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M. Sc. Dissertations
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ThesisItem Open Access Studies on gnotobiology, and effect of sowing dates and varieties of wheat on reproduction of root lesion nematode (Pratylenchus thornei Sher and Allen)(CCSHAU, 2011) K Kranti K V V S; Kanwar, R. S.Investigations were carried out on in vitro culturing of Pratylenchus thornei, and effect of sowing dates and wheat varieties on reproduction of this nematode. In gnotobiology study, carrot discs on 1% water agar and alfalfa callus on MS medium with 4 mg 2, 4-D were compared for culturing P. thornei, by inoculating 40 surface sterilized nematodes per flask. For callus production, 5-day-old seedlings grown aseptically from alfalfa seed sterilized in conc. sulphuric acid for 15-20 min were used. Carrot disc proved better than alfalfa callus technique in having higher nematode multiplication rate (18.01 in carrot disc, 13.25 in alfalfa callus). In date of sowing experiment, December 5 sowing resulted in maximum final nematode population (394) with reproduction factor 1.9. Reproduction factor was 1.0 and 0.6 in November 15 and October 30 sowings, respectively. The nematode reproduction rate increased with the delay in sowing from October 30 to December 5. Of the 20 wheat varieties/lines evaluated for resistance and tolerance, nine varieties/ lines namely AUS 15854, PBW 343, PBW 550, Raj MR1, Raj 3765, Turkey line (CROC 1/AE. SQUARROSA (224)//OPATA), WH 542, WH 896 and WHD 943 were rated as resistant (Rf <1) whereas 11 varieties viz., C 306, DBW 16, DBW 17, PBW 373, UP 2425, WH 147, WH 711, WH 912, WH 1021,WH 1025 and WH 1080 were rated as susceptible (Rf ≥ 1). Varieties PBW 550, UP 2425, Raj 3765, WH 147, WH 912, WHD 943 and WH 1025 were found tolerant and remaining varieties (AUS 15854, C 306, DBW 16, DBW 17, PBW 343, PBW 373, Raj MR1, Turkey line, WH 542, WH 711, WH 896, WH 1021 and WH 1080) were found intolerant. Bread wheat varieties PBW 550 and Raj 3765 and durum variety WHD 943 proved resistant as well tolerant to this nematode.ThesisItem Open Access Effect of some biotic and abiotic factors on the endospore adherence and development of bacterial parasite, Pasteuria penetrans on meloidogyne javanica(CCSHAU, 2011) Jasbit Singh; Walia, R. K.Influence of plant root exudates, and culture filtrates (CFs) of rhizobacteria and fungi on the attachment of Pasteuria penetrans endospores to J2 of Meloidogyne javanica was studied in lab experiments. Soil leachate did not affect the endospore attachment. Endospore attachment increased significantly with time. Brinjal, tomato, chickpea and onion root exudates enhanced endospore attachment. Garden bean root exudates promoted the growth and multiplication of protozoans that decreased the number of leftover endospores drastically. CFs of Azotobacter chroococcum, Gluconoacetobacter diazotrophicus and Providentia rettgeri promoted, while that of Enterobacter cloacae reduced the endospore attachment to J2 of M. javanica. The CFs (autoclaved or not) of soil fungi adversely affected the endospore attachment to nematode juveniles. Studies on attempted in vitro culture of P. penetrans in CFs of rhizobacteria revealed the growth of P. penetrans-like stages (tetrads, diads etc.) only in case of A. chroococcum, E. cloacae and G. diazotrophicus. In a screen-house experiment to study the development of P. penetrans during rabi season, it was observed that the life cycle of host nematode M. javanica was prolonged; although eggs were formed in the genital tracts of females but no egg masses were formed till the end of March. The sporogenesis phase of P. penetrans did not occur in any of the five sets inoculated on different dates. P. penetrans development was restricted to vegetative phase only (microcolonies and thallus stages). The nematode development continued albeit slowly; while that of bacterium failed to synchronize with that of nematode.ThesisItem Open Access Studies on bacterial parasite, pasteuria penetrans (Ex thorne) sayre & starr on root-knot nematode, meloidogyne javanica (Treub) chitwood(CCSHAU, 2012) Matcha, Udaya Kumar; Walia, R.K.Influence of root exudates (REs) and cell free plant root extracts (CFPREs) of brinjal, cowpea, cluster bean, garden bean, okra and tomato was tested on the germination of Pasteuria penetrans endospores. REs and CFPRE of all the plant species did not induce the germination of P. penetrans endospore. In garden bean CFPRE, cauliflower-like colonies similar to P. penetrans were observed in endospore suspension treated with alcohol. In all the treatments rod-shaped lumps of bacteria were evident; they may be contaminants because intact endospores of P. penetrans were also present. Based on a laboratory bioassay, a statistical model was developed for the estimation of P. penetrans endospores in field soils. The regression equation y= 0.045x + 3.779 with R2 = 0.89 was calculated for M. javanica. In a green-house experiment, the role of P. penetrans in the suppression of M. javanica was estimated in naturally infested field soil. Field soil was steam sterilised or treated with formalin/fungicides. Formalin and Bavistin proved to be phytotoxic. P. penetrans alone infected 87.5% nematode females and reduced egg production by ca. ten times. Native fungal and bacteria promoted attachment of P. penetrans endospores but inhibited parasitisation of nematode females by P. penetrans in untreated soil. The development of P. penetrans under open-field and poly-house (ordinary and hi-tech) conditions was studied in winter season. Poly-houses (both ordinary and hi-tech) enabled raising the maximum temperature by 10-12 °C during day time, but there were negligible differences in the minimum temperatures. M. javanica completed it life cycle (females with egg sacs) in 60 days in hi-tech, 70 days in ordinary poly-house and 80 days in open-field conditions. P. penetrans development could not keep pace with that of nematode and it stopped at thallus stage. No mature endospores were found. P. penetrans failed to curtail the reproduction of M. javanica even in hi-tech poly-house, although infection did take place.ThesisItem Open Access Bio-efficacy of bacterial parasite, pasteuria penetrans (ex thorne) sayre & starr application as seed coating and nursery soil treatment against root-knot nematodes, meloidogyne spp(CCSHAU, 2013) Vikaram; Walia, R. K.Four species of Meloidogyne i.e., M. arenaria, M. graminicola, M. incognita and M. javanica were tested for Pasteuria penetrans endospore adherence and development. After 48 hrs, endospore adherance was least in case of M. graminicola and M. arenaria but significantly more in M. javanica and M. incognita. This strain of P. penetrans did not infect M. graminicola, however, development on the other three species was very efficient, and the infection varied from 70-90%. Three experiments were conducted in summer, kharif and winter seasons using cotton, okra and chickpea to test the efficacy of P. penetrans as seed treatment against M. javanica. The results on cotton and chickpea were not significant. However, only during kharif season (mean maximum and minimum temperatures of 34.5° and 26.4° C, respectively) on okra, P. penetrans used @ 4 and 5% as seed coat resulted in significant reduction in root galling, egg production and infection of females. Efficacy of P. penetrans application as nursery soil treatment was tested in two seasons. In kharif season (mean maximum temperature of 34.5° C and the minimum temperature of 26.4° C) higher doses of P. penetrans (8 x 104 and 10 x 104 endospores per g soil) resulted in better seedling germination and improved fresh weight of seedlings. All the doses of P. penetrans caused significant reduction in galling and were also equally effective in suppressing nematode egg production. The two highest doses of 8 x 104 and 10 x 104 caused up to 70% nematode infection. Similar experiment during winter season did not produce any tangible results.ThesisItem Open Access Survey, pathogenicity and management of rice rootknot nematode, meloidogyne graminicola golden and birchfield on rice(CCSHAU, 2016) Puja Devi; Kanwar, R.S.Survey of rice nurseries in four districts viz; Sirsa, Fatehabad, hisar and Jind districts revealed 80.1% infestation of rice root-knot nematode. Light textured soil was more prominent for the rice root-knot nematode M.graminicola. Studies on the pathogenicity of different type soil i.e clay loam, loam and sandy loam soil revealed that clay loam soil resulted maximum plant growth and nematode reproduction and multiplication followed by loam and sandy loam soil, irrespective of the inoculum level. As the inoculum level increased from 1-4 J2/g soil. Plant growth decreased significanty and nematode reproduction and multiplication increased in all the soil types. Pathogenic level of rice root-knot nematode was found 1J2/g soil in all soil types. In host range studies all the varieties of rice, pearl millet, sorghum, weeds and brinjal were susceptible to different populations of M.graminicola, to varying degree on the basis of galling. However, tomato proved to be a non-host for all the populations. More galling and reproduction of Fatehabad and Jorhat populations were reportedon all susceptiblehosts as compared to other three populations. Studies on management of rice root-knot nematode revealed that maximum plant growth was found in sterilized soil as compared infested soil, irrespective of organic amendments. Castor cake @ 10g/kg soil was found to be best in increasing plant growth parameters and decreasing nematode reproduction which was followed by mustard cake @ 10g/kg soil and castor cake @ 5g/kg soil.ThesisItem Open Access Integrated management of root-knot nematode, meloidogyne javanica in hybrid tomato(CCSHAU, 2016) Gurpreet Singh; Verma, K.K.Studies were carried out on integrated management of root-knot nematode, Meloidogyne javanica in hybrid tomato under field conditions. This research endeavor consisted of three pronged strategies in which experimentation was done at nursery stage and under main field conditions at two stages of plant growth and nematode multiplication period. Individual practices of nematode management such as biocontrol agents (Trichoderma viride @ 50 g/m2, Paecilomyces lilacinus @ 50 g/m2, chemicals (carbofuran @ 7 g/m2, cartap hydrochloride @7.5 g/m2) and organic amendments (neem and mustard cake each @ 750 g/m2) were first evaluated in nursery beds and then treated nursery was transplanted in the main field to have integration of these components for better plant growth, yield and minimum nematode reproduction and multiplication. At nursery stage, there were 7 treatments of which the best two treatments, i.e., neem cake and T. viride along with untreated check were combined with three treatments in the main field.(T. viride @ 2.5 kg/ha, carbofuran @ 1 kg a.i./ha along with untreated check) making a total of nine treatments. Observations recorded at the time of transplanting revealed maximum plant growth of seedlings and minimum number of galls in neem cake followed by T. viride. When these treated seedlings were transplanted in M. javanica infested main field, the observations taken at 50 days after transplanting (mid season evaluation experiment) revealed that highest plant growth parameters were observed in neem cake treated nursery when transplanted in T. viride (main field application). Minimum nematode reproduction and multiplication was observed in neem cake (nursery) integrated with T. viride (field) followed by T. viride (nursery) + T.viride (main field). Similarly, when treated seedlings were transplanted in another field which was terminated at harvesting of tomato, it was observed that highest tomato yield was recorded in neem cake at nursery stage integrated with T. viride at main field followed by nursery application of T. viride combined with T. viride (field). Minimum nematode reproduction and multiplication in terms of number of galls, number of egg masses per plant, number of eggs per egg mass and final nematode population in the soil was observed in neem cake (nursery) + T. viride (field) followed by T.viride at nursery integrated with T. viride as main field application. Such studies will lead us to a scenario where management of M. javanica would be possible by integration of ecofriendly, economic and effective components starting from nursery upto main field application in hybrid tomato since the varieties have been replaced by hybrids.ThesisItem Open Access Studies on the nematicidal efficacy of extracts and essential oils of different plants against root-knot nematode, meloidogyne incognita (Kofoid & White) chitwood(CCSHAU, 2016) Neeraj; Sewak RamDetailed investigations were carried out to determine the nematicidal effect of few nematoxic plants viz., turmeric (Curcuma longa), marwatulsi (Origanum majorana), mentha (Mentha arvensis), aonla (Phyllanthus emblica) and jatropha (Jatropha curcas) in the management of Meloidogyne incognita infecting tomato at Department of Nematology ,CCSHAU, Hisar. Different extracts viz. aqueous and organic extracts and essential oils of plants were evaluated under in vitro and in vivo conditions for their nematicidal effect on egg hatching and larval mortality of root-knot nematode (Meloidogyne incognita). All the extracts and essential oils of plants inhibited egg hatchingof Meloidogyne incognita at all dilutions i.e. 1:5 and 1:20 and at each interval of exposure period i.e. 3, 6, 9 and 12 days. Maximum inhibition of egg hatching was found in essential oils followed by ethanolic and acetonic extracts as compared to aqueous extracts. Revival of egg hatching was minimum at 1:5 dilutions of all the extracts and oils .Extracts and oils of all the plants showed larval mortality of root-knot nematode (Meloidogyne incognita) at all the dilutions tested i.e. 1:5, 1:10, 1:20, 1:40 and 1:80 after 48 hours of exposure. However, highest larval mortality was exhibited by mentha oil at 1:5 dilution after 48 hours of exposure followed by turmeric oil at the same dilution. The immobilized larvae after 48 hours exposure to essential oils of plants at 1:5 dilution did not revive even after exposure to distilled water for 24 hours. Effect of chopped leaves and aqueous extracts of different plants on growth characters of tomato and multiplication of root-knot nematode (Meloidogyne incognita) was evaluated under screen-house conditions. The results revealed that in general chopped leaves of all the plants at both the doses i.e 20 g and 30 g/kg soil improved all the plant growth parameters of tomato viz., shoot length, fresh shoot weight , root weight , dry shoot and root weight . Number of galls , number of egg masses and final nematode population in soil were reduced significantly in treatments involving application of plants leaves @ 20 g and 30 g per kg soil as compared to untreated check. Mentha leaves @ 30 g per kg soil followed by turmeric leaves at same dose were found significantly effective in increasing plant growth parameters and reducing nematode reproduction factors. Aqueous extract of mentha @ 75 ml/pot followed by extract turmeric @75 ml/pot was found effective in enhancing plant growth parameters and reducing nematode reproduction factors.ThesisItem Open Access Life cycle and pathogenicity of root-knot nematode, meloidogyne graminicola (Golden and birchfield) on rice(CCSHAU, 2015) Vinod Kumar; Verma, K.K.Investigations were carried out on life cycle and pathogenicity of Meloidogyne graminicola on scented and non-scented rice varieties under screen house conditions. In life cycle and development experiment, life cycle of M. graminicola was studied on var. Pusa 1121 and. PR 114. The j 2 of M. graminicola penetrated into rice roots after 24 h in scented rice while in non-scented rice in 72 h. The third stage (j3 ) appeared on 7 DAI in scented rice as compared to non-scented rice in 9 DAI. Young females were first noticed on 11 DAI and 15 DAI for scented and non-scented rice, respectively. Further, egg deposition started as 19 DAI and 23 DAI inside the roots in scented and non-scented rice, respectively. Nematode completed its life cycle in 21 days in scented rice and 25 days in non-scented rice. The experiment on pathogenicity of M. graminicola was carried out in three different types of soil (clay loam, sandy loam and loamy sand) by using different inoculum levels such as 0 (non-inoculated check), 10, 100, 1000 and 10000 j 2 /kg soil. The results revealed that growth of both varieties of rice (scented and non-scented) was significantly improved in clay loam soil followed by sandy loam and loamy sand irrespective of inoculum levels. In case of inoculum levels, maximum plant growth was observed in non-inoculated check which was statistically at par with 10 j2. Plant growth parameters were decreased significantly as inoculum levels increased from 10-10000 j 2 irrespective of soil types. Significant reduction in growth parameters was observed from inoculum level of 100 j 2/kg soil onwards. Observations on nematode reproduction and multiplication revealed that maximum and significant reduction in number of galls, number of eggs per plant and final nematode population was observed in clay loam followed by sandy loam and loamy sand irrespective of inoculum levels. Nematode multiplication and reproduction was increased as inoculum levels increased from 10-1000 j 2 but decreased abruptly at 10000 j 2 . Maximum and significantly highest multiplication and reproduction was observed in 1000 j2 of M. graminicola. Nematode growth parameters at 100 j2 were significantly different from non-inoculated check and other inoculum levels which can be termed as pathogenic level of M. graminicola on rice.ThesisItem Open Access Life cycle and pathogenicity of Aphelenchoides swarupi Seth & Sharma on Agaricus bisporus (Lange) Singer(CCSHAU, 2015) Madhuri; Kanwar, R.S.Investigations were carried out on life cycle and pathogenicity of Aphelenchoides swarupi on U3 strain of Agaricus bisporus and relative susceptibility of its different strains to this nematode. The nematode took 5-6 days for completion of life cycle from J 2 to adult at 30 ºC which was prolonged to 13-14 days at 15 ºC. A total of 10 strains of A. bisporus viz., ABL2, ABL3, ABL4, ABL5, ABL6, B10, B13, DMR3, DMR7 and U3, and two strains (A1 and A6) of Pleurotus eous were evaluated against A. swarupi. Nematode multiplied on all the strains of A. bisporus and caused mycelial damage to them. Of these, ABL2 and DMR7 were found to be resistant and tolerant, respectively while remaining were susceptible. On these strains, nematode population and reproduction factor were higher after 30 days than after 15 days of nematode inoculation. Both the strains of P.eous were found immune to this nematode. In pathogenicity experiment, 10 nematodes per plate as well as per kg compost in bags were found pathogenic. In plates, reproduction factor and population growth rate were minimum at one and these decreased further when inoculum level increased from 10 to 40. In bags, spawn run was poor at 1000 nematodes per kg compost but moderate at 10 and 100 nematodes per kg compost in comparison to good in control. Yield and final nematode population were affected by inoculum levels and time of inoculation. Yield reduction was more in all inoculum levels when inoculation was done at spawning as compared to casing. Final nematode population was higher when inoculation was done at time of spawning as compared to time of casing. Reproduction factor decreased with increase in inoculum level and higher population growth rate was achieved when nematode inoculation was done at casing as compared to when it was done at spawning.