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ThesisItem Open Access Expression profiling of ADP-glucose pyrophosphorylase gene for thermo-tolerance in developing grains of wheat (Triticum aestivum L. Em. Thell)(CCSHAU, 2017) Mehndiratta, Ravi; Shikha YashveerThe present investigation was conducted to study the expression profile of ADP - glucose pyrophosphorylase (AGPase) large subunit for thermo-tolerance in developing grains of two genotypes (WH 711 and WH 730) of wheat under two different sowing conditions (normal and late sown). Gene expression and AGPase activity was studied at six different stages of grain filling (4, 8, 12, 16, 20 and 24DAA) along with physiomorphological and yield attributes. Under conditions of heat stress, there was a considerable reduction in grain yield/ plant, plant height, number of tillers and 1000 grain weight in thermosensitive genotype (WH 711) as compared to thermotolerant genotype (WH 730). The activity of AGPase was correlated with data obtained through expression profiling; the activity and expression of AGPase large subunit was highest at 20DAA for thermotolerant genotype (WH730) under both NS and LS conditions. In case of thermo sensitive genotype (WH711), the activity peaked at 20DAA under normal sown conditions and 16 DAA under late sown conditions; the activity was seized at 24 DAA after anthesis under NS and LS conditions. Putative cDNA obtained were sequenced for WH 711(1270bp) and WH 730 (1247bp). Cladogram generated using different AGPase large subunit sequences clustered WH 711 and WH 730 into different groups; WH 730 sequence was phylogenitically more similar to the sequence a cquired from AGPase large subunit of barley. This study supported the role of AGPase for thermotolerance in wheat by getting insights of the expression and enzyme activity at different stages of grain filling in LS as well under NS conditions; thus providing a deep understanding for the biologists working with ADP-glucose pyrophosphorylase in order to understand the functionality of AGPase .ThesisItem Open Access Marker-assisted selection of water-efficient aromatic lines from aerobic x lowland indica rice (Oryza sativa L.) crosses(CCSHAU, 2017) Meena, Rahul Kumar; Jain, R.K.Aerobic (MAS25, MAS26 and MAS-ARB25) x lowland Basmati (PUSA1121 and IB370) rice derived F3, F4, BC1F2 and BC1F3 populations displayed enormous variation for various physio-morphological (plant height, effective number of tillers/plant, panicle length, 1000-grain weight and grain yield per plant), quality (grain length/breadth ratio) and root (root length, thickness and biomass) traits under direct-seeded water-limited aerobic conditions in the field and net house. Frequency distribution curves for various physio-morphological and root traits were parabolic and in some cases, curves were partially inclined towards the respective aerobic rice parent. Phenotypic correlation coefficient analysis showed significant positive correlation between grain yield per plant and yield-related attributes. Positive correlation (r = 0.223 and 0.763) was observed between root length and dry root biomass in three populations and PUSA1121 x MAS26 F3 and F4 populations showed positive correlation (r = 0.429) between grain yield and root length/dry biomass. In these populations, plants had the allelic profile at BAD2A locus similar to the Basmati (585 and 257 bp) or non-Basmati (585 and 355 bp) parents or were heterozygous (585, 355 and 257 bp). A total of 221 of 604 SSR markers well distributed on 12 rice chromosomes, showed polymorphism in five parental rice genotypes; 51-71 polymorphic markers were used to assess the diversity in these populations. The NTSYS-pc UPGMA tree cluster analysis and twodimensional PCA scaling showed that Basmati and aerobic rice genotypes were quite divergent and segregating populations were interspersed between the two parents. Composite Interval Mapping analysis (WinQTL Cartographer 2.5) using SSR databases of various populations led to the mapping of 14 large-effect QTL for grain yield on chromosome 1, 2, 3, 5, 6, 7, 8, 9 and 12 (LOD: 2.5 - 8.4 and R2 0.8 – 72.2%). In addition 22, 13, 10, 7 and 2 QTL for grain L/B ratio, panicle length, effective number of tillers per plant, plant height and 1000 grain weight, were mapped, respectively. Notably, five QTL (qLB9.1, qLB11.1, qTGW10.1, qPL4.1 and qPL2.2) accounted for >70% variation for the respective traits. As many as 68 and 80 plants having higher or comparable grain yield, length/breadth ratio, root length/thickness/biomass than the parental genotypes with intact Basmati specific BAD2A allele in homozygous and heterozygous conditions, were selected from the four crosses in 2014 and 2015 Kharif seasons, respectively. These genotypes will serve as the novel material for the selection of stable aerobic Basmati rice varieties. It will also be interesting to investigate and confirm the role of QTL identified in this study towards improving the grain yield and quality under aerobic conditions.ThesisItem Open Access Marker-assisted selection for mineral (Iron and zinc) rich lines in Indica rice (Oryza sativa L.)(CCSHAU, 2017) Pippal, Amit; Jain, R.K.Molecular markers provide valuable tools to identify and quantify genetic diversity within and between species, population and available germplasm to accelerate the efficacy of breeding programs to improve mineral (iron and zinc) content in rice grains.F5, BC1F4 , F6, BC1F5 and RIL F6 populations derived from the cross between high-yielding (PAU201) and iron-rich (Palman 579) indica rice varietiesdisplayed large variation for various physio-morphological traits (plant height, effective number of tillers/plant, panicle length, grain yield per plant and 1000-grain weight) and mineral (iron and zinc) contents. Iron and zinc content varied between 4.6-312 μg/g and 2.2-117.5 μg/g (F5); 3.9-86.4 μg/g and 1.51-140.6 μg/g (BC1F4); 53-117.2 μg/g and 9.7-112.2 μg/g (F6);37.8-259 μg/g and 20-56 μg/g (BC1F5) and 23.6-383.4 μg/g and 13.6- 101 μg/g (RIL F6) respectively. Transgressive segregation for grain iron content was noticed in one RIL F6 plant (plant no.214A3-3) which had exceptionally higher iron (383.4 μg/g; Palman 579 326.5 μg/g. Phenotypic correlation analysis showed positive correlation between grain iron and zinc content in F5, BC1F4, F6 BC1F5 and RIL F6 populations. In F5 and BC1F4 populations, grain iron and zinc content showed positive correlation with grain yield.DNA fingerprint databases of PAU201 × Palman 579 F6 andRIL populations (30 plants each) were prepared using 62 and 61 polymorphic SSR markers. The NTSYS-pc UPGMA tree cluster analysis and two dimensional PCA scaling exhibited that two parental genotypes were quiet distinct and diverse whereas 30 F6 and RIL plants interspersed between the parental rice genotypes. QTL mapping using two SSR databases by Composite Interval Mapping (CIM) programme of WinQTL Cartographer 2.5, led to the identification of twelve QTL (six each in F6 and RIL population) for iron (8 QTL) and zinc (4 QTL) content in rice grain; ten of these QTL accounted for >30% variation. Out of 8 QTL for iron content, 6(qFE2.1, qFE2.2, qFE7.1, qFE9.1in F6 population; qFE2.1, qFE2.2from RIL population) were from “Palman 579” and rest of the two QTL (qFE10.1and qFE12.1) were from “PAU201”. Out of four QTL mapped for zinc content, one QTL, qZN7.1(F6)was from “PAU201” and three QTL, qZN2.1(F6), qZN2.1(RIL)and qZN10.1(RIL), werefrom Palman 579. In addition, eight QTL (qYP10.1, qYP10.2, qYP12.1, qYP12.2, qYP2.1, qYP2.2, qYP10.1 and qYP12.1) were mapped for grain yield; 7 of these QTL were from “Palman 579” and two QTL (qYP10.2 and qYP12.1) contributed >70% of phenotypic variation. As many as 30 F6 plants and 30 BC1F5 plants were selected on the basis of grain yield and mineral content for further progeny analysis and RIL population was advanced to F6 generation with 93.7% homozygosity.ThesisItem Open Access Molecular marker analysis for QTL identification in Indian mustard (Brassica juncea L. Czern & Coss.) for salinity tolerance(CCSHAU, 2017) Patel, Rekha; Yadav, Neelam R.Salinity tolerance is quantitative and complex phenomenon controlled by numerous interacting quantitative trait loci (QTLs). To find out genomic regions associated with terminal salinity tolerance a study entitled Molecular marker analysis for QTL identification in Indian mustard (Brassica juncea L. Czern & Coss.) for salinity tolerance was conducted with the objective to identify QTLs for salinity tolerance in RH 30 × CS 52 derived F2 and F2:3 populations of Brassica juncea (RH 30-Salinity sensitive and CS 52-Salinity tolerant) and to evaluate both generations for yield attributes such as plant height (cm), physiological maturity, main shoot length (cm), number of primary branches/plant, number of secondary branches/plant, number of siliquae/plant, number of seeds/siliqua, 1000-seed weight (g), seed yield/plant (g) and biological yield/plant (g); physiological traits such as relative water content (%), osmotic potential (-MPa), proline content (µmol/g) and membrane injury (%) under field and salinity treatments i.e. control, 6 dS/m and 8 dS/m. Out of 358 SSR markers 42 were found polymorphic and 153 were monomorphic. Linkage map was prepared using 41 SSR markers covering 8 linkage groups and 1 marker did not map to any of the linkage groups. A total of 91 alleles were identified in the CS 52 and RH 30 genotypes. The total length of the genetic linkage map was 2298.5 cM and PIC ranged from 0.427-0.730. Composite interval mapping revealed a total of 23 QTLs were identified for yield attributes and physiological traits in F2 population. Five QTLs identified for salinity tolerance on two linkage groups 4 and 8 under field and salinity treatments. QTLs identified in the present study firstly needs to be validated in other populations and then fine mapping of these salinity related QTLs have to be carried with them in marker assisted selection and breeding for salinity tolerant genotype in Brassica juncea.