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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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  • ThesisItemOpen Access
    Microsatellite markers based assessment of genetic diversity and construction of heterotic groups in maize
    (Dr. Rajendra Prasad Central Agricultural University, Pusa (Samastipur), 2016) Punya; Sharma, V. K.
    A study was undertaken to elucidate the nature and extent of differentiation and divergence among eighteen inbred lines of maize and to separate them into heterotic groups based on the analysis of targeted microsatellite sites. Experimental materials of the present study comprised fifteen inbred lines, three testers and forty five single cross experimental hybrids, which were evaluated in randomized block design with three replications under rabi and kharif seasons. Statistical analyses to derive inferences regarding divergence pattern of parental lines and heterosis manifestation in experimental hybrids were carried out using the data recorded on days to tasseling, days to silking, days to brown husk, plant height, ear height, ear length, number of kernels per ear and grain yield per plant. Numerical taxonomic approach of classification using these eight quantitative characters of agronomic importance based average taxonomic distance was done to discriminate the inbred lines into different clusters. Inbred lines were also raised in pots for extraction of genomic DNA from the young seedlings. Amplification of the genomic DNA was carried out using a panel of twenty eight microsatellite sites specific primer pairs covering all the chromosomes existing in the genome. Microsatellites based allelic diversity data were utilized for classification and formation of heterotic groups of the inbred lines. Analysis of variance revealed the existence of significant differences among the inbred lines and hybrids for all the characters recorded under both the seasons. Mean performance of parents and hybrids significantly varied for the eight quantitative characters under study. Among the inbred lines, CML467 recorded significantly higher grain yield per plant followed by tester CML165, which was statistically at par to inbred lines LM13, HKI586, CML465, HKI162 and HKI323B over the environments. Sixteen crosses recorded significantly higher mean performance and significantly positive heterosis for grain yield per plant. Among these sixteen crosses, the highest yielding cross HKI162 × CML161 was statistically at par to five crosses, namely, CML468 × CML161, CML471 ×CML161, HKI323B × CML161, CML469 × CML165 and CML471 × CML165 for grain yield per plant. Taxonomic distance based cluster analysis revealed ample genetic diversity among inbred lines, which occupied different clusters during rabi and kharif season and across the seasons. The results indicated a close correspondence with respect to clustering pattern of inbred lines during rabi and kharif seasons. A perusal of dendrogram generated on the basis of eight quantitative traits indicated that the inbred lines CML467, CML470 and DH2012 were relatively more diverse from the remaining inbred lines under evaluation. Altogether 296 allelic variants were detected amongst amplified products generated with 28 primer pairs. A total of 49 loci were assigned to 28 primer pairs with an average of 6.04 alleles per locus The number of alleles ranged from 6 in the cases of umc1136, phi072 and umc1179 to 16 in case of phi053 with a range between 56 to 352 bp. Considering the number of alleles generated by different primer pairs in conjunction with the level of polymorphism, the primers umc1297, phi053, umc1266, phi093, bnlg118, phi034, phi115, phi065 and phi084 appeared to be more informative primers. Among the inbred lines under molecular characterization, CML163 and CML467 appeared as the most diverse genotypes. A remarkably higher level of genetic polymorphism was revealed by the use of 28 microsatellite markers. Both quantitative and molecular data based cluster analyses revealed that inbred lines CML 468 and CML469 are closely related to each other. Remarkably greater extent of similarity was also noticed between inbred lines HKI323B and HKI586. All the three testers, namely, CML161, CML165 and CML163, were included in same cluster in both the cases and seemed to be closely related. However cluster analysis using quantitative characters and microsatellite markers based data did not exhibit a complete correspondence. Parental genetic distance based on quantitative data or genetic similarity based on molecular data did not exhibit significant association with grain yield heterosis in hybrids. Results clearly indicated that a high grain yield is not always obtained from hybrids that have high parental genetic distance value, but generally from those with moderate to high genetic distance. A comparative assessment of heterotic groups formed by using microsatellite markers based genetic similarity, hybrid index value and hybrid mean value, indicated that CML471 and CML115 or HKI323B and HKI586 or DH2012 and LM13 belonged to same heterotic group in all the cases. The distribution pattern of the entries into different heterotic groups formed on the basis of hybrid index value and hybrid mean value corresponded with the microsatellite markers based heterotic groups to the extent of 75% and 67% in the first heterotic group, 40% and 29% in the second heterotic group and 33% and 40% in the third heterotic group, respectively. Overall coincidences of inbred lines in heterotic groups formed on the basis of microsatellite markers with the hybrid index value and hybrid mean value based heterotic groups were considerably higher. Results, therefore, indicated that microsatellite markers may be effectively and efficiently utilized to assign the inbred lines to heterotic groups for grain yield and to establish heterotic groups of maize inbred lines.
  • ThesisItemOpen Access
    Molecular Characterization of Bipolaris sorokiniana Isolates Collected from Wheat by SSR Markers
    (Rajendra Agriculrural University, Pusa (Samastipur), 2016) Bharty, Archana; Kumar, Harsh
    Spot blotch of wheat caused by Bipolaris sorokiniana is one of the most important fungal diseases of wheat (Triticum aestivum). The pathogen is seed and soil borne. Pusa in Bihar is considered as a hot spot for the disease. 36 fungal isolates were collected from infected leaves and seeds of different wheat genotypes grown at the research farms of RAU, BISA, IARI Regional Station at Pusa and from Patna, Muzaffarpur, Nalanda, Madhepura, Mahua and Jamaui. They were characterized morphologically on the basis of colony colour, growth pattern and exudation, and molecularly through SSR markers. The isolates were tested on two wheat cultivars, susceptible Sonalika and resistant Chirya-3 for disease severity, AUDPC and aggressiveness under natural condition in field and controlled condition in polyhouse. The isolates were divided into five morphological groups on the basis of their colony colour namely black, grey, grey cottony knot, grey white and whitish black, among which the frequency of black was the maximum and showed high aggressiveness. To examine the molecular genotypic variability among the isolates, their genomic DNA was isolated and amplified using 10 Bipolaris specific SSR (microsatellite) primer pairs. A total of 110 allelic variants were detected including 35 unique alleles, 75 shared alleles (including 7 null alleles) at 18 loci with an average number of 6.11 alleles per locus during the amplification reaction conducted with the thirty six entries. The primer pairs BSO96 and BS065 appeared to be highly polymorphic and comparatively more informative primers for molecular characterization and differentiation of thirty six Bipolaris sorokiniana isolates. The dendrogram was generated following UPGMA and the clusters were identified at appropriate pennon level. There were nine clusters A to I at eighty five similarity units. Entries R.G.T-78 and R.G.T-79 were relatively more closely related with the highest similarity coefficient amongst the thirty six entries evaluated. Analysis of divergence pattern based on amplification profile allowed differentiation of thirty six isolates. The different clusters of isolates showed differentiation for aggressiveness. Bipolaris sorokiniana of cluster A were the highly aggressive and of clusters B, C, D, F and G were moderately aggressive. Those of clusters E, H and I were the least aggressive. Thus molecular marker can differentiate the Bipolaris sorokiniana fungal isolates in general and for their relative aggressiveness.
  • ThesisItemOpen Access
    Genetic Diversity Assessment In Relation To Submergence Tolerance In Rice Using Microsatellite Markers
    (DR. Rajendra Prasad Central Agricultural University, Pusa (Samastipur), 2016) Prakash, Nilmani; Kumar, Mithilesh
    A study was undertaken to initially characterize and evaluate 184 submergence tolerance related varieties of rice, but 38 were taken for further analysis in the present study for five submergence related characters. To investigate the SSR marker based polymorphism for characterization and differentiation, nineteen rice varieties were selected on the basis of maximum plant percent survival to estimate the extent of genetic diversity among these rice varieties using morphological characters and SSR markers. The experimental materials were evaluated in randomized complete block design with four replications at the research farm of Dr. Rajendra Prasad Central Agricultural University, Pusa during Kharif, 2014-15. Using standard formula for calculation of different submergence related traits in rice observations were recorded. The materials were also grown in pots for extraction of genomic DNA from the young seedlings and then targeted amplification of the genomic DNA using a panel of thirty four SSR primer pairs covering especially the chromosome 9 of rice. All molecular studies were conducted in the Rice Molecular Breeding Laboratory, Deptt. of PBG at Pusa. The statistical methods and parameters used for deriving inference were analysis of variance, range, mean values, coefficient of variation, taxonomic distance and similarity coefficient based dendrogram and numerical taxonomic analysis of divergence. High heritability values were observed for all the five submergence related characters percent plant survival, tolerance score, leaf senescence, total shoot elongation and relative shoot elongation. Considerable extent of variability existed for the values of genetic advance expressed as per cent of mean was observed to be high for all five characters relative shoot elongation, per cent plant survival, total shoot elongation, leaf senescence and tolerance score. By considering mean performance in conjunction with clustering pattern of entries based on quantitative attributes, hybridization involving CR 1009 with AnhHsungSeln (CI 1), Karnal Local and Suraha with Swarna, Jadhan and HsungTeing with Moroberekan, FR 13B and S-150 with Paiam, Zobgui and Rom Runtik with IR-36, Kala Bunde with AI Chih AI K Who 2, Swarna Sub-1 with Lunishreeappeared as superior combinations for their further utilization in genetic enhancement programme. A panel of thirty four primers generated 410 allelic variants ranging from four in the cases of RM 23843 to twenty seven in the cases of RM 23662. Altogether 410 allelic variants were detected at 42 SSR loci with an average of 9.76 alleles per locus. Out of 410 alleles, 177 were shared and 233 were unique alleles. The primer pairs RM 8300, RM 23901, RM 23958, RM 215, RM 257, RM 316 and RM 23662 generated amplified products due to amplification of more than one locus. In general, the SSR loci with di-nucleotide repeat motifs tended to detect greater number of alleles than the locus with tri-nucleotide, tetra-nucleotide or penta-nucleotide repeat sequence. The marker with a GA or AG repeat motif showed greater variability than marker with a CT or TG or TA repeat motif among the loci with perfect di-nucleotide repeat motifs. The number of unique allelic variants per primer ranged from 0 out of 5 amplified products in the case of SC 3 to 13 out of 15 amplified products in the case of RM 23901. The primer pairs RM 23901, RM 910, RM 23770, RM 23928, RM 23958, RM 24103 RM 23662, RM 23915 and RM 24005 generated considerably greater percentage of unique alleles. Considering the number of alleles in conjunction with the level of polymorphism detected, the primers RM 23662, RM 23958, RM 257, RM 23901, RM 8300, RM 215, RM 23928, RM 23915, RM 24005, RM 23902, RM 23679, RM 8303, RM 23770 and RM 23788 appeared to be more informative primers. Occurrence of null alleles for a particular repeat locus was noticed with primer pairs RM 23843, RM 23902, RM 23922, RM 24103, and RM 23865 in some of the varieties. The total repeat count of the di-nucleotide SSR loci seemed to be associated with the number of alleles, revealing that the larger the repeat number involved in the SSR locus, the larger was the number of identified alleles. Presence of stutter bands was detected generally for di-nucleotide SSR motif amplified by primer pairs RM 8303, RM 23770, RM 23805, RM 23831, RM 8300, RM 23901, RM 23915, RM 23917, RM 23922, RM 23928, RM 23958, RM 23996, RM 24103, RM 105, RM 215, RM 316, RM 23662, RM 24013, RM 910, RM 7175, ART 5, SC3, Sub 1BC2 and RM 23865 with generation of minor products. Among the rice varieties evaluated in the present study, AnhHsung Seln (CI 1) and HsungTeing exhibited maximum similarity in the divergence analysis using quantitative characters as well as SSR markers. Analysis of divergence pattern based on SSR markers allowed differentiation and classification of rice varieties into eight clusters. Use of SSR markers appeared more efficient in achieving unique and unambiguous characterization and differentiation of varieties used in the present study.The SSR analysis also revealed unique or variety specific allele, which could be useful as DNA fingerprints in the identification and preservation of rice varieties.