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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 GENETIC POLYMORPHISM ANALYSIS OF GENES INVOLVED IN LYSINE AND TRYPTOPHAN METABOLIC PATHWAYS AND HETEROTIC GROUPS OF QUALITY PROTEIN MAIZE INBRED LINES(Dr.RPCAU, Pusa, 2022) SNEHAL,SIKHA; SHARMA, V.K.A study was performed in order to establish a relationship between the genetic diversity of quality protein maize inbred lines and performance of hybrids thus obtained from these inbred lines, field experiment was conducted. The experimental hybrids were obtained from the 15-quality protein inbred lines when crossed with 3 tester inbred lines, which was observed in randomized block design having three replications. Parental lines and experimental hybrids were randomized separately in continuous plots at TCA, Dholi. Statistical analysis was performed based on 11 morphological characters was studied, including 50% tasseling in days, 50% silking in days, 75% brown husk in days, height of plant, cob girth, length of cob, ear height, kernels in a row, kernels in a cob, grain yield per plant and test weight during the Kharif and Rabi seasons. The tests were carried out in the Molecular Biology Laboratory of Dr. RPCAU, Pusa, Bihar, to assess the type and extent of divergence between eighteen QPM inbred lines based on the analysis of 20 SSR and 26 lysine and tryptophan metabolic pathways candidate gene-based marker targeted microsatellite sites-based polymorphism. The ANOVA was done independently for both the season and across two environments. 18 QPM inbred parental lines and 45 single crosses, were subjected to statistical analysis for determination of significant differences among 11 quantitative features used in the experiment. ANOVA values revealed significant variations and degree of reliability in the performance of inbred lines and crosses in Kharif and Rabi seasons, as well as utilizing pooled data. The 63 entries in this study were analyzed for 11 quantitative characters, and ANOVA thus obtained revealed significant variations among them. To study the relevance of variation in the sources, the mean sum of squares for parents, hybrids, and parents vs. hybrids. For all of the characters studied, including 50% tasseling in days, 50% silking in days, 75% brown husk in days, height of plant, cob girth, length of cob, ear height, kernels in a row, kernels in a cob, grain yield per plant and test weight during the Kharif and Rabi seasons, categorization of variance showed substantial differences among the inbred parental lines. As deduced from the results obtained the mean sum of squares due to parents, hybrids, and parent vs. hybrids was significant for all traits during both Kharif and Rabi seasons, as well as across seasons. For selecting economically important morphological traits, it involves taking into account the level of heterosis expression in grain yield, as well as hybrid performance per se, when evaluating the potential of hybrids. A careful examination of the related information for hybrid means performance and degree of heterosis expressed for grain yield per plant reveals that these 17 hybrids, namely: QPML-07×QPML-16, QPML-03×QPML-16, QPML-02×QPML-14, QPML-04×QPML-14, QPML-11×QPML-14, QPML-13×QPML-14, QPML-11×QPML-15, QPML-09×QPML-15, QPML-02×QPML-15, QPML-04×QPML-14, QPML-03×QPML-15, QPML-18×QPML-15, QPML-08×QPML-16, QPML-06×QPML-14, QPML-13×QPML-15, QPML-17×QPML-14, QPML-18×QPML-14, was observed to have higher mean performance as well as significantly positive heterosis for grain yield and appeared to be the most promising hybrids under consideration. All the 18 parental QPM inbred lines were further classified into different clusters based on the dissimilarity units as observed in the dendrogram. The 18 QPM inbred lines were arranged into 3 clusters, out of these 3 clusters, 2 clusters, i.e. Cluster A and Cluster B were containing multiple genotype, however cluster C was containing only 2 genotypes. According to the dendrogram, QPML-13 and QPML-14 were showing maximum similarity when the 11 morphological characters were taken into consideration. However, QPML-11 and QPMLI-16 were showing minimum similarity when the 11 morphological characters were taken into consideration, and thus, was considered as the most divergent. Thus, from the analysis of genetic diversity performed in the current study, it was observed that 18 QPM inbred lines revealed abundant genetic diversity amongst each other. An analysis of the genetic similarity, hybrid index value, and hybrid mean value of heterotic groups generated using microsatellite markers revealed that QPML-11, QPML-13, QPML-17, and QPML-18 all belonged to heterotic Group-1. In all cases, QPML-09 belonged to heterotic Group-2. In every case, QPML-01 belonged to heterotic Group-2. The overall coincidences were significantly high when inbred lines were classified on the basis of SSR markers in different heterotic groups according to hybrid index value and hybrid mean value. As a result, the values lead to the conclusion that microsatellite markers could be used for effective and efficient method for classification of inbred lines into diverse heterotic groups for minimizing the size of single crosses that is required for production and evaluation. Hence, enhancing the of hybrid maize breeding program. The genomic DNA of maize was amplified utilizing following 26 candidate gene-based SSR primers. Different number of alleles were obtained for each marker and it ranged from 3 in ZmOPQ-2 to 13 in ZmAKH-1. A sum total of 193 alleles were obtained with 26 SSR marker, with an average value of 7.4 alleles per locus for the 18 inbred lines. Utilizing 26 SSR markers 78 unique alleles and 117 shared alleles were obtained. No unique allele was obtained for ZmHSDH-3. Only 1 unique allele was obtained for ZmOPQ-1, ZmOPQ-2, ZmAS-1, ZmAS-3, ZmOM-2, ZmOM-4, ZmAKH-4 and ZmOM-8. Maximum number of unique alleles were obtained for ZmAKH-1 (10) and ZmCS-1 (7). This showed varying degree of polymorphism amongst the 18 QPM inbred lines utilized under the present study. Unique allele percentage is used for expressing the value Polymorphism Per cent (PP), thus maximum PP value was obtained for ZmAKH-1 (76.90%) and the minimum value was obtained for ZmHSDH-3 (0%), followed by ZmOPQ-01 and ZmAS-1 (14.20%). The mean value of PP thus obtained for all the 26 markers was 36.00%. Equally greater degree of PP was obtained for the following primers: ZmCS-1 (63.60%), ZmHSDH-5 (62.50%), ZmOM-3 (55.50%), ZmHSDH-1 (54.50%), ZmOPQ-4 (50.0%), ZmASK-1 (50.0%) and ZmHSDH-2 (50.0%). Maximum PIC value was obtained for ZmAKH-1 (0.902) and the minimum value was obtained for ZmOPQ-2 (0.500). The mean value of PIC thus obtained for all the 26 markers was 0.8. Thus, from the data obtained following are the primers pair having higher value of both PP and PIC as well as number of alleles: ZmOPQ-3, ZmOPQ-4, ZmASK-3, ZmCS-1, ZmAS-2, ZmOM-3, ZmAKH-1, ZmAKH-2, ZmHSDH-1, ZmHSDH-4, ZmHSDH-5 and ZmOM-6. The genomic DNA of maize was amplified utilizing following 20 SSR primers. Different number of alleles were obtained for each marker and it ranged from 3 in Bnlg-1666 and UMC-1809 to 14 in UMC-1413. A sum total of 160 alleles were obtained with 20 SSR marker, with an average value of 8 alleles per locus for the 18 inbred lines. Utilizing 20 SSR markers 72 unique alleles and 88 shared alleles were obtained. No unique allele was obtained for Bnlg-1666 and Phi-002. Only 1 unique allele was obtained for Phi-064, Phi-046, Phi-011 and UMC-1370. Maximum number of unique alleles were obtained for UMC-1413 (11) and UMC-1159 (5). This showed varying degree of polymorphism amongst the 18 QPM inbred lines utilized under the present study. Unique allele percentage is used for expressing the value Polymorphism Per cent (PP), thus maximum PP value was obtained for UMC-1413 (78.60%) and the minimum value was obtained for Phi-002, Bnlg-1666 and UMC-1809 (0%), followed by Phi-050 (16.60%), Phi-064 (20.0%) and Phi-046 (20.0%). The mean value of PP thus obtained for all the 20 markers was 32.42%. Equally greater degree of PP was obtained for the following primers: UMC-1159 (62.50%), UMC-1465 (50.0%), UMC-1587 (50.0%), UMC-1425 (40.0%), UMC-1117 (40.0%), UMC-1133 (40.0%), Phi-014 (40.0%) and UMC-1638 (40.0%). Maximum PIC value was obtained for UMC-1413 (0.908) and the minimum value was obtained for UMC-1809 (0.602). The mean value of PIC thus obtained for all the 20 markers was 0.77. Thus, from the data obtained following are the primers pair having higher value of both PP and PIC as well as number of alleles: UMC-1465, UMC-1117, UMC-1587, Bnlg-105, UMC-1413, Phi-014, UMC-1638, UMC-1370, Phi-050 and Phi-062, appeared to be more informative when the number of alleles generated by different primer combinations was compared to the level of polymorphism identified in the current study. Genetic profile of the 18 inbred lines utilizing 26 candidate gene-based primers showed the inbred lines along the two main axes showed the accommodation of all the inbred lines into 3 well distinguished major genotypic groups. Thus, cluster analyses and principal coordinate analysis showed the existence of wide variation at genetic level that was present among the 18 QPM inbred lines. Thus, with the utilization of 26 candidate gene-based primer, significantly high degree of polymorphism was obtained among the 18 QPM inbred lines at genetic level. Cluster analysis was performed for the evaluation of variability at genetic level amongst 18 QPM inbred lines utilizing 20 SSR primer pairs showed the accommodation of all the inbred lines into 3 well distinguished major genotypic groups. Thus, with the utilization of 20 SSR primer, significantly high degree of polymorphism was obtained among the 18 QPM inbred lines at genetic level. Thus, analysis showed the existence of wide variation at genetic level that was present among the 18 QPM inbred lines. Similarity coefficient was estimated based on the comparison of presence and absence of bands amplified utilizing 46 pairs of primers (genic and generic). All the 18 parental QPM inbred lines were further classified into different clusters based on the similarity units as observed in the dendrogram. Thus, with the utilization of 46 combined (genic and generic) primer pairs, significantly high degree of polymorphism was obtained among the 18 QPM inbred lines at genetic level. Cluster analysis was performed for the evaluation of variability at genetic level amongst 8 QPM inbred lines and 4 non-QPM inbred utilizing candidate gene-based primer pairs. Similarity coefficient was estimated based on the comparison of presence and absence of bands amplified utilizing 26 pairs of primers. The 8 QPM inbred lines and 4 non-QPM were further classified into 2 different clusters. Likewise, when cluster analysis was done utilizing the 4 designed primers, it was also able to differentiate the 8 QPM inbred lines and 4 non-QPM taken as experimental material. The 8 QPM inbred lines and 4 non-QPM were further classified into 2 different clusters based on the similarity units. Thus, cluster analysis and principal coordinate analysis showed the existence of wide variation at genetic level that was present among the 8 QPM inbred lines and 4 non QPM inbred lines. Cluster A comprised of the following 8 inbred lines: QPML-01, QPML-02, QPML-03, QPML-04, QPML-05, QPML-06, QPML-07 and QPML-08. Cluster B comprised of the following 4 non-QPM inbred lines: NQPML-01, NQPML-02, NQPML-03 and NQPML-04. The distribution pattern in 2-Dimension for inbred lines genetic profiles was in correspondence with the interrelationship exhibited by hierarchical classification on the basis of similarity coefficients (Fig. 4.). Genetic profile of the 8 QPM inbred lines and 4 non QPM inbred lines utilizing 26 candidate gene-based primers as well as the 4 designed primer pairs showed the inbred lines along the two main axes showed the accommodation of all the inbred lines into 2 well distinguished major genotypic groups. The genetic structure illustrated by 46 combined SSR primers (26 candidate gene specific and 20 SSR primers loci showed that the genetic structure of 18 QPM inbred lines are composed of three major ancestral components. The results can infer that the computational analysis of the genetic structure of the QPM inbred lines unambiguously reflected that the genotypes subjected high tryptophan and lysine content are having related molecular characterization are the admixture of three ancestral components present in different combinations in different genotypes.ThesisItem Open Access Microsatellite Marker Assisted Determination of Genetic Polymorphism and Heterotic Groups Among Baby Corn Inbred Lines(Dr.RPCAU, Pusa, 2022) KIRTI, SIMRAN; SHARMA, V. K.A study was conducted for characterization of baby corn inbred lines using agronomic characters and microsatellite markers to determine the molecular level genetic polymorphism and divergence among these baby corn inbred lines for construction of heterotic groups based on comparative analysis of amplification pattern of targeted microsatellite sites. The basic experimental material comprised twelve inbred lines and three inbred testers of baby corn, which were crossed to generate thirty-six single cross hybrids. The inbred lines and hybrids were evaluated in randomized block design with three replications during two seasons (rabi and kharif seasons). The degree of divergence and separation among inbred lines of baby corn was quantified and inbred lines were then divided into various heterotic groups based on the examination of focused microsatellite sites. Analysis of parental lines and heterosis in experimental hybrids was done statistically using nine quantitative agro-morphological characters based on the measurements and assessments of plant height, ear height, days to 50% tasseling, days to 50% silking, cob yield per plant, green fodder yield, baby corn length, baby corn girth and cob weight. Stastical analysis of nine quantitative characters using numerical taxonomic approach was employed and the inbred lines were classified into three clusters. Young seedlings were grown in almuninum container to extract genomic DNA for microsatellite primers based amplification of targeted genomic regions. Amplified products were produced using a set of 38 microsatellite sites specific primer pairs that covered every chromosome in the genome. Microsatellite-based allelic diversity was used for further classification and creation of various heterotic groups of the inbred lines. Statistically significant differences were revealed amongst inbred lines and crosses for nine quantitative morphological charactersrecorded during both the seasons. Among inbred lines, BCL-08 outperformed over all other inbred lines in respect of cob yeild per plant, followed by tester BCL-12, which was statistically equivalent to inbred lines BCL-05, BCL-07, BCL-10 and BCL-04 over the environments. Among 36 crosses, 14 crosses were found to have significantly greater mean performance as well as significantly positive heterosis. The cross BCL-10×BCL-12 was observed as high yielding cross followed by BCL-04 × BCL-03, BCL-09 × BCL-03, BCL-05 × BCL-03, BCL-07 × BCL-15, BCL-06 × BCL-15and BCL-14 × BCL-15 for cob yield per plant. By the use of 38 primer pairs, 279 allelic variant were investigated as amplified products. The value of the number of alleles varied from 4 for the primer phi011 to 11 in case of Nc130 having range between 75bp to 580 bp.The sum of total value of shared alleles was 166, which varied from 3 to 8 alleles per primer pairs. Similarly, the sum total of unique allels was 113, which ranged from 1 to 7 alleles per primer among the 15 inbred lines. Taking into the consideration the number of alleles and polymorphic status, the primers UMC1425, Phi024, UMC1407, UMC1159, Phi046, UMC1587, UMC1241, UMC1858, UMC1638 and UMC1370 seemed to be more informative on the basis of polymorphism level. On the basis of molecular level classification, BCL-15 and BCL-01 seemed to be highly diverse genotypes. Contrarily, BCL-03 and BCL-04 appeared most closely related to each other in same group. The inbred lines BCL-12 and BCL-13 were closely associated to each other. Both molecular and morphological attributes dependent cluster based data manifested that among the three testers, namely, BCL-03, BCL-12 and BCL-15 two testers excluding BCL-03 were included in same cluster. However, the cluster analysis used to group inbred lines using data from quantative characters and microsatellite markers did not show a perfect match. Results of genetic structure analysis based on repeat sequence length variation in primer specific genomic regions revealed that the genetic compositions with refrence to the targeted genomic regions of the inbreds are basically the admixtures of various combinations of three ancestral components. Analysis of the heterotic groups compositions using the hybrid index value and the hybrid mean value revealed that BCL-04, BCL-06, BCL-07, BCL-09, BC-L10, BCL-11 and BCL-14 were found in same group i.e in Group 1, BCL-01 and BCL-05 were found in Group 2 and BCL-02, BCL-08 , BCL-13 were found in Group 3.A comparative analysis of amplification pattern of targeted microsatellite sites was found effective in discriminating the inbred lines of various heterotic groups and therefore the employed panel of primer was found moderately efficient in discrimination of inbreds into heterotic groups.ThesisItem Open Access DETECTION AND VALIDATION OF EST-SSR MARKERS ASSOCIATED WITH SUGAR YIELD IN SUGARCANE(DRPCAU, PUSA, 2022) S, DIVAKAR; SINGH, ASHUTOSHSugarcane (Saccharum spp.) is a widely cultivated crop and fulfils around 75 % of sucrose demands worldwide. Due to its polyploidy and complex genetic nature, it is difficult to map the gene for sucrose content. But association mapping is one of the alternatives to identify the genes or markers for marker-assisted selection. In the present study total of 212 EST-SSR primers were collected from different literature. The functionality of each primer was tested by using Blast2Go software and 30 EST-SSR primers were selected that related with sugar content. These markers were validated by association mapping. Total 70 F1 diverse genotypes for sugar contents were phenotype with two check lines. All the parameters for sugar contents were recorded. The results showed a significant variation between the genotypes for sugar yield traits such as Brix value, purity, and sucrose content, etc. The correlation studies revealed that the Brix%, Sucrose content, and sucrose recovery were significantly correlated. The association analysis was performed by using MLM (Mixed Linear Model) and avoid false positive associations. The association analysis revealed that SEM 407 marker was significantly associated with Brix% and sucrose content. SEM 407 primers putative related with diphosphate-fructose-6-phosphate 1- phosphotransferase that are associated with Brix% and sucrose content. This functional marker can be used for marker assisted selection for sugar yield traits in sugarcane.ThesisItem Open Access Comparative Studies of Physio- Biochemical Traits and CAMTA Gene Expressions in Contrasting Genotypes of Chickpea (Cicer arietinum L.) Under Fusarium wilt STRESS(DRPCAU, PUSA, 2022) R.A, SUDHAN; BHUTIA, K. L.Chickpea is an important pulse crop and has high water use efficiency and largely grown as rain-fed on residual soil moisture. Chickpea has high protein content and it is a nutrient dense food with high vitamin and mineral content. Multiple abiotic tolerance and disease resistance factors were found to be associated with the Calmodulin-binding transcription activators (CAMTAs). The essential role of Cicer arietinum CAMTA (CaCAMTA) gene composition in the genome and their mechanism in resistance against the Fusarium wilt infection is not explored. The present study was conducted with contrasting genotypes of chickpea to assess the expression of phyiso-biochemical traits and CaCAMTA genes under Fusarium wilt stress. The comparative analysis of physio-biochemical traits showed differential expression of traits like chlorophyll, carotenoid, protein and phenol content in contrasting genotypes of chickpea under Fusarium wilt stress as compare to control plants. Similarly, the activities of antioxidant enzyme such as Guaiacol peroxidase, Catalase, Sodium dismutase and Ascorbate peroxidase were also found to be significantly varied among control and Fusarium infected plants. Likewise, for physiological traits such as relative water content, membrane stability index and root density, the six contrasting genotypes showed significant differences between control and Fusarium wilt treated plants. Similarly, out of seven CaCAMTA genes, three genes i.e., CaCAMTA-1.1, CaCAMTA-6.1, CaCAMTA-6.2 showed significant differential expression under Fusarium wilt stress as revealed by Real time PCR approach. When compared to RNA sequencing data, apart from these three CaCAMTA genes, three other CaCAMTA genes were found to be differentially expressed in KWR 108 (Resistant) and GL 13001 (Susceptible). Several proteins interacting with proteins encoded by CaCAMTA genes were identified among which, genes encoding 10 interacting proteins were found to be differentially expressed under Fusarium wilt stress along with CaCAMTA genes. Further, in silico survey revealed four proteins interacting with CAMTA proteins such as XP_004506228 (Enhanced Disease Susceptibility-1), XP_004494535 (Calmodulin binding, stress response protein), XP_004497845 (Myb-related protein Myb4-like), and XP_00459656 (Homeoboxleucine zipper protein ATHB-12-like) were reported of having important role in stress response mechanisms. Therefore, after comparative analysis of contrasting genotypes under Fusarium wilt stress, it can be concluded that different genotypes of chickpea have different approach of coping up against the wilt stress and after analysing at the expression profile of CaCAMTA genes and its interacting protein coding genes under Fusarium wilt stress, it can be concluded that some members of CaCAMTA gene family could be involved in imparting resistance against Fusarium wilt in chickpea. Therefore, these CaCAMTA genes may be targeted to design the markers for marker trait association studies in order to find its link to disease resistant through associated traits.ThesisItem Open Access Study of Marker Trait Association for Zn Fortification in Wild Rice(DRPCAU, PUSA, 2022) S, VIMANTH; Kumari, SaritaRice is an important source for food calories for world-wide population. World demand for both quantity and quality of rice. Zinc is an important mineral nutrient for human population. Fe and Zn is major cause of hidden hunger and malnourishment for the developing countries. Rice grain fortification using Zn will provide a cheap source for targeting problem around the world. Improvement of rice using breeding programme often compromised because of unavailability of suitable donor and marker. Present studies were conducted with objectives to study the diversity and population structure among wild rice accessions of Oryzanivara and Oryzarufipogon. The phenotypic variation was found 8.5% to 70% for different agronomically important trait and rice grain zinc content. Six accessions of wild rice were found for grain zinc content >45 PPM. Population structure analysis identified two subpopulations based on Structure, and PCoA and UPGMA clustering. The significant of two subpopulations were further estimated through AMOVA analysis. The diversity among population (14 %) was less than variation between populations (86%). The mean Fst value between population was 0.135. Marker trait analysis was studied through MLM (K+Q) model using total 93 different kind of SSR and SNP markers. A total of 16 genetic markers were discovered to be linked with grain zinc concentration.Annotation of linked loci identify the region coded for 589 different regulatory genes and 638 transporters. The identified loci may be used as an indicator of high zinc content after validations. Genes associated with the region may be use for transgenic program after studying their expression under concerned associated traits. Six accessions with high accumulator of zinc content may be used in breeding program for biparental mapping and marker assisted introgression.
