Loading...
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.
Browse
54 results
Search Results
ThesisItem Open Access Marker assisted selection for introgression of yellow rust resistance (Yr) genes in bread wheat (Triticum aestivum L. em. Thell)(CCSHAU, 2018) Reema Rani; Yadav, Neelam R.Wheat is major cereal crop that provides food to half of the world’s population. Yellow or stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst) is a devastating foliar disease that affects bread wheat (Triticum aestivum L.) in the major wheat growing regions of India. Development of host plant resistance is the most effective and efficient strategy for management of the disease. In the present study, 68 wheat genotypes were evaluated for Yr genes using tightly linked markers. Promising genotypes carrying multiple combinations of Yr genes were identified. Lassik, hard red spring wheat carries Yr36, Yr18 and Yr17 genes while WH711 and PBW343 are high yielding and yellow rust susceptible varieties. Therefore, Yr resistance genes (Yr36, Yr18 & Yr17) were introgressed from donor parent (Lassik) into the susceptible varieties (WH711 & PBW343) through marker assisted backcrossing approach. Foreground selection was carried out in the BC2F7 and BC2F8 plants derived from the crosses of (WH711 x Lassik) x WH711 and (PBW343 x Lassik) x PBW343 using markers linked to Yr genes Gpc/B1/Yr36, Yr18 and Yr17. In both the BC2F7 and BC2F8 generations of the cross (WH711 x Lassik) x WH711, five lines showed the presence of all the three yellow rust resistance genes. Two lines were found to be carrying Yr36 and Yr18 genes. One line had Yr36 and Yr17 genes and one line was identified with Yr36 only. In the cross, (PBW343 x Lassik) x PBW343, one line confirmed the presence of all the three Yr resistance genes. Two lines were found positive for Yr36 and Yr18 genes. One line had Yr36 and Yr17 genes and one line was identified with Yr36 gene only. These Yr gene positive plants were subjected to background selection using polymorphic SSR markers. A total of 575 SSR markers spanning throughout the genome were used to study the polymorphism between parental genotypes out of which 100 and 101 SSR markers produced polymorphic alleles between parents i.e. WH711 and Lassik and PBW343 and Lassik, respectively. Finally, the maximum genome recovery for pyramided genotypes was analysed by NTSYS and GGT softwares. The percentage recovery of recurrent parent genome in the BC2F7 plants of WH711 x Lassik and PBW343 x Lassik ranged from 94.0 % to 96.1 % and 93.6 % to 96.6 %, respectively. Similarly, the percentage recovery of recurrent parent genome in the BC2F8 plants of both crosses ranged from 94.0 % to 96.1 % and 95.5 % to 97.4 %. Based on agronomic evaluation, yellow rust reaction, and percentage recovery of recurrent parent genome, promising lines from both the crosses were identified. The present work demonstrates the successful pyramiding of yellow rust resistance gene i.e. Yr36, Yr18 and Yr17 into WH711 and PBW343 varieties using the marker assisted backcross breeding.ThesisItem Open Access Development and characterization of transgenic chickpea (Cicer arietinum L.) plants with OsRuvB gene for salt stress tolerance(CCSHAU, 2018) Preeti; Kharb, PushpaChickpea (Cicer arietinum L.) is a nutrient rich, second most important legume crop after soybean. Production and productivity of chickpea is affected by various abiotic stresses but salt stress is the most deleterious accounting for upto 100% yield losses. In the present study, transgenic chickpea plants (var. HC-1) carrying OsRuvB gene were developed for salt stress tolerance using Agrobacterium tumefaciens strain LBA4404 with pCAMBIA1301 (Kharb et al., 2012). Putative transformants were screened at early stage through PCR amplification using gene specific primers and a transformation frequency of 17% was observed. Southern blot and real time PCR analysis revealed stable and single copy insertion of the transgene in chickpea genome. Seeds from each of the PCR positive T0 chickpea plants were sown to raise T1 generation and a total of 74 plants (out of 170) showed the presence of OsRuvB gene. Physio-biochemical analysis of selected T1 transgenic plants subjected to 100 mM salt stress showed that transgenic plants were able to maintain higher chlorophyll content, relative water content, proline content, total soluble sugars, catalase and peroxidase activity compared to the wild type plants. Whereas, electrolytic leakage and lipid peroxidation were relatively less as compared to the wild type plants under 100 mM stress. Among all transgenic lines, line 8 performed well with respect to all the parameters studied and can be taken further for the development of transgenic chickpea plants for salt stress tolerance.ThesisItem Open Access Microsatellite analysis for targeting bacterial leaf blight resistance in cluster bean [Cyamopsis tetragonoloba (L.) Taub.](CCSHAU, 2018) Priti; Yadav, Neelam R.Cluster bean [Cyamopsis tetragonoloba (L.) Taub.], a drought tolerant and industrially important forage legume because of galactomannan (also known as guar gum) present in the endosperm of seeds. There is huge demand of guar gum but the production is less than demand. Bacterial leaf blight (BLB) is the major constraint which causes yield losses up to 60-80%. There is a need to develop high yielding and disease resistant varieties. Segregating populations F3-4 and F4-5 along with parents of interspecific cross (HG563 × C. serrata) and intraspecific cross (HG75 × PNB) respectively were evaluated for various morphological traits and bacterial leaf blight resistance under natural condition. A total 192 SSRs comprising EST SSR from cluster bean and SSRs from other legumes were used in molecular analysis. In interspecific cross, 38 SSRs were found polymorphic while 39 showed polymorphism in intraspecific cross. In intraspecific cross, the segregants grouped with PNB while the population of interspecific cross grouped with HG563. Linkage analysis using polymorphic markers in both the crosses revealed that nine markers formed a single linkage group in interspecific cross and only five markers formed a single linkage group in intraspecific cross while other markers remain unlinked. Four minor QTLs were identified in interspecific cross for plant height (qPH1.1), hundred seed weight (qHSW1.1), disease resistance (qPDI1.1) and gum content (qGC1.1) mapped on single linkage group at 39.41, 123.41, 112.41 and 102.41 cM, respectively. A few promising lines based on phenotypic and molecular analysis have been identified which can be further employed in cluster bean breeding program.ThesisItem Open Access Development and characterization of transgenic pigeon pea (Cajanus cajan (L.) Millsp.) with OsRuvB gene for salinity tolerance(CCSHAU, 2018) Rakshita Singh; Kharb, PushpaSalinity is a major stress which causes considerable loss in crop productivity. In this study, OsRuvB, a DNA helicase, under the control of CaMV35S promoter, is used to develop salinity tolerant transgenic pigeon pea plants (cv. Manak) using Agrobacterium-mediated transformation method. The transgene integration in 70 putative T0 plants was checked using PCR analysis and 25 plants showed amplification for the OsRuvB gene using specific primers, giving a transformation efficiency of 35.7 %. Out of these, 6 comparatively higher yielding plants were selected in the T1 generation for segregation analysis and to screen the plants for stable transgene integration. Further, Southern hybridization and real time PCR were done to determine the copy number of the transgene in these selected transgenic lines. To assess the efficacy of transgene, the wild type, and transgenic plants of T1 generation were subjected to 75mM NaCl stress and analysis was done based on various physiological parameters. The results showed that in addition to more chlorophyll and relative water content under salinity, the transgenic plants also showed higher activity of peroxidase and catalase. Level of proline and total soluble sugar was elevated in T1 transgenic plants when grown under salt stress, but the increase was lower than in wild type plants under salt stress. Moreover, membrane injury index and lipid peroxidation were found to be significantly reduced in transgenic plants in comparison to the wild type plants. These results indicate that transgenic pigeon pea lines were less affected by salt stress. The transgenic lines didn‘t have significant increase in osmolytes proline and total soluble sugar, which indicates that the tolerance is being imparted either by some other osmolytes or some entirely different mechanism yet to be uncovered might be working in these plants.ThesisItem Open Access Organogenesis and somatic embryogenesis for micropropagation of some elite cultivars of guava (Psidium guajava L.)(CCSHAU, 2018) Sunita Devi; Yadav, Neelam R.Guava (Psidium guajava L.), a member of Myrtaceae family, is hardy fruit tree and well adapted in wide range of tropical and subtropical regions. Vegetative and seedling derived plants are not suitable for large scale cultivation as such plants are not true to type and give lesser productivity. Providing quality planting material to farmers is limiting factor in adopting this crop. Micropropagation using in vitro approach can be instrumental in overcoming this bottleneck. An efficient and reproducible protocol is required for successful micropropagation. Four cultivars of guava viz. Hisar Safeda, Hisar Surkha, Lucknow-49 and Allahabad Safeda were used in the present study for micropropagation using nodal, shoot tip and mesocarp of immature fruits as explants. The explants collected during April-May, in solution of citric acid (75 mg/l), ascorbic acid (50 mg/l) and 2-3 drops of Tween 20 showed minimum browning. Sequential treatment of 0.2% each of bavistin and streptomycin (30 min.), 1 % NaOCl (5 min.), 0.1% HgCl2 (3 min.) and finally 70 % ethanol (30 sec.) was found best for initiating aseptic cultures. The four cultivars showed differential response in selected media.WPM medium was most suitable for micropropagation than MS basal medium. The percent shoot induction response was dependent on type of explant, composition of media and genotype. The nodal explants showed maximum percent shoot induction of 51.70% in Hisar Safeda on WBI (WPM, 2.5 mg/l BAP, 0.5 mg/l IAA) medium, 53.81 % in Hisar Surkha on WBN (WPM, 2.5 BAP mg/l, 0.2 mg/l NAA) medium, 56.69% in Lucknow-49 and 56.53% in Allahabad Safeda, both on WBNK (WPM, 2.5 BAP mg/l, 0.2 mg/l NAA and 1.0 mg/l kinetin) medium. In case of shoot tip explants, maximum percent shoot induction response of 52.28% (Hisar Safeda), 53.70% (Hisar Surkha), 54.39% (Lucknow-49) and 53.50% (Allahabad Safeda) was observed on same media as specified above. Among the four cultivars investigated, Lucknow-49 was found most responding. Higher root induction response (>64.10%) was observed in MS basal media (half strength) with 2.0 mg/l IBA. Rooted plants were transferred in potting mixture of cocopeat and vermiculite (1:1) with 40- 66.66% survival. MS basal medium supplemented with TDZ (1.5 mg/l) was best for callus induction from mesocarp tissue but 2, 4-D (5.0 mg/l) was most supportive for induction of embryogenic calli.ThesisItem Open Access Molecular breeding for drought tolerance in pearl millet [Pennisetum glaucum (L.) R. Br.] using microsatellite markers(CCSHAU, 2018) Jangra, Sumit; Yadav, Neelam R.Pearl millet is a small grain, 1.7Gb genome size, photosynthetically efficient, C4 monocot. It is a staple food and fodder crop for millions of people inhabiting hot and dry areas of arid and semi-arid tropics. Occurrence of terminal drought stress is very common in such regions, thus making stress tolerance an essential attribute in pearl millet breeding programmes. A major QTL associated with grain yield under terminal drought stress was mapped on linkage group 2, accounting for 32% phenotypic variation for grain yield under drought stress. Another QTL accounting for 14.8% phenotypic variation for grain yield was mapped on linkage group 5. In the present study, HBL 11 was introgressed with drought-tolerant QTLs from PRLT 2 and 863 B for improving a commercial pearl millet hybrid HHB 226 against terminal drought stress using marker-assisted backcrossing. Three SSR marker (PSMP2066, PSMP2077 and PSMP2059) linked to LG 2 and one SSR marker (PSMP2078) linked to LG 5 were used to carry out foreground selection in BC4F2 generation. The selected plants were undertaken for background analysis. Thirty-two and twenty-five polymorphic SSR markers used for background analysis in cross HBL 11 x PRLT 2 and HBL 11 x 863 B respectively. About 97% and 82% recurrent parent genome recovery was observed in one each of the selected lines in cross HBL 11 x PRLT 2 and HBL 11 x 863 B respectively. Plants positive in foreground selection and with high recurrent parent genome recovery were taken for analysis of morpho-physiological and yield-related traits. Thousand seed weight, relative water content and chlorophyll fluorescence were found to be increased and electrolyte leakage was decreased in comparison to recurrent parent. The introgression of drought tolerance QTLs from donor parents was established and recurrent parent genome was recovered. Improved versions of HBL 11 have been developed which can be crossed with ICMA 843-22 to develop improved HHB 226.ThesisItem Open Access Molecular mapping of bacterial blight resistance gene, drought tolerant QTL(s) and genetic diversity analysis in clusterbean {Cyamopsis tetragonoloba(L) Taub}(CCSHAU, 2011) Bajaj, Anshu; Boora, K.S.The present investigation was carried out to screen genetic diversity using RAPD markers and to identify and map the QTL for resistance to bacterial blight and drought tolerance in clusterbean .The plant material was procured from Forage section . For bacterial blight, the resistant (HG 75) & susceptible(PNB) genotypes were selected & crossed to generate F3 population. The F3 population was used to map QTLs. For drought tolerance studies genotypes HG 563 (tolerant) & PNB (susceptible) were used to generate the mapping population (F 2:3).The genetic diversity studies were carried out in 48 genotypes of clusterbean using 130 RAPD primers and 97 primers produced amplification of 705 alleles ;out of which 644 were polymorphic. The similarity ranged from 0.11 to 0.97.The dendrogram grouped the genotypes into two major clusters at a similarity coefficient of 0.68. A variety FS277 was out grouped which is a forage variety. The genotype GP96 and F5277 were found to be most diverse (89%).The QTLs were identified for bacterial blight resistance using parental genotypes (HG75 & PNB) & their F3 population. The F 3 population was screened artificially for disease resistance along with the parent genotypes. Polymorphic primers were used to screen the F3 population & the major QTLs were identified for disease resistance using parental genotypes, HG75, PNB and their F 3 population. A total of 83 RAPD primers were used to screen the parental genotypes out of which nine primers were polymorphic. These primers were used to screen the F 3 population and the QTLs found were L19 , D1, AB7 and QLTY 3. Parental genotypes HG563 and PNB and their F2-3 genotypes were screened at field and net house for resistance to drought. Out of fifty genotypes, 12 were highly tolerant, 23 were moderately tolerant and 15 were susceptible to drought. The parents were screened with eighty seven primers out of which, sixteen were found polymorphic and nine QTLs were found linked to drought tolerance. Loci OPQ 20,OPD10, OPD14,OPQ 12,OPAC 8 and OPF 9 were major contributing QTLs and OPAC2,OPL3 and OPF17 are minor QTLs The diverse lines may be used for breeding programs to improve the production of agronomically superior genotypes of guar. The major QTLs for bacterial blight and drought tolerance may be used for introgression into agronomically superior genotypes of guar.ThesisItem Open Access Molecular breeding for indica rice (Oryza sativa L.) genotypes with aerobic adaptation(CCSHAU, 2018) Bhoyar Pratik Istari; Jain, Rajinder KumarIn rice (Oryza sativa L.), HKR47 x MAS26, Pusa1121 x MAS25 and Pusa Basmati 1460 x MASARB25 derived F4, F5, F6, F7, F8, BC2F3, BC2F4 and BC2F5 populations were evaluated for various physio-morphological (plant height, grain yield and yield components), grain quality (L/B ratio of grain and aroma) and root (root length, biomass and thickness) traits under direct-seeded water-limited aerobic conditions in the field as well as net house during 2014, 2015 and 2016 kharif seasons. While HKR47 and Pusa1121 and Pusa Basmati 1460, respectively, are high yielding indica variety and premium Basmati rice varieties, MAS26, MAS25 and MASARB25 are aerobic/drought tolerant indica varieties developed at UAS, Bangalore and IRRI, Manila, Philippines. These aerobic rice varieties had 22 - 30 % higher yield and root biomass (15-20 %) than lowland indica/Basmati rice varieties under direct-seeded aerobic conditions. The filial and backcross populations showed variation for both agronomic and root traits. The grain yield/plant had positive correlation with root biomass in HKR47 x MAS26, Pusa1121 x MAS25 derived populations. A total of 186/500 SSR markers well distributed on 12 rice chromosomes, showed polymorphism in six parental rice genotypes; 83-87 polymorphic markers were used to assess the genetic diversity in these populations. NTSYS-pc UPGMA tree cluster analysis and two-dimensional PCA scaling showed that all the parental varieties were quite divergent and segregating populations were interspersed between the two parental rice genotypes. Composite interval mapping analysis (WinQTL Cartographer 2.5) using SSR databases of these populations led to the mapping of 81 QTL (LOD: 3.0-9.0 and R2% 5.3-53.4) for various traits promoting aerobic adaptation including grain yield/plant (21 QTL on chromosomes 1, 2, 5, 6, 8 and 10), plant height (13 QTL on chromosomes 1, 5, 6, 7, 10 and 11), effective number of tillers/plant (16 QTL on chromosomes 1, 2, 5, 6, 7, 8, 10, 11 and 12), panicle length (15 QTL on chromosomes 1, 2, 4, 5, 6, 7, 8, 9 and 10), test grain weight (5 QTL on chromosomes 2, 4, 8 and 12) and dehusked grain L/B ratio(11 QTL on chromosomes 1, 2, 6, 7 and 8). Notably, 36 of these were large effect QTL with r2 value >20%. In populations involving Basmati rice parent, plants had allelic profile at BAD2A locus like Basmati (585 and 257 bp) or non-Basmati (585 and 355 bp) parents or were heterozygous (585, 355 and 257 bp). Data on physiomorphological traits and status at BAD2A locus were used to select promising aerobic rice genotypes in each generation, advanced to next generation and evaluated further. Finally, in 2016, 23 plants were selected (five each from HKR47 x MAS26 F6, F7 and BC2F5; four from Pusa1121 x MAS25 F8 and four from PB140 x MASARB25 F7 populations) based on field and net house evaluation data. Plant no. H/M26/F4/56-1-1 had highest grain yield/plant (28.1 g; MAS26, 20.4±1.7) and root length (55.6 cm; MAS26, 46.1±3.4) followed by H/M26/F5/A15- 4-2 (25.8 g and 52.3 cm); H/M26/F4/56-1-1 had exceptionally higher fresh weight (33.3 g; MAS26, 13.7±1.4 g). The selected plants possessed 3-9 favorable QTL/alleles in homozygous state and 1-29 in heterozygous state. Plant H/M26/F4/56-1-2 had maximum (33) of favorable QTL/alleles either in homozygous (5) or heterozygous (28) state. Preliminary replicated RBD yield trails revealed that 11/15 selected promising genotypesoutperformed the respective check variety (HKR47, Pusa1121 or PB1460) under aerobic conditions. These genotypes will serve as the novel material for the selection of stable water efficient aerobic indica/Basmati rice varieties. It will also be interesting to investigate and confirm the role of these QTL in promoting aerobic adaptation.ThesisItem Open Access Genetic and molecular analysis of basmati x indica (Salt tolerant) derived segregating population(s) in rice (Oryza sativa L.)(CCSHAU, 2018) Mahavir; Jain, Rajinder KumarExperiments were carried out to develop and assess segregating population(s) for variation in salt tolerance and physio-morphological traits derived from two crosses, Pusa1121 x CSR10 and Pusa1121 x Pusa Basmati 1460 (PB1460). A novel selection strategy was used for screening segregating populations for salt tolerance at seedling stage; seven-day old germinated seedlings were subjected to 40 mM NaCl treatment in Yoshida nutrient solution for 3-7 days in the net-house, which allowed the selection of most salt tolerant segregating genotypes. Selected salt tolerant F2 (3 plants), F3 (64 plants), F4 (12 plants) and BC1F1 (21 plants) seedlings of the two crosses had values/scores for salinity tolerance related parameters including root length (7.6 – 13.6 cm), shoot length (13.0 – 24.4 cm), fresh weight (0.351 – 1.214 g), Na+ (4.14 – 6.16 μmoles/g DW) and K+ contents (37.35 – 75.35 μmoles/g DW), Na+/K+ ratio (0.72 – 0.124), physical appearance (score 1 - 3), yellowing of leaves (1 - 3), shoot tip burning (1 - 3) and leaf curling (1 - 3) which were comparable to the salt tolerant parent CSR10. Molecular analysis of BAD2A aroma locus revealed an overall ratio of 14:75:11 for homozygous fragrant, heterozygous fragrant and non-fragrant plants, respectively, in Basmati x indica derived F2, F3, F4 and BC1F1 segregating populations. DNA fingerprint databases of 40 Pusa1121 x CSR10 F3 plants, 24 PB1460 x CSR10 F3, 10 Pusa1121 x CSR10 F4 plants and 21 Pusa1121 x CSR10 BC1F1 plants were prepared using 99, 94, 97 and 99 polymorphic SSR markers, respectively, and used for linkage mapping via composite interval mapping (WinQTL Cartographer 2.5). As many as 100 QTL were mapped for various salt stress tolerance attributes (shoot length, 29; root length, 9; Na+, 10; K+, 22 and K+/Na+ ratio, 30). Eleven QTL associated with K+/Na+ ratio had percent phenotypic variance more than 20% while 11 QTL had LOD score more than 7.5. Ten QTL (qK/Na7.1, qK/Na10.2, qK9.1, qK11.1, qK6.1, qK10.1, qK10.2, qK2.1, qK10.3 and qK/Na2.1) had LOD score more than 9.0 and eight (qK2.1, qK10.2, qRL2.1, qK6.1, qK10.1, qRL2.1, qSL3.1 and qK/Na2.1) were identified with r2 values more than 50. Maximum number (28) of QTL were mapped on chromosome 2 followed by chromosome 1 (15 QTL). Notably, qK/Na2.1 on chromosome 2 (map position, 12.8 cM) accounted for 83.0% phenotypic variance (DPE - Pusa1121); qNa2.1 at same map position also contributed for Na+ content with 49.3% phenotypic effect. A number of promising plants were selected based on screening in the net house and field evaluation for grain yield, yield components and L/B ratio. In 2016, twelve F4 plants of the two crosses were selected through screening in the net house having low Na+ content (<5.12 μmoles/g DW) and high K+ (>47.97 μmoles/g DW) and Na+/K+ ratio between 0.084 - 0.108. The selected plants were homozygous fragrant, heterozygous fragrant and non-fragrant in the ratio of 4:6:2 based on BAD2A locus analysis. The selected plants possessed 78 - 93 favorable alleles/QTL for salt tolerance attributes either in homozygous or heterozygous state. Field evaluation in 2016 Kharif season also led to the selection of 55 plants (25 Pusa1121 x CSR10 and 15 PB1460 x CSR10 F4; 15 Pusa1121 x CSR10 BC1F2) having grain yield and grain length/breadth ratio in the range of 7.7 - 27.5 g and 4.17 - 5.14. Selected plants can be further used in salt tolerance breeding programs in Basmati rice.