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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 Tissue culture studies in Alfalfa (Medicago sativa Linn.)(CCSHAU, 2011) Rajesh Kumar; Kharb, PusphaIn the present investigation attempts were made to develop an efficient protocol for in vitro plant regeneration in Medicago sativa L. Initiation of callus formation from hypocotyl explants was observed within 13-15 days of culture. Initiation of callus formation from cotyledons explants was observed within 13-15 days of culture. Maximum callus formation was obtained in R3 medium containing MS basal + NAA (1.0 mg/l) + Kinetin (0.3 mg/l). Maximum number of shoots was obtained in R12 medium containing MS basal + NAA (1.0 mg/l) and Kinetin (0.5mg/l). MS basal medium was used for roots induction, roots were observed after 14 days with very good quality.In media R2 (MS basal + NAA 0.1mg/l + 2 IP 1.0 mg/l) and R8 (MS basal +NAA 2.0 mg/l), when shoots were left little longer rooting occurred. After transplantation, 73.3% plants survived and these plants looked normal with no morphological changes in leaf structure and plant type.Thus in the present study, regeneration protocol in Medicago sativa L variety T9 has been developed using hypocotyls and cotyledon explants.ThesisItem Open Access Molecular characterization of stripe rust resistance in wheat (Triticum aestivum L.) using DNA markers(CCSHAU, 2011) Admas Berhanu; Boora, Khazan SinghTriticum aestivum L., commonly called bread wheat, is the most common staple crop throughout the world. India is the second largest producer, next to china. Wheat is highly affected by rust diseases. Despite the natural resistance of wheat varieties to most stripe rust races, the highly variable nature of the pathogen and their time to time arise as a new strains put a major threat in wheat production. Hence, it becomes a major point of breeding interest in most part of the world. The present study was undertaken to study SSR polymorphism among 26 wheat genotypes using PCR. The 26 genotypes consisted of equal number of resistant and susceptible genotypes. Using 40 SSR primers, a total of 140 clear and reproducible bands were observed. For the genotypes studied, up to 9 bands (WMC 11) were produced with an average of 3.5 bands per primer. The size of amplified bands ranged from 100- 1517 bp. The similarity coefficients between different genotypes ranged from 0.46 to 0.88 with an average similarity value of 0.66. At an arbitrary cut-off at 27 per cent similarity level on a dendrogram, the wheat genotypes were categorized into two major clusters. PBW 590 and WH 896 were found to be genetically most divergent, while genotype Bijiga Yellow and WL 711 were least divergent. One unique band was selected for identification of resistant varieties for stripe rust. The size of unique band was around 400 bp. This marker WMC 206 has been found to have a tight linkage with resistant gene and present on chromosome 4D. Similar studies on gene identification by different researchers have revealed that chromosome 4D is a host for Yr 28 and Yr 32 genes, which are resistant to yellow rust. The unique band can be developed into Sequence Characterized Amplified Region (SCAR) marker after cloning and sequencing. The developed SCAR can be used for identification of yellow rust resistant genotypes from the susceptible ones. The present study successfully distinguishes yellow rust resistant genotypes from the susceptible ones using SSR Markers. The present study is of special value as it can play a key role in the introgression of a yellow rust resistant gene to the breeding programs.ThesisItem Open Access Micropropagation studies in important cultivars of bamboo (Bambusa bambos & Dendrocalamus asper)(CCSHAU, 2011) Ram Niwash; Chowdhury, V.K.The present investigation has been undertaken to: (i) To develop efficient micropropagation protocol in two important cultivars of bamboo (Bambusa bambos & Dendrocalamus asper). (ii) Standardization of protocol and transplantation of regenerated plantlets of both cultivars to green house. Maximum survival percentage of explants (91.7%) in Bambusa bambos and 83.3% in Dendrocalamus asper was obtained when the nodal explants were sterilized with 0.2% bavistin (45 minutes), 0.2% streptocyclin (15 minutes) and 0.1% HgCi2 (6 minutes) in series. Bambusa bambos was observed to be more responsive to shoot proliferation and multiplication as compared to Dendrocalamus asper. Maximum per cent shoot proliferation was observed (88.9%) on MS basal medium supplemented with 2.0 mg/l BAP, 0.5 mg/l kinetin and 0.25 mg/l NAA in Bambusa bambos and 77.8% in Dendrocalamus asper and average number of shoot proliferation per explant was 2.5 in Bambusa bambos and 1.5 in Dendrocalamus asper. Maximum shoot multiplication response was observed in Bambusa bambos with an average of 8.4 shoot regenerated per explants and 5.4 in Dendrocalamus asper on MS basal medium supplemented with 2.0 mg/l BAP, 0.5 mg/l kinetin, 0.20 mg/l NAA and 30.0 mg/l adenine sulphate in both the cultivars. Maximum root induction frequency was obtained when half strength MS media supplemented with 2.0mg/l IBA used and 77.8% rooting after 7 days in Bambusa bambos and 66.7% rooting after 10 days in Dendrocalamus asper respectively. Survival rate of regenerated plantlets transferred to potting mixture containing sand + soil + vermicompost (1:1:1) was 83.3% in Bambusa bambos and 75.0% in Dendrocalamus asper.ThesisItem Open Access Molecular Analysis of male and female genotypes of Simarouba glauca DC using RAPD markers(CCSHAU, 2011) Chauhan, Niharika; Dhillon, SantoshSimarouba glauca DC is a tropical oil-yielding tree belonging to the family Simaroubaceae, and known locally as “Oiltree”, “Paradise tree” & “Laxmitaru”. It is a native of Central America and was first introduced into India in 1966. It is a multipurpose evergreen oil-seed tree with the economically important feature that it can grow very well even in marginal & wastelands areas with degraded soils. The seeds contain 60-70% (w/v) oil of a quality suitable for edible & non-edible purposes. The knowledge of genetic variation and genetic relationship among genotypes is an important factor for classification, proper utilization of germplasm resources and crop breeding to increase both quality and quantity of crop production. Molecular markers are effectively and commonly being used in studying molecular polymorphism and genetic diversity in species or in populations. The present study was undertaken to assess molecular polymorphism among ten male and ten female genotypes of Simarouba glauca DC using RAPD markers. In the RAPD analysis of 20 S.glauca genotypes using 50 random primers, total 426 amplification products were obtained, out of which 3 were monomorphic and 423 were polymorphic. The number of amplified DNA bands varied between 2 and 17 with an average of 8.52 bands per primer. The polymorphism percentage ranged between 72.7% to 100%. The average polymorphism across all the 20 genotypes was found to be 99.4%. Overall size of PCR amplified products ranged between 200 bp - 3000 bp. The similarity indices between different genotypes ranged from 0.03 to 0.76 with the overall average genetic similarity of 0.28. The dendrogram mainly divided into two major clusters at a similarity coefficient of 0.10. Of all the twenty genotypes SG1 M and SG15 M were found to be most closely related having maximum similarity coefficient of 0.76 and PALEM 1 F was the most out grouped (diverse) genotype. These results indicate the presence of large genetic variation among the genotypes studied which can be attributed to high natural heterozygosity in this dioecious obligate cross-pollinated plant. Also as the genotypes were collected from different geographical areas (Gujarat, A.P. and Maharashtra), different geographical conditions allow some possible genetic changes which add to the genetic diversity. A very high polymorphism (99.4%) observed between the genotypes can be utilized for crop improvement in Simarouba glauca. The present study also demonstrates the utility and importance of RAPD markers in genetic diversity analysis and genotyping in Simarouba glauca.ThesisItem Open Access Variation in root morphology and molecular profile in aerobic × Basmati F2 population(s)(CCSHAU, 2011) Kharb, Anju; Jain, R.K.The present investigation has been undertaken to: (i) analyze the allelic profile of 60 F2 plants obtained from a cross between Pusa1121 (Basmati) and MAS25 (aerobic) rice varieties for five SSR markers and BAD2 aroma gene, and (ii) carry out greenhouse evaluation F2 plants for root length & biomass, shoot length & biomass, yield per plant, 1000 grain weight, length-breadth ratio, aroma, relative water content and photosynthetic efficiency. Polymorphism for five microsatellite markers (RM144, RM234, RM302, RM339 and RM440) and BAD2 aroma gene specific (Bradbury et al. 2005) markers with a base difference of >30 bp in the amplified products, could be clearly visualized on 2.5% w/v agarose gels. Of the 60 F2 plants, 20, 18 and 22 plants, respectively had Pusa1121, MAS25 and Pusa1121 + MAS25 specific alleles at BAD2 locus. The NTSYS-PC UPGMA tree cluster as well as PCA analysis using six marker dataset grouped the 60 F2 plants and parental rice genotypes into two major groups: major group I with Pusa1121 and 18 F2 plants and major group II with MAS25 and 42 F2 plants. Cluster as well as PCA analysis clearly indicated that the F2 population was scattered between the two parental rice genotypes, Pusa1121 and MAS25, with an inclination towards MAS25. F2 population also showed wide variation for various traits including root length and/or dry biomass, yield per plant and length-breadth ratio. Yield per plant showed a significant positive correlation with fresh root weight (0.370; p=0.01) and dry root weight (0.328; p=0.01). A total of 9 – 12 plants had root length and/or biomass greater than MAS25 and nine plants had yield per plant higher than Pusa1121. A number of F2 plants were identified with higher root length and/or dry biomass, yield per plant, lengthbreadth ratio and with Pusa1121 specific alleles in homozygous or heterozygous condition at BAD2 locus; these lines shall serve as the novel material for the selection of stable aerobic Basmati rice varieties.ThesisItem Open Access Variation for aroma, Iron and Zinc content in segregating populations of crosses between mineral-rich Indica and elite basmati variety(CCSHAU, 2011) Harsh Samar Ecka; Sikka, V.K.Experiments were conducted on 70 and 111 F2 plants derived from crosses between PB1 x Palman 579 and PB1 x HKR95-157 respectively and were evaluated for micronutrient content (Fe and Zn) and agronomical parameters. A large deviation was observed for plant height (65-130 and 65-102 cm), effective number of tillers per plant (3-15 and 3-21), panicle length (15.1-27.9 and 15.3-27.9), length/breadth (L/B) ratio (3.73-5.77 and 2.91-4.95), 1000-grain weight (14.08-25.36 and 16.00-26.65 g) (0.033-0.472), yield per plant (1.16-32.96 and 1.1-31.43 g) in F2 populations of PB1 x Palman 579 and PB1 x HKR95-157 respectively. Analysis of micronutrient content including Fe and Zn content revealed that the Fe and Zn content of the three parental rice genotypes namely PB1, Palman 579 and HKR95-157 was 33.6, 16.53 μg/g, 363.53, 22.16 μg/g and 422.13, 17.40 μg/g respectively. The Fe and Zn content among the 70 PB1/Palman 579 F2 plants ranged from 33.80-171.20 and 10.43-31.30 μg/g respectively whereas the micronutrient content in a population comprising of 111 PB1/HKR95-157 F2 plants was 8.63-221.50 μg/g (Fe content) and 3.67-48.80 μg/g (Zn content). An allelic profile was generated for 70 F2 plants of cross PB1×Palman 579 and their parental genotypes to ascertain their aroma trait. Thirty three of the segregants were adjudged aromatic from BAD2 marker analysis. The segregating population was analyzed and enabled us to narrow down certain transgressive segregants with the combination of desirable traits.ThesisItem Open Access Genotype identification of Pearl millet [Pennisetum glaucum (L.) R. Br.] hybrids and their parents using molecular markers(CCSHAU, 2012) Yadav, Sandeep; Yadav, Neelam R.Pennisetum glaucum (L.) R. Br., commonly called pearl millet, is the fourth most important world food crop. It is well adapted to drought, low soil fertility, salinity, low pH and high temperatures among tropical cereals and is grown on >29 million hectares in arid and semiarid regions of Asia (11 million hectares) and Africa (15 million hectares). This crop provides basic sustenance to resource poor farmers and can grow in the poorest soil. Fingerprinting can be used to identify and monitor germplasm after its release for commercial cultivation. The present study was undertaken to study RAPD polymorphism among 20 pearl millet genotypes using 35 decamer primers in PCR reaction. Out of 35 primers analysed, thirty-one primers showed amplification producing 144 bands, where 132 bands were polymorphic and 12 bands were monomorphic. Four primers (OPB-6, OPB-7, OPB-9 and OPB-14) gave no amplification. For the genotypes studied, upto 8 bands (OPB-8 and OPC-9) were produced with an average of 4.11 bands per primer. The size of amplified bands ranged from 350-3100 bp. The similarity coefficients between different genotypes ranged from 0.49-0.87 with an average similarity value of 0.70. At an arbitrary cut-off at 56 per cent similarity level on a dendrogram, the pearl millet accessions were categorized into two major clusters. HHB-197 and ICMA-97111A were found to genetically similar. HMS-37A was genetically most dissimilar as it has A4 source of cytoplasm. Hybrids are genetically more similar to female parent. Twelve unique bands were selected for identification of five hybrids and their parents of pearl millet. The size of unique bands ranged from 490- 2600 bp. These uniqe bands can be developed into Sequence characterized amplified region (SCAR) marker after cloning and sequencing. The developed SCAR can be used for identification of pearl millet hybrids from their parents. The present study successfully distinguishes five pearl millet hybrids and their parents using RAPD analysis. The study has special value since seed companies and national registration agencies have an interest in DNA fingerprinting because the technology can be used to protect intellectual property, establish identity and assess purity. Further, identification of particular variety/ hybrid with PCR is of particular interest in present day PPV&FR regime that gives immense legal powers to farmers for compensation if the suggested criteria by seed providers are not fulfilled.ThesisItem Open Access Impact of specific chromosomes on ADP–glucose pyrophosphorylase activity and grain growth in wheat (Triticum aestivum L.em. thell)(CCSHAU, 2011) Nishat; Sikka, V.K.The materials of present investigation consisted of two parents viz., Chinese spring and C591 and selected 8 disomic chromosome substitution lines of C591 in the background of Chinese Spring. The experiment was laid out in completely randomized block design with three treatments and replicated thrice. The data were recorded for different field traits and computational traits of selected 8 disomic lines, their parents and WH147, WH147M. The data for AGPase activity suggests the localization of gene on different chromosomes. It also suggests no. of copies for gene of AGPase enzyme are located at specific chromosome, active at different grain filling stages. Analysis of variance revealed the presence of significant variability in the materials under study. Observations recorded on yield and its attributing characters and macro and micronutrients contents as well as their plant uptake revealed genotype and chromosome specific variation. On the basis of genotypic performance under various treatments for different plant characters, macro and micro nutrients uptake and Azotobacter colony count, disomic chromosome substitution lines 1D, 5D, 6B, 7B figured promising. It could be postulated that the quantitative trait loci for high micronutrients uptake and response to Azotobacter inoculation were present on 1D, 5D, 6B, 7B chromosomes. This indicated good scope for wheat improvement through analytical breeding at chromosome level.ThesisItem Open Access Molecular characterization of multicut forage sorghum [Sorghum bicolor (L.) moench] using SSR markers(CCSHAU, 2012) Yadav, Anita; Boora, K.S.Sorghum is a diverse genus belonging to the tribe Andropogoneae and family Poaceae. (2n=20). The present study was undertaken to analyze genetic variability among F2 progenies of a cross between HJ541 (single cut variety) and SSG59-3 (multicut variety) by using SSR markers. Fifty SSR primers were used to study genetic variability among 32 F2 progenies along with the parents. A total of 78 amplified DNA products were observed out of which 26 were monomorphic and 52 were polymorphic. Average percentage polymorphism across the 32 progenies was 46.2%. The number of amplified DNA bands varied from 2 to 7 with an average of 1.81 bands per primer. The size of amplified alleles ranged from 110- 1000 bp respectively. Analysis of data grouped the 32 progenies into two major clusters. Cluster I constitutes of 29 F2 progenies and the parent SSG 59-3 whereas Cluster II had the remaining F2 progenies and parent HJ 541. Genetic similarity based on ‘Simqual’ l sub programme among various progenies ranged from 0.45 to 0.81 indicating moderate genetic variability among F2 population along with their parents. The average similarity between 32 progenies was found to be 0.66. Parental varieties were found to be most diverse followed by F2 progenies G18 and G7 with a coefficient of 0.45. F2 progeny G12 and G13, G9 and G11, G9 and G30, G29 and G30 were most similar with 0.81 coefficient of similarity. Two and three dimensional principle component analysis (PCA) also gives similar clustering of 32 F2 progenies. The highly diverse F2 progenies may be used for breeding of multicut varieties of sorghum in order to meet ever increasing demand of green fodder for the livestock. SSR would provide adequate coverage of genome for germplasm identification and pedigree validation.