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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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Subject: Biotechnology and molecular Biology × Start date: 2004 ×
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    ThesisItemOpen Access
    Investigating plausible role of AHL gene-family in drought stress tolerance in rice (Oryza sativa L.)
    (RPCAU, Pusa, 2023) AMBADAS, DHANORKAR ADITYA; Singh, Ashutosh
    The transcription factor AHL (AT-hook motif nuclear localized) are one of the largest and most conserved gene families in plants and they play critical roles in abiotic stress response. However very limited information available about the AHL family of genes in rice. In this context, a genome wide survey, was done to identify the AHL genes in rice (Oryza sativa L.) and total 20 members AHL family of genes were identified. The distribution of these 20 genes were on the 9 chromosome of rice. Mostly AHL genes transcription factors were located in nucleus. The phylogenetic analysis conducted for these 20 genes of AHL TFs and it divided into two major groups and four subgroups. To study the role of AHL group of genes in drought and salt stress at seedling stage of rice. The two genotypes IR64 & NL44 were chosen for gene expression study. The genotypes were grown in pot. The fourteen days old seedlings were exposed to 20 % PEG (2 hours and 4 hours) and 200 Mm NaCL (2 hours and 4 hours) for drought and salt treatments respectively. The mRNA of each treatment of 14 days old seedlings extracted in the replication and cDNA synthesized. Quantitative real time PCR based expression analysis of AHL genes in IR64 genotype of rice under drought and salinity stresses. The expression pattern of AHL family of genes was compared by one-way ANOVA in IR64 genotype. The result clearly showed that the OsAHL group of genes in drought and salt stress tolerance. Further, differential expression of AHL family gene was studied in IR64 and NL44 under drought and salt stress conditions. Total 8 genes (OsAHL1, OsAHL2, OsAHL3, OsAHL10, OsAHL14, OsAHL17, OsAHL18 and OsAHL19) were showed significantly more expression in NL44 as compared to IR64 after 2 hours of drought treatment. Similarly, all total 7 genes (OsAHL7, OsAHL10, OsAHL13, OsAHL14, OsAHL17, OsAHL18 and OsAHL20) showed significantly more expression in NL44 as compared to IR64 after 4hours drought treatment. Total 3 genes (OsAHL10, OsAHL13 and OsAHL20) showed significantly up-regulated after 4hours salinity treatment in cultivar NL44 as compare to IR64. Overall, these findings indicate that AHL genes might be involved in mediating drought and salt signaling transduction pathways when plants are exposed to drought and salt stress. The NL44 could be used as a potential donor for drought and salt stress tolerance.
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    ThesisItemOpen Access
    Morpho-physiological characterization and gene expression profiling of foxtail millet genotypes for waterlogging tolerance at seedling stage
    (RPCAU, Pusa, 2023) Borah, Bhabna; Anjani, Kumari
    The present study entitled “Morpho-physiological characterization and gene expression profiling of foxtail millet genotypes for waterlogging tolerance at seedling stage” was carried out to identify the best screening methodologies for early seedling stage waterlogging tolerance, identification of tolerant genotypes and identification of candidate genes responsible for ameliorating waterlogging tolerance. Altogether, 26 foxtail millet genotypes were used for morpho-physiological characterization and screening at three stages of growth under six different conditions (two in each stage) of waterlogging, to identify the best screening method and tolerant genotypes. The selected highly tolerant and highly susceptible genotypes were used for the gene expression study of the identified candidate genes, to determine their differential expression pattern in leaves under the identified short- and long-term stress conditions. The screening at germination stage revealed that foxtail millet is vulnerable to waterlogging stress. A significant reduction in the performance of the 26 genotypes evaluated in the present study was observed for both 3 days and 8 days stress conditions. A significant change in the performance of 26 genotypes under 12 h and 24 h waterlogging stress was observed at the seedling establishment stage. The waterlogging tolerance indices, waterlogging tolerance coefficient (WTC), root length stress index(RLSI), shoot length stress index (SLSI) and seed vigour wrt seedling length (SVI), significantly contributed to the variations observed among the genotypes, as revealed by Principal Component Analysis (PCA) and were found to be significantly correlated. A significant reduction in these parameters during 12 h stress suggests that even 12h of flooding at seedling establishment stage causes waterlogging stress in foxtail millet.The grouping of the genotypes based on WTC values, identified 4 genotypes (IIMR FxM-7, SiA-4245, GPUF-17) to be highly tolerant, 6 susceptible (CRS FxM-4, SiA-4243, ST.4, ST.5, ST.7 and RK.1) and 16 tolerant. The cluster analysis based on Euclidean distances using UPGMA method grouped the genotypes into five clusters at 40 phenon level. The genotypes RK.1 and ST.4 were present in mono-genotypic clusters. The screening of the selected 14 genotypes at seedling stage showed significant variations in all the six observed morphological characters for 3 days and 6 days stress conditions. A significant variation in physio-biochemical parameters were observed under both stress conditions. An overall reduction in chlorophyll A and total chlorophyll was observed, while value of chlorophyll B showed increase during 3-day stress and decrease 6-day stress. The proline content showed significant reduction while peroxidase activity showed an average increase during 5 day stress. It was notable that the changes in susceptible genotypes were more drastic than tolerant genotype. The performance of the genotype IIMR FxM-7 was found to be the best for both the stress conditions while genotype ST.5 showed poor performance was selected as highly susceptible genotype. The cluster analysis grouped all the susceptible genotypes in a single cluster while the highly tolerant genotype IIMR FxM-7 was present in single mono-genotypic cluster, indicating the effectiveness of this method of characterization in identification of waterlogging tolerant genotypes. The results of all the screening methods concluded that the 12 h waterlogging is short term stress for foxtail millet while 3 days is threshold for early seedling stage. The in-silico candidate gene search identified 14 candidate genes having probable role in waterlogging tolerance in foxtail millet. Most of the genes belong to ERF, WRKY and RBOHB gene family. The gene expression analysis of the selected five genes (SiRAP 2.2, SiRBOHB, SiRAP2.3, SiRAP2.12 and SiEREB180) in the leaves for tolerant and susceptible genotype under 12 h and 3 days stress revealed differential expression pattern suggesting a role of these genes in waterlogging tolerance mechanism in foxtail millet. The findings provide a fundamental clue for further cloning, characterization and functional analysis of other genes to help understand the molecular mechanism of waterlogging tolerance in foxtail millet.
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    ThesisItemUnknown
    Role of Iron Oxide Nanoparticles in alleviation of arsenic mediated phytotoxicity in rice (Oryza sativa L.)
    (RPCAU, Pusa, 2023) Murmu, Anju Kumari; Kumar, Shailesh
    The present study was done to study the effects of different concentrations of arsenic and iron oxide nanoparticles combinations on rice (Oryza sativa L.) seedlings. The objective for the study were: 1. To study the effects of different concentrations of arsenic and iron oxide nanoparticles on seed germination and seedling growth of rice. 2. To investigate the effectiveness of iron oxide nanoparticles in reducing the toxicity of arsenic based on physiological and biochemical changes in rice (Oryza sativa L.). Two rice genotypes (Rajendra Neelam & Gautam) were used for this experiment work. The experiment was carried out in two different ways: 1. In pots 2. In paper cups. In both the experiments the seeds and seedlings were subjected to different treatments and treatment combinations of arsenic and iron oxide (nanoparticles or bulk form). For paper cups experiment, the seeds were soaked in different concentrations of arsenic and iron oxide and the germination parameters were taken in 10-day old seedlings. For pot experiment, the seeds were first sown in soil filled pots and after it reached seedling stage, the plants were transplanted to plastic cups and were subjected to different combinations of arsenic and iron oxide. The treatments were as follows: T1: Control, T2: 25 µM As, T3: 50 µM As, T4: 25 mg/L FeNP, T5: 50 mg/L FeNP, T6: 25 mg/L Fe Bulk, T7: 50 mg/L Fe Bulk, T8: 25 µM As + 25 mg/L FeNP, T9: 25 µM As + 50 mg/L FeNP, T10: 50 µM As + 25 mg/L FeNP, T11: 50 µM As + 50 mg/L FeNP T12: 25 µM As + 25 mg/L Fe Bulk T13: 25 µM As + 50 mg/L Fe Bulk, T14: 50 µM As + 25 mg/L Fe Bulk and T15: 50 µM As + 50 mg/L Fe Bulk. The results showed that iron oxide reduced the germination %, shoot length, root length and also the seedling dry weight thereby, reducing the seedling vigor index I and II of seedlings exposed to Arsenic stress. The relative heavy metal injury rate increases on As treatment. The total biomass of the seedlings also reduces on As exposure. However, it gradually increases on iron oxide exposure. The relative water content (RWC) in leaves is reduced on exposure to As and it is increases on exposing it to different concentrations of iron oxide. Arsenic reduced the photosynthetic pigments, stomatal conductance and photosynthesis in leaves. On exposure of seedling to stress there is accumulation of reactive oxygen species (ROS) like hydrogen peroxide (H2O2) inducing oxidative stress, damaging the membrane lipids raising the electrolyte leakage (EL). Also there is peroxidation of lipid membrane causing rise in malondialdehyde (MDA). Iron oxide strengthen the antioxidant enzymes (Superoxide dismutase and Ascorbate peroxidase), thereby alleviating oxidative stress and reducing EL and MDA content. The results reveal that iron oxide (nanoparticles and bulk) had a positive influence on morphological, seed germination parameters, physiological and biochemical parameters. However, better results were seen in case of iron oxide nanoparticles. This shows that supplementation of iron oxide nanoparticles to arsenic stressed condition can help alleviate arsenic mediated phytotoxicity.
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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.