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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 CHARACTERIZATION OF CAMTA TRANSCRIPTION FACTOR FAMILY WITH RESPECT TO DROUGHT TOLERANCE IN CHICKPEA (Cicer arietinum L.)(DRPCAU, PUSA, 2021) KUNDU, SAYANTA; BHUTIA, K. L.Chickpea is the second most valued, protein rich annual legume that is abundant in dietary fibers, minerals, iron and phosphorus. It is cultivated in almost 50 countries and India is the largest producer of chickpea. However, the production is not enough to fulfill the demand. As chickpea is grown in arid and semi-arid region, the production faces huge loss under drought stress. So, the only solution of this problem is to generate drought tolerant varieties. CAMTA is a transcription factor family which specifically binds to Ca2+-calmodulin complex and regulates gene expression in response to various stress conditions including drought. Therefore, CAMTA family of chickpea was characterized using in silico and qRT-PCR based wet lab approaches under drought stress. Fifteen protein members basically encoded by 7 genes and their splice variants of CAMTA family were identified in chickpea and new names were assigned (CaCAMTA) to them as it is the first genome wide characterization of chickpea CAMTA gene family. The collected protein annotation and biological function depicts that the genes code for calmodulin-binding transcription activator proteins of different isoforms having role in gene regulation in response to stress tolerance i.e., freezing, salt and wounding etc. Among numerous cis-acting elements identified from 5’ upstream of CaCAMTA genes, MYBCORE, ABRELATERD1 and MYB1AT are the cis-acting elements having important roles during water stress conditions. The conserved domains present on polypeptide chains of each member of CaCAMTA family are CG-1 and IPT domain, ankyrin containing repeat domain, and IQ domain functioning as DNA binding, signal transducing and calmodulin binding domain. Dendrograms obtained for top 5 orthologs of 7 base genes suggested best hit orthologs gene were from Medicago truncatula and Vigna unguiculata. As the 15 proteins were basically the products of 7 CAMTA genes and their splice variants, products of the same CaCAMTA gene and their splice variants were clustered together in amino acid based phylogenetic analysis. Interaction analysis of CaCAMTA proteins revealed that each CaCAMTA proteins have network of interactions with various characterized and uncharacterized proteins. Under 15 days of drought treatment, growth and development of three chickpea genotypes namely RSG 888, Vijay and Pusa 256 were not much hampered, therefore were considered as drought tolerant genotypes. Similarly, the growth of chickpea genotype JG 14 was severely reduced during 15 days of drought treatment as compare to control plant, therefore, it was considered as susceptible genotype. The chlorophyll contents in treated plants were increased in RSG 888 and Vijay, but decreased in Pusa 256 and JG-14. The differential expression study of CAMTA genes using qRT PCR in chickpea under drought stress showed that CaCAMTA 1.1.2, 6.2.3, 7.1.1, and 5.1.1 are upregulated among the tolerant genotypes i.e., RSG 888, Vijay, and Pusa 256; and CaCAMTA 1.2.1 is downregulated in tolerant genotypes RSG 888 and Pusa 256 and up-regulated in susceptible genotype JG-14. The genome wide characterization of CAMTA family in chickpea will broaden the idea of drought stress mechanisms in Chickpea. The candidate gene-based markers could be designed targeting these differentially expressed genes to be utilized in marker trait association studies and if found significantly associated to yield attributing traits of chickpea, it can be utilized in marker assisted breeding of Chickpea for drought tolerance.ThesisItem Open Access EFFECTS OF GAMMA IRRADIATION ON FREE RADICAL PROCESSING ENZYMES AND YIELD ATTRIBUTING TRAITS OF SUNFLOWER (Helianthus annuus L.)(DRPCAU, PUSA, 2021) RAJEEV, RANJAN; BHUTIA, K. L.Sunflower (Helianthus annuus L.) is an important oilseed crop of India and world as a whole. Sunflower seed oil contains 90% unsaturated fatty acids and 10% saturated fatty acids.India produces 1.1 lakh tonnes of sunflower oil and imports 15.45 lakh tonnes. Total consumption of sunflower oil among all oil in India is 6.89%. However, many biotic and abiotic factors severely affects the production and productivity of sunflower along with some other constraints like overlapping of sowing time with other crop cultivation such as wheat, maize and minimum bee activity in growing areas in India. Therefore, the present investigation was conducted to investigate the performance of mutant lines (M3 generation) of sunflower obtained by treating the base sunflower (TNAUSUF 7) seeds with different doses of gamma radiations. M3 lines obtained from 100 Gy (T1) and 130 Gy (T2) gamma irradiated plants were used in the current investigation. Variations in yield and yield attributing traits between control, T1 and T2 plants were evaluated by taking field data from 5 randomly selected plants from each of four replicated plots. The germination % of control, T1 and T2 in M3 generation were 84%, 75%, and 72% respectively. The significant differences between the mean values of three groups i.e. control, T1 and T2 were observed in days to 50 % flowering, days to maturity, plant height (cm), no. of unfilled seeds per head and fertility % with P values of 0.00149, 0.0105, 0.00253, 0.00305 and 0.02648, respectively and Least Significant Differences of 2.08, 2.36, 3.51, 51.58 and 4.07, respectively. Oil from the seeds of control, T1 and T2 extracted using Soxhlet method showed 36.66 %, 37.26 % and 35.80 %, respectively. The antioxidant property of the oil was assayed using DPPH method and found that the antioxidant activity expressed in term of IC50 was best in oil derived from the seeds of T2 plants with IC50value of 184.53 mg/ml. In the present experiment, the effect of gamma irradiation on the expression of free radical processing enzymes was also assessed using qRT--PCR. Mn-Superoxide dismutase showed down regulation in T2 plants compared to both control and T1. Similarly, Alternative Oxidase and Catalase were significantly up-regulated in T1 plants as compared to both control and T2, respectively. These finding suggested that the treatment of sunflower seeds with different doses of gamma irradiations was successful in generating genetic variation of sunflower with positive response in several yield and yield attributing traits such as days to 50 % flowering, days to maturity, plant height (cm) and fertility %. These mutant lines after proper selection could be used as composite variety of sunflower and could be utilize as the source of genetic variation in sunflower breeding programs.ThesisItem Open Access Molecular characterization in relation to seed vigour related traits in rice(DRPCAU, PUSA, 2021) KUMAR, PRATIK; SHARMA, V. K.Seed vigour is important for crop establishment in rice. The present study was undertaken to characterize the rice genotypes using seed vigour influencing physio-biochemical traits, as well as, molecular profiling based on microsatellite markers to investigate molecular level genetic divergence and population structure pattern among a set of rice genotypes and to evaluate the marker trait association, if any, between marker phenotype and seed vigour related traits under consideration. Significant genotypic differences were observed for all physio-biochemical traits recorded in a set of locally adapted rice genotypes. On the basis of mean performance, six genotypes, namely, Rajendra Neelam, RAU1421-12-1-7-4-3, Vaidehi, Rajshree, RAU1415-35-76-9-5-3-4 and Rajendra Suwasini appeared to be superior entries based on the score of physio-biochemical traits. The correlation studies revealed, significant positive correlation of seed vigour with catalase, chlorophyll, carotenoid and amylose content. A significant positive correlation of higher order was observed among chlorophyll and carotenoid and between starch and amylopectin, although, correlation of starch with amylose was relatively weak. Significant positive correlation was also found between total anthocyanin, total flavonoids and total phenolics content. Catalase activity was positively correlated with superoxide dismutase activity. Negatively significant correlation was observed only between peroxidase activity and total flavonoid content, albeit of lower magnitude. First five principal component axes accounted for 82.6% of total variance between the sixteen recorded physio-biochemical traits. Starch, amylose, amylopectin, anthocyanin, phenolics and catalase appeared to be important classification variables, from this investigation. Average taxonomic distance based two-dimensional ordination identified three genotypes, namely, Rajendra Neelam, Sudha and RAU1451-66-1-1-5-1 to be relatively more diverse from rest of the genotypes based on physio-biochemical traits. Simple sequence repeats-based polymorphism survey in 18 genotypes of rice using 38 microsatellite markers detected a total of 275 allelic variants with an average of 7.2 alleles per primer. Polymorphism information content of 38 SSR primer pairs ranged from 0.593 to 0.877 with an average value of 0.766 and polymorphism per cent reflecting the proportion of unique alleles ranged from 0% to 57.14% with an average value of 34.40%. Out of 38 primer pairs, RM85, RM205, RM328, RM337, RM6275, RM7003, RM7364 and RM14978 appeared to be relatively more polymorphic and informative primers. Analysis of genetic divergence revealed ample molecular level genetic variation and allowed differentiation and classification of rice genotypes into four groups. Structure analysis revealed that the targeted genomic regions of eighteen rice genotypes are basically the admixtures of different combinations of three ancestral components. Single marker analysis detected the association of the traits, namely, amylose, total protein, total anthocyanin, total flavonoids, total phenolics, starch, carotenoids, total chlorophyll, chlorophyll a, chlorophyll b, seed vigour index I, superoxide dismutase, catalase and peroxidase with the molecular markers. Association of the trait, catalase with marker RM5310 located on chromosome 1, amylose with marker RM440 on chromosome 5, total anthocyanin with marker RM547 on chromosome 8 and total flavonoid with marker RM286 and RM547 on chromosome 11 and 8, respectively were also reported in earlier study and was validated in this investigation. The marker-trait association can be further validated and effectively utilized in marker-assisted selection program for improvement in marker-associated trait of interest.ThesisItem Open Access TARGET REGION AMPLIFICATION POLYMORPHISM BASED PROFILING OF SALT STRESS RESPONSIVE CANDIDATE GENES IN RICE(DRPCAU, PUSA, 2021) SUDARSAN, PARVATHY; SHARMA, V.K.The present study was undertaken to examine the molecular level genetic polymorphism in relation to salt tolerance ability among 18 genotypes of rice using 10 candidate gene specific a total of 20 TRAP markers. Experimental procedure adopted in this study was designed to ascertain the genetic variability and diversity with respect to salt tolerance amongst different rice genotypes. Using 20 TRAP primer pairs involving salt stress responsive ten candidate genes specific 10 fixed forward primers in combinations with 2 arbitrary reverse primers, reproducible amplification was successfully achieved in 18 rice genotypes. Molecular level genetic polymorphism was clearly recognized by the deviation in the physical position of the products during their resolution through electrophoresis as well as variation in the number of products generated between genotypes. Differential amplification pattern was observed amongst the genotypes and 610 polymorphic bands representing easily recognizable different types of amplified products were categorized into 291 unique products and 319 shared products. Polymorphism information content of 20 TRAP markers arrayed from 0.745 to 0.940 with an intermediate value of 0.891 across all the combinations of fixed and arbitrary primers. Arbitrary reverse primer ME02 in combination with candidate gene specific fixed forward primers CDMK produced the least number of polymorphic variants. Similarly, another arbitrary primer ME05 produced the least number of polymorphic variants in combination with the fixed forward primer OsHKT1; 5. However, the forward fixed primer OsHKT1; 5 produced the highest number of polymorphic variants in combination with the arbitrary reverse primer ME02. The reverse arbitrary primer ME05 yielded the highest number of polymorphic variants in combination with the forward fixed primer CCC. The primer combination SHMT1 with ME05, OsHKT2;3 with ME02, OsHKT1;3 with ME05 and OsHKT1;1 with ME02 yielded remarkably larger allelic size difference in descending order of magnitude. Numerical variation in polymorphism information content of the markers reflected their allelic diversity and distribution frequency among the genotypes. The primer combinations OsHKT1;5 with ME02, OsHKT1;1 with ME05, OsHKT1;3 with ME05, SNAC1 with ME05 and CCC with ME05 appeared to be highly polymorphic and informative with above average values for the number of polymorphic products, number of unique products, marker index, resolving power and polymorphism information content. These primer combinations also recorded higher values for polymorphism per cent. Experimental results clearly reflected the existence of considerable genetic diverseness at the molecular level amongst the salt tolerant and susceptible genotypes evaluated in the present study. Hierarchical classification pattern based on target region amplification polymorphism related genetic similarity among rice genotypes was in complete agreement with principal coordinate analysis dependent two-dimensional spatial distribution pattern of genetic profiles of rice genotypes. Using both the modules, the genotypes were discriminated into five multi-genotypic groups highly consistent with their salinity tolerance status. Salt responsive candidate gene based target region amplification polymorphism analysis was shown to be extremely efficient in differentiating rice genotypes based on salt tolerance status. As a result, these markers may be used to discriminate between different salt stress responsive rice genotypes and to choose genetically differentiated parental genotypes for the purpose of genetic enhancement in relation to salt tolerance capacity.ThesisItem Open Access GENOME WIDE CHARACTERIZATION AND EXPRESSION PROFILING OF AP2/EREBP TRANSCRIPTION FACTOR TO ABIOTIC STRESS IN MAIZE (ZEA MAYS L.)(DRPCAU, PUSA, 2021) KUMARI, MANISHA; Singh, AshutoshThe transcription factors APETALA2/ETHYLENE-RESPONSIVE ELEMENT BINDING PROTEIN (AP2/EREBP) 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 AP2/EREBP family genes in maize. In this context, a genome-wide survey was done to identify the AP2/EREBP genes in maize (Zea mays L.), and total 54 AP2/EREBP family of genes were identified. The distribution of these 54 genes were irregular on the 10 chromosomes of maize. The phylogenetic analysis conducted for these 54 genes of AP2/EREBP TFs and it divided into ten groups, namely groups I to X. To study the role of AP2/EREBP group of genes in drought and salt stress at seedling stage of maize. The two contrasting inbred lines (BTM-6 and BTM-14) were chosen for gene expression study. The inbreds were grown in pot. The fourteen days old seedlings were exposed to 20 % PEG (3h and 6h) and 200Mm NaCl (3h and 6h) for drought and salt treatments respectively. The mRNA of each treatment of 14 days old seedlings extracted in two replication and cDNA synthesized. The differential expression was performed using qRT-PCR and result clearly revealed that under drought stress GRMZM2G013657_P01, GRMZM2G021573_P01 and GRMZM2G022359_P01 were unregulated after 6 hour of drought treatment in cultivar BTM-14.GRMZM2G028151_P01, GRMZM2G076602_P01 and GRMZM2G086573_P01 were up-regulated after 6 hour salinity treatment in cultivar BTM-6. In case of GRMZM2G073982_P01 under drought treatment up- regulated in both 3 hour and 6 hour in cultivar BTM-14 and under 6 hour salinity treatment in cultivar BTM-6. GRMZM2G076602_P01 was up regulated after 3 j hour and 6 hour of drought treatment. GRMZM2G113078_P01 showed up regulation under drought treatment at 3 hour and 6hour in both cultivar (BTM-6 and BTM-14) and at 3 hour salinity treatment up-regulation was observed and at 6 h down regulation were observed in cultivar BTM-14. The expression study clearly showed the AP2 gene family have role in drought and salinity stress. The two inbreds are contrasting for heat and drought stress, So it respond differently.ThesisItem Open Access CANDIDATE GENE MARKERS BASED AMPLIFICATION PROFILING OF STERILITY MAINTAINERS AND FERTILITY RESTORERS FOR WILD ABORTIVE RICE CYTOPLASM(DRPCAU, PUSA, 2021) THAKUR, LOKESH; SHARMA, V. K.Identification of fertility restorers and sterility maintainers among the diverse rice germplasm lines is very important for the development of WA-CMS-based rice hybrids. A study was carried out to discriminate maintainers and restorers among the 24 entries using PPR9-782(M, I) and SF21 candidate genes-based panel of reported primer pairs, designed genic SSR and designed generic primer pairs and to investigate the genetic importance of these candidate genes-based panel of designed generic and microsatellite primers in discrimination of sterility maintainers and fertility restorers for validation of gene specific designed primers. The materials used in this investigation involved a set of 16 known fertility restorers and 8 known sterility maintainers of WA-CMS system. Employing a panel of 9 reported primers, 3 designed genic microsatellite primers and 12 designed candidate gene specific generic primers; amplification of targeted genomic regions was carried out. Statistical parameters utilized during present study included polymorphic per cent, polymorphism information content and similarity coefficient. Using two candidate genes based nine reported primer pairs, namely, RMS-PPR9-1, RMS-PPR9-2, RMS-PPR9-3, RMS-PPR9-4, RMS-PPR9-5, RMS-SF21-1, RMS-SF21-2, RMS-SF21-3 and RMS-SF21-5, the amplification profiling yielded 56 allelic variants with a total of 45 shared alleles and 11 unique alleles. Polymorphism per cent expressed in terms of the percentage of unique alleles of primers ranged from 0 to 42.9. The PIC value varied from 0.519 to 0.806, with an average of 0.709. The targeted amplification utilizing candidate gene based 3 designed genic SSR primers, namely, RRMS-PPR9-2, RRMS-SF21-1 and RRMS-SF21-2, yielded 16 allelic variants with 13 of them being shared alleles and 3 being unique alleles. Polymorphism per cent varied from 0 to 33.3. The PIC value for these three designed genic SSR primer pairs ranged from 0.660 for to 0.771 with an average of 0.721. The amplification using candidate gene based 12 designed generic primers, namely, RRCG-PPR9-1, RRCG-PPR9-2, RRCG-PPR9-3, RRCG-PPR9-4, RRCG-PPR9-5, RRCG-PPR9-6, RRCG-SF21-1, RRCG-SF21-2, RRCG-SF21-3, RRCG-SF21-4, RRCG-SF21-5 and RRCG-SF21-6 yielded 76 allelic variants, with 63 of them being shared alleles and 13 being unique alleles. While polymorphism per cent varied from 0 to 40, the PIC value ranged from 0.684 to 0.861, with an average of 0.775. Hierarchical cluster analysis as well as principal coordinate analysis using candidate gene based 9 reported markers enabled differentiation and classification of 8 sterility maintainers and 16 fertility restorers into two well separated groups with higher level of accuracy. Further, using candidate genes-based panel of three designed SSR primer pairs, reproducible amplification was successfully achieved. These three designed SSR primer pairs clearly differentiated 8 sterility maintainers and 16 fertility restorers into two distinct groups. Employing the two candidate genes based 12 designed generic primer pairs, the genotypes were differentiated into 3 groups with cluster A having 6 genotypes, cluster B having 8 genotypes and cluster C having 10 genotypes. Fertility restorers were accommodated into two groups, whereas sterility maintainers were conciliated into a separate group. Utilizing the two candidate gene based panel of five selected designed generic primer pairs, namely RRCG-PPR9-1, RRCG-PPR9-2, RRCG-PPR9-3, RRCG-PPR9-4 and RRCG-SF21-2, cluster analysis clearly differentiated the 16 fertility restorers from the 8 sterility maintainers, accommodating them into two well separated clusters. Analysis of the genetic polymorphism in the fertility restoration related candidate genes specific genomic regions using gene specific reported and designed primers provided the evidence to infer the existence of nucleotide sequence length variation among fertility restorers and sterility maintainers of wild abortive type cytoplasmic male sterility in rice. The differences appeared to be more pronounced between than within each of these two categories of genotypes, which contributed to their differential amplification profiles. Relatively more easily recognizable genetic polymorphism between sterility maintainers and fertility restorers was exhibited by reported primer pairs than designed primer pairs utilized for selective amplification in the present investigation. Experimental results established the genetic importance of fertility restoration related candidate genes-based panel of designed generic and microsatellite primers in discrimination of sterility maintainers and fertility restorers of wild abortive type cytoplasmic male sterility in rice. The usefulness of gene specific panel of three microsatellite primer pairs and gene specific selected panel of five generic primers was validated during the present experimentation. Hence, these markers can be further tested and effectively utilized for discrimination of sterility maintainers and fertility restorers among the rice genotypes.ThesisItem Open Access Molecular profiling of little millet genotypes using iron and zinc transporter based SSR markers of foxtail millet(DRPCAU, PUSA, 2021) KUMAR, KAUSHAL; Anjani, KumarThe present study entitled "Molecular profiling of little millet genotypes using iron and zinc transporter based SSR markers of foxtail millet" was carried out to identify useful microsatellite-based markers of foxtail millet, which can be used for characterization of little millet genotypes based on their iron and zinc content. Altogether, 2 genotypes of foxtail millet and 26 genotypes of little millet were used for molecular profiling using iron and zinc based designed genic SSR markers. The grain iron and zinc content were determined in 26 genotypes of little millet and they were categorized in 3 groups having high, medium and low iron and zinc content. For molecular profiling of little millet genotypes, genic SSR markers were designed using BatchPrimer3 against the 21 iron and zinc transporter genes of foxtail millet. The SSRs were detected in all the genes except five genes. A total of 39 SSRs were detected and 36 primers were designed. The trinucleotides repeats were found to be most common. The most common repeat motif was found to be GCG/CGC. Using the 36 designed genic SSR molecular markers, reproducible amplification was successfully achieved in foxtail millet. Out of 36 designed genic SSR molecular markers, 14 genic SSR markers showed successful amplification in 26 little millet genotypes. The 14 primers showed the polymorphic bands in the little millet genotypes (38.88 % transferability). Using a panel of 14 designed and transferable primers, 126 alleles were detected in foxtail and little millet. In little millet, 108 alleles were obtained, in which 34 were unique alleles. The molecular size of amplified product varied from 93 bp to 1083 bp. The polymorphism per cent was 50 in the primer SI-ZT-1A and SI-ZT-1B whereas the lowest value found in the primer SI-ZT-P29-E and SI-ZT-9A i.e. 0 with an average of 30.07. The PIC value varied from 0.245 to 0.827 with an average of 0.578. The DC of the primer ranged from 0.1826 in the primer SI-ITP-2 to 0.9947 in primer SI-ZT-1A with an average of 0.6888. The primers having dinucleotides repeats were the most effective in determining the allele in little millet and the maximum number of alleles was detected by the primer targeting gene zinc transporter 7. The Dice’s similarity coefficient was 0.8000 for the foxtail millet genotype pair whereas for little millet pair wise combination, the maximum similarity coefficient (0.7222) was found between the genotype WV-155 and WV-156 while 6 pair-wise combinations had shown minimum possible similarity (0.000). According to the cluster analysis and the principle coordinate analysis, the genotypes were differentiated into three groups which clearly differentiate high and low iron and zinc containing genotypes. Cluster I has the 5 genotypes, cluster II has 11 genotypes, cluster III has 12 genotypes. The experimental results amply emphasized that the panel of primers employed during experimentation was useful and adequate to discriminate the little millet genotypes in relation to iron and zinc content. These markers can therefore be used effectively and efficiently for further studies in relation with grain iron and zinc content.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.
