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Bidhan Chandra Krishi Viswavidyalaya, Mohanpur, Dist. - Nadia, West Bengal PIN - 741 252
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Bihar Agricultural University, Sabour
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ThesisItem Open Access Association of immune related genes with clinical mastitis, reproductive disorders and performance traits in Hardhenu cattle(LUVAS Hisar, 2024-04) Pushpa; Magotra,AnkitIn the present study, 1515 lactation records of Hardhenu cattle were screened from 2008 to 2021 to calculate the incidence of clinical mastitis and reproductive disorders. The data of affected animals was collected from treatment register maintained at cattle breeding farm, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana. For genetic profiling (CXCR1 and TLR4 gene) and association studies, blood samples of 181 Hardhenu animals were collected for DNA isolation. Overall incidence rate of 26.80 % and 37.24% were observed for clinical mastitis and reproductive disorder, respectively in our targeted Hardhenu population. Among reproductive disorder, retention of placenta (18.36 %) and repeat breeding (11.19) cases revealed high incidence compared to dystocia (0.60 %), metritis (6.27%), prolapse (0.37 %) and anestrous (0.45 %). Chi-square analysis and Logistic regression analysis revealed significant association (P<0.01) of year with incidence of clinical mastitis and reproductive disorder in our studied population. PCR amplified product of 592 bp was digested with the Bsa1 restriction enzyme to genotype targeted SNP rs211042414 (C>T) at g.106216468 loci of CXCR1 gene. The genotypic and allelic frequencies g.106216468T>C indicated that C allele (0.54) was highest in Hardhenu cattle. PCR amplified product (316bp) of TLR4 gene encompassing partial intron1, exon 2 and partial intron 2 digested with Hinf1 restriction enzyme to screen polymorphism at targeted locig.107062971 revealed all three genotypes in our resource population. The genotypic and allelic frequencies g.107062971A>B indicated highest frequency of A allele (0.93). Chi-square tests showed that mutation at g.107062971A>B loci did not meet with the Hardy– Weinberg equilibrium (p<0.05). Chi-square and logistic regression analysis showed significant association between (p<0.05) the g.106216468 T>C SNP of CXCR1 gene with incidence of clinical mastitis. Animals with CC genotype found to be 3.47 times more susceptible to mastitis compared to TT genotype and with CT genotypes 2.90 times more susceptible to mastitis compared to TT genotype. Chi square analysis also showed a significant association (p<0.05) between g.107062971A>B locus of TLR4 gene and susceptibility to reproductive disorders. Genotype AB found to be more susceptible to reproductive disorders compared to AA. The animals with genotype AA were found to be less susceptible to incidence of reproductive disorder as established by lower value of odd ratio. Parity and period of calving revealed significant association with clinical mastitis and reproductive disorders (p<0.01). Higher odds ratio indicated that animals in the second parity were more prone to clinical mastitis (p<0.01). Incidence of reproductive disorders was found to be highest during the third period (2017-21). Parity, period of calving and season of calving showed a highly significant association (p<0.01) with total milk yield (TMY), 305d MY, peak yield (PY) and Lactation length (LL). All these milk traits were highest in third parity. Period of calving showed a highly significant effect (p<0.01) on LL which is highest during first period (2008-13). Season of calving showed significant association (p<0.01) with TMY and PY. TMY and PY were highest during winter season. Season of calving also revealed significant effect (p<0.05) on AI/Conception. Period of calving showed a highly significant effect (p<0.01) on TMY, 305d MY and PY. All these milk traits were highest during third period (2017-21). These observations on identified polymorphism in CXCR1 and TLR4 gene and their association with studied traits can be incorporated in exiting selection criteria for genetic improvement of targeted traits in Hardhenu cattle. However, validation of results on large number of animals is warranted.ThesisItem Open Access CULTURAL AND MORPHOLOGICAL CHARACTERIZATIONS OF Metarhizium anisopliae.(College of Agriculture Nagpur, Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, 2019) SONUNE, BHUSHAN DEVIDAS.; Patil, Dr. M. J.Metarhizium anisopliae is characterized as green muscardine fungus due to green colour of the sporulating colonies. M. anisopliae is an entomopathogenic fungus, used as important natural biocontrol agent against many insect pests throughout the world. It causes the death of insect due to the infective peg penetrate in the layer of insects integument and obliteration of tissues also production of toxins and proteolytic enzymes by the fungus. The present study was aimed to isolate M. anisopliae from the infected insects of this region and evaluate the cultural and morphological characters of the fungus. Total ten isolates were collected from the infected insects and from market products. The isolates were identified by testing growth characters on media and morphological characters. The white, prominently raised mycelial growth became greenish on sporulation on the medium. The conidiophores were errect, branched covered by conidia, the conidia were apical long single celled ovoid to cylindrical with rounded end. These isolates were tested on Spodoptera litura larvae for pathogenicity of M. anisopliae fungus. All the isolates showed variable mortality percentage and maximum mortality was noticed by Ma2 (90%) in S. litura larvae. Further confirmation of efficacy of spore suspension was assessed by leaf dip method and it was observed that the mortality was rapidly increased upto 91.66 per cent in spore concentration of 1 x 109 spore/ml on 10th days of incubation. The rate of mortality was reduced with reduction in the spore concentration. Therefore Ma2 isolates was taken for further studies. Cultural and morphological studies of M. anisopliae was taken on four different media viz., Potato Dextrose Agar (PDA), Sabouraud Dextrose Agar (SDA), Sabouraud’s Dextrose Agar + Yeast Extract (SDAY) and Sabouraud Maltose Agar (SMA) medium. SDAY medium support maximum growth and sporulation of M. anisopliae at 250C temperature, 95 per cent relative humidity and 5.5 pH levels. Compatibility studies were assessed with four insecticides viz., Imidacloprid, Spinosad, Chloropyriphos and Indoxacarb. Out of these M. anisopliae slightly compatible with spinosad and imadacloprid at normal concentration dose of insecticide.ThesisItem Open Access CHARACTERIZATION OF Bradyrhizobium japonicum AND ITS COMPATIBILITY WITH FUNGICIDES.(College of Agriculture Nagpur, Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, 2019) DINKWAR, KU. GITANSHU TARENDRA.; Thakur, Dr. K. D.Soybean [Glycine max (L.) Merrill] is cultivated intensively in India. Legumes derive 5-83 per cent of their nitrogen (N) requirement by biological nitrogen fixation under field conditions. The use of various chemicals is imperative to achieve profitable yields. However, these chemicals used as plant protectants have been found to exert either synergistic or toxic effects on legume-Rhizobium symbiosis. Therefore, present studies on ‘‘Characterization of Bradyrhizobium japonicum and its compatibility with fungicides”, were planned and conducted with defined objectives, at Plant Pathology Section, College of Agriculture, Nagpur, during 2018-19. The test B. japonicum and Rhizobium sp. was isolated from functional root nodules of soybean plants, on congo-red yeast extract mannitol agar medium. The colonies of isolates were circular, convex, glistening, opaque and whitish pink colonies of 2-4 mm dia. with regular or entire margin with marked distinction from red colored colonies of Agrobacterium. The purified strains were aerobic, gram negative, non-spore forming and motile rods. Out of 25 purified isolates, 20 (80%) were slow grower Bradyrhizobium sp. (turn medium blue) and 5 (20%) were fast grower Rhizobium sp. (turn medium yellow) was found on YEMA media supplemented with Bromothymolblue. The optimum growth kinetics for B. japonicum and Rhizobium sp. was 35°C at neutral pH (7.0). Both fast and slow growing rhizobia were found to be positive for oxidase, KOH, starch hydrolysis, nitrate reduction and catalase activity. Among all the 25 isolates tested, 23 isolates showed positive reaction to gelatin hydrolysis except Bj-11 and Bj-21 while, 21 isolates were reacted positive to H2S production whereas, the four isolates viz., Bj-1, Bj-2, Bj-8 and Bj-23 were negative in reaction. While all the samples were found negative for IAA production. Pot culture experiment revealed that seed inoculation with different B. japonicum and Rhizobium sp. isolates recorded significant improvement in nodule number and plant height over uninoculated control. All the B. japonicum and Rhizobium sp. inoculants recorded higher plant height ranged from 35.00 to 65.00 cm at flowering stage. Interactions of all the inoculants ranged from 3.00 to 22.00 nodules per plant at maximum flowering period. Some of the isolates viz., Bj-9, Bj-10, Bj-14, Bj-17, Bj-19, Bj-23, Bj-25 could produce large and pink nodules indicating their effective symbiosis with soybean. The test of compatibility with systemic, non-systemic / contact, combi-fungicides at recommended field dosages 50%, 75%, 100% and 125% of the Reommended doses were evaluated in vitro against B. japonicum of soybean, by inhibition zone technique. The fungicides carbendazim 50% WP and mancozeb 75% WP at all four dosages were found compatible with B. japonicum, as they didn’t showed any zone of inhibition, at 72 hrs of incubation. Whereas, rest of the three fungicides viz., captan 75% WP, carboxin 75% WP, vitavax power 75% WP tested at four various concentrations were found inhibited by the test bacterium and produced average inhibition zone of around 00.00-19.66 mm, 8.56mm and 10.52mm respectively, at 72 hrs of incubation.ThesisItem Open Access COMPARATIVE EFFECT OF CARRIER BASED Bradyrhizobium japonicum AND PSB ON GROWTH AND YIELD OF SOYBEAN.(College of Agriculture Nagpur, Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, 2019) BORUDE, SANTOSH ANKUSHRAO.; Ingole, Dr. M. N.The present investigations entitled “Comparative effect of carrier and liquid based Bradyrhizobium japonicum and PSB on growth and yield of soybean was conducted in Plant Pathology Section, College of Agriculture, Nagpur during Kharif 2018-19.Effect of various soybean seed treatments on germination, shoot length, root length and seedling vigour index were recorded. Highest germination (93.33%), shoot length (16 cm), root length (16.66 cm) and maximum seedling vigour index (3048.15) were achieved in treatment of Carbendazim + Bradyrhizobium + PSB. Seed treatment with Carbendazim + Bradyrhizobium japonicum + PSB showed highest nodulation at 30 days (34.45), 45 days (39.19) and 60 days (41.33). In vivo evaluation of Carbendazim and biofertilizers seed treated with Carbendazim + Bradyrhizobium japonicum + PSB indicated maximum amount of available Nitrogen (180.24 kg / ha) and Phosphorous (15.1 kg / ha) as compare to control N (111.41 kg / ha) & P (8 kg / ha).Effect of various treatments on yield and dry matter were evaluated. Highest yield (2266 kg / ha) and dry matter (16.90 g / plant) was obtained in Carbendazim + Bradyrhizobium japonicum + PSB. Bradyrhizobium Japonicum and PSB were found compatible with all doses of carbendazim. Whereas Trichoderma viride was observed non compatible with test doses of carbendazim. Seeds treatment with Carbendazim + Bradyrhizobium + PSB achieved higher ICBR i.e. 1: 13.32 ratio as net return Rs. 17664 / ha.ThesisItem Open Access MANAGEMENT OF GLADIOLUS WILT BY USING LEAVES OF ALLELOPATHIC PLANTS.(College of Agriculture Nagpur, Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, 2019) GAVADE, VAIBHAV SUHAS.; Potdukhe, Dr. S. R.The present study entitled as "Management of gladiolus wilt by using leaves of allelopathic plants" was carried out during 2018-2019, at Plant Pathology Section, College of Agriculture, Nagpur, with two main treatment viz., Levels of allelopathic M1 (2g leaves/100g soil), M2 (4g leaves/100g soil) and M3 (6g leaves/100g soil) and five allelopathic plants viz., Allium sativum (Garlic clove), Ipomoea carnea (Besharm), Annona innoxia (Sitaphal), Eucalyptus globulus (Eucalyptus) and Azadirachta indica (Neem) were evaluated against Fusarium oxysporum. The test fungus was found pathogenic and resulted 80% wilt in Psittacenus Hybrid. In vivo experiment Azadirachta indica treatment was found significantly superior requires minimum days to emergence of corms (15 days) and minimize the wilt incidence (56.89%) and mortality (11.11%) and also increased shoot length plant-1 (35.10 cm/ plant-1), spike length plant-1 (50.22 cm), number of leaves plant-1 (5.73 number of flower/spike (7.21), number of corm plant-1 (6) and weight of corm (16.04g) over Fusarium. Treatment M3 showed minimum wilt incidence (55.01%) and mortality (22.22%) and maximum shoot length (35.45 cm/ plant-1), spike length plant-1 (49.41 cm), number of leaves plant-1 (5.74), number of flower/spike (6.96), number of corm plant-1 (6.29) and weight of corm (15.87 g/pl) than that of M2 treatment (4g leaves/100g soil) and M1 treatment (2g leaves/100g soil).ThesisItem Open Access MANAGEMENT OF TIKKA DISEASE OF GROUNDNUT USING BOTANICALS, BIOGENTS AND CHEMICALS.(College of Agriculture Nagpur, Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, 2019) PAWAR, VISHAL DEVENDRA.; Guldekar, Dr. D. D.Groundnut (Arachis hypogaea L.) is one of the most important oil seed crops of the world and Tikka disease or leaf spots of groundnut is very severe disease of it which is caused by Cercospora arachidicola (early leaf spot) and Phaeoisariopsis personata (late leaf spot). The present study was carried out to study the efficacy of botanicals, bioagents and fungicides on tikka disease of groundnut. The experiment was conducted in randomized block design with three replications and nine treatments during Kharif season 2017-18 at the farm of section of Plant Pathology, Nagpur. The results of present investigation revealed that, there was significant differences in percent disease incidence and intensity at 30, 60 and 90 DAS. Among the fungicides, three sprays of Carbendazim reduced the tikka disease significantly as compared to all other treatments followed by Mancozeb. Among the botanicals Azadirachta indica and Allium sativum showed good results in controlling the disease and incase of bioagents, Trichoderma viride proved effective for the same.ThesisItem Open Access MOLECULAR AND BIOCHEMICAL CHARACTERIZATION OF XANTHOMONAS Citri Subsp. citri.(College of Agriculture Nagpur, Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, 2019) ISOKAR, SHUBHAM SUBHASH.; Bramhankar, Dr. S. B.Citrus is one of the important tropical and subtropical fruits in the world as well as in India which belongs to the family Rutaceae and sub-family Aurantioedae. Among all factors responsible for decline in citrus production, citrus canker is one of the most serious problem which is caused by Xanthomonas citri subsp. citri. The bacterium infects the twigs, petioles, fruit stalks and fruits, resulting in both qualitative and quantitative loss in acid lime. The disease is characterized by raised corky necrotic lesions, surrounded by yellowish halo on the leaves. Xanthomonas citri subsp. citri was successfully isolated on the nutrient agar medium, from the fresh lesions on the leaves of naturally canker infected acid lime leaves collected from different agroclimatic zones of Maharashtra. The fourteen isolates were designated from Xcc-1 to Xcc-14 representing the agroclimatic zone. Pathogenic ability of all different isolates of Xcc were confirmed and found that isolate Xcc-6, Xcc-7 Xcc-10, Xcc11 and Xcc-12 showed highly pathogenic to initiate minute canker lesion and fully developed symptoms after 17-22 days. While Xcc-1, Xcc-2, Xcc-3, Xcc-4, Xcc-5, Xcc-8, Xcc-9 and Xcc-13 were found to produce very poor in virulence symptoms. The studies on physiological characteristics of Xanthomonas citri subsp. citri. showed the maximum growth of pathogen observe at temperature 300C and at pH level 7. A several bio-chemical test were conducted to differentiate the isolate. All isolates of bacteria were found negative test for gram’s staining while positive for starch hydrolysis, potassium hydroxide test, catalase test, hydrogen sulphide production, gelatin liquefaction, indole production, nitrate reduction acid and gas production tests to performed to characterized the Xanthomonas citri subsp. citri. Molecular detection by polymerase chain reaction with specific primers J-pth1/Jpth-2, J Rxg/Rxc2,Xac-01/Xac-02, M+/M- and Ms+/Ms- was performed on fourteen isolates. The patterns showed by PCR analysis were identical for all the isolates tested confirming their identity as Xanthomonas citri subsp. citri. The genetic variability was studied by using five inter simple sequence repeat and randomly amplified polymorphic DNA primers. A total of 16 amplicons were obtained with the 5 UBC series primers. Out of 16 bands 14 were found to be polymorphic and 2 were monomorphic and the level of average polymorphism was 87.50 per cent. Isolate Xcc-11 (Nagpur) had higher value of similarity coefficient (0.85) with Xcc-10 (Wardha), whereas Xcc-12 (Gadchiroli), had lower value of similarity coefficient (0.42) with Xcc-1 (Sindhudurg) with high degree of polymorphism while two primers OPF-4 and OPF-2 failed to produce scorable bands. A total of 28 amplicons were obtained with the 3 primers. All the bands produced were polymorphic with 100 per cent polymorphism. Isolate Xcc-11 (Nagpur) had higher value of similarity coefficient (0.81) with Xcc-10 (Wardha), whereas Xcc-2 (Dapoli), had lower value of similarity coefficient (0.41) with Xcc-13 (Gondia).ThesisItem Open Access CULTURAL AND MORPHOLOGICAL CHARACTERIZATIONS OF Beauveria bassiana.(College of Agriculture Nagpur, Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, 2019) WARGANE, VILAS SUKHADEO.; Parate, Shri. R. L.An investigation entitled "CULTURAL AND MORPHOLOGICAL CHARACTERIZATIONS OF Beauveria bassiana" was carried out during 2018-2019. Effects of nutrition, pH, temperature and humidity were assessed on the basis of radial growth. Influence of different spore concentrations viz., 105, 106, 107, 108 and 109 spores per ml were tested for the pathogenicity of B. bassiana against second instar larvae of Spodoptera litura. Similarly compatibility of B. bassiana was evaluated with commonly used insecticides viz., spinosad 45 SC, imidachloprid 200 SL, chloropyriphos 20 EC and indoxacarb 14.5 SC. The mummified bodies of lepidopteran pest larvae with symptoms of fungal infection were collected from field conditions from different places. Detailed morphological studies of effective isolate (Bb3) show that the fungus grew well on SDA (sabouraud dextrose agar). The mycelium on this medium, appeared white, dense and powdery. The colonies appeared raised in the center and developed radial furrows. The culture was turned yellowish white and powder type. Spore structures was globose to sub-globose. Two different semi synthetic media namely sabouraud dextrose agar and potato dextrose agar were evaluated for better growth of B. bassiana and it was observed that SDA medium (41.50 mm) support the growth of isolate. Significantly higher radial growth was however, seen in media on the basis of these results, pH value 6.0, temperature of 25°C and 95 per cent relative humidty was considered as optimum for growth and sporulation of B. bassiana. In terms of larval mortality, 107 spores per ml was recorded highest mortality of S. litura followed by 108 spores per ml, least larval mortality was observed at 105 spores per ml after 3, 5, 7 and 10 days of treatment. Compatibility of B. bassiana against commonly used insecticides in plant protection was tested. Imidachloprid were non-toxic to B. bassiana, as no significant reduction in radial growth was noticed. The insecticide chloropyriphos recorded 77.20 per cent inhibition of growth and was toxic to B. bassiana.ThesisItem Open Access SHELF LIFE OF Trichoderma asperellum in DIFFERENT OIL FORMULATIONS.(College of Agriculture Nagpur, Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, 2019) MANE, KU. KAJAL KISAN.; Guldekar, Dr. D. D.An investigation entitled “Shelf life of Trichoderma asperellum in different oil formulations” was carried out during 2018-2019. paraffin oil, soybean oil, groundnut oil, potato dextrose broth and talc was extensively used as carrier for Trichoderma asperellum. As compared to solid based carrier material the survival of Trichoderma asperellum in liquid based formulations is quite high and have the ability to limit the heat transfer high water holding capacity and maintain water around the cells for their metabolism. Shelf life studies clearly indicated significant differences at all the intervals. Population density of T. asperellum in the paraffin oil was 28.00 x 108 CFU/ml at 30 days whereas the population density of T. asperellum in the 180 days was 18.33 x 108 CFU/ml. It was found significantly superior over all other treatment. The per cent spore germination in formulation containing paraffin oil is 59.18 per cent. Maximum colonies was observed in pH range of 5.2-5.7 in paraffin oil treatment (T1) when the effect of liquid formulation on pH of Trichoderma asperellum was studied. It is significantly superior over the other treatment which we have under study. This ph range were found to be optimal for antagonistic potential of T. asperellum against Fusarium oxysporum f. sp. ciceri, Rhizoctonia bataticola and Sclerotium rolfsii. The effect on percent growth inhibition on Fusarium oxysporum f. sp. ciceri is 74.44 per cent, on Rhizoctonia bataticola is 81.85 per cent and Sclerotium rolfsii is 72.6 per cent. It was recorded maximum in formulation containing paraffin oil it was found significantly superior over all the treatments. The effect of liquid formulations on spore germination of T. asperellum was also recorded the maximum spore germination are found in formulation containing paraffin oil after 180 days of storage. The effect of Trichoderma formulation on seed germination, root length, shoot length, SVI of bengal gram, when seed inoculated with Fusarium oxysporum, Rhizoctonia bataticola, Sclerotium rolfsii by using paper towel method maximum germination percentage was 87.33, 74.33, 73.00 percent respectively in T1 (paraffin oil) . However root and shoot length was maximum in T1 (21.66 cm and 31.33 cm), (19.66 cm and 30.66 cm), (18.66 cm and 30.00 cm) respectively. The maximum seedling vigour index was found in paraffin oil seed treatment.
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