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Chaudhary Charan Singh Haryana Agricultural University, Hisar
Chaudhary Charan Singh Haryana Agricultural University popularly known as HAU, is one of Asia's biggest agricultural universities, located at Hisar in the Indian state of Haryana. It is named after India's seventh Prime Minister, Chaudhary Charan Singh. It is a leader in agricultural research in India and contributed significantly to Green Revolution and White Revolution in India in the 1960s and 70s. It has a very large campus and has several research centres throughout the state. It won the Indian Council of Agricultural Research's Award for the Best Institute in 1997. HAU was initially a campus of Punjab Agricultural University, Ludhiana. After the formation of Haryana in 1966, it became an autonomous institution on February 2, 1970 through a Presidential Ordinance, later ratified as Haryana and Punjab Agricultural Universities Act, 1970, passed by the Lok Sabha on March 29, 1970. A. L. Fletcher, the first Vice-Chancellor of the university, was instrumental in its initial growth.
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ThesisItem Open Access Linkage mapping of quantitative trait loci for root morphology and selection of desirable genotypes from aerobic x lowland indica crosses in rice (Oryza sativa L.)(CCSHAU, 2014) Kharb, Anju; Jain, Rajinder KumarIdentification of stable QTL for traits promoting aerobic adaptation using molecular markers can greatly enhance the efficacy of breeding programs to develop water-efficient, high yielding, aerobic rice varieties. Filial (F2 and F3) and backcross ( BC2F2 and BC1F2) populations derived from the crosses between high-yielding low-land (HKR47) and aerobic (MAS26 and MASARB25) indica rice varieties, displayed large variation for various physio-morphological traits including grain yield per plant and root traits (in case of net house evaluation). Phenotypic correlation analysis showed positive correlation (r = 0.279 and 0.351) between grain yield per plant and root length in both F2 and F3 population and between grain yield per plant and fresh & dry root weight (r = 0.232 and 0.269) in F2 population. Under aerobic field conditions, Grain yield per plant showed significant positive correlation with plant height, effective number of tillers/plant and panicle length in all the four populations. A total of 803 SSR markers, distributed on 12 rice chromosomes, were analyzed for polymorphism in two parental rice genotypes (HKR 47 and MAS 26); of these 125 (about 16%) displayed polymorphism. SSR fingerprint databases of three HKR47 x MAS26 F2 and F3 populations (94 F2 in net house, 42 F3 in net house and 52 F3 plants in field) were prepared using 125, 88 and 88 polymorphic SSR markers, respectively. NTSYS-pc UPGMA tree cluster analysis and two-dimensional PCA scaling showed scattering of the F2 and F3 populations between the two distinct parental genotypes; the populations were inclined towards MAS26. Composite interval mapping (CIM) analysis by Win QTL cartographer 2.5 revealed a total of six QTLs (qPN8.1, qPN8.2, qPN8.3, qTGW8.1, qYPP8.1 and qRL8.1) in HKR47 x MAS26 F2 population, grown in the net house on chromosome 8 (within a region of 24.9 cM) which individually explained 13.7-27.3% of the phenotypic variation. In case of HKR47 x MAS26 F3 population, based on field data and net house data, two (qTGW6.1 and qLB8.1) and six QTL (qPH3.1, qLB8.1, qTGW6.1, qTGW11.1, qRT8.1 and qRT8.2) were mapped, respectively. As many as 20 F3, 6 BC2F2 and 5 BC1F2 promising plants having higher grain yield and/or better root length/biomass were selected. Data on monitoring of putative QTL identified showed that out of twenty selected plants, four plants had 13 QTL, three plants had 12 QTL and two plants had 11 QTL in homozygous or heterozygous condition.ThesisItem Open Access Marker assisted selection in advanced segregating lines for drought tolerance in chickpea (Cicer arietinum L.)(CCSHAU, 2015) Summy; Boora, K.S.The present investigation on studies on Marker assisted selection in advanced segregating lines for drought tolerance in chickpea (Cicer arietinum L.) was conducted with the objective to validate QTL(s) for drought tolerance in HC-1(sensitive) and ICC4958 (tolerant) parental genotypes, to screen F4 and F5 progeny for presence of major QTL(s) for drought tolerance in chickpea and to evaluate both generations for yield and its component traits under drought. Fifty markers linked to QTLs scattered over all the eight chromosomes for drought tolerance in chickpea were used for validation of QTLs in the parents HC-1 and ICC4958. . Out of 50 QTLs, 20 QTLs distributed on six chromosomes were present in drought tolerant parent ICC4958. These markers were used for screening of QTLs in F4 and F5 progeny from cross HC1×ICC4958 of chickpea. Highest yielding F4 progeny lines G19, G20, G45, G42 and G6 had five common QTLs i.e. TAA170, TR24, TA118, NCPGR138 and TA180. Highest No. of pods/plant F5 progeny lines G14, G16, G17, G32, G101 and G115 had five common QTLs i.e. ICCM0249, TR24, NCPGR200, NCPGR138, and STMS11. Eight QTLs having maximum effect on drought tolerant traits are scattered on different chromosomes. One QTL (TR24) on chromosome No. 3, three QTLs (TAA170, ICCM0249, STMS11) on chromosome No. 4, one QTL (NCPGR200) on chromosome No. 6, one QTL (TA180) on chromosome No. 7 and two QTLs (TA118, NCPGR138) on chromosome No. 8. Majority of promising progeny lines had more than 10 QTLs linked to drought tolerance. Promising progeny lines also showed morpho-physiological traits similar/better to drought tolerant parent. Phenotypic correlation coefficient analysis of both generations showed significant correlation between morpho-physiological and yield related traits. Marker assisted selection was successfully utilized to introgress major drought tolerant QTLs in chickpea genotypes derived from cross HC-1×ICC4958. These genotypes (G6, G19, G20, G40, G42, G45 and G55) shall to be further evaluated in next generation for their yield attributes under normal and drought stress conditions.ThesisItem Open Access Molecular Dissection of AGPase to Improve Wheat Productivity(CCSHAU, 2014) Batra, Ritu; Sikka, V.K.The present investigation was conducted towards understanding role of ADP-glucose pyrophosphorylase (AGPase) in developing grains’ starch biosynthesis among diverse wheat lines including disomic chromosome substitution (DCS) lines of C-591 in the background of Chinese Spring, dwarf wheat varieties. isogenic lines and for WH-711 X WH 542. The experiment was laid out in completely randomized block design with three replications. The traits like grain growth rate (GGR) and AGPase enzyme activity were also measured for developing grains at 10th, 20th and 30th days after anthesis (DAA). Metabolites like total sugars, reducing sugars, non-reducing sugars, total starch, amylose and amylopectin were also quantified using wheat flour of the test lines. The recorded data was subjected to appropriate statistical analysis to arrive at inferences on the experiments in the study. Critical analysis of the data led us to conclude that determinants for AGPase expression are present across the genome over multiple chromosomes which are active at particular grain filling stages like during first half, chromosome 1B and 2D played major role whereas second half was mainly controlled by chromosome 6A and 5B. Hybridization efforts have improved grain attributes and yield components of WH-542 with ~28.6 % and 42 % enhancement in grain wt. per spike and yield per plant respectively. Maximum amylose (%) was observed in DCS line 2D which can be used to produce functional wheat flour. Grain yield was found to be significantly associated with grains/spike and plant biomass in case of DCS lines. Partitioning the variation by path analysis indicates grains per spike, though had less direct effect but its impact on grain yield through indirect effect via tiller no./plant and AGPase activity at 20th DAA was high suggesting their importance in yield improvement efforts. AGPase activity analysis suggests major contribution of chromosome 2D towards yield in a specific temperature window of 22±1.5oC during initial stages of grain filling which can be used as a screening tool for terminal heat tolerant genotypes.ThesisItem Open Access Mapping QTL(s) for drought tolerance in Indian mustard (Brassica juncea L. Czern & Coss.)(CCSHAU, 2015) Monika; Yadav, R. C.The present investigation on studies on Mapping QTL(s) for drought tolerance in Indian mustard (Brassica juncea L. Czern & Coss.) was conducted with the objective to identify QTL(s) for drought tolerance in RB50 x Kranti derived F 2 and F2:3 populations of Brassica juncea (RB50-Drought tolerant, Kranti-Drought sensitive)and to evaluate both generations for various phenological traits, growth related traits, physiological traits along with yield and its component traits under irrigated and drought conditions. RB50 × Kranti derived F 2 and F2:3 population showed significant variation for various observed agronomic traits. 200 SSR primers from various Brassica species were used to screen parental genotypes (RB50, Kranti) and 51 markers were found polymorphic. These polymorphic SSR primers were used to screen F2 population and were used to construct linkage map of Brassica juncea using MapmakerExp3.0 covering 10 linkage groups, 2 SSR markers did not show linkage with any of the marker groups and hence did not map to any of the linkage group. Composite interval mapping (CIM) analysis revealed a total of 30 QTLs in F 2 generation under irrigated conditions for various drought related, yield and other traits in Brassica juncea out of which four QTLs were identified for drought related physiological trait (Electrolyte leakage). A total of 19 QTLs were identified in F 2:3 under irrigated conditions for various drought related, yield and other traits in Brassica juncea including 4 QTLs for physiological trait (Relative water content). QTL analysis revealed a total of 7 QTLs accounting for different phenotypic variance in F 2:3 generation under drought conditions for various drought related, yield and other traits in Brassica juncea with one QTL identified for drought related physiological trait (Electrolyte leakage). There was clustering of QTLs on many LGs in linkage map. Most prominent clustering signifying multifunctional QTL region was observed in the LG 5 and 10. This multifunctional QTL region on the LG 5 contains at least one major QTL for various traits. Linkage groups 5 and 10 have been suggested to have major QTLs affecting drought tolerance. QTLs identified in the present study firstly needs to be validated in other populations and then fine mapping of these drought related QTLs have to be carried out them in marker assisted selection and breeding for drought tolerant genotypes in Brassica juncea.ThesisItem Open Access Selection of high-yielding iron-rich PAU201/Palman579 segregating rice (Oryza sativa L.) lines using conventional and molecular marker techniques(CCSHAU, 2014) Naveen Kumar; Chowdhury, V.K.Molecular markers provide novel tools for linkage mapping of QTLs of target traits and can greatly enhance the efficacy of breeding programs to improve mineral (iron and zinc) density in rice. F3, F4, BC1F2 and BC1F3 populations derived from the cross between high-yielding (PAU201) and iron-rich (Palman 579) indica rice varieties displayed large variation for various physio-morphological traits including grain yield per plant and mineral (iron and zinc) contents. Iron and zinc content varied from 0.9- 149.9 and 0-143.1 μg/g respectively in all the four populations (F3, F4, BC1F2 and BC1F3). Transgressive segregation for grain iron content was noticed in F3 population with one of the plants having exceptionally higher iron (746.8 μg/g) content. Phenotypic correlation analysis showed positive correlation (r=0.281) between grain iron content and zinc content in BC1F2 population but not in F3, F4 and BC1F3 populations. Grain yield per plant showed significant positive correlation with plant height and effective number of tillers/plant in all the four populations. A DNA fingerprint database of 33 PAU201 × Palman 579 F4 plants was prepared using 61 polymorphic SSR markers distributed on the entire genome of rice. NTSYS-pc UPGMA tree cluster analysis and two-dimensional PCA scaling showed scattering of the F4 population between the two distinct parental genotypes; the population was inclined towards Palman 579. The distribution of PAU201 and Palman 579 specific alleles among the PAU201 × Palman 579 F4 plants were determined and SSR data was used to identify QTLs for grain mineral content and various agronomical traits. A total of 128 alleles were identified in 33 PAU201 x Palman 579 F4 plants and three new recombinant alleles (different that those in parent rice varieties) were identified. Composite interval mapping (CIM) analysis by Win QTL cartographer 2.5 revealed a total of six QTLs for mineral content (five for Fe and one for Zn) in rice grains on chromosome 5, 6, 7 and 9; and sixteen QTLs for various agronomical traits (plant height, tillers per plant, yield per plant, Grains per panicle and 1000-grain weight). As many as 33 F4 and 25 BC1F3 promising plants having higher grain yield as well as mineral content have been selected for further progeny analysis.ThesisItem Open Access Marker assisted selection for drought tolerance in pearl millet (Pennisetum glaucum (L.) R. Br.)(CCSHAU, 2014) Asha Rani; Yadav, Ram C.Pearl millet (Pennisetum glaucum (L.) R. Br.) is the staple cereal of millions of poor rural families in the hottest and driest areas of the tropics and subtropics. Terminal drought stress is a regular occurrence and serious environmental constraint in these regions, making stress tolerance an essential attribute of new pearl millet cultivars. Marker assisted selection is an important molecular tool for improving yield and yield stability of the pearl millet. A number of genomic regions were found to be associated with drought tolerance in terms of both grain yield and its components. A major QTL associated with the drought tolerance of grain yield mapped on linkage group 2 and explained up to 32% of the phenotypic variation. Another QTL were observed on LG 5 with phenotypic variation of 14.8%. During the present investigation, HHB 226 pearl millet hybrid was improved for drought tolerance by marker assisted selection. For this, two crosses were made i.e. HBL 11 x PRLT 2 and HBL 11 x 863 B. PRLT 2 and 863 B were used as donor parent and HBL 11 (male parent of HHB 226) was used as recurrent parent. Eleven SSR primers specific to drought tolerance from LG2 and 5 were used for polymorphism analysis between recurrent and donor parent. Three SSR primers from LG 2 (PSMP2066, PSMP2077, and PSMP2059) and one from LG 5 (PSMP2078) were found polymorphic in both the crosses which were used for further screening. Recurrent and donor parents were crossed to raise the F1 generation. Selected plants were backcrossed with HBL11 three times and in each generation positive plants were selected. Finally, four plants were selected in BC3F1 generation in both crosses which were then used for background selection. 25 and 32 background primers were found polymorphic out of 64 primers in HBL 11 x PRLT 2 and HBL 11 x 863 B cross respectively. About 70 and 80% of the recurrent parent genome was found to be transferred in most of the selected plants in 1st and 2nd cross respectively till BC3F1 generation. During the drought screening test in BC2F1 generation in 1st cross, RWC and 1000 seed weight were found to be enhanced with 27.84% and 15% respectively, in drought tolerant BC2F1 generation plants than drought sensitive. In 2nd cross, RWC and 1000 seed weight were increased with 33.6% and 28% respectively, in drought tolerant plants as compared to drought sensitive.ThesisItem Open Access Phenotyping and molecular marker analysis of selected F3 segregating lines from aerobic x low land indica rice (Oryza sativa L.) crosses(CCSHAU, 2015) Kanika Rani; Jain, R.K.Experiments were conducted to evaluate F3 segregating populations derived from the five crosses (PAU201 x MAS25, MASARB25 x PAU201, PAU201 x MAS26, MASARB25 x HKR47 and MAS25 x HKR47) for various physio-morphological and/or root traits and microsatellite markers linked to the traits promoting aerobic adaptation. MASARB25, MAS25 and MAS26 are aerobic while PAU201 and HKR47 are low-land indica rice varieties. In all the five populations, wide variation was observed for plant height, panicle length, number of panicles per plant, number of effective tillers per plant, root length, root thickness, fresh and dry root weight, 1000 grain weight, grain length-breadth ratio and grain yield per plant. In these populations, significant positive correlation was observed between yield per plant with plant height, effective no. of tillers per plant, length-breadth ratio, 1000 grain weight, root biomass and/or root length. The NTSYS-pc UPGMA tree cluster analysis and 2-D PCA scaling of selected F3 plants derived from the five crosses clearly showed large variation among two parental genotypes and F3 plants were invariably interspersed between them. A number of promising F3 plants have been selected, which had higher grain yield, root length and biomass greater than MAS25, MAS26 and MASARB25 for further progeny analysis. Most of these selected plants had the desired allele for the markers reported earlier to be linked with the aerobic adaptation traits (RM234 and RM547 for root length on chromosome 7 and 8 respectively; RM525 for root thickness and root biomass on chromosome 2).ThesisItem Open Access Isolation of PHB Producing Rhizobacteria and Molecular Characterization of Gene(s) for PHB Synthase for Efficient Production of Polyhydroxybutyrate(CCSHAU, 2015) Mukesh Rani; Sikka, Virendra K.Polyhydroxybutyrate the bioplastic, producing bacteria expeditiously were isolated from fifty diverse locations and 45 of them were found PHB producing on the basis of Nile blue A colony screening method. Among PHB producing isolates, 6 isolates were further selected on the basis of maximum florescence on different carbon and nitrogen sources and these were evaluated for relative PHB production. Three bacterial isolates, B2, B3 and C6 produced maximum PHB by weight and crotonic acid production. Cultural, morphological and biochemical characterization identified them as Pseudomonas spp. After PHB production kinetics studies, strain B3 and B2 gave maximum PHB in three and four days respectively. Culture conditions were optimized using Response Surface Methodology for maximum PHB production. The strain B 2 gave 37mg/l PHB and B3 gave 45mg/l of PHB. Hence, B3 strain was selected for further studies. FTIR confirmed the properties of PHB in B3 strain. A gene phaC was PCR amplified, cloned and sequenced. Sequence analysis employing bioinformatics tools like BLAST, PHYRE2 evaluated the predicted models. Ecofriendly and renewable agriproducts based PHB production method developed in these investigations may in future proceed in the direction of adopting this at commercial scale towards making PHB the agroplastic.ThesisItem Open Access Expression profiling of soluble starch synthase I gene during grain development for thermotolerance in wheat (Triticum aestivum L. em. Thell)(CCSHAU, 2014) Gupta, Meenu; Dhillon, SantoshThe four genotypes of wheat i.e. two thermotolerant (WH 730 and WH 1021) and two thermosensitive (WH 147 and WH 711) were evaluated under two different sowing conditions (normal and late sown) to assess the effect of high temperature (HT) stress on grain yield traits and expression of soluble starch synthase I gene. Gene expression was studied by real time PCR analysis at 7 stages (2, 5, 8, 10, 15, 20 and 25 DAA) during grain development in developing grains and flag leaf. High temperature stress negatively affected all yield traits like number of tillers per plant, grain number per plant, thousand grain weight, biomass and grain yield per plant. Decreased grain filling duration under HT stress was not compensated by increased grain filling rate. The effect was more profound in case of thermosensitive genotypes. The order of thermotolerance based on heat susceptibility index (HSI) is: WH 730 (0.65) > WH1021 (0.79) > WH 711 (1.13) > WH 147 (1.52). In developing grains, temporal pattern of SSSI gene expression was altered and expression peaked earlier under LS conditions (15 DAA) than under NS conditions (at 20 DAA) in all genotypes studied. In flag leaf, peak expression under NS conditions was observed at 15 DAA in all genotypes while under LS conditions, expression peaked at 15 DAA in thermotolerant genotypes and at 10 DAA in thermosensitive genotypes. At late stages during grain development, SSSI gene expression fell more abruptly and to a higher level in case of thermosensitive genotypes in grains as well as flag leaf. Partial SSSI cDNA was sequenced from WH 730 (1916 bp), WH 1021 (1924 bp), WH 147 (1925 bp) and WH 711 (1924 bp) genotypes. Phylogram generated using different starch synthases isoforms clustered SSSI, SSSII, SSSIII and SSSIV into separate groups. WH 730 showed maximum tolerance to HT stress while WH 147 was least thermotolerant, based on the overall picture of physiological traits, yield traits and gene expression analysis studies.