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Dr. Rajendra Prasad Central Agricultural University, Pusa
In the imperial Gazetteer of India 1878, Pusa was recorded as a government estate of about 1350 acres in Darbhanba. It was acquired by East India Company for running a stud farm to supply better breed of horses mainly for the army. Frequent incidence of glanders disease (swelling of glands), mostly affecting the valuable imported bloodstock made the civil veterinary department to shift the entire stock out of Pusa. A British tobacco concern Beg Sutherland & co. got the estate on lease but it also left in 1897 abandoning the government estate of Pusa.
Lord Mayo, The Viceroy and Governor General, had been repeatedly trying to get through his proposal for setting up a directorate general of Agriculture that would take care of the soil and its productivity, formulate newer techniques of cultivation, improve the quality of seeds and livestock and also arrange for imparting agricultural education. The government of India had invited a British expert. Dr. J. A. Voelcker who had submitted as report on the development of Indian agriculture. As a follow-up action, three experts in different fields were appointed for the first time during 1885 to 1895 namely, agricultural chemist (Dr. J. W. Leafer), cryptogamic botanist (Dr. R. A. Butler) and entomologist (Dr. H. Maxwell Lefroy) with headquarters at Dehradun (U.P.) in the forest Research Institute complex.
Surprisingly, until now Pusa, which was destined to become the centre of agricultural revolution in the country, was lying as before an abandoned government estate. In 1898. Lord Curzon took over as the viceroy. A widely traveled person and an administrator, he salvaged out the earlier proposal and got London’s approval for the appointment of the inspector General of Agriculture to which the first incumbent Mr. J. Mollison (Dy. Director of Agriculture, Bombay) joined in 1901 with headquarters at Nagpur
The then government of Bengal had mooted in 1902 a proposal to the centre for setting up a model cattle farm for improving the dilapidated condition of the livestock at Pusa estate where plenty of land, water and feed would be available, and with Mr. Mollison’s support this was accepted in principle. Around Pusa, there were many British planters and also an indigo research centre Dalsing Sarai (near Pusa). Mr. Mollison’s visits to this mini British kingdom and his strong recommendations. In favour of Pusa as the most ideal place for the Bengal government project obviously caught the attention for the viceroy.
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ThesisItem Open Access Biotechnological development of Panama wilt resistance in local cultivars of banana(RAJENDRA AGRICULTURAL UNIVERSITY, BIHAR PUSA (SAMASTIPUR), 2017) Kumari, Anita; Kumar, HarshBiotechnological development of resistance to Panama wilt, the most important disease caused by Fusarium oxysporum f.sp. cubense, was explored in four important local cultivars of banana namely Robusta, Alpan, Malbhog and Kothia employing tissue culture studies and genetic transformation. Using tissue culture, efficient micropropagation protocol and disease resistance utilizing somaclonal variation were developed. Biological control and management of the disease was done utilizing endophytic association of nonpathogenic strain of Fusarium oxysporum. The culture of shoot apices and male flower buds resulted in their establishment and swelling, and callogenesis, caulogenesis, differentiation of shoots and roots from them. Elongation of existing shoots was observed from shoot apices and somatic embryogenesis from floral buds only. The best medium for caulogenesis was BM19 (MS+1.14 µM IAA+19.97 µM BAP) for cultured shoot apices and medium BM8 (MS+22.19 µM BAP) for cultured male flower buds. The best medium for somatic embryogenesis was BM20 (MS+18.10 µM 2,4-D+5.37 µM NAA+ 5.71 µM IAA+ 1mg Biotin). The cultivar Robusta was the best for all tissue culture responses except somatic embryogenesis followed by either cultivar Alpan or cultivar Malbhog. The cultivar Kothia showed the least tissue culture responses. However, the best response for somatic embryogenesis was observed in cultivar Kothia followed by cultivars Malbhog, Alpan and Robusta respectively. The subculture of in vitro developed multiple shoots on medium BM19 resulted in cent percent shoot multiplication with a high multiplication rate. An efficient micropropagation protocol was developed for banana cultivars. The Panama wilt resistance was developed in two susceptible cultivars Alpan and Malbhog exploiting somaclonal variation. Endophytic nonpathogenic Fusarium oxysporum strains were found effective in biocontrol of wilt disease in banana. Genetic transformation of callus and regenerating shoots was done using GUS and eGFP marker gene constructs and it was confirmed using histochemical GUS assay and fluorescence microscopy respectively, and by molecular validation using specific SSR primer pairs. Thus, the work established a suitable protocol of the genetic transformation, which can be used for the development of Panama wilt resistance in local cultivars of bananas.ThesisItem Open Access Gene Expression Analysis For Biosynthesis And Catabolism Of Glucosinolates In Rocket (Diplotaxis tenufolia L.) Under Abiotic Stress(Dr. Rajendra Prasad Central Agricultural University, Pusa (Samastipur), 2017) Mishra, Shubhi; Kumar, M.Glucosinolates are one of the important nutritive content of rocket (Diplotaxis tenufolia L.), a leafy vegetable salad crop. They are responsible for its pepper taste, pungent aroma and medicinal properties. In this experiment, content of chlorophyll and chlorophyll fluorescence parameters were measured and biochemical analysis of glucose and glucosinolates content was done. The experiment was further followed by expression profiling of the key genes of metabolic pathway of glucosinolate using real-time PCR. The results indicated that the plant had some adaptation mechanism to mitigate the adverse effects of heat and salt stress which resist the changes in the chlorophyll content and chlorophyll fluorescence of rocket leaves as non-significant changes in the chlorophyll content and chlorophyll a fluorescence of rocket leaves.An increase in glucosinolate content was observed due to heat and salt stress in agreement with the inference derived on the basis of gene expression analysis. However in the case of roots, it appeared that glucosinolates content started decreasing after reaching the saturation point which was amply supported by higher level of expression of glucosinolates catabolism gene DtMyrosinase as observed in roots.It appeared that glucosinolates content can be increased if grown under heat and salt stress and can also be limited if grown under controlled condition. Isolation of the full length of key genes involved in glucosinolates biosynthesis namely, DtCytochromeP79F1 and catabolism namely, (DtThiol-methyl transferase) was also done through RACE. Altogether ten primers including eight primers specific to target the eight genes of glucosinolate metabolic pathway and two RACE specific primers were designed and validated during the present study which may be further utilized as tools to examine target region amplification polymorphism and expression in relation to glucosinolate biosynthesis pathway in rocket plants.ThesisItem Open Access Genome wide association study for identification of genes related to ion accumulation in rice grain(Dr. Rajendra Prasad Central Agricultural University, Pusa (Samastipur), 2017) Kumar, Ajay; Sharma, V. K.A study was conducted to investigate and identify the candidate genes in relation to cadmium and zinc transportation and accumulation in rice. Using 281 diverse rice accessions and employing genome wide association analysis to relate 46294 single nucleotide polymorphic panel and cadmium and zinc ionomic data, mixed linear model was adopted to establish the association, if any, between genotypic and ionomic data. Statistical tools used and computational applications adopted during the analyses include Discriminant Analysis of Principal Components (DAPC), Principal Coordinate Analysis (PCoA), Neighbour Joining Tree, Structure, Haploview and R-tool. Differential accumulation of cadmium and zinc in different vegetative tissues of two diverse rice accessions was also investigated under cadmium induced stress condition. Finally, expression profiles of OsHMA2, OsHMA3 and OsPCSs genes under cadmium induced stress condition were examined. Differential cadmium accumulation was recorded in grains of rice accessions evaluated under alternate wetting and drying and permanent flooding conditions as well as across the conditions. While in the case of zinc, differential accumulation was observed between accessions but differences recorded between conditions were not so pronounced. PCoA based on SNPs data derived using 297 accessions clearly indicated that 12 entries were very diverse from rest of the entries under evaluation. Further, an analysis using neighbour joining tree based on distance matrix also revealed that these 12 genotypes were remarkably distant from rest of the entries. Cluster analysis applied to detect population genetic structure clearly indicated that 10 out of these 12 accessions belonged to a separate cluster. Finally, an analysis carried out after exclusion of 16 accessions including 12 accessions, which were found to be more diverse based on PCoA and neighbour joining tree and 4 very late maturing accessions, established that remaining 281 accessions belonged to three clusters. DAPC employed to find out the optimum number of clusters confirmed the results obtained through PCoA, neighbour joining tree and cluster analysis applied to detect population genetic structure in the present investigation. Association analysis revealed twelve and two markers; seven and one candidate loci; and 109 and 10 candidate genes associated with cadmium and zinc accumulation, respectively. Haplotype analysis of polymorphic markers identified in relation to cadmium and zinc accumulation, therefore, resulted in allocation of seven and one loci, respectively, in question to haplotype groups. Result of the present study led to the identification of candidate genes involved in cadmium and zinc accumulation under aerobic condition. Altogether, eight and five genes appeared to be involved in cadmium and zinc accumulation, respectively. Present study, therefore, provided the evidence based on which the candidate genes most likely to be involved in zinc accumulation have been proposed. Most probable involvement of MATE and PEZ family genes in cadmium accumulation in rice grain was confirmed on the basis of results of present study. Cadmium accumulation property appeared less dependent on its uptake by the plants but more dependent on the capacity of the accession to trap the cadmium into the roots by synthesizing two folds non-protein thiol content, thereby preventing its translocation or mobility from root to shoot. Antagonistic effect of cadmium accumulation was observed in respect of zinc accumulation in rice accessions. Expression analysis in relation to cadmium ion translocation reflected that OsHMA3 was more expressed in less efficient accession as compared to the efficient accession. Therefore, the overexpression of OsHMA3 most probably resulted in low accumulation of cadmium in the plants of less efficient accession due to lesser translocation.ThesisItem Open Access Microsatellite markers based characterization of rice genotypes in relation to drought tolerance(Dr. Rajendra Prasad Central Agricultural University, Pusa (Samastipur), 2017) Choudhary, Shuchi Shankar; Kumar, MithileshEighteen genotypes of rice were evaluated for various physio-morphological characters and to investigate the microsatellite markers based polymorphism for their characterization and differentiation using polymorphic and informative markers in order to estimate the extent of genetic diversity among these rice genotypes.Experimental materials were evaluated in randomized block design with three replications during three consecutive years. Molecular characterization was done by targeted amplification of the genomic DNA using a panel of thirty two microsatellite primer pairs covering nine chromosomes. Statistical methods and parameters used for deriving inference were analysis of variance, range, mean values, relative mean performance, drought tolerance indices, variability parameters, correlation andpath coefficients. An analysis of variance revealed significant differences among the genotypes for all the characters evaluated during present study. Considerable extent of variability existed for all the attributes recorded among the genotypes. The coefficient of variability was recorded moderate to high genetic variability for number of grains per panicle, root length and yield per plant under normal condition consistently over years, whereas for leaf rolling, number of tillers per plant, number of grains per panicle, plant height, harvest index and yield per plant under drought condition. Yield per plant recorded high variability amongst the genotypes evaluated under both conditions.The genotypes RAU-1426-4-3-2-57-2, RAU-1416-4-2-5-2-2 and Sahbhagi Dhan recorded remarkably higher mean performance fornumber of tillers per plant, number of grains per panicle, 1000-grains weight, harvest index and yield per plant, under both drought and normal conditions. Similarly, genotypes like RAU-1397-2-5-8-1-2-5-4, RAU-1453-12, RAU-1451-6-6-1-1-5-2 and Sahbhagi Dhan exhibited superior performance in respect of root length, plant height, dry root weight, relative leaf water content, canopy temperature and number of grains per panicle under both drought and normal conditions. The relative mean performance of genotypes, namely,RAU-1421-12-1-7-4-3, RAU-1415-3-5-7-6-9-5-3, RAU-1401-1-8-1-5, RAU-1478-5-2-2-4-6, RAU-1451-6-6-1-1-5-2, RAU-1416-4-2-5-2-2, RAU-1426-4-3-2-5-7-2, RAU-1453-12 and Sahbhagi Dhan to be relatively more drought tolerant than other genotypes under evaluation. Root length, plant height, dry root weight, spikelet fertility, number of grains per panicle, 1000-grain weight, harvest index and yield per plant exhibited moderate to high heritability in broad sense coupled with moderate to high estimates for genetic advance as per cent of mean under both drought and normal conditions over years.Correlation studies, revealed that root length, number of tillers per plant, relative leaf water content, spikelet fertility, 1000-grain weight and harvest indexshowed significant positive association with yield per plant under both conditions. However, positively significant association was also observed between yield per plant and number of grains per panicle under drought condition.Path coefficient analysis revealed that the characters, namely, numbers of tillers per plant, root length, dry root weight1000-grain weight and harvest index had considerable positive direct effects on yield per plant more or less consistently across the years under drought as well as normal conditions.Four tolerance indices viz. TOL, SSI, DTE and STI were used. According to TOL, SSI and DTE, 12 genotypes,namely, RAU-1421-12-1-7-4-3, RAU-1415-3-5-7-6-9-5-3, RAU-1401-1-8-1-5, RAU-1478-5-2-2-4-6, RAU-1477-9-7-2-2-5-7-3, RAU-1451-6-6-1-1-5-2, RAU-1416-4-2-5-2-2, RAU-1426-4-3-2-5-7-2, RAU-1463-16, RAU-1471-10, RAU-1453-12 and Sahbhagi Dhan appeared to be relatively more tolerant to drought stress. Among these 12 genotypes, 7 genotypes namely,RAU-1421-12-1-7-4-3, RAU-1415-3-5-76-9-5-3, RAU-1451-6-6-1-1-5-7-3, RAU-1416-4-2-5-2-2, RAU-1426-4-3-2-5-7-2, RAU-1453-12 and Sahbhagi Dhan recorded higher STI valueexhibiting high tolerance to drought condition. Using a set of thirty two microsatellite primers generated allelic variants ranged from 4 in case of RM 85 to 16 in case of RM 324 and RM518.The primer pair RM 3,RM 18, RM 72, RM 87, RM225, RM 262, RM302, RM 324, RM 327, RM 416, RM 518, RM 520, RM 521, RM1349, RM 5443 and RM 23679generated amplified products due to amplification of more than one microsatellite locus. Altogether 300 allelic variants were detected at 48 microsatellite loci with an average of 6.25 alleles per locus. A total of 174 shared and 126 unique allelic variants were generated. Considering the number of alleles in conjunction with the level of polymorphism detected, the primersRM 231, RM 225, RM 163, RM 555, RM 212, RM 72, RM 3549, RM 1349, RM 521, RM 5443 and RM 302 appeared to be more informative primers.Analysis of divergence pattern based on microsatellite markers allowed differentiation and classification of rice genotypes into two groups. The first multi-genotypic group consisted of twelve genotypes, namely, RAU-1428-6-7-3-6, RAU-1417-2-1-5-7-7,RAU-1421-12-1-7-4-3, RAU-1415-3-5-76-9-5-3, RAU-1401-1-8-1-5, RAU-1428-4-3-2-7-26, RAU-1478-5-2-2-4-6, RAU-1477-9-7-2-2-5-7-3, RAU-1451-6-6-1-1-5-2, RAU-1416-4-2-5-2-2, RAU-1397-2-5-8-1-2-5-4 and RAU-1451-3-5-7-6-9-5-1 whereas the second multi-genotypic group consisted of six genotypes, namely, RAU-1426-4-3-2-5-7-2, RAU-1428-3-1-5-4, RAU-1463-16, RAU-1471-10, RAU-1453-12 and Sahbhagi Dhan.The maximum similarity value of 0.46 was observed between the genotypes i.e. genotype RAU-1477-9-7-22-5-7-3 and RAU-1451-6-6-1-1-5-2 indicating that these were more closely related. The panel of microsatellite markers employed for molecular characterization in the present study showed very high degree of efficiency in discrimination of genotypes in relation to drought tolerance.ThesisItem Open Access Characterization of wheat (Triticum aestivum L.) genotypes for disease reaction against spot blotch pathogen(Dr. Rajendra Prasad Central Agricultural University, Pusa (Samastipur), 2017) Kumari, Priti; Kumar, RajeevWheat (Triticum aestivum L.) is the largest consumed cereals and most important element in the diet of the poor. Successful production of the wheat in the warmer regions of South Asia is constrained by several biotic and abiotic stresses. Spot blotch of wheat caused by Bipolaris sorokiniana is one of the most significant biotic stresses which limits the production and productivity of the crop in the region. Under present investigation eighty-eight genotypes of wheat were evaluated for the purpose of twelve morpho-metric traits and reaction against spot blotch disease in randomized block design (RBD) during rabi, 2015 Molecular characterization using 17 SSR markers (associated with tolerance against spot blotch disease) was also undertaken to as certain the genetic diversity among the studied genotypes Observations on 1000-grain weight, grain filling duration (GFD), no. of tiller per plant, canopy temperature, yield per plot, heading, days to greenness, disease severity and area under disease progress curve (AUDPC), lesion size, leaf tip necrosis, biomass per square meter were recorded. Significant differences were observed among the genotypes for all the traits under consideration. Genotype × environment interactions were found non-significant. Molecular characterization work revealed a total of 232 alleles in 19 genotypes using 17 SSR markers. The number of alleles per locus varied from four in Xgwm371 to thirty in Xgwm 445 with an average of 11.04 alleles per locus. Total 165 unique alleles were observed at 21 SSR loci, with an average of 7.85 unique alleles per locus. The number of unique alleles per locus ranged from one in Xgwm 371 to twenty six in Xgwm 445. Out of 17 SSR primers 10 showed null alleles. The maximum 4 genotypes (Ariana, Dharwad Dry, Ciano T 79 and HD 2967) exhibited null alleles for the primer pair Xgwm 120. Stutter bands were also detected in the present investigation. Such bands were observed in the case primer pairs, Xgwm-611, Xgwm-159 Xgwm 132, Xgwm 445, Xgwm 371, Xgwm146, Xgwm 425. The PIC values varied from 0.435 in the case of Xgwm120 to 0.959 in the case of Xgwm 146 with an average of 0.876 across the primer, indicating that the markers were highly informative (PIC>0.5). The use of SSR markers in the present analysis exhibited a remarkably higher level of genetic polymorphism and allowed unique genotyping of the genotypes included in the analysis. The nineteen genotypes classified into five different clusters or groups (I to V) at 22 phenon level based on cluster analysis using the UPGMA method based on Dice coefficients.
