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ThesisItem Open Access Effect of organic amendments on the efficacy of bacterial parasite, Pasteuria penetrans, for the management of root-knot nematode (Meloidogyne javanica) in tomato(CCSHAU, 2018) Sujata; Anil KumarInvestigations were carried out to evaluate the efficacy of Pasteuria penetrans (a bacterial parasite of root-knot nematode) and organic amendments (neem cake, mustard cake, castor cake and farmyard manure) in the management of root knot nematode Meloidogyne javanica infecting tomato. The experiments were conducted in laboratory and screen house of Department of Nematology, CCSHAU, Hisar. Leachates were collected after application of organic amendments to the soil and were tested in vitro, for their effect on the encumbrance of endospore of P. penetrans to second stage juvenile (J2) of root-knot nematode. Maximum spore adherence to J2 was observed in leachate collected from soil, where no organic amendment was applied, indicating that water along with soil chemical does not influence the attachment of endospores to juveniles. Among the tested organic amendments, higher attachment of endospores was observed in mustard cake and neem cake. Minimum spore encumbrance was observed in leachate collected from the pots where farmyard manure was applied. However, the number of spores adhered to J2 were sufficient to parasitize root-knot nematode. Another aspect of the present investigation was to study the effect of different organic amendments on the life cycle development of P. penetrans on the M. javanica infecting tomato. The life cycle of P. penetrans on root-knot nematode was completed in all the organic amendments but, it was well synchronised where mustard and castor cakes were applied. There was a slight delay in life cycle of bacterium in neem cake and FYM. All life cycle stages of bacterium viz., microcolony, thallus, quartets, diads, immature spore and mature spore were observed. Integration of P. penetrans @ 1x104 spores/g soil with the organic amendments improved plant growth parameters and reduced nematode multiplication in tomato infected with M. javanica. Minimum number of galls (20.7), eggs per root system (656.0) and final nematode population per 200 cc soil (92.1) and improved plant growth parameters were recorded in combined application of P. penetrans with mustard cake @ 10 g/kg soil. The application of P. penetrans at different spore load (1x103, 1x104, and 1x105 spores/g soil) in combination mustard cake at the rate of 10 g/kg soil was also tested for the management of M. javanica in tomato nursery. The nematode reproduction reduced with the increase in spore load of bacterium. P. penetrans @ 1x105 spores/g soil was found best in increasing plant growth parameters and decreasing nematode reproduction followed by spore load 1x104 spores/g soil.ThesisItem Open Access Studies on rice root-knot nematode, meloidogyne graminicola golden and birchfield, 1965 with emphasis on its management in rice nursery(CCSHAU, 2018) Matcha, Udaya Kumar; Kanwar, R.S.Survey of rice nurseries for M. graminicola in Ambala, Kaithal, Karnal, Kurukshetra, Panchkula and Yamunanagar districts of Haryana showed that 55.0% samples in in situ and 80.0 % samples after bioassay were infected with rice root-knot nematode. Nematode infestation was more in sandy loam than loamy sand and loam soils. In the experiment on the effect of M. graminicola on absorption and translocation of nutrients from the soil, at all three levels of 0, 2, and 4 J2/g soil, maximum plant growth parameters were observed in the treatment with all nutrients (shoot weight 2.4 g, root weight 2.2 g and shoot length 21.9 cm) and minimum in treatment where no nutrient was applied (shoot weight 1.3 g, root weight 1.0 g and shoot length 15.1 cm). Nematode population per plant was more (2650.0) in treatment where no nutrients were applied and minimum in potassium treatment (1895.6). The nutrient content of N, P, K, Zn, Fe and Cu were more in soil, in their respective treatments where the individual nutrients were applied alone. The data on uptake of nutrients in rice seedlings revealed the maximum uptake of nutrients (N 23.0%, P 12.7 %, K 19.4 %, Zn 14.3 ppm, Fe 20.5 ppm, and Cu 6.0 ppm) in rice seedlings was recorded in control (0 J2/g soil). Among the treatments maximum nutrients uptake was observed in treatment where all nutrients were applied in soil (N 15.1 %, P 11.7 %, K 14.9 %, Zn 13.6 ppm, Fe 21.0 ppm and Cu 4.1 ppm). Amount of nutrients was more in soil at 4 J2/g soil (N 151.5 mg, P 33.7 mg, K 125.2 mg, Zn 37.5 ppm, Fe 78.2 ppm, and Cu 20.4 ppm per kg soil) than 2J2/g soil (N 140.0 mg, P 32.9 mg, K 123.7 mg, Zn 31.5 ppm, Fe 74.5 ppm, and Cu 19.4 ppm per kg soil). Among five bacterial isolates evaluated for nematoxicity, Bacillus strain (RKB-91) showed best results on nematode mortality as well as in inhibiting egg hatching at S/2 concentration in both cell cultures and cell free culture filterates. This strain also suppressed the nematode population as well as promoted plant growth. Among neem, cabbage and cauliflower, neem showed best results in terms of nematode mortality at 100 % concentration and > 50% hatching inhibition at 1:10 dilution. It reduced the nematode population and improved plant growth in pots. In vitro results on synthetic chemicals showed that the Coumarin carbamate series, Coumarin I and Coumarin III gave > 50% on mortality of nematode as well as hatching supression at 1000 and 500 ppm concentration. In planta study revealed that Coumarin carbamate III and Coumarin Carbamate V were more effective in reducing the nematode population and improving plant growth. Integration of different treatments for the management of M. graminicola in rice nursery showed that application of Bacillus sp. (RKB-91) + Neem extract + Coumarin carbamate III ranked first in reducing the galls as well as the nematode population and also in improving the plant growth.ThesisItem Open Access Studies on cereal cyst nematode, heterodera avenae woll. on wheat (Triticum aestivum) with emphasis on nutrients(CCSHAU, 2018) Yadav, Saroj; Kanwar, R.S.Investigations were carried out on absorption and translocation of nutrients in wheat infected with Heterodera avenae, its management through organic amendments, crop rotation and additional fertilizers as soil application and spray. Effect of fertilizers on emergence of larvae from cyst was also studied. Absorption and translocation of nutrients in wheat was hampered by nematode. Combination of all four nutrients promoted plant growth and reduced nematode population. N, P, K and Zn contents in soil were more, in their respective treatments, where the individual nutrients were applied. Amount of nutrients was more in infested soil than uninfested soil. Combination of all the four nutrients increased the nutrient uptake in wheat and nutrient content in straw and grain was more in uninfested soil. Organic amendments reduced nematode populations and increased plant growth both with and without nematicide. Castor cake at 20g/kg soil increased plant growth but neem cake at same dose gave maximum population reduction. All the organic amendments performed better in combination with nematicide than alone. Recommended doses of fertilizers with urea (2.5 %) + zinc (0.5 %) spray at 30 days after sowing followed by kinetin (100 ppm) spray at 40 days after sowing improved maximum plant growth but 30 % higher dose of fertilizers reduced maximum nematode population. Fenugreekwheat rotation increased plant growth more than other rotations but mustard-wheat and cabbage-wheat rotations reduced maximum nematode population. Fenugreek and gram increased N content in soil but maize reduced the N and P content and cabbage reduced the K content in soil. Nematode penetration occurred in susceptible wheat, resistant wheat and maize but not in gram, fenugreek, mustard, cabbage, garlic and onion. Under laboratory conditions, all fertilizers suppressed larval emergence from cysts, except zinc, which stimulated it. In pots, minimum nematodes in soil and roots were found at higher dose of urea. In zinc larvae in roots and soil were less than untreated check (water) but higher than other fertilizer treatments.ThesisItem Open Access Investigations on resistance in barley against cereal cyst nematode, heterodera avenae(CCSHAU, 2018) Chhachhia, Anshul; Kanwar, R.S.Study was done to find out the sources of resistance in barley against Heterodera avenae and to study the nature of resistance in resistant varieties. Experiments were done during 2016-17 and 2017-18, under screen house conditions at CCS HAU, Hisar. In total 95 genotypes of barley were screened against Heterodera avenae population from Haryana (Hisar). Out of these 14 were found resistant (DWRB 137, DWRB 163, DWRB 170, DWRB 73, DWRB 91, RD 2948, RD 2957, RGT PLANET, TRAVELLER, KWS IRINA, PL 895, PL 897, PL 898, PL 751) and three susceptible (DWRB 127, BK 1508, RD 2903) and remaining were found highly susceptible. Life cycle of Heterodera avenae was studied in BH 393, BH 959, RD 2035, DWRB 91 (resistant) and BH 902 (susceptible) varieties. Penetration and number of cyst formed were less in resistant varieties than susceptible and no cyst development on RD 2035. In susceptible as well as resistant varieties, development was similar. Average cyst content was also higher in susceptible variety than resistant varieties. Phenols and total sugar contents increased after infection in roots and shoots of both susceptible and resistant varieties. Both pre- and post- infection resistance were found in resistant varieties of barley.ThesisItem Open Access Studies on physiological changes induced by cereal cyst nematode (Heterodera avenae) in wheat (Triticum aestivum)(CCSHAU, 2017) Sindhu, Ajay Singh; Kanwar, R.S.Present study was conducted to investigate the the sequential effect of Heterodera avenae on physiology of wheat, the effect of sowing time on physiology of H.avenae infected wheat and the effect of different inoculum levels of H.avenae on physiology of wheat. The experiments were conducted in screenhouse of Department of Nematology, CCSHAU, Hisar during Rabi-2016. For evaluating the sequential effect of H.avenae on wheat cultivar WH 1105 was sown on 7th Nov. and observations on physiological parameters were recorded 30, 60 and 90 days after sowing (DAS). Total chlorophyll, carotenoid and chlorophyll fluorescence were significantly lower in inoculated plants as compared to uninoculated plants at 90 DAS. They were reduced to the tune of 37.78%, 36.73% and 10.46%, respectively. Total chlorophyll, carotenoid and chlorophyll fluorescence increased till 60 DAS and decreased at 90 DAS. Photosynthetic rate, transpiration rate and stomatal conductance decreased with the increase in age of plant. Inoculated plants when compared with uninoculated plants showed significant decrease at 60 DAS by 38.17, 47.39 and 42.85%, respectively. For comparing the physiology of H.avenae infected wheat sown on 1st, 15th and 30th Nov., observation were taken 30 DAS. Total chlorophyll (25.00%), carotenoid (28.94%), chlorophyll fluorescence (6.27%), photosynthetic rate (57.33%) and stomatal conductance (62.50%) showed maximum reduction in inoculated plants of 15th Nov. sowing when compared with uninoculated plants while transpiration rate decreased significantly (56.19%) in inoculated plants of 30th Nov. sowing. Inoculated plants showed significant reduction in these parameters when compared with uninoculated plants at all the three sowing dates. Biomass of 15th Nov. sown plants was greatly hampered by nematode parasitism as compared to plants sown on 1st and 30th Nov. Maximum nematode population was obtained in 15th Nov. sown plants. Effect of different inoculum levels viz. 5, 10 and 15 eggs and juveniles/g soil of H.avenae on physiology of wheat was studied by recording observations 30 DAS. With the increase in inoculum level of H.avenae, there was significant decrease in total chlorophyll, carotenoid, chlorophyll fluorescence, photosynthetic rate, transpiration rate and stomatal conductance at each level. Maximum reduction in these parameters was observed at 15 eggs and juveniles/g soil that was 39.71, 30.55, 7.90, 39.75, 51.58 and 64.86% respectively. Maximum nematode population build up and least biomass were obtained at highest inoculum level.ThesisItem Open Access Studies on the incidence, interaction and management of root- knot nematode, Meloidogyne incognita and soil borne fungus infesting cucumber in polyhouse(CCSHAU, 2017) Jaydeep Ashok, Patil; Sewak RamSurvey of polyhouse crops revealed 68.4 %, and 45.8 % frequency of occurrence of root-knot nematode, Meloidogyne incognita, respectively, in Rewari and Jhajjar districts. The fungus, Fusarium oxysporum f. sp. cucumerinum which was found most prevalent during survey of the polyhouse crops proved pathogenic on cucumber. Concomitant and sequential nematode and fungus inoculation resulted in a significant reduction in number of galls per plant. Maximum galling was observed when the nematodes were inoculated 7 days prior to the fungus. Soil treatment with formalin @ 30 %, followed by NSKP as soil amendment @ 30 g per kg soil proved significantly effective in pots against M. incognita and F. oxysporum f. sp. cucumerinum in improving plant growth parameters and reduced nematode reproduction and wilt severity. Seed treatment with liquid formulation of bio-agents @ 15 ml per kg seed followed by seed treatment with neem oil @ 20% v/w in pots proved most effective against M. incognita and F. oxysporum f. sp. cucumerinum. Combinations of soil fumigation with formalin @ 30 % + seed treatment with Paecilomyces lilacinus @ 20 g per kg seed was found highly effective against M. incognita and F. oxysporum f. sp. cucumerinum. Integration of most effective management practices of soil and seed treatment in cucumber against M. incognita and F. oxysporum f. sp. cucumerinum under polyhouse conditions revealed that soil amendment with neem cake @ 200 g per sq meter + seed treatment with P. lilacinus @ 20 g per kg seed in cucumber proved most effective in significantly improving plant growth parameters while soil fumigation with formalin @ 30 250 ml per sq meter + seed treatment with P. lilacinus @ 20 gm per kg seed was significantly better in suppressing nematode reproduction and fungal wilt incidence.ThesisItem Open Access Efficacy of entomopathogenic nematodes (EPNs), Steinernema spp. and Heterorhabditis spp. on tobacco caterpillar, Spodoptera litura F.(CCSHAU, 2017) Babita kumari; Sewak RamOut of 120 soil samples collected from different habitats, 9 samples (7.5%) were found to be positive for EPN. The highest frequency of occurrence of EPNs (58.3%) was recorded in Ber orchard with 7 samples infected for EPNs followed by citrus and cactus (8.3%). Amongst them, the frequency of occurrence of Steinernematid and Heterorhabditid nematodes were 41.7 and 16.7 per cent, respectively. As the inoculum level increased, there was a gradual increase in progeny production of both S. abaasi and H. indica. The present study revealed that both the species strain of EPNs were found to be virulent and could cause 50 per cent mortality at 10 infective juveniles/larva under in vitro conditions. Among all the isolates of EPNs, S. abaasi isolate HAR-EPN-Sa-3 was highly virulent against S. litura. It was followed by other isolates in the order of HAR-EPN-Sa-2, HAR-EPN-Sa-5 and HAR-EPN-Sa-4. The strain HAR-EPN-Sa-1 recorded the least virulence of all the isolates. However, nearly 50 per cent mortality of the larvae was observed at an inoculum level of 10IJs per larva after 24h. In case of H. indica, isolate HAR-EPN-Hi-2 was highly virulent and HAR-EPN-Hi-1 recorded the least virulence against S. litura. Mass multiplication of EPNs, highest yield of S. abaasi isolate HAR-EPN-Sa-3 followed by HAR-EPN-Sa- 2, HAR-EPN-Sa-5 and HAR-EPN-Sa-4. The strain HAR-EPN-Sa-1 recorded the least multiplication of all the isolates. In case of H. indica, highest yield of HAR-EPN-Hi-2 followed by HAR-EPN-Hi-1. Laboratory studies revealed that S. abaasi in combination with H. indica had been more virulence than S. abaasi and H. indica when applied alone. S. abaasi in combination with H. indica caused 72.5, 87.4, 97.3 and 99.7 per cent mortality, respectively against S. litura as compared to virulent strain of Steinernema spp alone. Mass production of IJs of tested EPNs from S. litura larva showed highest yield of EPNs (H. indica+S. abaasi) followed by S. abaasi alone and H. indica.ThesisItem Open Access Predatory behaviour of Fictor composticola Khan et al. and its potential for the management of nematode pests of button mushroom(CCSHAU, 2016) Keshari, Nishi; Kanwar, R. S.Investigations were carried out on prey range, prey preference, strike rate, predation behaviour, effect of prey density on predation rate of Fictor composticola, survival of F. composticola in agar plates and spent mushroom compost and management of mycophagous nematode, Aphelenchoides swarupi (mushroom pest) in button mushroom. F. composticola preyed upon all the twelve nematode species tested including fungal feeders (Aphelenchus avenae, A. swarupi and Ditylenchus myceliophagus), microbivorous (Panagrolaimus sp., Bursilla sp., Tylencholaimus sp., Rhabdolaimus sp. and Aerolaimid), plant parasitic (Heterodera avenae males and Hoplolaimus sp.) and predatory nematodes (Aporcelaimium sp. and Nygolaimus harishi). F. composticola preferred mycophagous nematodes over microbivorous nematodes and D. myceliophagus among the fungal feeders. Female F. composticola was more voracious feeders than males. Strike rate of female F. composticola was 78.6 and 48.2 in males. The myceliophagous nematodes have more prey susceptibility than the microbivorous nematodes. The feeding duration of female and male F. composticola was 8 min 31 sec and 4 min 11 sec, respectively. It preferred juveniles over adults and posterior part of preys over other parts. Predation efficiency of F. composticola increased with increase in prey density but, the per cent consumption was minimum at highest prey density level (1600 per plate). The optimum per cent prey consumption was at 200 and 400 prey density levels. F. composticola could survive in agar plates up to two months. In spent mushroom compost, its survival was better in polythene bags than in cloth bags (75 days in polythene bags v/s 60 days in cloth bags). In compost heap stored in open, F. composticola survived in active stage during off-season (April to September). No anhydrobiotic survival was seen under moisture stress conditions. In mushroom bags, population of A. swarupi was found minimum when F. composticola was inoculated at spawning.