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ThesisItem Open Access Genome wide association mapping for stem water soluble carbohydrates in bread wheat (Triticum aestivum L.) under terminal water stress(CCSHAU, Hisar, 2020-11-26) Gaur, Arpit; Jindal, YogeshGenomic assisted breeding is one way to improve drought stress tolerance in wheat. To uncover the genomic regions for grain yield, its attributing traits, water soluble carbohydrates (WSCs) and its components mainly fructans, a comprehensive multidisciplinary study was conducted with a diverse set of 302 bread wheat genotypes. All the genotypes were evaluated under complete irrigation, rainfed and complete water stressed condition for two consecutive crop seasons at three locations (Hisar, Karnal and Baramati). The study indicated significant effect of different water regimes and locations on the expression of traits and as much as 80% of yield reduction was reported under DT. Seven promising drought tolerant verities were identified on the basis of drought susceptibility index. In this study, correlation between yield and studied traits largely varied with environmental and water conditions. However, shorter plant height, longer grain filling duration, days to heading, thousand kernel weight (35-40 g) is the most favorable trait combination for obtaining high grain yield under studied conditions. Positive correlation of WSCs with grain yield was reported only for drought tolerant genotypes. High values of all the genetic components were reported for all the traits within environments, however in pooled over management these values ranged between low to moderate for most of the studied traits due to genotype x environment. Traits like plant height, peduncle length, spikelet per spike and thousand kernel weight were least influenced with genotype x environment interaction thus found suitable for selection of grain yield. Substantial genetic diversity was reported in the studied material which was attributed by each of the studied trait equally. With 35K Axiom array and applying compressed mixed linear model, 431 stable SNPs were identified for 20 studied traits in 1288986.92 cM genomic region of bread wheat. These SNPs explained ca. 30-53% of phenotypic variations. Of 431 stable SNPs 77 were showing pleiotropy for various traits. Five SNPs (AX-94583229, AX-94879209, AX-94759517, AX-94887053 and AX-94964616) for fructans and total non-structural carbohydrates, three (AX-94508292, AX-95257620 and AX-94456473) for days to heading and one (AX-94412521) for plant height were most significant. In functional analysis of significant SNPs, 249 genes were identified which were mainly involved in carbohydrate metabolism and repair mechanism. Only few were reported with direct involvement in stress responses. In an in silco study 104 genes (TaFruc) transcribing into stable protein associated with fructans metabolism were identified. All of these were involved in either hydrolase or transferase activities of glycosyl groups of soluble sugars. These genes were found expressive under different stress conditions, mainly water stress. To facilitate the future breeding programs a total of 189 SSR molecular markers and 60 micro RNAs have been predicted successfully for TaFruc genes.ThesisItem Open Access Association Mapping of Seed Yield Related Traits in Mungbean (Vigna radiata (L.) Wilczek)(CCSHAU, Hisar, 2021-07) Versha; Yadav, RajeshThe experimental material consisting of 80 released and advanced mungbean genotypes were evaluated for 30 morphological and yield traits during kharif and summer seasons of 2018-19 and 2019-20 to ascertain genetic variability, correlations, genetic divergence and to detect loci controlling yield and yield attributes by association analysis. Significant amount of variability was observed for all the traits except stem pubescence, leaflet lobes, leaf shape, premature pod color, pod pubescence and seed color and the genotypes were characterized and categorized as per DUS guidelines. The narrow differences between PCV and GCV revealed that the existing variation was mainly due to the genetic factors indicating scope for selection. Most of the characters exhibited high heritability and low genetic advance indicating control of non additive gene action; therefore, intermating in early generations of the breeding programme will be helpful. Seed yield per plant was found positively associated with pod length, number of branches per plant, number of pods per plant and number of seeds per pod and negatively with yellow mosaic disease.The hierarchical cluster analysis grouped the 80 mungbean genotypes in 10 clusters. The maximum inter cluster distance was observed between cluster II and IV followed by IV and IX and I and VI and Cluster II was found best for most of the traits. Stability analysis revealed differential response of genotypes to the environments. The estimates of environmental additive effects revealed E-4 (Summer, 2020) as the best for all the seed yield related traits and the genotypes MH 421, MH 1767, MH 318, MH 560, MH 1706 and MH 1452 were found stable for seed yield per plant. Out of 164 SSRs, 139 markers amplified with allele size between 100-310 bp and among these 89 were found polymorphic which generated 317 polymorphic alleles with an average of 3.562 alleles per SSR while the number of alleles at each locus varied from two to seven. Both the Weighted Neighbour Joining and Population Genetic Structure analyses categorized genotypes in a similar manner suggesting that all the advanced genotypes may have common ancestral history at some level. On the basis of pure and admixture model, sub population 1, 2 and 3 were found pure and SP-4 contained admixture. Polymorphic SSRs, used to estimate the average degree of Linkage Disequilibium (LD) present at the genome level in 80 mungbean genotypes, disclosed significant LD blocks. Over the four seasons, 38 markertrait associations were observed significant for seed yield related traits by GLM method and ten marker-trait associations were found significant for five seed yield related traits viz., days to flowering, days to maturity, plant height, yellow mosaic disease and number of pods per plant by MLM method which indicated low phenotypic and genotypic variations in association mapping which may be due to low diversity among the genotypes and exploitation of less number of markers and genotypes in investigation.ThesisItem Open Access Morphological, biochemical and molecular diversity analysis of upland cotton (Gossypium hirsutum L.)(CCSHAU, Hisar, 2021-02) Rathi, Meenakshi; Dahiya, G.SThe present investigation “Morphological, biochemical and molecular diversity analysis of upland cotton (Gossypium hirsutum L.)” was carried out at Chaudhary Charan Singh Haryana Agricultural University, Hisar in kharif 2018-2019 and 2019-20 with the objective of estimating diversity of 150 genotypes of cotton. Variation among genotypes was present for hypocotyl pigmentation, pubescence on stem, stem pigmentation, leaf color, pigmentation of petiole, flower petal color, boll shape, prominence of tip, plant height, growth habit, seed index, ginning percentage, fiber length, fiber strength and fiber fineness. Analysis of variance revealed presence of variability for all the traits among the genotypes other than number of monopods and number of locules. GCV and PCV also shows that number of locules, days to first flower, ginning percentage and number of seeds per locules had low variability whereas as highest PCV and GCV values were possessed by number of bolls per plant and seed cotton yield. Additive gene action was dominant in plant height, number of sympods per plant, boll weight, seed index, lint index, number of bolls per plant and seed cotton yield per plant as high heritability coupled with high genetic advance was reported for these traits. Correlation studies showed that seed cotton yield per plant was significantly and positively correlated with all the traits highest among them was of number of bolls per plant but days to first flower, number of locules per boll and fiber quality traits exhibited negative association with yield. Path analysis revealed that maximum contribution is from number of bolls per plant and boll weight. Genetic diversity analysis was done using 118 SSR markers out of which 26 were found polymorphic with average number of alleles as 2.1 per locus. UPGMA clustering analysis formed 13 groups of genotypes. The biochemical profiling of the genotypes showed variability among the genotypes. Weather affected biochemical parameters but gossypol remained unaffected. Seed cotton yield did not show any relation with biochemical parameters but was in negative association with CLCuDThesisItem Open Access Quantitative and molecular characterization for heat tolerance in wheat (Triticum aestivum L. em. Thell).(CCSHAU, Hisar, 2020-02) Anil Kumar; Dhanda, S.SThe present study was carried out to evaluate 96 advanced recombinant inbred lines derived from a cross WH 730 (heat tolerant) and WH 147 (higher yielder, heat susceptible) to find out physio-morphological traits and quantitative trait loci (QTLs) associated with heat tolerance in bread wheat. The experiment was conducted in augmented design and randomized complete block design (RBD) during Rabi season of 2014- 15, 2015-16 and 2016-17 in the Department of Genetics and Plant Breeding, CCS Haryana Agricultural University, Hisar. Analysis of variance revealed significant differences for all traits studied among the RILs. Medium to high values of GCV, PCV, heritability and genetic advance as percent of mean for grain yield per plant, number of grains per spike,100-grain weight, biological yield, canopy temperature depression and cell membrane thermostability under either one or both environments over the years indicating a high scope of selection for these traits for heat tolerance. Grain yield was significantly and positively correlated with number of tillers per plant, harvest index, biological yield per plant, 100-grain weight, canopy temperature depression, cell membrane thermostability and osmotic potential, whereas negative and significantly correlated with days to heading and days to maturity under timely and late sown conditions over the years. Biological yield and harvest index had high direct and indirect effects on grain yield per plant over environment and years, indicating the true relationships with grain yield, in this set of material. Nine promising RILs namely RIL No.46, 22, 40, 43, 57, 19, 48, 81, 71 and 88, out of 96 were identified for the traits i.e., 100-grain weight, harvest index, grain yield, canopy temperature depression and cell membrane thermostability related to heat tolerance under heat stress condition. Two RILs 22 and 40 showing better performance over the environment and year. Twenty-six, out of 81 SSR markers used were found polymorphic in parental genotypes and in RILs. Composite interval mapping (CIM) identified quantitative trait loci (QTL) with significant phenotypic variation for number of spikelets per spike, spike length and biological yield per plant along with heat susceptibility index (HSI). QTL identified for biological yield per plant reported maximum phenotypic variation (50.0%) followed by spike length (46.37%) and number of spikelets per spike (12.59%).ThesisItem Open Access Population Structure Analysis and Genetic Diversity Studies for Yield Contributing Traits in Upland Cotton (Gossypium hirsutum L.)(CCSHAU, Hisar, 2020-11) Pawan Kumar; SomveerThe present study was undertaken with the objectives i) investigate the phenotypic and genetic diversity of promising genotypes of G. hirsutum and ii) to estimate the population structure, linkage disequilibrium (LD) using SSR markers. The experimental material comprised of 190 genotypes of upland cotton out of which 96 genotypes were used for population structure and further analysis. The test enteries were evaluated for various quantitative traits under three environments (early, normal and late sown conditions) during kharif-2018 and kharif-2019. Among the qualitative characters studied leaf color, leaf hairiness, leaf appearance, plant stem hairiness, plant height, flower petal color, flower stigma position, flower pollen color, boll shape, boll prominence of tip, and boll opening proved to be useful and stable diagnostic characters which proved useful in the classification of test genotypes. Analysis of variance revealed significant differences among the genotypes for all traits under investigation. Highly significant positive association of seed cotton yield per plant with number of bolls per plant, boll weight, lint yield per plant, and seed index was recorded. All 96 genotypes were grouped into two clusters by population structure and analysis of molecular variance showed that most of the variation was among the genotypes as compare to among the sub-population. A total of 46, 48 and 46 significant marker-trait associations were identified in early, normal and late sown plantings, respectively. Out of these 21, 19 and 21 significant marker-trait associations were novel in early, normal and late sown environment, respectively.ThesisItem Open Access Molecular and morphological characterization of rice (Oryza sativa L) genotypes under aerobic and irrigated conditions(CCSHAU, Hisar, 2020-10) Susmita Dey; Khushi RamThe present study was conducted for evaluation and identification of rice genotypes under aerobic and irrigated conditions. The experiment was carried out at Rice Research Station, Kaul, CCS Haryana Agricultural University, during kharif 2018 and kharif 2019. The thirty (30) rice genotypes were grown in randomized block design (RBD) under both the conditions. Data recorded for 22 traits viz., days to 50% flowering, plant height (cm), leaf length (cm), leaf width (cm), number of tillers / plant, panicle length (cm), number of grains/panicle, grain yield (g) / plant, days to maturity, biological yield (g)/ plant, 1000 seed weight (g), harvest index (%), root length (cm), root volume (ml), root fresh weight (g)/ plant, root dry weight (g)/ plant, kernel length (mm), kernel breadth (mm), kernel length - breadth ratio, brown rice (%), milled rice (%) and head rice recovery (%).Genotypes IR14L521, NVSR-2017 and RDR 1158 were identified as superior genotypes, suitable for both aerobic and irrigated conditions on the basis of grain yield performance over the seasons and conditions. Under both the conditions, analysis of Variance (ANOVA) revealed significant difference for all the traits except leaf width, kernel length, kernel breadth and kernel length-breadth ratio. High GCV was observed for characters like root dry weight/plant and grain yield / plant in both the conditions. High heritability along with high genetic advance were recorded for the characters viz., leaf length, number of grains /panicle, 1000 seed weight, root volume, root fresh weight/plant, root dry weight/plant, kernel length-breadth ratio, grain yield / plant, biological yield /plant and harvest index under both the conditions. Correlation studies under both the conditions revealed that grain yield / plant exhibited positive significant correlation with plant height, leaf length, 1000 grain weight, root length, root volume, root fresh weight/ plant, root dry weight/ plant, kernel length, biological yield/ plant and harvest index. The path coefficient analysis under both conditions indicated that biological yield/plant and harvest index had major positive direct effect on grain yield/plant. The 30 genotypes were also subjected to genetic diversity study through morphological traits under both the conditions and molecular markers. Out of 60 SSR markers, 39 markers showed polymorphism used for diversity analysis. From diversity analysis study, we recommended crossing between RP 5594- 410- 53-4-2 and HKR-48 for hybridization programme.ThesisItem Open Access Assessment of genotype x environment interactions for seed cotton yield & quality traits in desi cotton (Gossypium arboreum L.)(CCSHAU, Hisar, 2020-07) Jangid, Kuldeep; Sangwan, OmenderThe present study comprised of thirty Gossypium arboreum genotypes grown in two locations during the kharif seasons in the year 2016 and 2017 in a randomized block design (RBD) at CCS Haryana Agricultural University, Hisar and cotton research station, Sirsa, CCS Haryana Agricultural University. The observations on five randomly selected plants from each replication in each environment were recorded for fifteen quantitative and qualitative characters viz., days to first flower, plant height, number of monopods per plant, number of bolls per plant, boll weight (g), ginning out turn (%),seed index (g), lint index (g), lint yield (g), protein (%), oil (%), gossypol (%), sugar (%), seed cotton yield per plant (g). The data were subjected to examine variability parameters, such as coefficient of variation, association among seed cotton yield and component traits and to study stability of various traits, biochemical parameters and genetic diversity also analysed using SSR markers. Seed cotton yield per plant showed highly significant positive correlation with lint yield per plant, number of bolls per plant, boll weight and gossypol content. The path-coefficient analysis suggested the importance of number of bolls per plant, seed index, boll weight, lint yield and number of seeds per boll as they exhibited positive direct effects on seed cotton yield. The estimation of environmental additive effects (Ij) revealed that environment 2 was most suitable for number of monopods per plant, number of bolls per plant and number of seeds per boll. Environment 3 was found favourable for days to first flower, plant height, number of bolls per plant and oil content. Similarly, environment 4 was best for plant height number of seeds per boll, lint yield per plant and seed cotton yield plant. Significant diversity was found among all 30 genotypes even at molecular level. And all 3o genotypes were groups intro 4 clustered by polymorphism information of 22 SSR markers.ThesisItem Open Access Studies on induced mutations in mungbean [Vigna radiata (L.) Wilczek] and identification of MYMIV resistant mutants using molecular makers(CCSHAU, Hisar, 2020-11) Anuragi, Hirdayesh; Yadav, RajeshMungbean (Vigna radiata L. Wilczek) is an economically important pulse crop of South-East Asia It is mostly liked in Indian diets due to its good palatability, easy digestibility and high nutritional quality. Unavailability of sufficient genetic variability is a major challenge for mungbean breeders. In this study, dry seeds of two mungbean varieties viz., MH 2-15 and MH 318 were mutagenized by 300, 400, 500 Gy of γ-rays and 0.2, 0.4 % EMS as well as their combination treatments and variability for various parameters were studied in M1, M2 and M3 populations during 2017-18 at CCS HAU, Hisar. In M1 generation, seed germination parameters like seed germination per cent, shoot and root length, seedling length, seedling fresh and dry weight, SVI-I and II, mean germination rate, synchronization index, germination index, coefficient of velocity of germination and speed of germination decreased whereas seed/seedling mortality, mean germination time and days to 50 % germination increased with the increase in concentration of both the mutagens under controlled lab and natural field conditions. LD50 values for γ-rays ranged from 488.47 to 694.28 Gy in MH 2-15 and 519.77 to 715.15 Gy in MH 318 and 0.35 to 0.69 % in MH 2-15 and 0.37 to 0.74 % in MH 318 for EMS under controlled lab and natural field conditions. The M2 population of 24,464 mutants were screened and a broad spectrum of chlorophyll mutants (1.08 %) like xantha>viridis>chlorina>albina and morphological mutants (1.98 %) with compact, bushy, erect, spreading, tall, dwarf nature, tetra-foliate, penta-foliate, elliptic, lanceolate, lobbed, wrinkled, round leaves, early maturing, late maturing, sterile plants and large, small, lobbed pods were observed. Mutagen exhibited non-linear reduction in effectiveness and efficiency with increase in concentration. Mutagenesis generated significant variability in seed yield and attributing traits. Based on correlation and path coefficient studies, number of pods plant-1, number of seeds pod-1 and 100-seed weight were observed to be important parameters for increasing seed yield plant-1. Overall, MH 2-15 was found to be more sensitive for mutagenic changes as compared to MH 318, whereas among mutagens, the combination treatments (γ-rays + EMS) have exhibited more shift in magnitude of all the parameters followed by EMS and γ-rays. In M3 generation, 64 MYMIV resistant mutants were identified with <1 % PDI through phenotypic screening which were further confirmed by amplification of YMD linked SSR markers viz., YR4, CYR1 and CEDG180. Twenty-two mutants were identified with true resistance based on marker assisted screening.ThesisItem Open Access Identification of salinity tolerant chickpea (Cicer arietinum L.) genotypes using morpho-physiological and molecular markers(CCSHAU, Hisar, 2020-09-30) Janghel, Dharmendra Kumar; Chhabra, A.K.In the present study entitled “Identification of salinity tolerant chickpea (Cicer arietinum L.) genotypes using morpho-physiological and molecular markers,” conducted three experiments in 25 Desi chickpea genotypes along with 7 ICRISAT accessions under different labs and field conditions i.e. (1) Laboratory/ net house experiments: petri -dish experiment to study seed and seed germination indices in 0, 40, 60, 80, 100, 120, 140 & 160 mM salinity levels of both NaCl and chloride dominated salts (2017 & 2018), between paper experiment to evaluate seed and seeding characteristics in 0, 40, 60 & 80 mM salinity levels of chloride dominated salt (2018) & net/ pot house experiment in 0, 40, 60, 80 & 100 mM salinity levels of chloride dominated salt (2018-19); (2) molecular characterization using 100 SSR markers (2018-19) and, (3) field experiments under natural existing salinity stress and normal field condition (2018-19). ANOVA showed that the 25 Desi chickpea genotypes differed significantly for all morpho-physiological traits studied for salinity tolerance, indicated the ample amount of genetic variability for salinity s tress tolerance had present in these material. The chickpea genotypes identified and screened across 0 to 120 mM salinity levels of chloride dominated salt for salinity tolerance were HC 3, HC 5, C 235, CSG 8962, ICCV 92944, ICCV 4958, H 07-120, H 09-96, H 08-18, RSG 931, ICC 12964, ICC 6306, ICC 6263, ICC 2242 and ICC 1915 at seed and seedling stage. Based on morpho-physiological traits across 0 to 80 mM salinity levels of chloride dominated salt, HC 3, H 07-120, H 08-18 and CSG 8962 had shown better mean performances, among them HC 3 genotype performed superior than CSG 8962 which need to be confirmed by further studies. The highest (0.516) discriminating power (D) displayed for SSR primers (ICCeM 0086, CaM 0015 and CaM 1469) in all 32 chickpea genotypes. The SSR molecular markers divided 32 chickpea genotypes into more clusters than few clusters by morpho - physiological clustering which indicated more informativeness of SSR markers than the morpho-physiological markers. So the molecular markers capable of detect narrow genetic base more efficiently. Based on both morpho-physiological and molecular genetic diversity analysis, the genotypes viz., HC 3, CSG 8962, H 07-120, H 08-18 and C 235 showed sufficient amount of genetic diversity for salinity tolerance wh ich may be used in chickpea breeding programmes for further improvement in salinity toleran.
