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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 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.ThesisItem Open Access Molecular Characterization of Bipolaris sorokiniana Isolates Collected from Wheat by SSR Markers(Rajendra Agriculrural University, Pusa (Samastipur), 2016) Bharty, Archana; Kumar, HarshSpot 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.ThesisItem Open Access Morpho-Molecular Characterization of Elite Germplasm of Sweet Potato (Ipomoea batatas L.).(Rajendra Agricultural University, Pusa (Samastipur), 2015) Singh, Chandra Prakash; Kumar, HarshThe present investigation was carried out to characterize 17 elite genotypes of sweet potato(Ipomoea batatasL.) using morphological features and molecular markers. Genotypes were evaluated for eleven traitsnamelynumber of branch, vine length, number of leaves/plant, leaf area, leaf area index, length of tuber, girth of tuber, neck length, average weight/tuber, yield/hectare and skin and flesh colour.The mean square due to genotypes was found significant for all the characters. The mean performance of genotypes has been compared with the check Shree Bhadra for each character as it is one of the commonly cultivated genotypes of Bihar.The observations were also recorded based on descriptors coding in the Descriptors of Sweet Potato (Human, 1991) for grouping of the seventeen genotypes of sweet potato under study on the basis of their similarity and differences with respect to vine length, neck length, flesh colour and skin colour. The exploitable extent of genetic variability amongst the entries was present as revealed by considerably higher estimates of GCV, PCV, h2 and GA as percentage of mean. For GCV the highest value was recorded for leaf area index (51.57) and the lowest for girth of tuber (20.56). For PCV the highest value was recorded for leaf area index (53.76) and the lowest for girth of tuber (21.79). For heritability the highest value was recorded for leaf area (97.19) and the lowest for length of tuber (78.50). For GA as percentage of mean the highest value was recorded for leaf area index (97.56) and the lowest for length of tuber (36.21). Sixteen sweet potato microsatellite markers (SSR)were used for molecular characterization, which werecapable of detecting 276 alleles with an average of 17.25 alleles per locus. The highest number of alleles per locus was observed for Ib3/31 SSR primer. The number of alleles per locus ranged from 14 to 28 and allelic polymorphism information content (PIC) value ranged from 0.79 for the primer Ib-242 to 0.92 for Ib2/55 with an average of 0.88.A perusal of similarity coefficients clearly reflected that a very high degree of similarity exists between sweet potato genotypesDOP-MIX-93-13 and RS-5 (0.29), on the other hand, the two most distantly related genotypes were DOP-MIX-97-6 and 85-X-04 (0.02).The result of the present study clearly established that the utilization of sixteen selected SSR markers was sufficient for discrimination and unambiguous identification of all the seventeen sweet potato genotypes used.ThesisItem Open Access In vitro propagation of Orchids and Clonal Fidelity Analysis(Rajendra Agricultural University, Pusa (Samastipur), 2015) Kumari, Kamni; Kumar, MithileshThesisItem Open Access Genetic purity assessment of hybrid rice using microsatellite markers(Rajendra Agricultural University, Pusa (Samastipur), 2015) Prakash, Kumar Satya; Sharma, V. K.A study was carried out to evaluate the microsatellite markers based polymorphism for identification of polymorphic and informative markers in order to characterize two WA type CMs lines, four fertility restorer lines, eight experimental hybrids and four genotypic mixtures containing seeds of experimental hybrid and respective fertility restorer male parent and to validate the usefulness of these markers for assessment of genetic purity of hybrid rice genotypes. The experimental hybrids were evaluated on the basis of pollen and spikelet fertility to assess the extent of fertility restoration. Using a set of 18 microsatellite primer pairs, molecular profiling was conducted and amplification pattern based polymorphism was recognized on the basis of presence or absence of bands, besides variation in number and position of bands. Altogether 73 allelic variants were detected at 29 microsatellite loci with an average of 2.51 alleles per locus. A total of 33 shared and 40 unique allelic variants were generated. The number of unique alleles per primer pairs ranged from one in the cases of RM 114, RM 250, RM 276, RM 280, RM 427, RM 524, RM 538, RM 1108 and RM 5359 to five alleles in the case of RM 17 and RM 201. Remarkably higher polymorphism per cent was exhibited by the primer pairs RM 17, RM 201, RM 558, RM 337, RM 206 and RM 171, since these primer pairs generated considerably greater percentage of unique alleles in descending order of magnitude. Considering the number of alleles generated in conjunction with the level of polymorphism detected, the primer pairs RM 17, RM 171, RM 201 and RM 206 appeared to be highly polymorphic and comparatively more informative for the purpose of molecular characterization of entries under evaluation. These primers generated considerably greater number of allelic and polymorphic variants due to variation in the length of simple sequence repeats. In general, the microsatellite specific primer pairs flanking di-nucleotide and complex repeat motifs tended to detect lesser number of allelic variants than loci with tri-nucleotide and tetra-nucleotide repeat motifs. Among the microsatellite loci having di-nucleotide repeat motifs, the markers with CT, GA and AT repeat motifs tended to detect greater number of allelic variants than primers targeting microsatellite loci with TG, AG and TC di-nucleotide repeat motifs. Considerably greater ability to differentiate pair-wise combinations of entries was observed in the cases of primer pairs RM 17 followed by RM 152, RM 201, RM 280, RM 171, RM 206, RM 341, RM 276, RM 337, RM 524 and RM 8146 in decreasing order of magnitude. Contrarily, the primer pairs RM 538 RM 558, RM 5359, RM 114, RM 250, RM 427 and RM 1108 exhibited lesser ability to differentiate pair-wise combinations of entries. Appearance of more than one band in the same entry was noticed revealing the existence of the duplicated region in the genome. The primer pairs RM 17, RM 152, RM 171, RM 201, RM 206, RM 276, RM 280, RM 337, RM 524, RM 558 and RM 8146 generated amplified products due to amplification of only two locus. However, further investigation under more stringent condition is required to confirm it because the intensity of bands was comparatively lower in some of the cases. The total repeat count of microsatellite loci did not appear to be strongly associated with the number of alleles generated by the primers. Results did not reflect that the larger the repeat number involved in the microsatellite locus, larger was the number of identified alleles. Presence of stutter bands mostly for di-nucleotide repeat motifs indicated the presence of minor products amplified that had lower intensity than the main allele and normally lacked or had extra repeat units. Such bands were observed to be present in the case of di-nucleotide repeat sequences detected by primer pairs RM 171, RM 427, RM 524, RM 538, and RM 558. Analysis of divergence pattern allowed relative assessment of genetic diversity amongst the two WA type CMS lines and four fertility restorer lines. A combination of only five microsatellite primer pairs, namely, RM 17, RM 171, RM 201, RM 206 and RM 558 was equally effective in differentiation of two WA type CMS lines from four fertility restorer lines and genetic purity assessment of their hybrids. Experimental results finally led to validation of five microsatellite primer pairs, which allowed easily recognizable differentiation of WA type CMS lines and fertility restorer lines and provided a basis for genetic fidelity assessment of experimental hybrids. These microsatellite primer pairs, namely, RM 17, RM 171, RM 201, RM 206 and RM 558 may be further validated and utilized for the purpose of genetic purity assessment of three-line hybrid rice. Amongst these five primer pairs, RM 17, RM 171, RM 206 and RM 558 appeared to be more important in differentiating WA type CMS lines from their fertility restorer lines. Results of the present study also revealed that use of RM 17 and RM 171 along with RM 206 or RM 558 was equally effective in differentiation of two WA type CMS lines from four fertility restorer lines and evaluation of genetic purity of their hybrids. Experimental hybrids were not differentiated from the genotypic mixtures containing seeds of experimental hybrid and respective fertility restorer male parent because of the reason that microsatellite primer pairs directed genomic profiling and further analysis was based on bulk seedlings. Therefore, single plant based molecular profiling and analysis appeared to be an essential requirement for ascertaining the genetic purity of three-line hybrid rice.ThesisItem Open Access Molecular characterization of fertility restorers for wild abortive rice cytoplasm(Rajendra Agricultural University, Pusa (Samastipur), 2015) Kumar, Alok; Sharma, V. K.A study was conducted to evaluate the microsatellite markers based polymorphism for identification of polymorphic and informative markers in order to characterize two WA type CMS lines, six fertility restorer lines, twelve experimental hybrids and F2 individuals of cross combination IR80559A x PRR78 to assess the genetic effects of fertility restorer genes in relation to fertility restoration and to validate the association of molecular markers with fertility restoration of cytoplasmic male sterile lines. The experimental hybrids and F2 individuals were evaluated on the basis of pollen and spikelet fertility to assess the extent of fertility restoration. Using a set of 24 microsatellite primer pairs for molecular characterization of CMS lines, fertility restorers and experimental hybrids, amplified products were generated and polymorphism was recognized on the basis of presence or absence of bands, besides variation in number and position of bands. Altogether 142 allelic variants were detected at 37 loci with an average of 3.83 alleles per locus. A total of 46 shared and 96 unique allelic variants were generated. The number of unique alleles per primer pairs ranged from one out of three alleles in the case of RM 315 to eight out of nine alleles in the cases of RM 216 and RM 6100. Remarkably higher polymorphism per cent was exhibited by primer pairs RM 216, RM 6100, RM 17, RM 591, RM 5359, RM 206, RM 171, RM 3233, RM 443, RM 1108 and RM 10318. Considering the number of alleles generated in conjunction with the level of polymorphism detected, the primer pairs RM 216, RM 6100, RM 17, RM 206, RM 171, RM 1108, RM 3233 and RM 10318 appeared to be highly polymorphic and comparatively more informative for the purpose of molecular characterization of entries under evaluation. These primers generated considerably greater number of allelic and polymorphic variants due to variation in the length of simple sequence repeats.ThesisItem Open Access Genetic purity assessment of scented rice using microsatellite markers.(Rajendra Agricultural University, Pusa (Samastipur), 2015) Gopaliya, Madhu; Shahi, V. K.A study was undertaken to examine the microsatellite markers based polymorphism for identification of polymorphic and informative markers in order to characterize four aromatic rice varieties, two non-aromatic rice varieties, twelve inter-varietal seed mixtures and to validate the utilization of these markers for assessment of genetic purity of scented rice. The experimental materials were evaluated on the basis of biochemical test to ascertain the presence of aroma. Amplification of the targeted genomic DNA was done using a panel of 24 microsatellite based primer pairs targeting all the chromosomes in the rice genome. All molecular studies were conducted in the Molecular Biology Laboratory at Pusa. The statistical methods and parameters used for deriving inference were polymorphism per cent, polymorphism information content, discrimination coefficient, similarity coefficient and numerical taxonomic analysis of divergence. The amplification was successfully achieved with all the microsatellite primers used in the present study. Molecular profiling was conducted and amplification pattern based polymorphism using microsatellite primer pairs was recognized on the basis of presence or absence of bands, besides variation in number and position of bands. Altogether 105 allelic variants were detected at 28 microsatellite loci with an average of 3.75 alleles per locus. A total of 42 shared and 63 unique allelic variants were generated. The number of unique alleles per primer pairs ranged from zero out of three alleles in Aro 7 to eight out of ten alleles in the case of RM 7049. Remarkably higher polymorphism per cent was exhibited by the primer pairs RM 42, RM 7049, BAD 2a, RM 85, RM 444, RM 1109, RM 505 and RM 330, since these primer pairs generated considerably greater percentage of unique alleles in descending order of magnitude. Considerably greater ability to differentiate pair-wise combinations of entries was observed in the cases of primer pairs RM 42, RM 444, RM 7049, BAD 2a, RM 223, RM 252, RM 284, RM 8264, RM 23097, E03_92.0, F05_103.5, and B03_127.8 in decreasing order of magnitude. Contrarily, the primer pairs SCUSSR 1, RM 505, E11_44.5, and BAD 2b exhibited lesser ability to differentiate pair-wise combinations of entries. Considering the number of alleles generated in conjunction with the level of polymorphism detected, the primer pairs RM 42, RM 85, RM 223, RM 252, RM 284, RM 444, RM 1109, RM 23097, Aro 7, BAD 2a, B03_127.8, F05_103.5 and E03_92.0appeared to be highly polymorphic and comparatively more informative for the purpose of molecular characterization of entries under evaluation. These primers generated considerably greater number of allelic and polymorphic variants due to variation in the length of simple sequence repeats. Appearance of more than one band in the same genotype was noticed revealing most probably the existence of the duplicated region in the genome. The primer pairs RM 330, RM 505, RM 7049, and RM 8264 generated amplified products due to amplification of two loci. However, further investigation under more stringent condition is required to confirm it because the intensity of bands was comparatively lower in some of the cases. Presence of stutter bands indicated the presence of minor products amplified that had lower intensity than the main allele and normally lacked or had extra repeat units. Such bands were observed to be present in the case of primer pairs RM 7049 and RM 8264. The total repeat count of microsatellite loci did not appear to be associated with the number of alleles generated by the primers. Results did not reflect that the larger the repeat number involved in the microsatellite locus, larger was the number of identified alleles. In general, the microsatellite specific primer pairs flanking di-nucleotide and tri-nucleotide repeat motifs tended to detect lesser number of allelic variants than loci with tetra-nucleotide and complex repeat motifs. Among the microsatellite loci having di-nucleotide repeat motifs, the markers with AG, AT and CT repeat motifs tended to detect greater number of allelic variants than primers targeting microsatellite loci with TA, GA, and TC di-nucleotide repeat motifs. Analysis of divergence pattern allowed relative assessment of genetic diversity amongst the four aromatic rice varieties and two non-aromatic rice varieties. A combination of only seven microsatellite primer pairs, RM 42, RM 223 and RM 8264 in combination with RM 85, RM 23097, BAD 2a, or E03_92.0 was purposefully effective in differentiation of four aromatic rice varieties, two non-aromatic rice varieties and genetic purity assessment of their seed mixture. Experimental results finally led to validation of three microsatellite primer pairs, which allowed easily recognizable and unambiguous differentiation of scented rice varieties from non-scented rice varieties evaluated in the present study. The molecular profile based on these three microsatellite markers served as distinct molecular tags for distinguishing the genotypes and inter-genotypic mixtures and provided a basis for genetic purity assessment of scented rice genotypes with at least with a single marker allele difference. These microsatellite primer pairs, namely, RM 42, RM 223, and RM 8264 may be further validated and utilized for the purpose of genetic purity assessment of scented rice. Results of the present study also revealed that use of RM 42 along with RM 223 or RM 8264 was equally effective in differentiation of four aromatic rice varieties from two non-aromatic rice varieties and evaluation of genetic purity of scented rice.ThesisItem Open Access Characterization of wheat (Triticum aestivum L.) genotypes with respect to heat stress tolerance and spot blotch resistance(Rajendra Agricultural University, Pusa (Samastipur), 2015) Chaurasia, Surendra Kumar; Kumar, RajeevTwelve wheat (Triticumaestivum L.) genotypes were characterized with respect to heat stress and spot blotch diseaseunder four environmental conditions viz. no stress (E1), stress due to spot blotch (E2), heat stress (E3) and combined stress (E4) alongwith molecular characterization using 28 SSR markers(associated with tolerance against heat stress and resistance against spot blotch disease). The experimental materials were evaluated in randomized block design (RBD) factorial. Observation on 1000-grain weight, grain filling duration (GFD), plant height, peduncle length, spike length, disease severity and area under disease progress curve (AUDPC) were recorded. Significant differences were observed among the genotypes for all the traits under consideration. Genotype × environment were found non-significant. The lowest performance of the genotypes under combined stress condition (E4) was recorded. The individual impact of heat stress with respect to performance of the genotype was found significantly higher than that caused by spot blotch disease. Raj 3765 for 1000-grain weight, WH 760 for GFD, Yangmai-6 for plant height, PBW 343 for peduncle length and HD 2733 for spike length, showed stable performance across the environment. On the basis of stress susceptibility index (SSI) the genotypes WH 760, DBW 14 and Raj 3765 were found better for the characters, 1000-grain weight and GFD. The overall disease severity was found maximum under heat stress condition. Similarly genotypes were having relatively more AUDPC under heat stress condition. The twenty eight SSR markers detected total 244 alleles in 12 genotypes, the number of alleles per locus varied from four in Xgwm133a and Xgwm456 to twenty three in Xgwm293with an average of 6.5 alleles per locus. A total 158 unique alleles were observed at 37 SSR loci, with an average of 5.6 unique alleles per locus. The number of unique alleles per locus ranged from one in Xgwm273 to twenty two in Xgwm293. In set of 12 varieties, 6 SSR loci showed null alleles, in case of primer pair Xbarc1047, Xgwm133a, Wmc168,cfd44, Xbarc147 and Xgwm626.The polymorphism information content(PIC) values revealing allele diversity and frequency among the entries varied from 0.292 in the case of Wmc273 to 0.902 in the case of Xgwm356 with an average of 0.763 across the primer. Pair-wise genetic similarity coefficient, widely varied from 0.72 to 0.82 indicating a considerably greater extent of variation among the wheat genotypes. High degree of similarity coefficient exists between wheat genotypes Raj 3765 and PBW 343 (0.828) or WH 760 and DBW 14 (0.828), whereas Sonalika and HD 2967 (0.721) weremost distantly related. Cluster analysis grouped twelve wheat genotypes into four different clusters (I to IV) at 35 phenon level, the use of twenty eight SSR markers allowed unique genotyping of twelve genotypes included in the analysis.ThesisItem Open Access In vitro screening and induction of resistance against brown spot disease of rice caused by Helminthosporium oryzae(Rajendra Agricultural University, Pusa (Samastipur), 2015) Agarwal, Ruchi; Kumar, HarshCopious callus formation from cultured dehusked seeds and high frequency and number of regenerated plantlets from subcultured seed callus through caulogenesis followed by rhizogenesis or through somatic embryogenesis were achieved in six selected rice genotypes namely Pankaj, Rajendra Bhagwati, Rajshree, RAU-Aer03, RAU-Aer04 and Sugandha using eleven different media with MS basal salts supplemented with various concentrations and combinations of plant growth regulators NAA, 2,4-D, BAP, KIN and TDZ. The work resulted in efficient protocol for callus formation and plant regeneration. In vitro screening of selected genotypes for resistance/tolerance to brown spot disease through callus growth and shoot differentiation in presence of culture filtrate ranked the selected genotypes for their relative resistance to disease. The callus development and shoot differentiation in presence of high concentration of culture filtrate indicated the induction of resistance / tolerance to disease. Molecular marker studies were also done for assessment of genetic variability with respect to brown spot disease resistance. Utilizing a panel of ten SSR primers linked to brown spot disease namely, RM 21, RM 206, RM 229, RM 257, RM 263, RM 566, RM 1367, RM 3515, RM 3907 and RM 6499 in the study revealed remarkably higher level of genetic polymorphism which allowed unique genotyping of the entries and somaclonal variants. Clustering of genotypes based on amplification profiles differentiated resistant and moderately resistant entries from highly susceptible with respect to brown spot disease. The molecular marker study graded the six selected genotypes of rice for their resistance/tolerance to brown spot disease in the order: RAU-Aer04, Rajshree, Rajendra Bhagwati, RAU-Aer03, Sugandha and Pankaj which was the same as observed during in vitro screening.
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