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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 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.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.