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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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20211
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Subject: Genetics and Plant Breeding × Author: Kumar, Abhay × End date: 2023 × Start date: 2020 × Type: Thesis × Thesis Type: Ph.D ×
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    ThesisItemOpen Access
    CANDIDATE GENE BASED MICROSATELLITE PROFILING IN RELATION TO FRAGRANCE OF LAND RACES AND IMPROVED VARIETIES OF AROMATIC RICE
    (Dr.RPCAU, Pusa, 2021) Kumar, Abhay; SHARMA, V.K.
    A study was performed to evaluate the genetic polymorphism using candidate gene based microsatellite specific reported and designed primers and to validate them in relation to fragrance in 18 rice genotypes. Molecular-level genetic divergence along with the importance of candidate genes based microsatellite markers was assessed in relation to fragrance in these landraces and improved varieties of aromatic rice used as experimental material during molecular profiling. The experimental materials were grown in aluminum containers to extract genomic DNA from young seedlings. The targeted amplification of genomic DNA was achieved using candidate genes based microsatellite specific 24 reported primer pairs and candidate genes based microsatellite specific 38 designed primer pairs. Based on the information available in the literature, a total of eleven genes, namely, Os03g0327600, Os04g0352400, Os04g0434800, Os04g0401700, Os04g0438300, Os04g0445700, Os04g0468600, Os04g0474800, Os04g046970, Os08g0424500 and OsNPB_0504555500, which are known to be associated with aroma in aromatic rice, were searched through the internet. The fragrance related these 11 candidate genes are located on the short arm of four different chromosomes (3, 4, 8 and 5) existing in the rice genome. A total of 47 SSR sequences were detected for all eleven candidate genes by using BatchPrimer3 v1.0 software under investigation. Most of the SSRs were tri-nucleotides followed by di-nucleotides; only few were tetra- nucleotides. As a result, altogether investigated 38 microsatellites contained 10, 22, and 06 microsatellites with di-nucleotide, tri-nucleotide and tetra-nucleotide repeat motifs, respectively. The gene Os08g0424500 possessed the highest number of SSRs, while gene Os04g0445700 and Os04g0474800 were not observed to possess any SSR. Differential amplification profile was ascertained amongst the genotypes subjected to molecular characterization by employing 24 reported and 38 designed microsatellite specific primer pairs. The migration of amplicons resulted in appearance of bands representing amplification products localized at different positions on the gel, reflecting the variation in product length derived by primer directed amplification of genomic templates. By using 24 reported primers, a survey of the molecular profiles generated on the basis of the evaluation of the amplified products clearly indicated that a total of 160 alleles were detected with an average number of 6.66 alleles per primer. Similarly, by using 38 designed primers, a total of 248 allelic variants were detected with an average number of 6.52 alleles per primer. Null allele was also noticed in the specific combinations between genotypes and primers. Statistical measure of polymorphic information content (PIC) of the 24 reported microsatellite primer pairs, which reflected allelic diversity and frequency among the genotypes, varied from 0.638 to 0.870. Similarly, for the 38 designed primers, polymorphic information content (PIC) varied from 0.692 to 0.880. Polymorphism per cent generated by these 24 reported primer pairs varied from 0.00 to 62.50 with an average of 24.26 per primer. Similarly, by using 38 designed primers, polymorphism per cent varied from the 0.00 to 66.66. The reported primers CHR 8_34, CHR 3_20, CHR 3_24, CHR 4_3, CHR 8_10, CHR 3_10 and CHR 8_5 appeared to be highly informative primers with above average polymorphism per cent and polymorphic information content. Similarly, the designed primers, CHR4.1_1, CHR4.1_5, CHR4.1_6, CHR4.1_10, CHR4.1_11, CHR4.1_13, CHR8.1_4, CHR8.1_6, CHR8.1_9, CHR8.1_10, CHR8.1_11, CHR8.1_12, and CHR8.1_21 appeared to be highly informative primers with above average polymorphism per cent and polymorphic information content. The results indicated a relatively higher magnitude of discrimination coefficient for the reported primer pairs CHR3_16, CHR3_10, CHR3_20, CHR3_24, CHR4_3, CHR4_25, CHR4_26, CHR8_29, CHR8_6, CHR8_49, CHR8_10, and CHR8_5. Similarly, for the designed primers the relatively higher magnitude was found for the primers CHR3.1_2, CHR4.1_1, CHR4.1_5, CHR4.1_6, CHR4.1_8, CHR4.1_10, CHR4.1_11, CHR4.1_12, CHR4.1_13, CHR8.1_4, CHR8.1_7, CHR8.1_9, CHR8.1_10, CHR8.1_11, CHR8.1_13, CHR8.1_15, CHR8.1_17, CHR8.1_20, CHR8.1_21, and CHR8.1_22. Differential amplification pattern revealing molecular level genetic polymorphism among the genotypes subjected to molecular characterization provided a basis for deducting that the sequence length variation observed in candidate genes may be a role player in differential fragrance in landraces and improved varieties of aromatic rice. Sizable variation was clearly recognized in the molecular size of the genomic region targeted by the primer pairs. The values of similarity coefficient ranged from 0.00 to 0.641, 0.024 to 0.481 and 0.015 to 0.545 for pair-wise combinations of genotypes by using 24 reported primers, 38 designed primers and 62 combined (reported and designed) primers based binary matrices data. Wide genetic differentiation and divergence at the molecular level was revealed among the genotypes. Hierarchical pattern of classification based on similarity coefficient matrix of pair-wise combinations of genotypes was in extremely good agreement with principal coordinated analysis based spatial distribution pattern of genetic profiles of genotypes. Hierarchical cluster analysis, as well as principal coordinate analysis using microsatellite-specific markers derived from candidate genes made it possible to differentiate aromatic landraces and improved varieties of rice. The landraces were discriminated from improved aromatic varieties and accommodated into different cluster. The computational analysis of the genetic structure of the aromatic landraces and improved varieties unambiguously reflected that the genotypes subjected to fragrance related molecular characterization are the admixture of three ancestral components present in different combinations in different genotypes. The Practical utility of the fragrance related candidate gene specific designed primers in discrimination of aromatic landraces and improved varieties was amply demonstrated. The results led to validation of designed primers by amply demonstrating their efficiency to discriminate aromatic landraces from improved varieties of aromatic rice. These reported and designed candidate genes based SSR primers could be utilized for molecular characterization, identification, genetic divergence analysis, parental selection and biological individuation in relation to fragrance of landraces and improved varieties of aromatic rice.