species
fruit per
seed
O**
R
O
R
560-700
O
R
I
seed type
560-700
50-200
100-200
avge. fruit per no. kg seeds per fruit
Species
viable
4-8 million #3
repli-
0.01
3500050000 #4
21 - 28
100000400000
recomm.
35
avge time
5
14
21 - 28
42
22
avge time
14
35
42
56
42
38-57
avge time avge time start germiend gernation (days) mination (days)
recomm. temp for germination deg C
replicate weight
viable seed per kg
coarse sand to well drained friable soils
sand and peat moss
well drained soil or potting mix, can also use sand or sandy loam soil mixed with well composted organic matter
well drained, sandy soils
substrates
pre-treatments
none required
substrates
vermiculite
none required for freely draining fresh seed sown loam is preferred within several days of collection
soak 12 -48 hrs;
no pre-treatment is required
soak 12 -48 hrs;
soak seeds in fresh water for 24 hours before sowing
crack shells or shell seeds entirely using a mallet, pliers, or hammer
pre-treatments
comments
storage
#1
#1
#1 & #3
#1 & #4
#1
reference
comments
reference
#1&#6
initial viability highly # 1 & #4 variable & often low due to wasp larvae damage, wasp infested fruit float in water & should be discarded at collection. fresh seed moisture content 51%. expected germination of fresh seed 35-50%.
store up to 1 yr
not advisable to store seednuts longer than necessary, if they are to be kept for longer period then they are to be picked at 11 months of age when the epicarp is starting to turn brown
store up to 1 yr
seeds to be sown immediately after collection
in hermetic storage at seed/chaff contain inhibi3-5°C (38–41°F) and tors also at -20°C (-4°F)
not long - suggest moist, well-aerated medium (e.g. slightly moistened peat moss, sawdust or plain cotton or open plastic bag @ 10-15°C [50-59°F]). can also try refrigeration at 3-5°C (38–41°F).
dried seed stored in cool, dry conditions
store in a cool dry place
dried seed stored in cool, dry conditions
seeds can only be stored for a few weeks - outer flesh should be removed and nut in shell stored in a shady, cool dry place. can also try refrigeration at 3-5°C (38–41°F).
fresh dried seeds can also grow in clay, stored in cool and dry rocky, or calcareous soils conditions as well
storage
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
5-21
38,000 44,000
4-6
5-15
1
2-8
2-8
1
Flueggea flexuosa
Hibiscus tiliaceus
Inocarpus fagifer
Intsia bijuga
Pandanus tectorius
Paraserianthes falcataria
Pometia pinnata
150
10-20
30,00035,000
300,000
avge. fruit per no. kg Seeds per fruit
Species
R
O
R
O
R
O
O
seed type
160
300-500
recomm. temp for germination deg C
replicate weight
viable seed per kg
7-10
5-10
7
7-11
7
14-28
14-35
60-70
70
84
avge time avge time start germiend gernation (days) mination (days)
loose and well drained soil with a surface layer of mulch
keep the fruit intact (nutin-shell) and store them in a shady, cool area with low humidity If storage is necessary
comments
moisture content is about 35-55% so need good air ventilation around fruits and, at 10-15°C (50-59°F), seeds can be stored to up to 6 weeks with coat intact.
air dry storage at 4-8°C (39-46°F)
#1 & #4
#1
#1
#1
reference
#6
seed is recalcitrant so #1 & #4 loses viability if dried down. fresh seed moisture content 35-55%. expected germination of fresh seed 90%.
before storage, seeds are air dried for 24 hours and then packed into bags
seeds are probably recalci- #1 trant, meaning that they will lose viability if dried
store with a dessicant fresh seed moisture in an airtight concontent 10%. tainer in a refrigerator at 3-5°C (38–41°F) or freezer.
shady, cool (19–25°C [66–77°F]) area with low humidity (<20%)
dried seed stored in refrigerator at 3-5°C (38–41°F)
keep in airtight containers under refrigeration at 3-5°C (38–41°F)
storage
well drained, storage of clean keys sandy, or coralline (seed) for weeks medium or months may be possible
well drained high pH soil particularly limestone
seeds should be buried in the media at a depth of 3–5 cm (1.2–2 in)
one part peat to 3 parts perlite or vermiculite
well drained, neutral, fertile loamy soils
substrates
no pre-treatment standard potting necessary if mix - more fertile seeds are sown and loamy types immediately after cleaning
soak in boiling water for 1-3 mins or immerse in concentrated sulphuric acid for 10 mins followed by subsequent washing & soaking in water for 18 hours
soak in cool tap water for 5 days change the water daily
nick the seed coat at the pointed end then soak in cool water for 24 hours
no pre-treatment is required
soak in water;
soak seeds in water overnight
pre-treatments
5-22
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
30,00035,000
one fruit can weigh up to 0.45 kg
2,400
1
1
1-5
Pterocarpus indicus
Santalum spp
Spondias dulcis
Swietenia macrophylla
fruit per
Species
avge.
2,000
Tectona grandis 0-4
3,00013,000
avge. fruit per no. kg Seeds per fruit
Species
seed
O
O*
R
O
O
seed type
viable
repli-
17,000
recomm.
25-28°C
28–31°C (82–88°F).
recomm. temp for germination deg C
replicate weight
viable seed per kg
avge time
10
10-17
28
30-40
3-4
avge time
60-90
40-120
15-30
avge time avge time start germiend gernation (days) mination (days)
pre-treatments
put seeds in oven for 24 hrs @ 80°C or soak fruits for 48 hrs before sowing or char (half burn) fruits by covering them with a thin layer of grass or coconut leaves and lighting it.
no pre-treatment is required
no pre-treatment required
nick the seed coat at the pointed end of seed using a sharp knife
no pre-treatment required for germination
pre-treatments
substrates
vermiculite
storage
#7
#1
# 1 & #4
reference
comments
reference
#6
2.5% of seeds germinated #6 following 2 yrs of hermetic storage of dry seeds at 3-5°C
can also be propagated by large hardwood cuttings, or air-layers. It can be grafted on its own rootstock but is often grafted onto S. pinnata in India
different varieties have different methods of germination, substrate mix, treatment, etc
Fresh seed moisture content 15-17 %. expected germination of fresh seed 10-20%.
comments
hermetic air dry stor- viability can be mainage at 0-4°C (32-39°F) tained for at least 7 yrs in with 12% MC hermetic , air dry storage at room temperature or even for 10 yrs in hermetic dry storage at 2°C.
seeds can be stored up to 7 months at 12°C (54°F ) with 9% MC
fertile alluvial soil
remove flesh, store surface dried seeds in airtight containers in the refrigerator (at 3-5°C [38–41°F])
store de-winged fruits at low temperature, 3-5°C (38–41°F), and low moisture content.
storage
fertile, forest loam (67%), and river sand (33%) plus 2 kg NPK fertilizer per cubic meter
a freely-draining, standard nursery potting mix
substrates
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
5-23
1
Terminalia catappa
Gunn B, Agiwa A, Bosimbi D, Brammall B, Jarua L, Uwamariya A (2004) ‘Seed Handling and Propagation of Papua New Guinea’s Tree Species. CSIRO: Canberra)
Forest, Farm, and Community Tree Network (FACT Net) - http://www.winrock.org/page/fact-net-archive (verified 13 December 2013)
World Agroforestry Organisation Species Profiles - http://www.worldagroforestry.org/treedb2/speciesprofile.php (verified 13 December 2013)
Purdue University Centre for New Crops & Plants Products http://www.hort.purdue.edu/newcrop/morton/ambarella_ars.html (verified 13 December 2013)
4
5
6
7
Recommended that a con- #1 trolled release, complete fertilizer (12g/l) is added if potting mix is used.
#1 & #4
reference
Australian Tree Seed Centre Database (2013) Germination Standards available at: http://www.csiro.au/ATSCOrdering/GetSpeciesStandard.aspx (verified 13 December 2013)
seeds can tolerate temperatures of 1417°C (57-62°F) and/or as low as 7°C (45°F)
It is recommended that seed storage behavseeds are sown within 4-6 iour is unknown but seeds tend to lose weeks after collection. viability rapidly under storage. try s storing at 3-5°C (38–41°F)
comments
3
standard potting mixture or in fertile loam to clay loam soils with good drainage and some organic matter.
standard potting mixture or in fertile, freely draining sandy loam or loam soils with some organic matter.
storage
Orwa C, Mutua A, Kindt R, Jamnadass R, Simons A (2009) ‘Agroforestree Database: a tree reference and selection guide version 4.0 (http://www.worldagroforestry.org/resources/databases/ agroforestree ) (verified 13 December 2013)
Short lived at room temperature
**
no pre-treatment needed
no pre-treatment is required
substrates
Elevitch CR (ed.) (2006) ‘Traditional Trees of Pacific Island’s. (Permanent Agriculture Resources: Hawaii)
Short lived orthodox
*
germination can go on for several months germination is not uniform
56
pre-treatments
2
Intermediate
I
germination is slow and goes on for several months
21
avge time avge time start germiend gernation (days) mination (days)
1
Recalictrant
R
References:
Orthodox
O
Key:
I
5002000
O?
recomm. temp for germination deg C
replicate weight
2600
viable seed per kg
seed type
Terminalia richii
70-150
avge. fruit per no. kg Seeds per fruit
Species
5.10.2
GERMINATION TEST SHEET
5
FOREST AND TREES PROGRAM PACIFIC ISLANDS TREE SEED CENTRE Germination Test Sheet Species
Seedlot
Origin
Alt (m)
Supplier
Collection Date ......./......./........
Pre-treatment
Date received ......./......./........
Test Type
individuals
bulk
Seed Mass
Test Conditions Rep. Mass (g)
OR Number of Seeds
Replications
Start of Test ......./......./........
Germination Began ......./......./........
Th mu twe ma rep ran
Nu i
End of Test ......./......./........ Average Germination for species (/10g)
±
No. of Tests Germination occurred on day
to seedlot + rep number; or seedlot + individual number
Date Examined
Test Period (days)
Mouldy Seeds Mass of seed Total germinants Squash test1 soft
hard
IST
2
Germination %
Germination /10g3
A = Albino; (Abnormal C = cotyledon, R = radicle, H = hypocotyl); M = mouldy seedling4
For bulk
Average germination % =
seedlot
Average viability % =
Average viability /10g
1
If squash test >25% seed hard or soft, re-test using different method/conditions.
2
When number of seed is known: total germinants ÷ (number of seed in test ) x 100
3
total germinants ÷ mass of seed ) x 10
4
Record the number of germinants that are not healthy using these codes.
5
Viability includes un-germinated healthy seed
Cut test of Seed Tester
5-24
Average germination /10g
5
seeds
Potentially Viable
seeds = Date
% ......./......./........
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
5.10.3
TOLERANCE TABLES
5.10.3.1 A. Maximum tolerated range between weighed replicates This table based on the Poisson distribution indicates the maximum range (i.e. maximum difference between the highest and the lowest) in germination data tolerable between weighed replicates, allowing for random variation at 0.05 probability. To find the maximum tolerated range, calculate the sum of the numbers of seeds germinated in all replicates. Locate the sum in column 1 of the table and read off the maximum tolerated range in column 2. Number of seeds germinated in the total weight of seed
Maximum range
Number of seeds germinated in the total weight of seed tested
Maximum range
1
2
1
2
0-6 7-10 11-14 15-18 19-22 23-26 27-30 31-38 39-50 51-56 57-62 63-70 71-82 83-90 91-102 103-112 113-122 123-134 135-146 147-160
4 6 8 9 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26
161-174 175-188 189-202 203-216 217-230 231-244 245-256 257-270 271-288 289-302 303-321 322-338 339-358 359-378 379-402 403-420 421-438 439-460 >460
27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45
ISTA (2011)
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
5-25
5.10.3
TOLERANCE TABLES (CONCLUDED)
5.10.3.2 B. Maximum tolerated ranges between replicates of 25 seeds Assuming a probability level of 2.5% calculated using the Binomial distributions for three, four, twelve and sixteen replicates of 25 seeds. To find the maximum tolerated range, calculate the average percentage to the nearest whole number. Locate the average percentage and read off the maximum tolerated range against the appropriate replicate number. Av. germination %
5-26
99 2 98 3 97 4 96 5 95 6 94 7 93 8 92 9 91 10 90 11 89 12 88 13 87 14 86 15 85 16 84 17 83 18 82 19 81 20 80 21 79 22 78 23 77 24 76 25 75 26 74 27 73 28 72 29 71 30 70 31 69 32 68 33 67 34 66 35 65 36 64 37 63 38 62 39 61 40 60 41 59 42 58 43 57 44 56 45 55 46 54 47 53 48 52 49 51 50 Williams et al. (1992).
3 reps 1 2 2 3 3 4 4 4 4 5 5 5 5 5 6 6 6 6 6 6 7 7 7 7 7 7 7 7 7 7 7 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8 8
4 reps 1 2 3 3 4 4 4 5 5 5 5 6 6 6 6 6 7 7 7 7 7 7 7 7 8 8 8 8 8 8 8 8 8 8 8 8 9 9 9 9 9 9 9 9 9 9 9 9 9
12 reps
16 reps
2 3 3 4 4 5 5 6 6 6 7 7 7 8 8 8 8 8 9 9 9 9 9 9 10 10 10 10 10 10 10 10 10 11 11 11 11 11 11 11 11 11 11 11 11 11 11 11 11
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
2 3 3 4 5 5 6 6 6 7 7 7 8 8 8 8 9 9 9 9 9 10 10 10 10 10 10 10 10 11 11 11 11 11 11 11 11 11 11 11 11 11 11 12 12 12 12 12 12
M do
for
est ted
PACIFIC ISLANDS TREE SEED CENTRE Manual of operating procedures for seed collection, documentation, treatment and storage
6. Storage
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
6-0
6
Se ran nee
The term Ho nee
In-c tha in o als
The ed cou can
Se que
The for The
Ro age
Ort aw bei
Re be cal
Wit ma
6 Storage Seed storage can be required for a short or long period of time and can be needed for a range of reasons from managing the difference between seed availability and planting needs to conservation storage (Schmidt 2007). The PITSC expects to operate as an active seed store, in most cases providing short term storage for research quantities of seed for dispatch to other member countries. However, long term storage will be enacted when there are conservation and research needs. In-country seed stores may have a range of reasons for needing to store seed rather than sow it immediately. This may be to have seed available for planting programmes in other regions, to bring together a range of provenances for provenance/progeny trials, to store seed collected opportunistically and so on. The ability of PITSC to store seed of many of the current priority species will be restricted due to their recalcitrance (‘desiccation intolerance’). PITSC will work with member countries to conduct storage trials on these species to identify those where shelf life can be extended with the correct handling and storage techniques. Seedlots will be held in stock until exhausted through consignment or as a consequence of viability loss over time. The storage life of seed is strongly influenced by the type and condition of the seed for storage, environmental conditions leading up to seed maturity and during storage. These factors are briefly discussed as follows: Roberts (1973) classified seed into two broad physiological categories based on storage characteristics: Orthodox – these seeds tolerate desiccation to low moisture contents (4-10% #w/w, on a wet weight basis), are comparatively long lived if handled appropriately and tolerate being stored at sub-zero temperatures. Recalcitrant – by contrast, these seeds are desiccation sensitive, short lived and may be intolerant of low temperatures (sub-zero for temperate species and <18oC for tropical species). Within these two categories of orthodox and recalcitrant, further sub-divisions can be made. Bonner (1990) refined the categories to comprise:
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
6-1
true orthodox
•
sub-orthodox; otherwise referred to as ‘intermediate’ between orthodox and recalcitrant in which the seed can tolerate drying to some extent, but not low temperatures (Ellis et al. 1991)
Th hav
•
temperate recalcitrant
1)
•
tropical recalcitrant
Hong and Ellis (1996) and Hong et al. (1998) reported that studies have shown that seed of certain species do not fall in with the above definitions. For this reason, there are those who avoid using definitions to describe seed characteristics of species preferring, instead, to define a species’ actual level of tolerance to desiccation and temperature. The following points may have an influence on the longevity of seed in storage: •
environmental factors leading up to seed maturation – if sub-optimal, environmental factors may have adverse effects on seed quality. Hot dry conditions, for example, may cause seed development to be impaired.
•
maturity of seed at time of collection – seed collected when immature tends to lose viability more rapidly than mature seed.
•
handling of the seed between collection and processing – adverse conditions such as high temperatures, humidity and development of fungi will damage seed.
• •
6-2
6.
•
injury of the seed during processing – for example, accidental cracking of the seed coat may reduce storage life. This has been discussed under the sections dealing with seed collection and seed processing seed coat structure – seeds with hard seedcoats are tougher than seeds with thin seedcoats.
•
seed chemistry – oily seeds tend to be harder to store than starchy seeds (Bonner et al. 1994, Stubsgaard, 1992).
•
insects and fungi – if not controlled, these can destroy the seeds.
•
storage conditions – the most important factors are to control seed moisture content and storage temperature while gaseous environments may also influence seed longevity.
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
2) 3)
Pro lec tem lan eve
Ar for fac ran mo nor be
Se rou tem (EM as see cha dry sph (ab tur
Wh suf ing per can the ser see al.
hat ere reem-
.
6.1 Principles of Storage The main factors associated with loss of seed viability in storage are given below. All have an influence on the rate of respiration: 1) Moisture content of the seed 2) Storage temperature 3) Storage atmosphere (oxygen) Protection of seed against pests and diseases is also critical at all stages from collection to storage. For recalcitrant seed, the safe minimum levels of moisture content, temperature and oxygen are all considerably higher than those for orthodox seed (Willan 1985). As seed ages this leads to deterioration in viability and vigour leading to eventual death of the seed.
6.1.1 MOISTURE CONTENT
A reduction in seed moisture content (MC) slows down the rate of respiration and therefore reduces the rate of physiological aging. MC is probably the single most important factor in determining seed longevity. The rule of thumb for orthodox seed is - within the range of 4-14% seed storage life is approximately doubled for each 1% decrease in moisture content. To reach an optimum moisture content (4-8% for orthodox seed) it is normally required that the seed be dried down. By contrast, recalcitrant seeds should be stored fully imbibed (Bonner et al. 1994). Seeds with permeable seed coats either lose or absorb moisture to or from the surrounding atmosphere until the MC reaches a point of equilibrium with the humidity and temperature of the surrounding air. This is known as the equilibrium moisture content (EMC) or equilibrium with the humidity (equilibrium relative humidity (eRH)). As long as the atmospheric humidity remains constant once the EMC has been reached in the seed, it will be maintained. Over time, should the surrounding atmospheric humidity change this will also cause the MC of the seed to adjust accordingly. The process of drying relates to the loss of moisture through evaporation of moisture to the atmosphere (desorption) and is in contrast to seed taking up moisture from the atmosphere (absorption). Thirty percent relative humidity is approximately equivalent to 8% moisture content in seed. When drying down seed, it is necessary, therefore, that the relative humidity of the air is sufficiently low enough to enable the seed to reach the desired moisture content. Drying facilities should allow for the control of humidity (dehumidified conditions) and temperature. The speed of drying is determined mainly by the speed at which the moisture can migrate to the surface of the seed for evaporation, the air velocity around the seed, the temperature and the relative humidity. For long term seed storage (Genebank conservation) the International Plant Genetic Resources Institute (IPGRI) recommend that seed should be dried down under conditions of 10 â&#x20AC;&#x201C; 15% RH and 10 â&#x20AC;&#x201C; 25oC (Rao et al. 2006). PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
6-3
Indicator silica gel can be used to dry down small quantities of seed as an alternative to dehumidified conditions and a weight of silica gel equal to one tenth the weight of seed is recommended (Harrington 1972). Refer to Stubsgaard and Poulsen (1995) for a more accurate calculation of the amount of silica gel required.
6.1.2 TEMPERATURE
The lower the temperature of the storage room, the lower the rate of respiration and the longer the life-span of the seed in storage. The rule of thumb is; between 0 â&#x20AC;&#x201C; 50oC, seed storage life is approximately doubled for each 5oC reduction in storage temperature. The choice of storage temperature varies considerably according to species and the period for which the seed is to be stored.
6.1.3 ATMOSPHERE
Excluding oxygen from the atmosphere is the third method for checking the rate of respiration. Even though this method may be beneficial to orthodox seed which has a low metabolic rate of exchange, it can be damaging to recalcitrant seed which requires oxygen. The method is commonly achieved by replacing oxygen with carbon dioxide, nitrogen or forming a vacuum. Shrestha et al. (1985) reported on the effects of controlled atmosphere storage on storage life of seed of Pinus radiata. Germination capacity, energy and seed vigour were best maintained by storage in nitrogen followed by carbon dioxide. Tests carried out in storage in a vacuum or air showed these treatments to be least effective irrespective of storage temperature. Refer to Bonner et al. (1994), FAO (1993), Justice and Bass (1978), Stubsgaard (1992), Stubsgaard and Poulsen (1995), Willan (1985) for more detailed information on seed storage.
6.2 Storage procedures at PITSC for orthodox Seed
6.2.1 FUMIGATION
All seedlots (orthodox) that come into the lab are routinely fumigated for two weeks (14 days) in carbon dioxide prior to storage in order to control insects. This method is both effective and safe to both the viability of the seed and user in contrast to a number of other methods of fumigation (e.g. carbon disulphide, methyl bromide). The CO2 method is based on the use of laminated gas-impermeable plastic bags. The simplest method is to place the seedlots in the bag and partially seal the neck using a heat sealer (threequarters of the neck width) then compressed industrial CO2 gas is then fed into the bag using a hose placed in the bottom until the bag is fully inflated. The gas is then turned off and the rest of the bag neck is sealed taking precautions to minimise loss of gas. Refer to Sary et al. (1993) for more information on the method of CO2 fumigation.
6-4
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
Th pre wa sib fro drie
6
Lon tur wit in n tem
Se
Co Se see als (RH and
Fre Fo und plu lon fre ran Ba mi pra
6
Sta wh tive hel of s an
In a ten intr
ive of for
and o C, raand
of has rebon cts ion wed at-
2), eed
(14 oth r of hod hod eebag ned s.
The PITSC does not routinely treat seeds for pathogens (fungi, bacteria and viruses) preferring to adopt preventative measures that ensure the seed is handled in such a way so as to keep damage to a minimum. To achieve this and also minimise the possibility of fungal infection, seeds, where possible, are kept dry, cool and well aerated from the time of collection through to storage. Prior to storage, orthodox seed is well dried and stored in a dry, cool environment under hygienic conditions.
6.2.2 SEED STORAGE 6.2.2.1 Temperature
Longevity of seed of orthodox species can be improved with storage at low temperatures and moisture content. Hong et al. (1998) found that orthodox seeds can be dried with no damage to low moisture content, lower than what they would normally achieve in nature. The seed longevity increases with the reduction in both moisture content and temperature over a wide range of storage environments. Seed storage will be available under the following temperature regimes at PITSC: Cool room (3-5oC) Seeds that do not store well at room temperature are kept in the cool room along with seedlots which are regarded as ‘irreplaceable or of high genetic value’ which should also be stored in the cool room. Since there is no coolroom-control of relative humidity (RH~90%), it is important that seed be dried down to a moisture content of below 8% and stored in laminated plastic bags in airtight containers. Freezer (-15 to -18oC) (not yet available) For species or seedlots considered ‘irreplaceable or of high genetic value’ as discussed under cool room storage, it may be wise to store in the freezer that portion of seed surplus to anticipated requirements in the next five years. Seeds specifically set aside for long term genetic conservation purposes and storage trials are other candidates for freezer storage. Seeds stored at this temperature must have moisture content in the range of 5-7% and be kept in sealed laminated plastic bags. (The Millennium Seed Bank [MSB] protocol of slowly drying seed down to the desired MC at 15% humidity and 15oC prior to being stored at -20oC in sealed foil packets is a desirable practice) 6.2.2.2 Control of seed moisture and atmosphere Standard practice at ATSC is for processed seed to be placed in airtight containers while in storage without further drying down. This method has been found to be effective in maintaining seed viability and vigour at an acceptable level for most seedlots held in the seed store for up to approximately 10 years. The loss of vigour and viability of seed beyond this time, however, has accelerated reducing the quality of the seed to an unacceptable level. In an attempt to maintain the viability and vigour of seed at an acceptable level beyond ten years, a policy of drying seed down to a moisture content of below 8% has been introduced. A method used for attaining the required seed MC is to use a cupboard
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
6-5
dryer with an electric fan and thermostatically controlled heater mounted at the bottom and vent at the top. Seeds contained in standard calico bags or paper envelopes are placed on racks. The fan forced air dryer located in the air-conditioned seed store (19-22oC, RH 25-45%) runs for an initial period of approximately one week to bring the seed down to a moisture content of about 8-9%. The dryer’s heater is then turned on to a temperature of 24-26oC for a further period until the moisture drops below 8%. The seed is then placed in vacuum sealed laminated plastic bags to maintain this MC. The control of MC of soft seeded orthodox species entering the store at PITSC will be an important consideration due to the high temperatures and humidity’s typical of PICs. The MSB protocol described earlier (6.2.2.1) would be a desirable aim but until the necessary equipment is available practises similar to those used by ATSC will be adopted.
6.3 Storage Procedures at PITSC for Recalcitrant Seed
A number of tree species, particularly those found in the rainforest, have fleshy or moist seed with a relatively high moisture content at maturity (>20%) making the seeds sensitive to moisture loss. These seeds have a comparatively high metabolic rate and are difficult to store for any length of time (even a few weeks in some cases).
6.
Pri and is p on bag con ma of t see an cat
On the cor stil
Th sto son
Given the variable nature of recalcitrant seed and limited experience in their handling, it is not possible to provide clear procedures for their handling and storage. A protocol had been developed by DFSC-IPGRI (1999) for assessing seed characteristics which will provide information on the storage life and method of storing the seed. The following points are provided as a baseline approach to handling recalcitrant fleshy fruit.
•
• Determine the initial moisture content. Cut seed into 5 mm thick slices and determine MC using the low temperature oven method. • Immerse in water for 24 hours to kill insects. • Determine whether the seed can be dried down safely without significant loss of viability (>10%). If drying does not have a detrimental effect, it may be possible to store the seed for longer compared with seed stored in the fully imbibed state. • Test for germination. This may take several months. • Keep seed in plastic bags that allow free air exchange (not laminated plastic). Moist seed that is likely to dehydrate under these conditions should be stored in a moist substrate (moist vermiculite or sawdust). • Seed should be tested for viability every 3-6 months. • Seed should be distributed as soon as possible following collection and processing.
•
•
• • • • • • •
6-6
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
botpes ore the on The
be Cs. eced.
oist nsiare
6.4 Maintaining Seed Identity in Storage Prior to storage, all seedlots must be clearly labelled with at least the seedlot number and collector’s number for individual tree lots. For large bulk lots (over 60 kg) the seed is placed directly in containers with a label placed both inside the container and another on the outside whilst for all other orthodox seedlots, the seed is packaged in calico bags or paper envelopes, sealed in laminated plastic and placed in containers. These containers (18 l) are filled with individually identified seedlots to a weight of approximately 6 kg with the seedlot number recorded on both the package and the outside of the container. All containers are also numbered sequentially and the location of the seedlot is recorded on the seed card (and later in the seed database). Where there are a number of packages or containers involved for a single seedlot, this should be indicated (e.g. 1 of 4, 2 of 4, 3 of 4, 4 of 4 and so on). Once the seed has been exhausted from the store, the seedlot weight will show ‘0’ on the seed card. The seedlot number is removed from the container and the Seed Record Card is placed in the ‘Dead Card System’. However, the record of the seedlot is still maintained in the system. The following is a summary of the steps that must be taken when documenting and storing seed. A summary diagram of this process is given in Appendix 6.5.1. The person responsible for each task is indicated in brackets:
ng, col ich ow-
• Ensure the seedlot is clean and supported by appropriate source information (seed collector for own collections, otherwise seed tester)
and
• Write out a seed record card using information from provenance data sheets (seed collector for own collections, otherwise seed tester)
ant be ully
• Weigh seed and record weight on card and provenance data sheets (seed collector for own collections, otherwise seed tester)
asbe
ro-
• Enter the seedlot in the register and allocate seedlot number to all related documentation and seed (seed collector for own collections, otherwise seed tester)
• Conduct seed germination tests (seed tester) • Fumigate (orthodox) seed for two weeks with CO2 (seed tester) • Seed placed in storage with seedlot number securely attached to the seed packaging (calico bag or envelope) and storage container (seed tester) • Provenance data sheets filed once completed (seed tester or seed collector for own collections) • Record card placed in system (seed tester) • Completion of germination test – information on viability and treatment placed on card and provenance data sheet (seed tester)
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
6-7
6.5 Appendix to Section 6
6.5.1
DRAFT SEED CENTRE FLOWCHART
M do
6-8
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
PACIFIC ISLANDS TREE SEED CENTRE Manual of operating procedures for seed collection, documentation, treatment and storage
7. Quarantine Procedures
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
7-0
7
Th and in sto som sur res me
Ne dur sio
Ne ecu of t
Ev mo is f
7 Quarantine Procedures
The exchange of seed between countries needs to minimise the risk of spreading weeds, and pests and diseases of plants. This is why there is a strong emphasis on cleanliness in all processes described in this manual, from seed collection, through processing to storage. The accidental importation of weeds, pests and diseases or weedy species is something that all countries want to avoid and this is why they have biosecurity measures in place. The National Plant Protection Organisations in each country have the responsibility for managing this risk and are the contact point for information on requirements for seed importation and exportation. Negotiations are currently in process with Fiji Biosecurity with regard to required procedures for the import and export of seed for PITSC. These will be detailed in a later version of this document once they have been developed and agreed. Negotiations will also be required with participating Pacific Island Countries (PICâ&#x20AC;&#x2122;s) Biosecurity agencies as well, to develop agreements and protocols to enable the exchange of tree germplasm with other PICâ&#x20AC;&#x2122;s through PITSC. Even within countries some basic quarantine checks should be done on seed being moved between regions (e.g. cleaning seed to remove weed seeds and ensuring seed is free of live insects).
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
7-1
M do
PACIFIC ISLANDS TREE SEED CENTRE Manual of operating procedures for seed collection, documentation, treatment and storage
8. Documentation for Seed Exchange
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
8-0
8
An me
8.
Th Co Ev
Th of suc
Th lot kno to k the
8.
Fo 201
8.
Ind
Se sea En
8 Documentation for Seed Exchange An important part of the PITSC activities will be the supply of high quality, well-documented seed to partner countries.
8.1 The Process Three key documents that will be supplied with seed shipped to partner countries are a Consignment Note, Explanation of Codes used on the Consignment Note and an Seed Evaluation Form for completion and return to the PITSC. The consignment note provides details of the seed being shipped and the explanation of codes used on the consignment note provides expanded information on key data such as country codes and seed pre-treatment recommendations. The seed evaluation form information is important to complete and return for each seedlot obtained from the centre under the seed sharing arrangements. Countries need to know who has accessed seed they have provided for exchange and the centre needs to know of any issues that arise with a particular shipment and to be able to report on the use of seed supplied to countries.
8.2 Material Transfer Agreement Four member countries have signed the Material Transfer Agreement as of December 2013. The copy of the MTA is shown in Appendix 8.4.4
8.3 Other Requirements for Seed Movement Individual countries may have their own requirements for seed movement. Seed movement out of Vanuatu for research purposes requires payment of a â&#x20AC;&#x2DC;Research Bondâ&#x20AC;&#x2122;. This is refundable upon completion and submission of results to Vanuatu Environment Department. Check with the department for current information.
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
8-1
8-2
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
Tectona grandis Swietenia macrophylla Flueggea flexuosa
00001 00002 00003
No. of Parent Trees
10 1 12
Tree No
bulk T10 bulk
Location
Qty (g)
Kolombangara, Western Province Colo-i-Suva, Vanua Levu Vailima, Upolu
100 50 10
FJI WSM
Deg
Latitude
SLB
Country
18 13
07
03 52
58
Origin
Quotation number: 00001 File reference: Import Permit No.: IP12345 Dispatch Date: 11/11/2013
8.4.2 DRAFT EXPLANATION OF CODES USED IN SEED CONSIGNMENTS
Species
Seedlot No
Attention:
Australian Tree Seed Centre CSIRO Plant Industry GPO Box 1600 Acton ACT 2601 Australia
Consignment Note and Seed Certificate
Telephone: Int +679 3370 73; Facsimile: Int +679 3370 73; Email cenonp@spc.int or elinay@spc.int
Secretariat of the Pacific Community CePaCT Building, Narere Complex, Narere, Suva, FIJI
FOREST AND TREES PROGRAM
PACIFIC ISLANDS TREE SEED CENTRE
8.4.1 DRAFT CONSIGNMENT NOTE
8.4 Appendix to Section 8
S S
S
Min
157 178 171
Deg 4 27 46
Longitude
E E W
NE/W
28 165 140
Alt (m)
21 18 2658
Viable Seeds/10g
T A A
Pretreatment
8.4.2 DRAFT EXPLANATION OF CODES USED IN SEED CONSIGNMENTS
Vailima, Upolu
WSM
10
13
52
S
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
8-3
T A A 21 18 2658 28 165 140 E E W
8.4.3 SEED EVALUATION FORM FOR INCLUSION WITH PITSC SEED SHIPMENTS
SECRETARIAT OF THE PACIFIC COMMUNITY
SECRÉTARIAT GÉNÉRAL DE LA COMMUNAUTÉ du PACIFIQUE
CABLE ADDRESS: “SOUTHPACOM” FIJI TELEPHONE: (679) 3370 733 CETC 3391 033 FAX: (679) 3370 021
ADDRESSE TÉLÉGRAPHIQUE: “SOUTHPACOM” FIDJI TÉLÉPHONE: (679) 3370 733 CETC 3391 033 TÉLÉCOPIEUR: (679) 3370 021
PRIVATE MAIL BAG, SUVA FIJI
4.
PRIVATE MAIL BAG, SUVA FIDJI
5.
S S 03 52 18 13 50 10
58
FJI WSM
1.
2.
bulk T10 bulk
10 1 12
Kolombangara, Western Province Colo-i-Suva, Vanua Levu Vailima, Upolu
SLB
100
07
S
Deg
Please fill in one of these forms for each seedlot you have received from the PITSC. Completion of this form will provide the PITSC with very valuable information. If you received seedlots from different shipments, please fill separate form for each. For an electronic copy, please contact cenonp@spc.int or elinay@spc.int
SPC PITSC Request No: Country: City: Institution/Organisation:
Date Seeds Received: Species:
Name of Researcher:
# of seedlings:
6.
7.
Amount of seeds:
Were any additional quarantine treatments applied when the seeds arrived in your country (e.g. methyl bromide fumigation) _____________________________________________________________________ _____________________________________________________________________ _____________________________________________________________________
8.
9.
Did you sow the seeds immediately after you received them or did you keep them for a while? _____________________________________________________________________ _____________________________________________________________________
Tectona grandis Swietenia macrophylla Flueggea flexuosa
_____________________________________________________________________ 3.
If kept, for how long before sowing? Also provide details of the storage conditions (e.g., fridge, coolroom, airconditioned room, non-airconditioned conditions), & what type of container it was kept in (was it airtight)? _____________________________________________________________________ _____________________________________________________________________ _____________________________________________________________________
00001 00002 00003
Deg
SCREENHOUSE evaluation of seeds/seedlings
Min
PITSC Evaluation Form for Distributed Seeds
8-4
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
10.
4.
_____________________________________________________________________
LA E
_____________________________________________________________________ _____________________________________________________________________ 5. What was the germination percentage? _______________________________________________________________________ _______________________________________________________________________ 6.
Did you use a screen house and if so, how did you maintain humidity in the screen house? _______________________________________________________________
8.4.2 DRAFT EXPLANATION OF CODES USED IN SEED CONSIGNMENTS
_____________________________________________________________________ _____________________________________________________________________ 7.
How long were the seedlings in the screen house or nursery prior to planting out? _____________________________________________________________________ _____________________________________________________________________ _____________________________________________________________________
8.
Were there any losses\mortalities in the nursery? _____________________________________________________________________ _____________________________________________________________________
9.
Were there any problems (pest, disease, mineral disorders, stunting etc) in the nursery â&#x20AC;&#x201C; if so, describe. _______________________________________________________________________ _____________________________________________________________________ _____________________________________________________________________
10. Please provide any other relevant information about the success or failure of the seedlings in the nursery. _____________________________________________________________________ _____________________________________________________________________ _____________________________________________________________________
3
When sowing, was any pre-treatment used? If so, please describe? Also provide details about the substrate the seeds were sown in (eg. if a mix, what was the composition?
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
8-5
8.4.4
DRAFT MATERIAL TRANSFER AGREEMENT
MATERIAL TRANSFER AGREEMENT FOR FOREST GENETIC RESOURCES TO THE SECRETARIAT OF THE PACIFIC COMMUNITY
PREAMBLE WHEREAS Parties agree on the importance of conserving, researching, and the use of forest genetic resources identified in Annex 1 of this Agreement for food security, adaptation to climate change and to conserve them for further generations; Secretariat of the Pacific Community will at all times uphold the traditional knowledge and intellectual property rights of providers of the forest genetic resources identified in Annex 1 of this Agreement and ensure the provider has the opportunity to partake in accessing information, capacity building and research programmes of forest genetic resources;
the fur und me
— its mo
AR
Th Ag mu
AR
Th ing
The providers of the forest genetic resources identified in Annex 1 of this Agreement agree that the Secretariat of the Pacific Community stores the materials for research on food security, adaptation to climate change and to conserve them for further generations. 1.2 This Agreement is: BETWEEN: (name and address of the provider or providing institution, name of authorized official, contact information for authorized official ) (hereinafter referred to as —the Provider“), AND: Secretariat of the Pacific Community; 3 Luke Street, Nabua, Private Mailbag, Suva, Fiji Islands
AR 1.3 The parties to this Agreement hereby agree as follows: ARTICLE 2 – DEFINITIONS In this Agreement the expressions set out below shall have the following meaning: —Genetic material“ means any material of plant origin, including reproductive and vegetative propagating material, containing functional units of heredity. —Forest Genetic Resources“means any genetic material of plant origin of actual or potential value for forestry and related application. —Forest Genetic Resources under Development“means material derived from the Material, and hence distinct from it, that is not yet ready for commercialization and which
8-6
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
5.1 use com
5.2 res bre
5.3 ert
gen to
dge d in e in etic
ent rch ra-
e er
g,
or
Maich
the developer intends to further develop or to transfer to another person or entity for further development. The period of development for the Forest Genetic Resources under Development shall be deemed to have ceased when those resources are commercialized as a Product. 2
—Product“means Forest Genetic Resources that incorporate the Material or any of its genetic parts or components that are ready for commercialization, excluding commodities and other products used for food, feed and processing. ARTICLE 3 – SUBJECT MATTER OF THE MATERIAL TRANSFER AGREEMENT The Forest Genetic Resources specified in Annex 1, Passport Data Form, to this Agreement hereby transferred from the Provider to the Secretariat of the Pacific Community subject to the terms and conditions set out in this Agreement. ARTICLE 4 – RIGHTS AND OBLIGATIONS OF THE PROVIDER The Provider undertakes that the Material is transferred in accordance with the following provisions: a) access shall be accorded expeditiously, without the need to track individual accessions and free of charge, or, when a fee is charged, it shall not exceed the minimal cost involved; b) subject to applicable law, any other associated available non-confidential descriptive information, shall be identified in Annex 1; c) access to Forest Genetic Resources under Development, including material being developed, shall be at the discretion of its developer, during the period of its development; d) access to Plant Genetic Resources for Forestry protected by traditional knowledge, intellectual property and other property rights shall be consistent with relevant international agreements, and with relevant national laws. ARTICLE 5 – RIGHTS AND OBLIGATIONS OF THE SECRETRIAT OF THE PACIFIC COMMUNITY 5.1. The Secretariat of the Pacific Community undertakes that the Material shall be used or conserved only for the purposes of research, breeding, training and other non commercial use. 5.2. The Secretariat of the Pacific Community undertakes to develop collaborative research programmes so that the Provider may actively partake in the researching, breeding and training programmes. This should be expressly identified in Annex 2. 5.3. The Secretariat of the Pacific Community shall not claim any intellectual property or other rights that limit the facilitated access to the Material provided under this
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
8-7
Agreement, or its genetic parts or components, in the form received from the provider. 5.4. Where there has been a request from a third party to the Secretariat of the Pacific Community to transfer Material supplied under this Agreement, the Secretariat of the Pacific Community shall notify and expressly attain the consent from the Provider. Upon receiving the Provider’s response, the Secretariat of the Pacific Community shall act accordingly.
AN
5.5. Where the Provider has consented to the transfer to a third party, the Secretariat of the Pacific Community will transfer to the third party and have no further obligations regarding the actions of the third party or the Provider.
Th dat des
5.6. Where the Secretariat of the Pacific Community intends to obtain intellectual property rights on any Products developed from the Material or its components that is obtained from the Provider, the Secretariat of the Pacific community shall enter into benefit-sharing obligations with the Provider.
PA
Th the
Lis
ARTICLE 6 – APPLICABLE LAW The applicable law shall be the laws governing where the Provider is from and the Fiji legal system. ARTICLE 7 – DISPUTE SETTLEMENT 7.1. ner:
Any dispute arising from this Agreement shall be resolved in the following man-
a) Amicable dispute settlement: The parties shall attempt in good faith to resolve the dispute by negotiation. b) Mediation: If the dispute is not resolved by negotiation, the parties may choose mediation through a neutral third party mediator, to be mutually agreed. c) Arbitration: If the dispute has not been settled by negotiation or mediation, any party may submit the dispute for arbitration under the Arbitration Rules of an international body as agreed by the parties to the dispute. ARTICLE 8 – WARRANTY 8. The Provider makes no warranties as to the safety of or title to the Material, nor as to the accuracy or correctness of any passport or other data provided with the Material. Neither does it make any warranties as to the quality, viability, or purity (genetic or mechanical) of the Material being furnished. The phytosanitary condition of the Material is warranted only as described in any attached phytosanitary certificate. The Secretariat of the Pacific Community assumes full responsibility for complying with the Secretariat of the Pacific Community nation‘s quarantine and biosafety regulations and rules as to import or release of genetic material.
8-8
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
AN
RE
er. Pat of er. hall
ANNEX 1
riat ons
The following information is included for each Material listed: all available passport data and, subject to applicable law, any other associated, available, non-confidential descriptive information.
ual hat nto
PASSPORT DATA FORM This Annex contains a list of the Material provided under this Agreement, including the associated information referred to in Article 4b.
List
Accession number or other identifier
Associated information, or source from which it may be obtained
an-
lve
any an
ANNEX 2 RESEARCH PROGRAMMES WITH THE PROVIDER
as ial. mel is riat riat s to
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
8-9
9
Mo et a
Ab
Are
Ari
Bip
Ca
Ca
Ch
Co
De
De
De
Do
Do
Dru
Em
En
Ep
9 Glossary Most entries have been taken from Boland et al. (1980)1, Eldridge et al. (1993)2, Doran et al. (1983)3, Willan (1985)4 and Hong et al. (1998)5. Absorption (of seed) Uptake of moisture until the seed comes into equilibrium with the moisture of the surrounding air. See also desorption and equilibrium relative humidity Areole (of seed) The area encompassed by the pleurogram. The differences between the areole and the remainder of the face may be slight differences in colour, surface texture or fracture lines(3) Aril (of seed appendages) A pulpy structure which grows from some part of the ovule or funicle after fertilisation and covers part or the whole of the seed(3) Bipinnate (of compound leaves) Twice pinnately divided; twice compound(3) Capsule Dry, usually many-seeded fruit composed of two or more fused carpels that split at maturity to release their seeds as in Eucalyptus(4) Carabiner A metal safety clip, used by climbers with ropes, which can be locked in the closed position as an insurance against accidental opening during climbing and fruit harvesting(4) Chaff In eucalypts, sterile particles derived from infertile or nonfertilised ovules (1) Cotyledon Seed leaf or primary leaf of the embryo(1) Deciduous Of leaves, bark, etc. falling regularly at the end of the growth period(1) Dehiscence Opening of the fruit by splitting along definite lines(1) Desorption (of seed) Loss of moisture from the seed until it comes into equilibrium with the moisture of the surrounding air. See also absorption and equilibrium relative humidity Dormancy (of seed) A resting or quiescent condition. In acacias dormancy is frequently imposed on a non-dormant embryo by the â&#x20AC;&#x2122;hardâ&#x20AC;&#x2122; seed coat which prevents water from reaching the embryo(3) Dormancy (embryo) Dormancy as a result of conditions within the embryo itself; inhibiting substances, incompletely developed embryo. Syn: internal dormancy(4) Drupe A stone-fruit such as a plum; the pericarp fleshy or leathery, containing a stone with one or more seed(4) Embryo The rudimentary plant formed within the seed. It consists of an axis bearing an apical meristem or plumule, radicle and one or more cotyledons(5) Endosperm The nutritive tissue contained in some seed in addition to the embryo; not present in eucalypts(1) Epigeal Germination in which the cotyledons are forced above the ground by the elongation of the hypocotyl as in Eucalyptus (4)
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
9-1
Equilibrium relative humidity Seed will desorb or absorb water until it reaches equilibrium moisture content with the relative humidity of the surrounding air. This relative humidity, where the moisture content of the seed is stable, is called the equilibrium relative humidity. Fermentation The process of chemical changes in organic substances caused by the catalytic action of a “ferment”, which may be an independent plant such as yeast or bacteria, or an enzyme. May be accompanied by the production of heat and of toxic substances, hence the fermentation of fleshy fruits may adversely affect the seeds which they contain(4)
Lan
Me
Mic
Mo
Follicle A dry dehiscent fruit formed from a single carpel, dehiscing along the ventral side only(4) Funicle = funiculus (of seed appendages) The ‘umbilical cord’ of the seed, attaching it to the pod. When detached from a mature seed near the seedcoat it leaves a scar (the hilum) (3) Germination Growth of the embryo in the seed until the emergence of the embryonic radicle through the seedcoat. In seed testing, the capacity of the embryo to emerge from the seedcoat with those essential structures which indicate a potential to produce normal plants(1) Germination capacity Proportion of a seed sample that has germinated normally in a specified test period, usually expressed as a percentage. Syn: Germination percentage. It should be noted that in some earlier literature the term “Germination Capacity” has been used to express the total of the seeds which germinate plus the ungerminated but sound seeds (on cutting test), as a percentage of the seeds sown(4) Germination energy That proportion of germination which has occurred up to the time of peak germination, or the time of maximum germination rate, or up to some pre-selected point, usually 7 test days. (The critical time of measurement can be chosen by several means) (4) Germinative capacity Percentage of seed that germinate during the whole of the germination test period(1) Hard seeds Seeds with thick and tough testas which delay water penetration and germination(3) Hypocotyl That part of the axis of a germinating embryo which is between the cotyledon and the radicle (4) Hypogeal (germination) Germination in which the cotyledons remain in the seed below the ground while the epicotyl elongates(4) Indehiscent (of fruit) Not opening at maturity
(3)
Intermediate seed storage behaviour A category of seed storage behaviour intermediate between those defined as orthodox and recalcitrant. Mature whole seeds are able to tolerate desiccation to seed moisture contents in equilibrium at 20°C with about 40-50% relative humidity. Further desiccation, however, often reduces viability and always results in more rapid deterioration in subsequent hermetic storage the more the seeds are dried below this value(5)
9-2
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
Nu
Ort
Ort
Pe
Ph
Ph
Ph
Plu
Plu
Po
Pre
Pro
Pu
quielalib-
the ast d of the
tral
g it car
nic rge ro-
ally ion ion lus eds
the me be
Land race A land race develops when exotic trees are introduced in a new environment: Genetic changes take place in the population of trees over one or more generations of selection by natural or human agencies; a land race of poor quality develops when the first planting was from a poorly adapted provenance or, worse, from the seeds of a single tree(2) Mesocarp Middle layer of the pericarp; the pulp of berries and drupes(4) Micropyle (of seed) In mature seeds, a plugged opening(3) Moisture content The amount of water present in a material eg. wood, soils or seeds. May be expressed in terms of weight of moisture as a % of the material’s oven-dry weight (“dry-weight basis”) or, preferably in the case of seeds and fruits, as a % of the material’s wet weight including water (“wet-weight” or “fresh-weight basis”) (4) Nut Dry, indehiscent, one-seeded fruit with a woody or leathery pericarp developing from an inferior compound ovary(4) Orthodox Term used to describe species of which the seeds can be dried down to a low moisture content of around 5% and successfully stored at low or sub-freezing temperatures for long periods(4) Orthodox seed storage behaviour Mature whole seeds not only survive considerable desiccation (to at least 5% moisture content) but their longevity in air-dry storage is increased in a predictable way by reduction in seed storage moisture content and temperature(5) Periodicity The tendency, in an individual, stand or species, to produce seed at more or less regular intervals of more than one year(4) Phenology (Study of) relations between seasonal climatic changes and periodic biological phenomena such as flowering, fruiting, leaf flushing and dormancy(4) Phenotype All characteristics of a plant, morphological, anatomical and physiological as determined by the interaction between genotype and environment(4)
the
Phyllode A leaf whose blade is much reduced or absent, and whose whole petiole and rhachis have assumed the functions of the whole leaf(3)
and
Plumule Primary bud of a plant embryo situated at the apex of the hypocotyl; portion of the seedling axis above the cotyledons, consisting of leaves and an epicotyl, which elongates to form the primary stem(4)
yle-
be-
ereds 0°C ces or-
Plus tree A tree appearing distinctly superior to the average on a similar site. The superior character(s) are specified as plus for volume, quality, disease resistance etc. (1) Pod A superior, one-celled, one- or many-seeded dehiscent fruit of two valves. Resembles the follicle in being dehiscent and formed from a single carpel but differs from it in dehiscing on both sides(4) Precuring The deliberate storage and slow air drying under shade of fruits and contained seeds in order to tender them more suitable for subsequent operations, eg. kiln drying, extraction and storage(4) Provenance The original geographic source of seed or propagules(1) Pure seed That component of a seedlot which consists of seeds of the designated
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
9-3
species. According to ISTA rules, it includes not only mature, undamaged seeds but also undersized, shrivelled, immature and germinated seeds provided they can be positively identified as the designated species, and pieces of seed resulting from breakage which are more than half their original size. Excludes seeds of other species, wings of coniferous seeds, seeds of coniferous or leguminous species with seedcoats entirely removed, broken seed particles less than half the original size and other matter such as stones, twigs and leaves(4) Purity Proportion of clean, intact seed of the designated species in a seedlot, usually expressed as a percentage by weight(4) Radicle The rudimentary root of the embryo(1) Recalcitrant seed storage behaviour Mature whole seeds that are unable to tolerate more than a limited amount of desiccation, for example to moisture contents in equilibrium at 20°C with about 96-98% relative humidity(5) Relative humidity (of air) Amount of water vapour present as a percentage of the maximum amount of water vapour air can contain at a given temperature Scarification Disruption of seed coats, usually by mechanical abrasion or by brief chemical treatment in a strong acid, to increase their permeability to water and gases, or to lower their mechanical resistance(4) Seed The dispersal or germination unit of a fertilised ovule(3) Seed orchard A special plantation of highly selected trees, isolated to minimise contamination with pollen from outside sources, and managed for maximum seed production(2) Seedlot An indefinite quantity of seed having uniform quality, produced at a specific location and collected from a single crop(1) Serotinous Fruit or cones that remain on the tree without opening for one or more years (eg. Allocasuarina verticillata) Squash test A simple, indirect test of viability, by which seeds are first allowed to imbibe water and are then squashed with a pair of forceps to reveal the condition of the embryo. The number of seeds appearing fresh and healthy per unit weight of seed plus chaff (in eucalypts) or per 100 (in larger seeds) provides a rough estimate of viability(4) Stratification A pre-germination treatment to break dormancy in seed and to promote rapid uniform germination; the seed are exposed to moisture at a temperature just above freezing point (1-5°C) for a specified time(1) Testa The outer coat of the seed; usually hard and tough, but may be soft in some species(4) Thresh To separate, by any mechanical means, eg. rubbing, shaking, trampling, stamping, beating or intermittent pressure, the grains of any cereal from the husks and straw, especially by beating with a flail. Applied also to the separation of other than cereal seeds from their fruits(4) Viable of seed, able to germinate(1)
9-4
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
Vig
Wo
eds can om pewith ize
Vigour Those seed properties which determine the potential for rapid, uniform emergence and development of normal seedlings under a wide range of field conditions(4) Working sample A reduced seed sample taken from the submitted sample in the laboratory, on which some test of seed quality is made(4)
ally
ers in
the
rief and
eed
ific
ore
imn of t of ate
ote ust
me
ng, sks her
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
9-5
10
Ba gen tion ton http sou 11
Be Se Dic http sou 11
Bo ent
Bo line Wa (ve
Bo Ge Sta 11
Bo tion
Bo tor ern fs.f
DF ate Hu
Do aca me
Eld and
10 Bibliography Baskin JB, Baskin CC (2003) ‘Classification, Biogeography and Phylogenetic Relationships of Seed Dormancy’ [Online]. In: ‘Seed Conservation: Turning science into practice’. (Eds RD Smith, JB Dickie, SH Linington, HW Pritchard, RJ Probert) (Royal Botanic Gardens: Kew). Available at http://www.kew.org/science-research-data/kew-in-depth/msbp/publications-data-resources/technical-resources/seed-conservation-science-practice/index.htm (verified 11 December 2013) Berjak P, Pammenter NW (2003) ‘Understanding and Handling Desiccation-Sensitive Seeds’ [Online]. In: ‘Seed Conservation: Turning science into practice’. (Eds RD Smith, JB Dickie, SH Linington, HW Pritchard, RJ Probert) (Royal Botanic Gardens: Kew). Available at http://www.kew.org/science-research-data/kew-in-depth/msbp/publications-data-resources/technical-resources/seed-conservation-science-practice/index.htm (verified 11 December 2013) Boland DJ, Brooker MIH, Turnbull JW (1980) ‘Eucalyptus Seed’. (Commonwealth Scientific and Industrial Research Organisation: Melbourne) Bonner FT (1974) ‘Seed testing’. In: ‘Seeds of Woody Plants in the United States’ [Online]. Agriculture Handbook No. 450. (Ed CS Schopmeyer) (Forestry Service. USDA: Washington DC). Available at https://archive.org/details/seedsofwoodyplan00fore (verified 11 December 2013) Bonner FT (1984) ‘Glossary of seed germination terms for tree seed workers’ [Online]. General Technical Report SO-49. (USDA Forest Service, Southern Forest Experiment Station: New Orleans, LA). Available at http://www.srs.fs.usda.gov/pubs/1235 (verified 11 December 2013) Bonner FT (1990) Storage of seeds: potential and limitations for germplasm conservation. Forest Ecology and Management 35, 35-43. Bonner FT, Vozzo JA, Elam WW, Land Jr SB (1994) ‘Tree Seed Technology, Instructor’s Manual’ [Online]. General Technical Report SO-106. (USDA Forest Service, Southern Forest Experiment Station: New Orleans, LA). Available at http://www.treesearch. fs.fed.us/pubs/68 (verified 11 December 2013) DFSC-IPGRI (1999) The project on handling and storage of recalcitrant and intermediate tropical forest tree seeds. Newsletter 5th April, 1999. (Danida Forest Seed Centre: Humlebaek, Denmark) Doran JC, Turnbull JW, Boland DJ, Gunn BV (1983) ‘Handbook of seeds of dry zone acacias. A guide for collecting, extracting, cleaning and storing the seed and for treatment to promote germination of dry-zone acacias’. (FAO: Rome) Eldridge KG, Davidson J, Van Wyk G, Harwood CE (1993) ‘Eucalypt Domestication and Breeding’. (Oxford University Press: Oxford)
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
10-1
Ellis RH, Hong TD, Roberts EH (1991) An intermediate category of seed storage behaviour? II. Effects of provenance, immaturity and imbibitions on desiccation-tolerance of coffee. Journal of Experimental Botany Vol. 42, 238, 653-657. FAO (1993) Ex situ storage of seeds, pollen and in vitro cultures of perennial woody plant species [Online]. FAO Forestry Paper 113 based on work of Wang BSP, Charest PJ, Downie B (FAO: Rome). Available at http://www.fao.org/documents/en/docrep.jsp (verified 11 December 2013) Gunn, BV. 2001. ‘Australian Tree Seed Centre Operations Manual’. (Commonwealth Scientific and Industrial Research Organisation: Canberra) Gunn B, Agiwa A, Bosimbi D, Brammall B, Jarua L, Uwamariya A (2004) ‘Seed Handling and Propagation of Papua New Guinea’s Tree Species’ . (Commonwealth Scientific and Industrial Research Organisation: Canberra) Harrington JF (1972) Seed Storage and Longevity. In ‘Seed Biology. Vol. III’. (Ed. TT Kozlowski) pp.145-245 (Academic Press Inc.: New York) Hong TD, Ellis RH (1996) A protocol to determine seed storage behaviour. In IPGRI Technical Bulletin No. 1. (Eds. JMM Engels, J Toll) (International Plant Genetic Resources Institute: Rome) Hong TD, Linington S, Ellis RH (1998) ‘Seed Storage Behaviour: a Compendium’ [Online]. Handbooks for Genebanks: No. 4. (International Plant Genetic Resources Institute: Rome). Available at http://www.bioversityinternational.org/e-library/publications/ detail/seed-storage-behavior-a-compendium/ (verified 11 December 2013)
Mu Se
NA tion
No tion
Po and
Po coa ma
Pro [On ie, http sou 11
Howcroft NHS (2002) The Balsa Manual. Techniques for Establishment & the Management of Balsa (Ochroma lagopus) Plantations in Papua New Guinea. ITTO East New Britain Balsa Industry Strengthening Project PD 7/99 REV.2(F). (Papua New Guinea National Forest Service: Keravat, Papua New Guinea)
Pro tur SH http sou 11
ISTA (2011) ‘International Rules for Seed Testing: Edition 2011/1’. (The International Seed Testing Association: Zurich).
Qu tific
Justice OL, Bass LN (1978) ‘Principles and practices of seed storage’. Agriculture Handbook No. 506. (Forestry Service, USDA: Washington DC)
Ra Pla
Köppen W (1923) ‘Die Klimate der Erde’. p. 31 (De Gruyter: Berlin)
Ra Ha nat lica
Langkamp PJ (1987) ‘Germination of Australian Native Plant Seed’. (Inkata Press: Melbourne). Lauridsen EB, Wellendorf H, Keiding H (1987) Evaluation of an international series of Gmelina provenance trials. Danida Research Report. (Danida Forest Seed Centre: Humlebaek, Denmark) Laurie MV (1974) Tree planting practices in African savannas. FAO Forestry Development Paper No. 19. (FAO: Rome). Marunda CT (1990) Effects of seed pretreatments on the development of Acacia auriculiformis and A. holosericea seedlings. In ‘Tropical Tree Seed Research’. Proceedings of an international workshop held at the Forestry Training Centre, Gympie, Qld, Australia, 21-24 August 1989. (Ed. JW Turnbull) ACIAR Proceedings No. 28, 33-36. (Australian Centre for International Research: Canberra)
10-2
Mc Lan
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
Ro 1, 4
Sa tion De Sc
Sh Pin
bence
ody est jsp
alth
anen-
TT
GRI Re-
Onstins/
geew nea
McDonald RC, Isbell RF, Speight JG, Walker J, Hopkins MS (1998) ‘Australian Soil and Land Survey Field Handbook (2nd edition)’. (Goanna Print: Canberra) Mueller-Dombois F, Fosberg FR (1998) ‘Vegetation of the Tropical Pacific Islands’. Series: Ecological Studies; v. 132. (Springer-Verlag: New York) NAS (1980) ‘Firewood Crops. Shrubs and Tree Species for Energy Production’. (National Academy of Sciences: Washington DC) Northcote KH, (1979) ‘A Factual Key for the Recognition of Australian Soils (4th edition)’. (Rellim Technical Publications: Glenside, South Australia). Poulsen KM, Parratt MJ, Gosling PG (Eds) (1998) ‘ISTA Tropical and Sub-tropical Tree and Shrub Seed Handbook’. (International Seed Testing Association: Zurich). Poulsen K, Stubsgaard F (1995) Three methods for mechanical scarification of hardcoated seed. Technical Note No. 27. (Danida Forest Seed Centre: Humlebaek, Denmark) Probert RJ (2003) Seed Viability under Ambient Conditions, and the Importance of Drying [Online]. In: ‘Seed Conservation: Turning science into practice’. (Eds RD Smith, JB Dickie, SH Linington, HW Pritchard, RJ Probert) (Royal Botanic Gardens: Kew). Available at http://www.kew.org/science-research-data/kew-in-depth/msbp/publications-data-resources/technical-resources/seed-conservation-science-practice/index.htm (verified 11 December 2013) Probert RJ, Keith RM, Adams (2003) Non-destructive Measurement of Seed Moisture. In: ‘Seed Conservation: Turning science into practice’. (Eds RD Smith, JB Dickie, SH Linington, HW Pritchard, RJ Probert) (Royal Botanic Gardens: Kew). Available at http://www.kew.org/science-research-data/kew-in-depth/msbp/publications-data-resources/technical-resources/seed-conservation-science-practice/index.htm (verified 11 December 2013)
nal
Quayle S, Gunn B (1998) ‘Tree Nursery Manual for Namibia’. (Commonwealth Scientific and Industrial Research Organisation: Canberra)
ure
Ralph M (1994) ‘Seed Collection of Australian Native Plants: For Revegetation, Tree Planting and Direct Seeding (2nd Edition)’. (Imprenta: Melbourne)
ss:
ies tre:
op-
auedQld, 36.
Rao NK, Hanson J, Dulloo ME, Ghosh K, Nowell D, Larinde M (2006) ‘Manual of Seed Handling in Genebanks’[Online]. Handbooks for Genebanks: No. 8. (Bioversity International: Rome, Italy). Available at http://www.bioversityinternational.org/e-library/publications/detail/manual-of-seed-handling-in-genebanks/ (verified 11 December 2013) Roberts EH (1973) Predicting the storage life of seed. Seed Science and Technology 1, 499-514. Sary H, Yameogo CS, Stubsgaard F (1993) The CO2 method to control insect infestation in tree seed. Technical Note No. 42. (Danida Forest Seed Centre: Humlebaek, Denmark) Schmidt L (2007) ‘Tropical Forest Seed’. (Springer-Verlag: Heidelberg) Shrestha KB, Shepherd KR, Turnbull JW, (1985) Controlled atmosphere storage for Pinus radiata seed. Commonwealth Forestry Review 64 (2), 141-150.
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
10-3
SPC (2009). Forest and Tree Genetic Resources Conservation, Management and Sustainable Use in Pacific Island Countries and Territories: Priorities, Strategies and Actions, 2007-2015. Forests and Trees Programme of the Land Resources Division. (Secretariat of the Pacific Community: Suva, Fiji) Stubsgaard F (1992) Seed Storage. Lecture Note No. C-9. (Danida Forest Seed Centre: Humlebaek, Denmark) Stubsgaard F (1997) Tree climbing for seed collection techniques and equipment. Technical Note No. 44. (Danida Forest Seed Centre: Humlebaek, Denmark) Stubsgaard F, Poulsen KM (1995) Seed moisture and drying principles. Lecture Note No. C-5. (Danida Forest Seed Centre: Humlebaek, Denmark) Thomsen K (2000) Handling of Desiccation and Temperature Sensitive Tree Seeds. DFSC Series of Technical Notes. TN56. (Danida Forest Seed Centre: Humlebaek, Denmark) Thomson, LAJ. 1995. Collecting woody perennials [Online]. In ‘Collecting Plant Genetic Diversity: Technical Guidelines’ (Eds. L Guarino, V Ramanantha Rao, R Reid) pp. 485-509 (International Plant Genetic Resources Institute: Rome). Original and updated versions available at http://cropgenebank.sgrp.cgiar.org/index.php?option=com_conte nt&view=article&id=390&Itemid=557 (verified 11 December 2013) Willan RL, (compiler) (1985). A guide to forest seed handling [Online]. FAO Forestry Paper 20/2. (Food and Agriculture Organisation: Rome). Available at http://www.fao. org/docrep/006/ad232e/ad232e00.htm (verified 11 December 2013) Williams ER, Gunn B, Reynolds D, Westcott M (1992) Germination tests for small collections of Acacia seed. Seed Science and Technology 20, 321-326. Yuan QZ, Old KM, Midgley SJ (1990) Investigations of Mycoflora and Pathology of Fungi Present on Stored Seeds of Australian Trees. In ‘Tropical Tree Seed Research’ Ed JW Turnbull). Proceedings of an international workshop held at the Forestry Training Centre, Gympie, Qld., Australia, 21-24 August 1989. ACIAR Proceedings No. 28. (Australian Centre for International Agricultural Research: Canberra)
10-4
PACIFIC ISLANDS TREE SEED CENTRE Manual of Operating Procedures for Seed Collection, Documentation, Treatment and Storage
and and on.
en-
ent.
ote
ds. ek,
Gepp. ted nte
stry ao.
col-
of châ&#x20AC;&#x2122; ain28.