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1 Available online at Anusha et al Int. J. Pure App. Biosci. 6 (1): (2018) ISSN: DOI: ISSN: Int. J. Pure App. Biosci. 6 (1): (2018) Research Article Evaluation of Different Methods of Inoculations of Bioinoculants With Reference to Paecilomyces (Thom) Sams Against Meloidogyne Incognita in Tamato Anusha * B. G., Shripad Kulkarni and Lingaraju, S. Dept of Plant Pathology College of agriculture science Dharwad University of Agriculture Science Dharwad *Corresponding Author abgindi@gmail.com Received: Revised: Accepted: ABSTRACT In vivo studies were conducted by various methods such as seed treatment, seedling dip method and soil application. Seed treatment with 6 g/kg resulted in more than 82 per cent germination. Shoot and root were d with enhanced vigour index of after seven. It also d root and shoot by 125 and per cent the compared to other treatments. Tomato seedling were free from root knot infestation when seedling were dipped in P. and Lecanicillium lecaniii at 10 g/lre concentration and reduced 100 per cent RKN index in both the treatments. However, root knot index was 4 in 0-5 scale in indicating the presence of sufficient inoculums to cause the disease. Key words: P., Lecanicillium lecaniii, Vegetable, Crop INTRODUCTION Tomato (Lycopersicum esculantum L.) is one of the most popular vegetable crops grown in the world, next to potato. It is used as a fresh vegetable and processed and canned as a paste, juice, sauce, powder or as a whole 5. The ripe fruits are good source of vitamin A, B and C which add wide varieties of colour and flavour to the food. Recently, it started gaining more medicinal value because of the antioxidant property because of ascorbic acid and lycopene content. Hence tomatoes are called as poor man s apple. Tomatoes offer significant nutritional advantages, including providing a significant source of dietary lycopene, β-carotene, carotenoids, vitamin C, potassium, fiber, color, flavor and antioxidant properties in a low energy dense food 7. Several human studies indicated a relationship between a high intake of tomato products and a decreased risk of several types of cancer, atherosclerosis and cardiovascular diseases 9. Recently, this crop is recognized as a model for plant-pathogen interactions 2. Many factors operate in successful cultivation as well as marketing of quality tomatoes of which diseases play an important role. Cite this article: Anusha, B.G., Kulkarni, S. and Lingaraju, S., Evaluation of different methods of inoculations of bioinoculants with reference to Paecilomyces (Thom) sams against meloidogyne incognita in tamato, Int. J. Pure App. Biosci. 6(1): (2018). doi: Copyright Jan.-Feb., 2018; IJPAB 599

2 Several diseases appear on tomato caused by complete destruction of the crop. Infection on fungi, bacteria, viruses, nematodes and a biotic older plants results in slight effects or they factors. Among the nematode diseases in may reduce yield considerably. tomato, root knot nematode (Meloidogyne There are numerous micro organisms species), sting nematode (Belonolaimus that prey or parasitize on nematodes. They fall longicaudatus), stubby root nematode into two broad categories- such as predators (Paratrichodorus and Trichodorus spp.) and and parasites. Apart from that opportunistic lesion nematode (Pratylenchus sp) are fungus are soil fungi capable of colonizing important. Root knot nematode Meloidogyne nematode reproductive structures causing incognita is one of the world s most disease to females, cysts and eggs. Most catastrophic diseases resulting in loss at common opportunistic species belong to the different growth stages of tomato. Root knot genera Cylindrocarpon, Exophiala, Fusariun, nematodes are distributed throughout the Gliocladium, Paecilomyces and Verticillium. world, especially in areas with warm or hot Biological agents developed climates and short or mild winter. And they on the basis of mycelium and spores of fungi appear in nursery, open field, poly house and belonging to the genus Paecilomyces Pers. even in green house. ExFr., under phylum Ascomycota, class Root knot nematode (M. incognita) is Eurotiomycetes Order Eurotioales has gained the most dominant species accounting for 64 immense importance since last four to five per cent of total population which is widely decades due to its biological ability prevalent inflicting serious loss to tomato fruit against nematodes. yield. Root knot nematodes are reported to Ability of bio agent to capture cause losses ranging from 15 to 60 per cent in nematodes depends on development of many vegetable crops such as brinjal, okra, structures by the fungal mycelia. The french bean and cowpea. Apart from M. predacious fungus forms hyphal structures, incognita, other major nematode species such as adhesive hyphal nets, knobs, branches causing diseases include M. javanica, M. and constricting or non constricting rings, in arenaria and M. hapla. which nematodes are captured mechanically or They attack more than 2000 species of through adhesion. Fungal hyphae then plants, infecting almost all cultivated plants penetrate into nematode and utilize its content and reported to reduce world crop production as a food. The nematode parasitic fungi on the by five per cent in individual fields loss may other hand infect nematodes with their spores be even higher. In general, the vegetables and which either adhere to the surface of pulses are the good hosts whereas, cereal crops nematodes or are swallowed by them. Ultimate are considered to be the poor hosts of this result of nematode infection in any way is nematode. Root knot nematodes are a serious always the death of the host. problem on vegetables, such as brinjal, okra, Paecilomyces is a common tomato, cucurbits, potato and chilli etc. These saprophytic, filamentous fungus. It has been diseases can be managed by cultural, physical, isolated from a wide range of habitats chemical and biological methods. Among including cultivated and uncultivated soils these methods biological is eco from forests, grassland, deserts, estuarine friendly and cheapest method. sediments and sewage sludge. It has been Root knot nematodes devitalize root found in nematode eggs and occasionally from tips and cause the formation of swellings on females of root knot and cyst nematodes. In roots. These effects not only deprive plants of addition, it has been frequently detected in the nutrients but also disfigure and reduce the rhizosphere of many crops. These species can market value of many root crops. When grow at a wide range of temperatures from 8 C susceptible plants are infected at the seedling to 30 C with optimal growth in the range of 20 stage, losses are heavy and may result in to 25 C. It has a wide ph tolerance and can Copyright Jan.-Feb., 2018; IJPAB 600

3 grow on a variety of substrates. P. has shown promising results as a bio agent to the growth of destructive root knot nematodes. Looking into the severity of root knot nematode infestation on various crops and farmers inclination towards bio agents for the management of nematode diseases, present study has been taken up with specific objectives mentioned above. MATERIAL AND METHOD Present investigations were carried out in kharif season during at the Main Agriccultural Research Station (MARS), University of Agriccultural Sciences, and Dharwad. Laboratory experiments were conducted at the Institute of Organic Farming, College of Agricculture, University of Agriccultural Sciences, Dharwad, and Karnataka. Various studies were undertaken with the objectives such as, isolation of Paecilomyces sp. and maintenance of pure culture, studies on mass production of Paecilomyces and their shelf life, in vitro and in vivo evaluation of talc based formulation of the bioagent Paecilomyces against root knot nematode. Details of the materials used and the methodology adopted during the course of investigation are presented in this chapter. General procedure Glassware and Cleaning For all the laboratory studies Corning and Borosil glassware were used. The glass ware were kept submerged night in the cleaning solution prepared by dissolving 60 g of potassium dichromate (K 2 Cr 2 O 7 ) and 60 ml of concentrated sulphuric acid (H 2 SO 4 ) in one litre of distilled water. Then, they were washed with vim powder followed by rinsing several times in running tap water and finally used when needed for laboratory studies. Sterilization All the glassware used in the studies were sterilized in an autoclave at 1.1 kg cm 2 pressure for 20 min. and then dried in a hot air oven at 60 C. Media were sterilized at 1.1 kg/cm² pressure for 15 min. Collection of cultures and maintenance of Meloidogyne incognita nematodes Root knot infected tomato plants were collected from the farmer s fields during the survey in northern Karnataka in polythene bags and kept in the freezer. Root portion was carefully removed from the soil and washed gently under running tap water. Egg masses were picked and kept for hatching in watch glass with water. After hours, hatched juveniles were used to inoculate tomato grown in sterilized soil:sand (1:1) mixture in greenhouse. These plants served as culture plants. After completion of 3-4 generations of the nematode, the plants were depotted carefully. The root system was washed free of soil, the knots containing egg masses were used to get inoculum of the nematode for further studies. Extraction of nematodes Soil samples of 200 g was washed thoroughly and processed using combined Cobb s sieving and Bauermann s funnel method as given. The root knot nematode and other plant parasitic nematodes present in the suspension were identified to genus level by observing different morpho anatomical characters. Their number present in the suspension was determined by taking the average number of nematodes present in five different one mililitre aliquot of nematode suspension. POT CULTURE STUDIES THROUGH SEED TREATMENT Seed treatment is the common and general method followed by farmers to manage the diseases. Procedure: Earthen pots (15 cm diameter) were taken and disinfected with 4% formalin and filled with sterilized soil and known no of nematodes (400 second stage juveniles / pots) were added to each pot. Tomato seeds were coated uniformly with 0.1, 0.2 and 0.4% Paecilomyces commercial formulation powder (containing 2 x 10 6 cfu/g) and dried them for 15 min under shade. Copyright Jan.-Feb., 2018; IJPAB 601

4 Similar procedure was followed to treat the seeds with other organisms and Carbofuran. Ten seeds of tomato were sown in each pot and number of seeds germinated was recorded after 5-7. Bioagents and their concentrations used for invivo studies Bioagents Concentation (g/kg seed) Paecilomyces 2 P. 4 P. 6 Trichoderma harzianum 2 T. harzianum 4 Lecanicillium lecanii 2 Carbofuran (treated check) 3 Control (untreated) ---- Bioefficacy studies with bioagents through seedling dip method This is another method commonly followed in the nursery field to reduce the disease intensity. Procedure Earthen pots (15 cm diameter) were taken and disinfected with 4% formalin and filled with sterilised soil and known no of nematodes (400 second stage juveniles) were added. Seedlings were dipped in solution of different organisms and concentrations for 15 minutes. Five seedlings were planted in each pot and three replications were maintained. Following are the different treatments and their concentrations used to study their bioefficacy through seedling dip method. Bioagents Concentration (g/l of water) Paecilomyces 2 P. 5 P. 10 Trichoderma harzianum 5 T. harzianum 10 Lecanicillium lecanii 10 Observations on fresh shoot and root mass, root knot index and number of plants with knots etc. were recorded 60 after transplanting and converted into percentage Bioefficacy studies with bioagent as a soil application Earthen pots (15 cm diameter) were taken and disinfected with 4% formalin and filled with sterilised soil and known no of nematodes (400 second stage juveniles) were added to each pot. The different bio agents, manures and their combinations mentioned below were added to soil. Ten seeds of tomato were sown in each pot and number of seeds germinated was recorded. Treatment Concentration (g/pot ) Paecilomyces alone 0.25 P. alone 0.50 P. alone 1.00 Neemcake alone 25 Neemcake 25g + T Neemcake 25g+T Neemcake 25g +T FYM alone 50 FYM 50g + T FYM 50g + T FYM 50g + T Carbofuran(treated check) 0.3 Control - Observation on per centgermination, vigour index, fresh shoot and root mass, root knot index and number of plants with knots were recorded 90 after germination and converted into percentage. Root knot index was calculated by following the scale suggested by Taylor and Sasser as given below. Gall index (Taylor and Sasser, 1978) (Fig. 1) Grade Description 0 No galls or egg masses/plant 1 1 to 2 galls or egg masses/plant 2 3 to 10 galls or egg masses/plant 3 11 to 30 galls or eggmasses/plant 4 31 to 100 galls or eggmasses/plant More than 100 galls or 5 eggmasses/plant Pseudomonas fluorescence 10 RESULTS Carbofuran (treated check) 10 The results of the present investigation on Control --- Mass production of Paecilomyces and bioefficacy against root knot nematode Copyright Jan.-Feb., 2018; IJPAB 602

5 caused by Meloidogyne incognita infecting Seed treatment with 3g /kg also tomato conducted at Department of Plant resulted in enhanced growth of shoot, root Pathology, College of Agriculture, UAS, (36.00 and 9.13cm) ultimately resulting in Dharwad and laboratory experiments fresh and dry of and 7.47 g conducted at Institute of Organic farming respectively with least root knot index of 1.00 UAS, Dharwad during kharif are compared to (5.00) presented here under. Treatment involving 6 Pot culture studies g/kg seeds was most effective in enhancing the Evaluation of Paecilomyces on shoot and root by and per growth and root knot development in cent, respectively. It was also effective in tomato through seed treatment increasing the fresh root and dry root In pot culture studies, various P. by and per cent campared isolates were tested against root knot with other treatment. nematodes of tomato with different Evaluation of Paecilomyces on concentrations through seed treatment. Their growth and rootknot nematode results of the bioefficacy test through seed development through seedling dip treatment are similar to that of roll towel Observations on effect of P. on method and there was significant difference growth and root knot development in tomato among all the treatments with respect to shoot, through seedling dip method indicate that there root, fresh and dry of roots. Seed was significant difference among the treatment with 6g/kg seed treatments with respect to root and showed maximum germination (82 %) and shoot and fresh and dry. Treatment involving P. and T. produced more shoot and root with harzianum showed enhanced root and enhanced fresh and dry of seedlings produced more shoot with higher fresh and dry compared to after 90 (Table 1 and of seedlings compared to other Plate 1). treatments and after 60 (Table 2 Maximum shoot and root and Plate 2). (43.33 cm) and (11.80 cm) was recorded when Maximum shoot and root of tomato seeds treated with P. 6g/kg cm and cm and maximum fresh after 90. It recorded maximum fresh and and dry of g and g were dry root of and 8.30 g recorded with P. 10g/l treatment. and respectively. It also d shoot and root root and shoot of and by and per cent per cent the were recorded respectively. Tomato seeds at in tomato seedlings when treated with P. treated with P. 4g/kg resulted in 10g /lre after 60 followed by shoot and root of cm and tomato seedling treated with Lecanicillium cm along with enhanced fresh and dry root 10g /lre with shoot and root of and 5.90 g respectively of cm and cm and fresh and dry compared to shoot and root of of 21.00g and 9.23g respectively. In and 5.38 cm in. However, fresh and Untreated shoot and root dry of and g were recorded were and 5.41 cm whereas registered in untreated (Table 6) fresh and dry were g and g Among various treatments tested P. 6g/kg and 4g/kg Treatment involving seeds were most effective against Meloidogyne 10g/l was most effective in enhancing the with root knot index of 1.00 in 0-5 scale and it shoot and root by and per was reduced by per centin both the cent, respectively. It was also effective in treatment the. increasing the fresh root and dry root Copyright Jan.-Feb., 2018; IJPAB 603

6 by and per cent the There was 100 per cent reduction of RKN. The disease was completely managed index recorded in seedling when treated with in seedlings when treated with FYM + P. and Neem cake + P. 10 g/l and Lecanicillium 10 g/l compared to. compared to. Effect of Paecilomyces on growth DISCUSSION and root knot nematode development in Tomato (Solanum lycopersicum Mill) is one of tomato through soil application the most important crop and out ranks all other Among various bioagents tested P. vegetables except the potato crop in popularity was most effective either alone or in and value in the world. combination with different organic The modern intensive Agricultural amendments at different concentrations. There practices have considerably raised the output was significant difference among all the but created problems like land and treatments tested (Plate 3 and 4). environmental degradation and pollution. The maximum shoot and root Besides, this contamination of food products (47.00 and cm) with an in root and pesticidal residue in food has caused much and shoot of and per cent panic among the consumers and also the the were recorded in tomato producers. Hence, it is necessary to find out plants applied with FYM + P. (50 g + innovative and suitable alternative technology 1.00 g). Maximum fresh and dry ( like biological of pests and diseases and 9.70 g ) were also recorded in this Recently, Paecilomyces based treatment without any root knot incidence different formulations are used for the followed by treatment tomato seedlings were of nematode diseases, which is economical, treated with neem cake + P. ( 25 g + ecofriendly and sustainable in the long run g ) which recorded shoot and root Among the different bioagents used in the of and cm and fresh and dry tomato ecosystem, P. is extensively of g and 8.30 g respectively. used as seed dressing bioagent against many Shoot and root of and 5.72cm and soil borne diseases, however, its usage against fresh and dry of and 4.03 g were foliar diseases is limited. recorded in untreated with an root knot Among several nematodes of index of 4.00 in 0-5 scale (Table 3). economic importance, root knot nematodes are Treatment involving neem cake + P. most widely studied and are commonly 25 g g resulted in root and involved in synergistic interactions with wilt shoot of 8.25 and cm respectively inducing fungi and bacteria. enhancing the fresh and dry up to Microbial of soil borne plant and 4.20 g, respectively. Treatment involving diseases is economically viable and 50 g showed root and shoot of environmental friendly method aimed at 8.10 and cm with fresh and dry sustainable Agriculture. Among the micro of and 4.20 g. Root knot incidence in organisms, bioagent are having great promise both the treatments was least with root knot with the dual advantage of plant growth index of 1.33 and 1.66 respectively compared promotion and plant disease suppression. to. Recently, P. spp. has received greater Treatment involving FYM+ P. attention of the scientists working was most effective in management of plant parasitic nematodes. enhancing the shoot and root by Hence, in the present study, an attempt was and per centrespectively. It was also made to isolate, to screen the different isolates effective in increasing the fresh root and dry and to study the bioefficacy and antagonistic root by 40.0 and per effect of P. against Meloidogyne centrespectively compared to other treatments. incognita and to study their effect against plant Copyright Jan.-Feb., 2018; IJPAB 604

7 growth parameters through different methods Cadioli et al. 8 also reported that isolates of P. in tomato. reduced the population of eggs and J2 In the present study, all the seven of M. paranaensis in the root system as well as isolates of P. were further examined in the soil and favoured the growth of the for nematicidal activity against M. incognita. coffee plants. Similar studies have been conducted by Tomato seedling were free from root various workers who have randomly screened knot infestation when seedling were dipped in Paecilomyces for their antagonistic P. and Lecanicillium lecaniii at 10 activity against nematodes and evaluated them g/lre concentration and reduced 100 per cent for their plant growth promoting activity also RKN index in both the treatments. However, Goswami and Kumar. root knot index was 4 in 0-5 scale in Pot culture studies indicated that the indicating the presence of sufficient inoculums efficacy of the talc based formulations of P. to cause the disease. Similar observations were was the most effective in enhancing made by many scientists while working with the plant growth as they exhibited higher P. and M. incognita. Thakur and Devi antagonistic activity against Meloidogyne found that Arthospora oligospora alone and in incognita egg (89.14% egg hatching combination with P significantly inhibition) masses. Bioformulation containing suppressed the root galling and nematode Mudhol isolate (PL-6) was most effective in population of M. incognita on okra thereby reducing the disease incidence (root knot improving the plant growth parameters. infestation) in tomato compared to all other In the present study it was evident treatments and untreated in glasshouse. from results that nematode multiplication Results of the experiments also significantly reduced when P. was revealed significant variation in plant growth applied along with organic amendments such characters, viz. shoot and root and as FYM and Neem cake to soil. Apart from as well as yield per plant in tomato nematode management in root when treated with different concentration of and shoot was significantly higher in Paecilomyces strains. P. treated plants than the plants in In vivo studies were conducted by untreated. Similar results were various methods such as seed treatment, recorded by Khan by appling seedling dip method and soil application. Seed 0.5, 1.0, 2.0, 4.0, 5.0, 6.0, 7.0 and g/pot treatment with 6 g/kg resulted along with neem cake against M. incognita in in more than 82 per cent germination. Shoot okra and recorded an in plant growth and root were d with enhanced parameters. vigour index of after seven. It also Total reduction of RKN index was d root and shoot by 125 and recorded in tomato seedlings grown in soil per cent the compared to applied with FYM + P. and Neem other treatments. cake + P..Similar results were Management of insect pests and observed by El-Shanshoury et al. 14 and they nematode diseases by different Paecilomyces determined that P. chlamydosporia, P. strains and different formulations have been and Arthrobotrytis dactyloides reported by many workers. Similar studies reduced population densities of M. incognita were conducted by Dhawan et al. 12 where they upto 98.9% and were as effective as tested the efficacy of P. against M. Carbofuran and improved the root and shoot incognita on okra as seed treatment at 10, 15 growth of faba bean. Reduction in galls and and 20 g/kg seed and found significant in egg masses per root system and soil population improvement in plant growth character as well of M. incognita and consequent improvement as reduction in number of galls, egg masses in growth of tomato due to P. were and eggs per egg mass. also recorded by Pathan. Copyright Jan.-Feb., 2018; IJPAB 605

8 Sharma also recorded similar results in the respectively. Similarly Sharma also observed management of M. incognita on okra with P. similar results such as P. along with alone and in combined application addition of neem cake reducing number of with carbofuran, phorate and neem cake and galls, eggs per egg mass by 64 per cent each observed that P. alone reduced and soil population by 77 per cent. number of galls, eggs per egg mass by 32 per The pot experiments conducted in the cent each and soil population by 77 per cent. present investigation also revealed a Many scientists observed that P. not significant in plant growth only reduced the disease but also d the parameters, viz. plant height, shoot and root yield of crop. They opined that it is having, number of branches as well as root dual role such as disease suppression and plant yield per plant in tomato treated with growth promotion activity. The results of the Paecilomyces strains. present study are in tune with study conducted For better understanding about by Xiao and their observation reveal that nematophagous fungi ie P. and its cucumber plants when treated with P. mode of action. It is necessary to focus led M.incognita by per cent, attention on the following future line of work. resulting in 19.7 per cent in yield. Survey may be conducted for occurrence Similar results were recorded by many and prevalence of different species of workers while working with different crops. Paecilomyces in other areas and to record Kiewnick and Sikora reported that application incidence of root knot nematode disease. of P. to soil infested with M. Further studies on Identification of different incognita prior to planting reduced galls, egg species of Paecilomyces effective against masses and the final nematode population in nematode diseases and their utilization in the roots by 66, 74 and 71 per cent biological. Seed treatment Copyright Jan.-Feb., 2018; IJPAB 606

9 Seedling dip method Soil application Copyright Jan.-Feb., 2018; IJPAB 607

10 Growth parameter observation Treatment Conc. (g/kg seed) Table 1: Evaluation of Paecilomyces on growth and root knot development through seed treatment germination Shoot at 90 (cm) in shoot Root at 90 (cm) in root Fresh root at 90 (g) in fresh Dry root at 90 (g) in dry Mean root knot index (0-5 scale) reduction in RKN index Paecilomyces Paecilomyces Paecilomyces Trichoderma 76 2 harzianum Trichoderma 78 4 harzianum Lecanicillium 74 2 lecanii Carbofuran 3 g/kg Control _ S.Em ± CD at 1% Treatment Conc. (g/l) Table 2: Evaluation of Paecilomyces on growth and root knot nematode development through seedling dip Shoot at 60 (cm) in shoot Root at 60 (cm) in root Root fresh at 60 (g) in fresh Mean root dry at 60 (g) in dry Copyright Jan.-Feb., 2018; IJPAB 608 Mean Root knot index (0-5 scale) reduction in RKN index Paecilomyces Paecilomyces Paecilomyces Trichoderma harzianum Trichoderma harzianum Lecanicillium lecanii Pseudomonas fluorescence Carbofuran Control S.Em ± CD at 1%

11 Table 3: Effect of soil application of Paecilomyces on growth and root knot nematode development in tomato Sl. No. Treatment Conc. (g/pot) Mean shoot at 90 (cm) in shoot Mean root at 90 (cm) in root Fresh (g) in fresh Dry (g) in dry Mean root knot index (0-5 scale) reduction in RKN index T1 Paecilomyces T2 Paecilomyces T3 Paecilomyces T4 Neem cake T5 Neem cake+ T T6 Neem cake +T T7 Neem cake +T T8 Farm yard manure T9 FYM + T T10 FYM +T T11 FYM +T T12 Carbofuran T13 Control S.Em ± CD at 1% REFERENCES 8. Cadioli, M. C., Santiago, D. C., Oliveira, 1. Archana, U., 2012, Studies on the disease A. D. D., Paes, V. D. S., Arieira, G. D. O. complex incidence by Meloidogyne and Baida, F. C., Effect of isolates of incognita and Fusarium solani on poi, Paecilomyces on the Basella rubra. Arch. Phypath. Pl. Prot., development of coffee plantations and on 45(16): (2012). the population of Meloidogyne 2. Arie, T., Takahshi, H., Kodama, M. and paranaensis Ciencia. Agrotechologia, 33: Teraoka, T., Tomato as a model plant for (2009). plant pathogen interactions. Pl. Biotech., 9. Cecilia, A. S., Evelina, A. T. and Lilia, M. 24(1): (2007). A., Processing of tomato: impact on in 3. Bainer, G., Mycotheque de 1 ecole de vitro bioaccessibility of lycopene and Pharmacie, XI. Paecilomyces genere textural properties. J. Sci. Food Agric., nouveae de Mucedinees. Bull. Trimmest. 90(10): (2010). Soc. Medit., 16: (1907). 10. Cobb, N. A., Estimating the nematode 4. Baki, A. A. and Anderson, J. D., Vigour population of soil. U.S. Department of determination in soybean seed by multiple Agriculture Circulation No. 1, p. 48 criteria. Crop Sci., 13: (1973). (1918). 5. Barone, A. and Frusciante, L., Molecular 11. Daisy, M., In vitro production of marker assisted selection for resistance to entomopathogenic fungi P. farinosus and pathogens in tomato, marker assisted P. lilainus using by product of sugar selection, current status and future industry and other industrial by products perspectives in crops, livestock, forestry and wastes. J. Sugar Tech., 5(4): and fish. Ed. Sharma, H. C., Oxford and (2003). IBH Publishing Co. Pvt. Ltd., New Delhi. 12. Dhawan, S. C., Narayana, R. and Babu, N. pp (2007). P., Bio-management of root knot 6. Berkaley, M. J., First report of root knot nematode, Meloidogyne incognita in okra infestation in cucumber. Gardeners by Paecilomyces. Ann. Pl. Prot. chronicle, April, p. 200 (1855). Sci., 12(2): (2004). 7. Britt, B. F. and Kristin, R., Tomato 13. Duan, Effect of storage conditions on the consumption and health: emerging survival of two potential bio agents benefits. American. J. Lifestyle Med., 8: of nematodes the fungus Paecilomyces (2011). and Pochonia chlamydosporia. Copyright Jan.-Feb., 2018; IJPAB 609

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