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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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Author: Kisku, Anima × Start date: 2003 ×
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    ThesisItemOpen Access
    Exploration and characterization of candidate genes for race 4 Fusarium wilt resistance in chickpea (Cicer arietinum L.)
    (RPCAU, Pusa, 2023) Kisku, Anima; Bhutia, Karma L.
    The production and productivity of chickpea is significantly hampered by numerous factors of biotic and abiotic nature. Among the biotic stress, wilt disease caused by fungal species i.e. Fusarium oxysporum f. sp. ciceris is a devastating one. Several races of the pathogens already been reported and the researchers are working continuously to develop the robust resistance mechanism against the wilt disease of chickpea. Marker assisted selection is employed in developing chickpea plants resistant against Fusarium wilt using the markers like TA59 and TR19. The TA59 and TR19 are reported to be linked particularly with race 4 (Foc4) of Fusarium oxysporum f. Sp. ciceris, however, the region between TA59 and TR19 in linkage group 2 is significantly wide and the region is largely unexplored for the identification of potential candidate genes that is actually imparting resistance against Foc4 wilt resistance. Therefore, the present study was conducted to explore and characterize the genes present within the region flanked by TA59 and TR19 markers using in silico as well as gene specific marker based wet lab approach. A total of 225 genes were identified to be present in the targeted region among which 51 were found to be showing differential expression under Fusarium wilt stress when assessed from Fusarium wilt specific shoot transcriptome data of two contrasting chickpea genotypes available in NCBI SRA database. Further in silico analysis was carried out for these selected 51 genes. Several cis acting elements such as BIHD1OS, WRKY71OS, SEBFCONSSTPR10A, WBOXATNPR1 etc., which are reported to be involved in response against biotic stresses, were found to be present in the 1kb 5’UTR of the 51 selected genes. Most of the proteins encoded by 51 selected genes were hydrophilic in nature having good solubility and were mostly localized to nucleous and cell membrane. These proteins were found to be interacting with 398 other proteins among which only 30 proteins interacting with query proteins encoded by 15 selected genes of the region were found to be showing differential expression under Fusarium wilt stress when assessed from Fusarium wilt specific shoot transcriptome data of two contrasting chickpea genotypes available in NCBI SRA database. Simultaneously, 246 diverse set of chickpea lines were sown in a Fusarium wilt sick plot and data on different field traits including disease incidence % were recorded. The PCR was done with TA59 and TR19 markers in 246 lines of chickpea and 40 highly resistant and 40 highly susceptible lines of chickpea were selected based on amplification pattern of TA59 and disease incidence% observed in 246 lines of chickpea. The gel based polymorphism survey using primers specific to 51 selected candidate genes revealed low level of polymorphism mostly in term of presence or absence of the amplified products with only 27 gene specific primers showing amplification in 80 selected lines. The polymorphism information content of the gene specific primers ranged from 0.049 to 0.554 with the mean Nei’ Gene Diversity and Shanon Index of 0.2162 and 0.3403 respectively, explaining low level of gene diversity in 80 selected lines of chickpea. Regression analysis revealed 8 out of 27 gene specific primers had R2value of >0.1 with the traits like disease incidence, no. of pods/plant, plant height, shoot dry weight and 100 seed weight. Among 8 gene specific primers showing R2value of >0.1, the amplification of primers specific to 101502928, 101495508, 101505289, 101496712, 101499005 and 101488582 showed positive association with the desired traits and the amplification of primers specific to 101505077 and 101510207 showed negative associations with desired traits. Regression analysis also revealed non-significant association of TA59 and TR19 with the disease incidence %. The comparison of cistron and promoter (5’UTR) sequences of the two potential candidate genes namely 101502928 (WRKY transcription factor 55) and LOC101488582 (CBL-interacting serine/threonine-protein kinase 2-like) in 4 contrasting chickpea genotypes revealed several single nucleotide polymorphisms (SNPs), nucleotide deletions and undetermined nucleotide present in the 5’UTR, exonic and intronic regions. Further studies could be done to understand the pathways of protein-protein interaction, impact of SNPs or nucleotide deletions in the genes leading to susceptibility or resistantance against Fusarium wilt in chickpea. From the findings of the present experiment, it could be concluded that the use of TA59 and TR19 alone is not sufficient in marker assisted selection for developing wilt resistant chickpea lines. Therefore, the use of markers specific to candidate genes like 101502928 (WRKY transcription factor 55) and LOC101488582 (CBL-interacting serine/threonine-protein kinase 2-like) could be more effective in developing robust resistance mechanism in chickpea plants against Fusarium wilt disease.