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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 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 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 Molecular Characterization of Transgenic Tomato (Lycopersicon esculentum Mill.) for fruit borer [Helicoverpa armigera Hübner] resistance(CCSHAU, 2015) Wadhwa, Zeenat; Yadav, Ram C.Tomato (Lycopersicon esculentum M.) is the most imperative vegetable crop cultivated across the globe. In India, the tomato productivity is stumpy as one of the major yield constraint is the tomato fruit borer (Helicoverpa armigera Hübner). At present there is no source for genetic resistance in tomato germplasm against this pest. The conventional methods for management of this particular pest are often futile. Therefore, Bt transgenic technology provides a secure and consistent means for management of this pest. Bt transgenic tomato cv. Hisar Arun carrying cry1Ac gene was developed through Agrobacterium- mediated transformation for providing resistance against fruit borer in our laboratory. In the present study, the developed transgenics were evaluated in three generations for transgene integration, expression and bioefficacy against Helicoverpa armigera. Transgenic plants carrying cry1Ac gene were analyzed by PCR for both the cry1Ac and nptII gene (selectable marker). Out of 96 T1 plants, 68 plants were found positive for cry1Ac and nptII gene confirming the presence of transgene in transgenic tomato plants. Similarly, in T2 and T 3 generations, screening of transgenic plants showed amplification of nptII and cry1Ac gene with amplification product of 1050 bp and 533 bp, respectively. Semi-quantitative RT-PCR analysis also revealed transcripts of 533 bp for cry1Ac and 1050 bp for nptII verifying the expression of respective genes in the transgenic plants in T 1 and T 2 generations. The study proved the stability of introduced gene in all three generations. Further, quantitative assessment of Cry protein using enzyme-linked immunosorbent assay (ELISA) proved the expression of Cry protein with the average value 357.5 ng/g, 231.5 ng/g and 216.9 ng/g FW in leaves of 60 days old transgenic plants in T 1 , T2 and T 3 generations respectively. The efficacy of transgenic tomato plants expressing cry1Ac gene against the target pest was required to judge the performance of these plants. Therefore, insect bioassay was performed with in vitro reared freshly hatched second instar larvae of Helicoverpa armigera. Transgenic plants showed resistance to Helicoverpa armigera showing significant mortality of H. armigera after 72 hours in all the three generations. Few promising tomato plants were selected on the basis of expression and bioefficacy analysis, which could be further used for developing Helicoverpa armigera resistant tomato lines/cultivars.ThesisItem Open Access Molecular breeding for pyramiding bacterial leaf blight resistance genes in rice (Oryza sativa L.)(CCSHAU, 2015) Baliyan, Nikita; Boora, K.S.Rice (Oryza sativa) is an important cereal food crop that serve as a major carbohydrate source for half of the world’s population. Bacterial leaf blight (BLB), caused by the bacterium, Xanthomonas oryzae pv. oryzae (Xoo) is one of the most serious threat to the rice production. The present work was undertaken to introgress the major BLB resistance genes (Xa21, xa13 and xa5) into BLB susceptible rice variety CSR-30, from BLB resistant donor varieties Pusa Basmati -1460 (having two BLB resistance genes Xa21 and xa13) and IRBB-60 (having three BLB resistance genes Xa21, xa13 and xa5) through marker assisted selection. Foreground selection was carried out in the F2 plants of cross CSR-30 x Pusa Basmati-1460 and BC2F1 plants of cross CSR-30 x IRBB-60 using specific STS markers pTA248, RG136 and RG556 linked to Xa21, xa13 and xa5 genes, respectively. In cross CSR-30 x Pusa Basmati-1460, a total of six F2 plants were found to have both Xa21 and xa13 genes and in cross CSR- 30 x IRBB-60, three BC2F1 plants were found to have all the three resistance genes Xa21, xa13 and xa5.These positive F2 (CSR-30 x Pusa Basmati-1460) and BC2F1 (CSR-30 x IRBB-60) plants were backcrossed with recurrent parent CSR-30. BC1F1 and BC3F1 seeds of these crosses were harvested separately. Foreground selection of BC1F1 and BC3F1 was carried out to confirm the presence of two and three BLB resistant genes using specific markers. Plants having two (CSR-30 x Pusa Basmati-1460) and three genes (CSR-30 x IRBB- 60) were selected for background selection. Four hundred and twenty eight SSR markers representing whole of the genome were used to identify polymorphism between parental genotypes out of which 111 and 131 SSR markers produced polymorphic alleles between parents CSR-30 and Pusa Basmati-1460 and parents, CSR-30 and IRBB-60, respectively. Finally, the maximum genome recovery for pyramided genotypes was analyzed by NTSYS and GGT softwares. The percentage recovery of recurrent parent genome in two-gene pyramided BC1F1 genotypes ranged from 62.7% to 83% and in three-gene pyramided BC3F1 genotypes, RPG ranged from 72.9% to 97.1%. Based on agronomic evaluation, BLB reaction, percentage recovery of recurrent parent genome, grain quality evaluation and aroma, four genotypes, C-70, C-123, C-142 and C-149 from cross CSR-30 x Pusa Basmati-1460 and four genotypes, R-28, R-68, R-32 and R-42 from cross CSR-30 x IRBB-60 were found promising. The four genotypes of cross CSR-30 x Pusa Basmati-1460 were backcrossed with CSR-30 and BC2F1 seeds were harvested. In cross CSR-30 x IRBB-60, the genotypes R-28, R-68, R-32 and R-42 were selfed and BC3F2 seeds were obtained. The present work demonstrates the successful pyramiding of bacterial leaf blight resistant genes Xa21, xa13 and xa5 into CSR-30 Basmati rice variety.