Issuu on Google+

International Journal of Agricultural Science and Research (IJASR) ISSN 2250-0057 Vol. 3, Issue 3, Aug 2013, 91-98 Š TJPRC Pvt. Ltd.


ABSTRACT Nematicidal activity of extracts from plants was assayed against Meloidogyne graminicola. In laboratory assayas extracts from Neem (Azadirachta indica), Bael (Aegle marmelos), Jatrofa (Jatropha curcas), Eucalyptus (Eucalyptus globus), Sahjan (Moringa oleifera), Ber (Ziziphus mauritiana), Sarifa (Annona reticulate), Congress grass (Parthenium argentatum) were most effective in killing the nematode. In vitro studies 25%, 50% and 100% concentrations of leaf extracts significantly reduced second stage juvenile of mortality after 24 and 48 (hrs.). The plant leaf extracts of Bael (88.53%) and Neem` (80.31%) exhibited highly promising mortality 80-88% after 48 (hrs.) exposure. In pot experiment the Neem, Bael, Jatropha, Eucalyptus and Congress grass leaf extracxts were found significantly increased Shoot length, Root length (cm.) and Root weight (gm.) and reduced root gall in different concentration 100% and 50%. The 100% plant leaf extracts of Neem and Bael showed highest significantly increased plant growth parameters of shoot length 30.23 and 29.50 (cm.), root length 54.00 and 51.33 (cm.), root weight 0.66 and 0.43 (gm.) and highest significantly reduction of root gall 22.66 and 35.33 as compared to treatment (T1 - Nematode alone). The study organic amendment which will act as an asset to control environmental pollution.

KEYWORDS: Meloidogyne graminicola, Botanical Extracts, Rice INTRODUCTION Rice (Oryza sativa L.) is the world’s most important food crop of Asian origin. It is an indispensable source of calorie for almost half of the world population with in Asia Nematodes are small unsegmented worms live in water, soil, plants and animals. Plant parasitic nematodes are mostly microscopic. It causes significant damage for almost all crops. Global crop loss caused by plant parasitic nematodes is more than $100 billion annually [1]. Root knot nematodes, Meloidogyne spp, are the major plant parasitic nematodes attacking most of the crops. In UAE it is one of the main pests in vegetables and orchards. It is a major problem in green houses and nurseries. Many date palm trees are also infected with root knot nematodes. Many plants are known to have nematicidal properties which may be utilized as organic amendments or biopestcides. Many scientists have carried out the research on plant extracts for the management of root knot nematodes.The essential oil of A. marmelos showed remarkable brine shrimp lethality.Antimicrobial and antifungal activity of the leaf oil was negligible. Most of the observed biological activity was apparently due to the relatively high level of limonene (64.1%) in the essential of the leaf essential oil from A. marmelos [4].

MATERIALS AND METHODS Culture of Meloidogyne graminicola Root knot nematode Meloidogyne graminicola was maintained in rice plants in the bucket filled in the infested soil inside the glasshouse and from the field infected plants were also uprooted, carefully washed in running tap water and


Mukesh Dongre & Sobita Simon

egg- masses and J2 larvae were collected in to Petri dishes containing distilled water and inoculated to the bucket for mass culturing of Meloidogyne graminicola. Preparation of Botanical Extracts The leaves of different selected plants viz. Neem (Azadirachta indica), Eucalyptus (Eucalyptus globus), Ber (Ziziphus mauritiana), Sarifa (Annona reticulate), Sahjan (Moringa oleifera), Jatropha (Jatropha curcas), Bael (Aegle marmelos) and Congress grass (Parthenium argentatum). Leaves of 25 gram each were washed under running tap water and added 100 ml distilled water and grinded in blender for 3 minutes. The mixture was allowed to stand 3 h. filtered through Whatman filter paper then the extracts of all leaves kept for centrifugation in centrifuge machine at 1000 rpm for 10-15 minute. The supernatant was removed for further research trail. Screenings of Eight Botanicals against Meloidogyne graminicola Larval Mortality (In Vitro) The evaluation was carried out in counting Petri dishes. There were nine treatments and replicated three times. The Petri dishes with distilled water was taken as control. Freshly hatched 100 J 2 of M. graminicola were suspended in 100%, 50% and 25% of each leaf extracts. All the Petri dishes were kept at ambient temperature 25 0 (±) C. in BOD incubator. After 24 and 48 hrs. of incubation, all dead and alive 2nd stage juveniles (J 2) were counted with the aid of counting dish under stereomicroscope. The dead juveniles attained the shape of straight line and the mortality was ensured by touching the juvenile with a fine needle. The ratio of dead nematodes/number of total nematodes expressed the percentage mortality. Larval Mortality was Calculated by Following Formula [5] Recorded Number of larvae killed Mortality (%) = …………………………. × 100 Total No. of larvae Evaluation of Botanical Leaf Extracts on Root-Knot Gall Formation and Growth in Seedling Stage of Rice (In vivo) Approximately 250 rice seeds were washed well with 0.5 mg l-1 potassium permanganate and then rinsed thoroughly with distilled water. They were equally spread wetted filter papers. Seeds were maintained until they are germinated in the laboratory. After germinated the seeds were sown in thirty nine small pots. One week germination in the pots 5 seedlings were allowed to remain in each pot and others were removed and 2 ml. of nematode J2 suspension was added (approximately 500 J 2) each/pots of the following treatments including control (with out leaf extracts) were given separately into three containers and the treatments were replicated three times. After 48 hour of inoculation with Meloidogyne graminicola. The botanical leaf extracts was added @ 15 ml. into each inoculated pot. Nematicides carbofuran 3G @ 2 kg. a.i./hac was amended to the 2 days before inoculation of the soil. After 30 days all the inoculated pots were dipotted pots and observation were recorded for plant growth parameters (plant height, root length and weight of root) and number of root gall from each pot. Statistical Analysis The data from the two experiment were subjected to analysis of variance (ANOVA) and standard deviation and critical differences (CD) at 0.05were calculated.

Efficacy of Certain Botanical Extracts in the Management of Meloidogyne graminicola of Rice


Plate 1: Effects of Plant Extracts on Growth Parameters (Plant Height, Root Weight, Length of Root) and Root Gall. T0 (Plant Alone), T1 (Nematode Alone), T2 (Carbofuran 3G), T3 (Neem), T4 (Bael), T5 (Jatropha), T6 (Congress Grass), T7 (Eucalyptus)

Plate 2: Effects of Plant Extracts on Growth Parameters (Plant Height, Root Weight, Length of Root) and Root Gall. T0 (Plant Alone), T1 (Nematode Alone), T2 (Carbofuran 3G), T3 (Neem), T4 (Bael), T5 (Jatropha), T6 (Congress Grass), T7 (Eucalyptus)


Mukesh Dongre & Sobita Simon

RESULTS AND DISCUSSIONS Screenings of Eight Botanicals against Meloidogyne graminicola Larval Mortality (In Vitro) Percentage of M. graminicola viable J2 was decreased with increased in exposure time from 24 to 48 hrs. in each concentration. All the treatments significantly increased the percentage of mortality at 24 and 48 hrs. of exposure in different plant extracts concentration (100%, 50% and 25%) as compared to control (Nematode alone). At 48 hrs. (100% concentration) the maximum mortality percent was found in Bael (Aegle marmelos) 88.53% as compared with Neem (Azadirachta indica), Jatropha (Jatropha curcas), Eucalyptus (Eucalyptus globus), Sahjan (Moringa oleifera), Sarifa (Annona reticulate), Ber (Ziziphus mauritiana), and Congress grass (Parthenium argentatum) and control (Nematode alone) 2.09%. Whereas both the treatments Bael and Neem was found non significantly increased to each other but they are significantly increased mortality percent from themselves. At 48 hrs. (50% and 25%concentration) was also found increased the maximum mortality percent of M. graminicola juvenile (J2) respectively (70.59% and 43.60%) as compared to others. Using plant extracts in controlling plant parasitic nematodes has shown by several authors [4]. The essential oil of A. marmelos showed remarkable brine shrimp lethality. Antimicrobial and antifungal activity of the leaf oil was negligible. Most of the observed biological activity was apparently due to the relatively high level of limonene (64.1%) in the essential of the leaf essential oil from A. marmelos. Earlier antimicrobial studies using essential oils from A. marmelos. [6] and [7] reported that antifungal components of essential oil from A. marmelos. [8] also reported The toxicity of the leaf essential oil to the nematode C. elegans is comparable to Cymbopogon martinii essential oil. [9] also reported Limonene has been shown to be relatively inactive against the root-knot nematode, Meloidogyne incognita. Many essential oils have relatively high LC50 values against this nematode [9,10]. Other related study the broad insecticidal activity against termites, fruit flies, and fire ants undoubtedly also results from the limonene, a compound known to be toxic to insects, including termites [11] and weevils [12]. The current study was initiated to mortality percent explore the composition of the essential oil in leaves of A. marmelos plants and to broaden the biological activity profile of this understudied taxon. Table 1: Effect of Mean Mortality Percent at Different Concentration on J2 of Meloidogyne graminicola S. N.

Treatment Name

T0 T1 T2 T3 T4 T5 T6 T7 T8

Distilled water Neem (Azadirachta indica) Eucalyptus (Eucalyptus globus) Ber (Ziziphus mauritiana) Sarifa (Annona reticulate) Sajan (Moringa oleifera) Jatropha (Jatropha curcas) Congress Grass (P.argentatum) Bael (Aegle marmelos) Overall Mean F - test S. Ed (Âą) C. D. (P = 0.05%)

100% Concentration 24 hrs 48 hrs 0 2.09 27.57 54.18 51.55 80.31 32.44 49.24 42.06 50.42 26.28 41.07 26.93 42.22 23.35 37.57 66.78 88.53 296.96 445.63 S S 5.936 7.479 12.472 15.71

50% Concentration 24 hrs 48 hrs 0 2.67 22.17 37.01 35.58 68.82 18.51 23.74 31.42 46.34 21.06 35.37 16.12 24.25 25.61 44.89 37.72 70.59 208.19 353.68 S S 2.83 4.201 5.95 8.82

25% Concentration 24 hrs 48 hrs 0 1.83 22.46 27.95 33.70 41.83 16.06 20.50 27.05 33.84 19.63 24.19 13.59 16.46 17.70 21.26 33.73 43.60 183.92 231.46 S S 2.54 3.66 5.34 7.69

Evaluation of Leaf Extracts on Root-Knot Gall Formation and Growth in Seedling Stage of Rice (In vivo) The root-knot nematode multiplied well without plant extracts in treated rice plant (T 1 control). All botanical extracts amended in the soil suppressed the development of M. graminicola root galls, size of galls and of nematode population density. There were increased in the growth variable shoot length, root length (cm.) and weight of root (gm.) in the all plant extracts treated seedlings at doses of 100% and 50%. The maximum number root galls was reduced by


Efficacy of Certain Botanical Extracts in the Management of Meloidogyne graminicola of Rice

Azadirachta indica (22.66) followed by Aegle marmelos (35.33), Carbofuron 3G (38.66), Eucalyptus globus (43.66), Jatropha curcas (51.33), Parthenium argentatum (56.33) and over control Nematode alone (114.66) control and increased the length of shoot, root (cm.) and root weight (gm.) were found increased significantly in all the plant extracts treatments as compared with control (nematode alone). Our results are agreement with the reported by various scientist [2] found that neem cake and Rakshak gold (neem-based product) were effective against hatched eggs of Meloidogyne incognita. [3] Who reported that extract of neem seed was more effective against juvenile of M. javanica than bark and leaf. Pot experiment with all the extracts gave significant suppression in root galling and population of the nematode. [13] observed that aqueous neem leaf extracts found effective root-knot nematode Meloidogyne incognita in laboratory and greenhouse condition. [14] Reported 62% and 75% larval mortality at 100% concentration of root extracts of neem. [15] also recorded that the application of neem products as soil amendments significantly reduced the number of plant parasitic nematode before subsequent cropping. [16] observed that the most mortality percent of M. incognita (J2) in leaf extracts of neem (Azadirachta indica).

Figure 1: Effect of Botanicals on Plant Height (cm.) in 100% and 50% Concentration in Seedling Stage on Rice (In vivo)

Figure 2: Effect of Botanicals on Length of Root (cm.) in 100% and 50% Concentration in Seedling Stage of Rice (In vivo) Table 2: Effect of Botanicals on Mean of Growth Parameter and Root-Gall in 100% and 50% Concentration in Seedling Stage

S. N.


T0 T1

Plant alone Nematode alone Carbofuron + J2 Neem(Azadirachta indica) Bael (Aegle marmelos)

T2 T3 T4

Shoot Length(cm.) Concen Concent tration ration (100%) (50%) Mean Mean 22.96 22.96 16.16 16.16 17.16 17.16

Root Length(cm.) Concen Concen tration tration (100%) (50%) Mean Mean 55.50 55.50 37.66 37.66 47.00 47.00

Root Weigth(gm.) Concent Concent ration ration (100%) (50%) Mean Mean 0.6 0.6 0.16 0.16 0.33 0.33

Root Galls Concent Concentr ration ation (100%) (50%) Mean Mean 0 0 114.66 114.66 38.66 38.66


















Mukesh Dongre & Sobita Simon

Table 2: Contd., T5 T6


Jatropha (Jatropha curcas) Congress grass (Parthenium argentatum) Eucalyptus (Eucalyptus globus) Overall Mean F- test S. Ed. (Âą) C. D. (P = 0.05)

























21.85 S 1.97 4.1

19.38 S 1.7 3.61

46.64 S 5.43 11.51

42.14 S 3.74 7.93

0.392 S 0.125 0.266

0.317 S 0.102 0.217

38.91 S 9.82 20.83

45.16 S 9.56 20.28

Figure 3: Effect of Botanicals on Weight of Root (Cm.) in 100% and 50% Concentration in Seedling Stage of Rice (In Vivo)

Figure 4: Effect of Botanicals on Mean of Root Knot in 100% and 50% Concentration in Seedling of Rice Root (In Vivo)


Khan, I. A., Sayed, M., Shaukat, S. S. and Handoo, Z. A. (2008). Efficacy of four plant extracts on nematodes associated with papaya in Sindh, Pakistan. Nematologia Mediterranea., 36: 93 - 98.


Randhawa, N., Sakhuja, P. K. and Singh, I. (2001). Management of root knot nematode Meloidogyne incognita in tomato with organic amendment. Plant Disease Research, 16: 274-276.


Yasmin, L., Rashid, M. H., Uddin, M. N., Hossain, M. S., Hossain, M. E. and Ahmed, M. U. (2003) Use of neem extract in controlling root knot nematode (Meloidogyne javanica) of sweet gourd. Pakistan Journal of Plant Pathology, 2(3): 161-168.

Efficacy of Certain Botanical Extracts in the Management of Meloidogyne graminicola of Rice



Satyal, P., Woods, K. E., Dosoky, N. S., Neupane, S. and Setzer, W. N. (2012). Biological activities and volatile constituents of aegle marmelos (L.) CorrĂŞa from Nepal. Journal of Medicinally Active Plants 1(3): 114122.


Ahmad, M. S., Mukhtar, T. and Ahmad, R. (2004). Some studies on the control of citrus nematode (Tylenchulus semipenetrans) by leaf extract of three plants and their effect on growth variable. Asian Journal of Plant Science, 3(5): 544-548.


Rana, B. K., Singh, U. P. and Taneja, V. (1997). Antifungal activity and kinetics of inhibition by essential oil isolated from leaves of Aegle marmelos. J. Ethnopharmacol. 57: 29-34.


Balakumar, S., S. Rajan, T. Thirunalasundari, and S. Jeeva. (2011). Antifungal activity of Aegle marmelos (L.) Correa (Rutaceae) leaf extract on dermatophytes. Asian Pac. J. Trop. Med. 1: 309-312.


Kumaran, A. M., Souza, P. D, Agarwal, A., Bokkolla, R. M. and Balasubramaniam. M. (2003). Geraniol, the putative anthelmintic principle of Cymbopogon martinii. Phytother. Res. 17: 957.


Ntalli, N. G., Ferrari, F., Giannakou, I. and Menkissoglu-Spiroudi U. (2010). Phytochemistry and nematocidal activity of the essential oils from 8 Greek Lamiaceae aromatic plants and 13 terpene components. J. Agric. Food Chem. 58: 7856-7863.

10. Ntalli, N. G., Ferrari, F., Giannakou, I. and Menkissoglu-Spiroudi, U. (2011). Synergistic and antagonistic interactions of terpenes against Meloidogyne incognita and the nematocidal activity of essential oils from seven plants indigenous to Greece. Pest Manag. Sci. 67: 341-351. 11. Raina, A., Bland, J., Doolittle, M., Lax, A., Boopathy, R. and Folkins, M. (2007). Effect of orange oil extract on the Formosan subterranean termite (Isoptera: Rhinotermitidae). J. Econ. Entomol. 100:880-885. 12. Papachristos, D. P., Karamonoli, K. I., Stamopoulos, D. C. and Menkissoglu-Spiroudi, U. (2004). The relationship between the chemical composition of three essential oils and their insecticidal activity against Acanthosceloides objectus (Say). Pest Manag. Sci. 60: 514-520. 13. Kaur, H., Kaur, H. and Rishi, P. (2012). Effect of neem leaf extract on the mobility, egg hatch and infestation rate of Meloidogyne incognita on tomato under greenhouse condition. Advance in Life Science, 1(2): 141-144. 14. Adegbite, A. A. and Adesiyan, S. O. (2005). Root extractsofplants to control root-knot nematode on edible soyabean. World J. Agri. Sci., 1(1): 18-21. 15. Javed, N., Gowen, S. R., Fyaz, S. and Khan, M. M. (2008). Effect of neem formulation applied as soil drenching on the development of root-knot nematode Meloidogyne javanica on roots of tomato. Pakistan Journal of Botany, 40(2):905-910. 16. Chedekal, A. N. (2013). Effect of four leaf extracts on egg hatching and juvenile mortality of root knot nematode

Meloidogyne incognita. International journal of adv anced life science 6(1):68-74.

11 efficacy of certain full