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International Journal of Agricultural Science and Research (IJASR) ISSN 2250-0057 Vol. 3, Issue 3, Aug 2013, 1-12 © TJPRC Pvt. Ltd.

VARIATIONS IN LEAF DUST ACCUMULATION, FOLIAGE AND PIGMENT ATTRIBUTES IN FRUITING PLANT SPECIES EXPOSED TO PARTICULATE POLLUTION FROM MULTAN UZMA YOUNIS, TASVEER ZAHRA BOKHARI, SAEED AHMAD MALIK, SHAKIL AHMAD & RAMIZ RAJA Botany Division, Institute of Pure and Applied Biology, Bahauddin Zakariya University, Multan, Pakistan

ABSTRACT The dust accumulation capacity of fruiting plants was evaluated from eight different sites in and around Multan.

The impact of dust accumulation/deposition was observed via various foliage (leaf area, fresh and dry weights, stomatal clogging) and biochemical attributes (chlorophyll contents, carotenoids & ascorbic acid) from leaves of these species. The maximum dust accumulation/deposition was occurred in the plants growing at Road sides while, the minimum dust was found on plants growing at Bahauddin Zakariya University. Dust accumulation has caused a significant effect on almost all foliage and biochemical attributes of all the plant species. Foliage attributes of all the plant species showed species specific responses because in some plants a positive correlation was observed between foliage attributes and dust load but in other plants it is negative. Biochemical responses had shown an inconsistency as chlorophylls (b & total) contents decreased and ascorbic acid contents increased with an increase in dust accumulation/deposition in all the plant species, except chlorophyll a and carotenoid contents which give species specific response to dust pollution. A negative correlation was found between dust deposition and chlorophyll b and total contents. Whereas, accumulation of ascorbic acid was associated with a decline in pigment contents.

KEYWORDS: Dust Accumulation, Fruiting Plants, Foliage and Biochemical Attributes INTRODUCTION Dust is a general name for solid particles having diameters >500 µm but particles of 2.5 -10 µm in atmospheric are of great concern for health of local public (Borja-Aburto, et al., 1998; Beckett, et al., 1998). Dust particles arise from natural sources such as soil dust lifted up by wind and volcanic eruptions but sometimes dust also contains small amounts of pollen grains, human and animal hairs, stuff and paper fibers and many other materials (Kathleen Hess-Kosa, 2002).

Windblown dust is a common feature of arid ecosystems as soils remain dry and serve as a major source of small particulate matter (Sharifi, et al., 1997). In addition, agricultural activities and fast moving traffic also generate high dust concentrations (Leys, et al., 1998; Manins, et al., 2001). A report has been revealed that approximately 30 millions of tons of dust go into air all over the world (Van Jaarsveld 2008). Considerable attention is given to particulate matter pollution in the recent years because it caused severe health harms (Jafary, et al., 2007). Worldwide particularly in urban areas almost 600 million persons suffered from a variety of ailments due to dust pollution (Cacciola, et al., 2002). Accumulation and deposition of gaseous pollutants and particulate matter depends upon the vegetation type (Bunzl, et al., 1989; Fowler, et al., 1989). Vegetation makes contribution in reducing dust concentration in environment by acting as a sink for air pollutants. Due to surface characteristics of twigs, bark and foliage of the plants particulate matters are captured by them and remain there for extended time period. Generally exposed areas of a plant especially leaves act as constant absorbers for particulate matters (Samal and Santra, 2002). Thus air quality in urban/ arid areas can be improved by planting trees along road sides and agricultural lands (Beckett, et al., 2000; Freer-Smith, et al., 2005; Raupach, et al., 2001). However,

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dust accumulation capability of plants depends on their range of characteristics which include outside geometry, phyllotaxy and leaf attributes (cuticle and pubescence of leaves), tallness and canopy of plants.

On the other hand, morphology and internal structure of leaves is altered by heavy load of dust pollutants (Somashekar, et al., 1999; Gostin, 2009; Sukumaran, 2012). Particulate pollutants can cause many lethal effects on plants like stomatal clogging, reduced photosynthetic activity, leaf fall and death of tissues (Farooq, et al., 2000; Shrivastava and Joshi, 2002; Garg, et al., 2000). Physical or chemical changes take place due to the deposition of dust on aerial parts of plants (Grantz et al., 2003). Large proportion of stomata may cover by dust which reduces the rate of transpiration and rate of evaporative cooling (Sharifi et al., 1997). Leaf chlorosis also caused by dust pollution due to its effect on chlorophyll biosynthesis (Seyyednejad, et al., 2011). However, many plant types are able to stable their populations despite significant dust fall/deposition on their exposed surfaces because harmful effects of particulate matter are scavenged by carotenoids and ascorbic acid production which are the expression of non-enzymatic resistance in plants to numerous abiotic stresses (Prajapati and Tripathi, 2008). Multan (Plate-I) is the third largest city of the province Punjab, Pakistan. The city is famous for dust storm. Urbanization of the city particularly during the last decade had resulted in deforestation and significant increase in the number of automobiles has aggravated the problem of particulate matter pollution.

Plate-I: Map Showing Study Site i.e. Multan City Keeping in view the above situation, the present study aimed to evaluate the variation in dust accumulation capacity of fruiting plants because cultivated populations of these fruit species were present in a variety of habitats in Multan. The foremost objective of the study was to assess the role of foliage of the species in reducing particulate matter pollution. In addition, biometric and biochemical attributes were studied to suggest the possible mechanism of plants to combat stress induced by dust pollution.

MATERIALS AND METHODS Study Material Seven fruiting plant species (Ziziphus jujuba Mill., Punica granatum L., Morus alba L., Vitis vinifera L., Mangifera indica L. , Syzygium cumini (L.) Skeels, Grewia asiatica L.) were selected for study. All species have different morphology and phyllotaxy. The selection of these species was made because of their contrasting leaf characteristics which are represented in Table 1.

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Variations in Leaf Dust Accumulation, Foliage and Pigment Attributes in Fruiting Plant Species Exposed to Particulate Pollution from Multan

Table 1: Description of Fruiting Plants at Study Sites with their Leaf Characteristics Plant Species

Family

Leaf form

Phyllotaxy

Habit

Ziziphus jujuba Mill Punica granatum L. Morus alba L. Vitis vinifera L. Mangifera indica L. Syzygium cumini (L.) Skeels Grewia asiatica L.

Rhamnaceae Punicaceae Moraceae Vitaceae Anacardiaceae Myrtaceae Tiliaceae

Ovate to acute Oblong or obovate dentate, ovate to broad ovate orbicular, cordate, dissected Lanceolate to oblong Oblong Cordate to ovate

Alternate Opposite Alternate Opposite Alternate Serrate Alternate

Tree Shrub Tree Vine Tree Tree Shrub

Plant Height (m) 5-10 2-3 8-15 Girth 1.5 20 6-20 4-5

Study Area Selection Eight main areas of Multan city (map reference) were selected for sampling of plant tissues from cultivated population of two fruit species. Multan region was selected for this study because it is the third largest city of the Punjab province in Pakistan. These sites were selected on the basis of traffic density, dust concentration, urbanization and industrialization. The sampling was carried out during may-June, 2011 because weather remains dry during these two months and the frequency of incidence of dust storm, which is typical to the city, is greater during these months. The detail of these areas is summarized in Table 2 and site is marked on the map.

Table 2: Study Sites with Source of Dust in Environment Site/ Population I II III IV V VI VII VIII

Area

Source of Particulate Matter Pollution

Bahauddin Zakariya University Cricket Stadium area Pak Arab Fertilizers Industrial Estate Parks Multan Railway Station

Dust storm, wind erosion Barren land, dust storm, wind erosion Barren land & heavy loaded trucks Cement, Ceramic, and pesticides industries Barren land & wind blown dust & sand Barren land and fast moving trains, smoke from diesel of locomotives Busy road sides, fast moving automobiles/vehicles Plouging & Thrashing

Bosan Road Cultivated wheat field

Sampling Protocols for Dust and Leaves Fruiting plant species were selected at each sampling site and 20 leaves were randomly marked on each plant at 23 m height. The leaves were carefully removed from the branches and were wrapped in pre-weighed glazed papers then put in polythene bags and brought back to the laboratory. The wrapped leaves were carefully taken out from the bags, glazed paper was spread in a glass chamber and dust present on the leaves was cleaned using a fine brush. All the dust was collected on the glazed papers.

They were reweighed and the amount of dust collected from each leaf was recorded. Leaf area (cm 2) measurements were taken using leaf area meter (Delta-T-Device area meter) and capacity of dust accumulation was calculated following Prajapati and Tripathi (2008) as under: W = (w2 – w1)/a Where W is dust content (g m–2), w1 is initial weight of glazed paper, w2 is final weight of glazed paper with dust, and “a” is total area of the leaf (m2).

The leaves samples were then used for various foliage and pigment parameters. Fresh weights were taken and leaf material was oven dried at 70o C for 72 h for dry weight measurements using a digital balance (AC 220-V, Tokyo, Japan)

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Uzma Younis, Tasveer Zahra Bokhari, Saeed Ahmad Malik, Shakil Ahmad & Ramiz Raja

Chlorophyll Determination Chlorophyll was extracted in 80% acetone from leaves and the amount of chlorophyll a, b, and total, were quantified spectrophotometrically (UV-1900 Japan) following Arnon (1949).

The carotenoids were calculated by the following Krik and Allen formulae (1965).

Ascorbic Acid For determination of ascorbic acid 5 g of fresh leaf sample was homogenized in 20ml extracting solution (0.75 g sodium salt of EDTA and 5 g oxalic acid dissolved in 100ml distilled water). Homogenized leaf sample was centrifuged for 15 min at 628.2 rads–1. Then 1ml homogenate was mixed thoroughly with 5ml Dichlorophenol indophenol (DCPIP) (0.02 g L–1) with constant shaking. Optical density of pink colored solution was determined at 520nm (Es) with the help of a spectrophotometer. After this one drop of 1% ascorbic acid solution was added to decolorize the pink solution, then optical density of turbid solution (Et) was determined at the same wave length. A calibration curve was plotted using ascorbic acid standard (Camlab U.K.). Keller and Schwager (1977) were followed to calculate Ascorbic acid content. The formula is as follows: Ascorbic Acid (mg/g F.W)

=

(Eo-Es-Et) V ×100 100 W

where, Eo, Es, and Et are optical densities of blank sample, plant sample, and sample with ascorbic acid, respectively, V = volume of extract, and W = leaf sample weight (g).

RESULTS The percentage dust accumulation on fruiting plants under study at different sites of Multan, Pakistan is presented in Figure 1. It is evident from the Figure that all the plants showed higher dust deposition on leaves at road sides followed

by Railway station, cultivated wheat field, Pak Arab Fertilizer - Industrial Estate, parks, cricket stadium area and lowest at Bahauddin Zakariya University. It showed that Z. jujuba has highest and G. asiatica has lowest capacity of accumulating dust. The capacity of dust accumulation among the different species was Z jujuba > P. granatum > M. indica > V. vinifera > S. cumini > M.alba > G. asiatica.

Figure 2 A represented that higher dust accumulation had affected foliage attributes of all the fruiting plants and leaf area of P. granatum, M. indica, V. vinifera,, S. cumini, and G. asiatica increased with increasing dust accumulation whereas leaf area of M.alba and Z. jujuba decreased with increasing dust accumulation because it had negative correlation with dust load on leaves of these plants (Table 3). Whereas, significant increased of fresh and dry weights of leaves of Z. jujuba and G. asiatica was observed with increasing dust accumulation.

In most of the plants under study maximum leaf fresh and dry weights were observed in the plants growing at road sides with more dust accumulation and minimum were observed in the plants growing in Bahauddin Zakariya University with minimum dust accumulation. (Figure 2. B & C). The Z. jujuba and G. asiatica had the maximum leaf fresh weight (0.975 g & 1.74 g) at more dust accumulation site. Although, a gradual reduction of leaf fresh weight (0.180 g & 1.39 g) was observed in the plants of Z. jujuba and G. asiatica growing at lowest dust accumulation site (Figure 1 B). The leaf dry weight also exhibited a similar trend as shown by fresh weight and a noticeable inhibition in dry weight was observed with increasing dust accumulation where leaves of most of the plants (Punica granatum, Morus alba, Vitis vinifera, Mangifera indica) produced less dry weight as compared with leaves of the plants growing at minimum dust accumulation site (Figure 1 C).

Variations in Leaf Dust Accumulation, Foliage and Pigment Attributes in Fruiting Plant Species Exposed to Particulate Pollution from Multan

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Stomatal opening is greatly affected by dust load in the environment. As represented in Figure 1 D that increased in dust accumulation cause more and more stomatal clogging. Increased in dust accumulation on leaves enhance stomatal clogging and more stomatal clogging is observed at Road sites whereas less clogging was observed at Bahauddin Zakariya University site which had minimum dust load. M. indica had more clogged stomatal percentage than V.vinifera. A profound increased in chlorophyll a contents with increasing dust accumulation was observed in V. vinifera, Z. jujuba and P. granatum (Figure 2 A). The chlorophyll b and total chlorophyll contents were comparatively higher in the plants growing at lowest dust accumulation site but dust load induced a profound decline in chlorophyll b and total chlorophyll contents (Figure 2 B, C). The pigments declined progressively with increasing dust accumulation. A dramatic decline in carotenoids content was also observed in some plants (V. vinifera, G.asiatica & Z. jujuba), whereas, other plants showed increased in carotenoids contents with increasing dust accumulation. (Figure 2 D).

The responses of ascorbic acid contents of selected plant species under study are presented in Figure III. It is evident from the figures that there is an increase in ascorbic acid content in leaves with a decrease in chlorophyll content.

The concentration of ascorbic acid in all plant species was highest at more dust accumulation site (Road sides). ANOVA (significant at P < 0.05) showed that there is a significant variation in pigment content between plant species and collection sites for both chlorophyll and ascorbic acid (Figure 3).

The study showed changes in the levels of foliage attributes (Leaf area, leaf fresh and dry weight, Stomatal clogging) and of pigment (Chlorophyll a, b, total, carotenoids and ascorbic acid) content in different fruiting plant specie exposed to atmospheric dust fall. Chlorophyll b and total contents decreased and ascorbic acid contents increased with increased dust deposition on leaves. The Pearson correlation coefficient values of dust accumulation with foliage attributes and pigment contents in plant species is presented in Table 3. The table clearly shows highly significant negative correlations between dust load and chlorophyll contents (b, total) and highly significant positive correlations between dust load and ascorbic acid content. But foliage, carotenoid contents and chlorophyll a showed inconsistency in correlation results because species specific responses were observed with dust load.

DISCUSSIONS The current work illustrates the significant variation in dust accumulation in fruiting plants at different locations inside and around Multan. Different kinds of factors, such as leaf uniqueness i.e. form and size, orientation, surface texture, occurrence/lack of leaf hairs, petioles length etc., air current and its speed, climatic conditions and anthropogenic actions affect the dust interception/accumulation capacity of different plants. Z. jujuba and P. granatum owing to their short petioles had shown the utmost dust accumulation capacity as compared with G. asiatica which had long- leaves stalks. In addition, degree of pubescence and large surface area allow the species to capture more dust particles which is an important sign of particulate pollution. The influence of foliage characteristics on dust accumulation has also been studied by many workers (Somashekar et al., 1999; Garg et al., 2000). Morphological attributes such as cuticular waxes, raised epidermal cell margins, trichomes, ledges of stomata and epicuticular lignin and waxes give roughness to leaves (Pal et al., 2002). Plants growing at road sides have additional chances of dust accumulation which may be due to high dust concentration which consequences by the vehicles action and capturing dust, with a mild wind. The plants of B.Z.University locality have minimum dust accumulation due to more cultivated area and less bare land. Hence, Z. jujuba can one of the chief contributors for clean-up up dust pollution from the surroundings.

Farmer (1993) declared that adequate amount of dust particles may form a smother layer on foliage, reduced light capturing ability of plants, thus lower photosynthetic rates, clogged stomata, increased foliage temperature, effect fruit set,

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Uzma Younis, Tasveer Zahra Bokhari, Saeed Ahmad Malik, Shakil Ahmad & Ramiz Raja

leaf development, pollen growth, reduced plant growth, caused chlorosis and necrosis of leaf, bark peeling and the same results were seen in the present study that dust accumulation had significant effect on leaf area of almost all the fruiting plants because there was positive correlation between dust accumulation and leaf area of some plants while, negative correlation between dust accumulation and leaf area of others was observed. Hence, it is accomplished that dust accumulation on leaves has differential effect on leaf area of selected fruiting plant species. Same is the case with leaf biomass.

Number of ecological factors such as soil, water and air are responsible for control of development and growth of plants (Katiyar and Dubey, 2000). These factors may cause variation in leaf pigments (chlorophyll a, b, total, carotenoids and ascorbic acid). Dust particles bear numerous polycyclic hydrocarbon and metals with them which inhibits the manufacturing of enzymes necessary for chlorophyll synthesis and results in reduction of chlorophyll molecules with increasing dust accumulation on plants (Prajapati and Tripathi, 2008). Photosynthetic activity and gaseous exchange in plants is affected by pressure exerted due to dust pollution which block stomata, (Hope, et al., 1991; Keller and Lamprecht, 1995; Anthony, 2001). Same case was observed with fruiting plants because more and more stomatal clogging was observed with increasing dust accumulation. From the present study it is cleared that both chlorophyll (a, b, total) and ascorbic acid content showed different responses to dust accumulation. Dust accumulation caused alkalinity in leaves by chemicals dissolution of dust particles in cell sap which results in chlorophyll damaged. To accommodate these stresses leaves ascorbic acid contents increased which decreased the leaf senescence (Garg and Kapoor, 1972). Several scientists demonstrated that chlorophyll content of dust polluted leaves is lower than that of control leaves (Somashekar, et al., 1999; Mandal and Mukherji, 2000; Samal and Santra, 2002). The study clearly suggested that plantation of Z. jujuba and G. asiatica in dusty areas can control particulate pollution which may cause hazardous consequences for human health.

SUGGESTIONS 

Red colour marked is for deletion

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Blue colour marked for addition

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Red colour bold mark with under line is for clarification

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26. Manins, P., Allan, R., Beer, T., Fraser, P., Holper, P., Suppiah, R., & Walsh, K. (2001). Atmosphere. Australia State of the Environment Rep. 2001 (Theme Rep.). CSIRO Publ., Melbourne.

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28. Prajapati, S. K., & Tripathi, B. D. (2008). Seasonal Variation of Leaf Dust Accumulation and Pigment Content in Plant Species Exposed to Urban Particulates Pollution. J Environ Qual. 37, 865-870 29. Raupach, M.R., Woods, N., Dorr, G., Leys, J. F., & Cleugh, H. A. (2001). The entrapment of particles by windbreaks. Atmos. Environ. 35, 3373–3383.

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APPENDICES

Figure 1: Dust Accumulation per Day in Fruiting Plants under Observation (g m–2 Leaf Area)

Variations in Leaf Dust Accumulation, Foliage and Pigment Attributes in Fruiting Plant Species Exposed to Particulate Pollution from Multan

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Figure 2: A. Leaf Area, B. Leaf Fresh Weight, C. Leaf Dry Weight, D. Percentage Stomatal Clogging of Zizyphus Jajuba Jujuba, Punica Granatum, Morus Alba, Vitis Vinifera, Mangifera Indica, Syzygium Cumini, Grewia Asiatica Collected from Different Sites of Multan, Pakistan

Variations in Leaf Dust Accumulation, Foliage and Pigment Attributes in Fruiting Plant Species Exposed to Particulate Pollution from Multan

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Figure 3: A. Chlorophyll a, b & Total) Contents, B. Chlorophyll b, C. Total Chlorophyll, Carotenoid Contents of Zizyphus Jajuba Jujuba, Punica Granatum, Morus Alba, Vitis Vinifera, Mangifera Indica, Syzygium Cumini, Grewia Asiatica Collected from Different Sites of Multan, Pakistan

Figure 4: Ascorbic Acid Contents of Zizyphus Jajuba Jujuba, Punica Granatum, Morus Alba, Vitis Vinifera, Mangifera Indica, Syzygium Cumini, Grewia Asiatica Collected from Different Sites of Multan, Pakistan

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Table 3: Correlation of Dust Load with Foliage and Biochemical Attributes of Selected Fruiting Plants under Study Attributes

Biometric Attributes

Biochemical Attributes

Leaf Area Clogged stomata Leaf Fresh Weight Leaf Dry Weight Chlorophyll a Chlorophyll b Total chlorophyll Carotenoids Ascorbic Acid

V. vinifera 0.514 0.732 -0.336 -0.216 0.668 -0.462 -0.657 -0.186 0.863

S. cumini 0.619 0.84 -0.405 0.261 -0.634 -0.732 -0.756 0.133 0.845

Plant Species Z. jajuba M. M.alba jujuba indica -0.434 0.315 -0.829 0.9 0.935 0.541 0.5 -0.764 -0.6 0.286 -0.581 -0.65 0.083 -0.507 -0.57 -0.219 -0.375 -0.71 -0.536 -0.745 -0.67 -0.3 0.049 0.561 0.775 0.932 0.955

G. asiatica 0.49 0.392 0.405 0.286 -0.225 -0.369 -0.375 -0.811 0.781

P. granatum 0.007 0.567 -0.068 -0.24 0.834 -0.891 -0.681 0.563 0.883