International Journal of Bio-Technology and Research (IJBTR) ISSN 2249-6858 Vol. 3, Issue 1, Mar 2013, 11-16 TJPRC Pvt.

. Ltd.

COMPARATIVE EFFICACY OF NEMATICIDES WITH VAM FUNGI AGAINST HETERODERA AVENAE INFECTING WHEAT
AMEETA SHARMA Department of Biotechnology, The IIS University, Jaipur, Rajasthan, India

ABSTRACT
The comparative efficacy of locally available systemic nematicides with VAM fungi was evaluated against Heterodera avenae infecting Triticum aestivum, L. var.wh-147 under pot trials. An experiment was set up with nematicides like Carbofuran, Aldicarb, Phorate and Rugby along with urea application on wheat. Glomus fasciculatum, Glomus mosseae, Gigaspora gigantea and Gigaspora margarita were applied in 4 g dosage to the treated plants, individually and in combination. Finding of this study established that Glomus species, Carbofuran and Phorate were more potent and compatible to mitigate the nematode infection as well as endomycorrhizae enhanced wheat growth and grain yield.

KEYWORDS: Wheat, Heterodera Avenae, VAM Fungi, Nematicides, Biocontrol INTRODUCTION

The success of mycorrhizal evolution has been attributed to the role that mycorrhizal fungi play in the capture of nutrients from the soil of all ecosystems (Brundrett, M. C. 2002). The VAM fungal partner played a larger role in protecting the root from a pathogen in the fine-rooted plant compared to the coarse-rooted plant. However, in addition, our data also support the hypothesis that the identity of the AM fungi influences the ability of the mycorrhiza to reduce pathogen infection as previously demonstrated by Maherali and Klironomos (2007). The host-parasite relationship and interaction of nematode and VAM fungi has been comprehensively reviewed (Smith, 1988; Arya and Saxena, 2009). The interaction between VAM and plant parasitic nematodes has been studies by many workers (Bagyaraj et al., 1979 Hussey and Rencadori, 1982). Systemic nematicides and fumigants have been reported to control H.avenae by earlier workers (Yadav and Singh, 1975; Johnson 1985a,b,) Mycorrhizae are common form of symbiosis between plants and fungi (Harley and smith, 1983). The interaction between VAM and plant parasitic nematodes has been studies by many workers (Bagyaraj et al., 1979 Hussey and Ren cadori, 1982). Cyst nematode, Heterodera avenae dominates in the wheat growing fields of Rajasthan state, India causing 'Molya' disease of wheat. In the native language 'Molya' means deformed and hampers wheat production which is the major cereal of Rajasthan state. Preceding survey done earlier unveiled, association of VAM with rhizosphere of wheat. Frequency distribution comprehended the predominance of G.fasciculatum, inhabiting in most of the wheat cultivated fields, with greatest 92.34 percent occurrence. Percent loss in the surveyed areas ranged from 15 to 45 percent. Therefore an experiment was undertaken to study the interaction of VAM and H.avenae on wheat, as shaped by the time of inoculation.

MATERIALS AND METHODS

Fifteen local fields were surveyed and soil samples were collected. Samples were than proceed to determine the endomycorrhizal infection, cyst of H.avenae and other soil attributes. VAM was isolated and pure cultures were maintaned on onion, Cenchrus ciliaris and wheat. VAM colonization was a more conspicuous on onion. The identification was made by using standard key by N.C.Schenck and Yvohne Pe'rez, (1988). Spores were isolated from the soil by wet sieving and

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decanting technique (Gerdemann and Nicolson, 1963) and cysts were collected from the soil and roots of wheat by simple sieving and floatation process. The VAM was applied in 4 g dosage according to no. of spores present/g of soil. The treatments were replicated five times and untreated pot served as control. Mycorrhizal inoculums comprised of chlamydospores, azygospores, infective fungal hyphae and fungal roots, which were then placed just below the seeds during sowing. They were surface sterilized by 0.1% Hgcl2 and then sown in 15cm earthen pots containing steamed soil. One week old seedlings then were inoculated with 1000 freshly hatched juveniles of H.avenae, by pouring larval suspension. The pots were arranged in randomized complete block design. Most of the organophosphates and dithiocarbamates are much less phyto-toxic and were applied at the time of seed sowing. All these chemicals were used in granular form and were applied in the following treatments: G. fasciculatum (4g) + Nematode. G. mosseae (4g) + Nematode. G. gigantea (4g) + Nematode. G. margarita (4g) + Nematode. G. fasciculatum(1g) + G. mosseae(1g) + G. margarita(1g) + G. gigantea (1g) + Nematode. Carbofuran @ 1.5 kg a i / ha + Nematode. Phorate @ 2 kg a i / ha + Nematode. Aldicarb @ 1.5 kg a i / ha + Nematode. Rugby @ 2 kg a i / ha + Nematode. Urea @ 110 kg a i / ha + Nematode. Un-inoculated control test plant (C). Nematode alone inoculated plant (N).

Following 90 days, at the time of annihilation of experiment, the plants were uprooted and diverse growth parameters were noted in terms of shoot-root length, fresh and dry weight of plants, ear length and its fresh and dry weight, total cyst per plant, total spores per gram of soil, percent VAM colonization. Data obtained were statistically analyzed. The significance of difference between length and weight of host, number of cyst and other quantitative data were calculated from original figures by analysis of variance. Critical difference at 5% and 1% level of significance were calculated for significant comparison. Root infection was assessed by using staining technique (Phillips and Hyman, 1970) and slide method (Giovanneti and Mosse, 1980).

RESULTS AND DISCUSSIONS

The results showed that mycorrhizal fungi increased the plant growth significantly in comparison to nonmycorrhizal plants. A significant reduction in disease incidence and better plant growth and yield were also obtained by the treatment with nematicides. All VAM species which were tested have capacity of root colonization on wheat but when comparison was made, Glomus species showed to have greater potential to mitigate the nematode infection in comparison to Gigaspora species. Carbofuran and phorate gave compatible results in reducing H.avenae population per plant. The maximum weight of 21.86g was observed in control, followed by 19.72g in carbofuran, 18.94g in phorate, 18.67g in

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rugby, 17.66g in aldicarb, 16.93g in GF+GM+GiG+GiM+N, 16.90g in GF+N and 16.71g in GM+N, with minimum of 12.3g in N alone treated plants and ranged between 16.39g and 15.71g in other treatments. Dry shoot weight was maximum in control (5.21g) and minimum in N alone (1.30g). Per cent VAM colonization varied from 30.2% in GF+GM+GiG+GiM+N to 63% in GF+N treatment. It was 55% in GiM+N and 34.65% in GM+N whereas in GiG +N it was 60% (Table : 1). As evidenced from fore mentioned observations that all nematicides and VAM fungi alone and in combination reduced cyst multiplication per plant significantly as compared to untreated N alone inoculated plants systemic nematicides and fumigants have been reported to control H.avenae as reported by earlier workers( Bhatti and Dalal, 1972; Brown and rovira, 1975; Steele, 1976; Sharma, P. and Trivedi, P.C. 2010). Total nematode population was per plant was reduced with nematicides and VAM application. Less cyst were found in carbofuran (8), followed by rugby (15). GF+N (21.00) was more effective than GiM+N (27.00). Total cyst content was not fixed and it varied from one treatment to other. Eggs/cyst was maximum in GF+N(228.65), followed by 216.00 in N alone treated plants, 210.00 in GiG+N, 197.29 in Rugby, 197.23 in GM+N, 189.53 in GiM+N and 178.28 in GiG+N. The plants heavily colonized with mycorrhizal fungi are able to grow well instead of presence of damaging levels of nematodes and in addition VAM has reported to enhance overall growth and vigor of various crop and vegetable plants (Smith, 1987; Thomas et al., 2000; Chawla et al., 2009). Sethi and Kaul (1987) showed similar results that cyst content of the newly formed nematode were almost similar except in nematicides and VAM treated plants, where they were less. Tolerance to attack by plant nematodes is the most frequently reported benefit of VAM fungi (Smith, 1987; Ganwar and Dutt, 2009). Mycorrhizal fungi benefit the plant by increasing the absorption of nutrients and water and by protecting the root from soil-borne diseases (Jalali and Chand, 1990). The present study showed that VAM fungi cause an increase in the wheat plant growth and resistance to H.avenae infection, development and population. Conclusively it could be inferred from the study that treated plants showed better growth, enhance ear formation and resistance whereas untreated plants showed less vigour, chlorosis of leaves and stunted growth. Thus in natural habitat, endomycorrhizae may have significant tortuous impacts on plant and nematode population dynamics and ecosystem level. On the whole, VAM should be exploited as a bio-control agent for controlling H. avenae as it is eco-friendly and gives very promising results. Table 1: Comparative Effect of Nematicides and VAM Fungi on Heterodera Avenae Infecting Wheat Fresh Dry Ear Ear Weight Total % VAM Total Weight (g) Weight (g) Length (g) Fresh Cysts ColonizS.No Treatments Eggs/Cyst Shoot Root Shoot Root (cm) Dry /Plant ation 2 3 4 5 6 7 8 9 10 11 12 1 1 GF+N 16.9 5.48 2.12 0.70 12.9 4.59 0.64 21.00 228.65 63.00 (4.62) (15.14) (7.98) 2 GM+N 16.71 5.08 2.03 0.63 12.83 4.47 0.63 19.33 197.23 34.65 (4.42) (14.07) (5.77) 3 GiG+N 15.97 4.51 1.99 0.62 12.46 4.43 0.59 23.66 210.00 60.00 (4.92) (14.51) (7.74) 4 GiM+N 15.71 4.39 1.85 0.58 11.76 4.43 0.57 27.00 189.53 55.00 (5.26) (13.79) (7.46) 5 GF + GM+ 16.93 5.50 2.34 0.73 13.53 4.67 0.69 23.00 143.70 30.2 GiG+GiM+N (4.89) (11.99) (5.57) 6 Carbofuran +N 19.72 5.94 3.55 0.88 14.70 5.48 0.81 8.00 153.00 0.00 (2.96) (12.30) (1.00) 7 Aldicarb +N 17.66 5.78 2.78 0.74 13.86 4.82 0.71 14.00 178.28 0.00 (3.81) (13.35) (1.00) 8 Phorate+N 18.94 6.42 3.02 0.81 14.73 5.15 0.76 7.00 137.67 0.00 (2.72) (11.76) (1.00)

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Ameeta Sharma

S.No 9 10 11 12

Treatments Rugby+N Urea+N Nematode Control SEM CD at 5% CD at 1%

Fresh Weight (g) Shoot Root 18.67 6.13 16.39 12.30 21.86 0.96 1.99 2.71 4.06 1.98 6.96 0.25 0.52 0.71

Table 1 Contd., Dry Ear Ear weight Weight (g) Length (g) Fresh Shoot Root (cm) Dry 2.97 0.77 14.26 4.93 0.74 1.77 1.30 5.21 0.14 0.3 0.41 0.48 0.27 1.00 0.03 0.07 0.10 11.46 8.13 15.16 0.54 1.12 1.52 4.17 0.85 5.76 0.17 0.36 0.50 0.51 0.19 0.87 0.04 0.09 0.12

Total Cysts /Plant 15.00 (3.98) 127.00 (11.31) 264.6 (16.29) 0.00 (1.00) 1.97 4.08 5.56

Total Eggs/Cyst 201.33 (14.18) 197.29 (14.07) 216.00 (14.71) 0.00 (1.00) 0.75 1.57 2.14

% VAM Colonization 0.00 (1.00) 0.00 (1.00) 0.00 (1.00) 0.00 (1.00) 0.57 1.19 1.61

Values are means of five replications. Figures in parenthesis are

n + 1 transformed values. N=Nematode, GF=Glomus

fasciculatum, GM=Glomus mosseae, GiG=Gigaspora gigantea, GiM=Gigaspora margarita.

ACKNOWLEDGEMENTS
We thank the Indian Council of Agricultural Research (I.C.A.R.), New Delhi for financial assistance and Head, Department of Botany, U.O.R, Jaipur for providing necessary facilities.

REFERENCES
1. Arya, R. & Saxena, S. K. (2009). Efficacy of rhizosphere fungi against root-knot nematode (Meloidogyne incognita) and Rhizoctonia solani in tomato crop. Indian J Nematol. 39 (1)113-6. 2. Bagyaraj, D.I., Manjunath, A. & Reddy, D.D.R. (1979). Interaction of vescicular arbuscular-mycorrhiza with root-knot nematode in tomato. Plant and Soil. 51: 397-403. 3. Bhatti, D.S. & Dalal, M.R. (1972). Yield responses to wheat to nematicidal treatment in a soil highly infested with wheat cyst nematode. HAUJ. Res. 2:231-232. 4. 5. Brundrett, M. C. (2002). Coevolution of roots and mycorrhizas of land plants. New Phytologist 154(2) 275304. Chawla A. K., Raja G., Bharathidasan Kumari B.D.R. & Ramachandran, A. (2009). Influence of VAM fungi and microbial inoculants on growth, nutrients, and biochemical constituents in Jatropha curcas L. Indian Journal of Plant Physiology, 14 (2)181-185. 6. Gangwar, M. & Dutt, D. ( 2009). Effect of Azorhizobium and VAM inoculation on fertilizer economy and yield of maize (Zea mays.L ). Journal of Research Punjab Agricultural University (India).46(1-2) 55-59. 7. Gerdemann, J.W. & Nicolson, T.H. (1963). Spores of mycorrhizal Endogone species extracted from soil by wet seiving and decanting. Trans. Br. Mycol. Soc. 46: 235-244. 8. Giovannetti, M., & Mosse, B. (1980). An evaluation of techniques for measuring Vescicular-Arbuscular Mycorrhizal infection in roots. New phytologist. 84:489-500. 9. Harley, J.L., & Smith, S.E. (1983). Mycorrhizal Symbiosis. Academic Press, New York. 368p.

10. Hussey, R.S., & Roncadori, R.W. (1982). Vescicular Arbuscular Mycorrhiza my limit nematode activity and improve plant growth. Plant Disease. 66:9-14. 11. Jalali, B.L. & Chand, H. (1990). Current trends in mycorrhizal research. Proc of the national conference on Mycorrhiza, HAU, Hisar, 210.

Comparative Efficacy of Nematicides with Vam Fungi against Heterodera Avenae Infecting Wheat

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12. Johnson, AW. (1975a). The role of nematicides in nematode management. In: An advance treatise on Meloidogyne. Vol:1 Biology and control(Eds- Sasser, JN and carter CC) NC State university, USDA. 283-301. 13. Johnson, AW. (1975b). Specific crop rotation effects combined with cultural practices and nematicides. In: An advance treatise on Meloidogyne. Vol:1 Biology and control(Eds- Sasser, JN and carter CC) NC State university, USDA. 283-301. 14. Maherali, H. & Klironomos, J.N. (2007). Influence of Phylogeny on fungal community assembly and ecosystem functioning. Science, 316(5832) 17461748. 15. Phillips, J.M. & Haymann, D.H. (1970). Improved Procedure for clearing roots and staining parasite and vescicular-arbuscular mycorrhizal fungi for assesment of infection. Transactions of the British Mycological Society. 55:158-161. 16. Schenck, N.C. & Y.Perez. (1988). Manual for identification of VA mycorrhizal fungi. Second edition. Gainehville Florida. University of Florida. 241. 17. Sharma, P. & Trivedi, P.C. (2010). Microbes in the management of plant diseases. In Plant disease and its management (ed. P.C. Trivadi) Pointer Publisher Jaipur. 318. 18. Smith, G. S. & Kaplan, D. T. (1988). Influence of mycorrhizal fungus, phosphorus and burrowing nematode interactions on rough lemon seedlings. Journal of nematology, 20 (1): 49-52. 19. Smith, G. S. (1987). Interaction of nematodes with mycorrhizal fungi. In vistas of nematology (eds-Veech, J.A.Dickson, D.W.) Published by Society of nematologist. Hayattville, Maryland, 292-300. 20. Steele, A. E. (1976). Effect of Ozime carbamates and organophosphates on development of H.schachtii on sugarbeet. J of Nematol. 8:137-141. 21. Thomas Forage, Andrea Muehlchen, Clemens Hackenberg, Gerry Neilsen & Thierry Vrain, (2000). VAM fungi increase nematode resistance, growth, and nutrient uptake of .eld grown apple trees. Agriculture and Agri-Food Canada, Pacific Agri-Food Research Centre, Summerland, BC, VOH 1ZO, Canada. Presented at the 25th International Nematology Symposium, Herzliya, Israel, 2-7 April. 22. Yadav, B.S. & Singh, H. (1975). Effect of certain chemicals for the control of Molya disease of wheat caused by Heterodera avenae. Indian journal of Mycology and plant pathology. 5(1): 17.