Thumbnail Image

Theses

Browse

20196
Reset filters
Subject: Biotechnology and molecular Biology × End date: 2019 × Type: Thesis ×
Reset filters

Search Results

Now showing 1 - 6 of 6
  • Thumbnail Image
    ThesisItemOpen Access
    Evaluation of advanced wheat lines for stripe rust resistance and grain quality
    (CCSHAU, Hisar, 2019-08) Bhawna; Bhawna; Yadav, Neelam R.; Yadav, Neelam R.
    Biofortification of staple foods like wheat without any yield penalty is of utmost importance. Advanced lines introgressed with Gpc-B1 allele were used in the study. The Gpc-B1 allele co-segregated with Yr36 yellow rust resistance gene in all the advanced lines. All WH711-derived advanced wheat lines confirmed for the Yr36/Gpc-B1 allele. Seven advanced lines (WL-8-1, WL-8-2, WL-8-3, WL-8-5, WL-8-6, WL-8-7, WL-8-8) were confirmed for Yr18 while six advanced lines (WL-8-1, WL-8-2, WL-8- 6, WL-8-7, WL-8-8, WL-8-9) were positive for Yr17 gene. All five PBW343-derived advanced lines confirmed for the Yr36/Gpc-B1 allele presence. Three advanced lines (PL-8-2, PL-8-3, PL-8-4) displayed the presence of the Yr18 gene. Two advanced lines (Pl-8-3, PL-8-5) confirmed presence of the Yr17 gene. All the advanced wheat lines derived from WH711 as well as from PBW343 displayed very high levels of resistance against yellow rust and better agronomic characteristics than the parents. Positive correlation was observed in yield and yield related traits. The hectoliter weights of PBW343-derived lines indicated their superiority in terms of good flour recovery than PBW343 and comparable to the checks. WH711- and PBW343- derived advanced wheat lines show better gluten quality and high quantity in sedimentation test. Gpc-B1 allele introgression provided high GPC in WH711 derived advanced wheat lines. Iron and zinc contents of both WH711-derived and PBW343-derived advanced wheat lines were at par when compared to either parent and checks, indicating successful bio fortification of both micronutrients. Two of the WH711-derived lines are being further tested in IPPSN/station trials.
  • Thumbnail Image
    ThesisItemOpen Access
    ―Phenotypic and molecular characterization of HD 2967 x Kharchia 65 F5 and backcross generations for salt tolerance in wheat (Triticum aestivum L. em. Thell”
    (CCSHAU, Hisar, 2019-08) Saini, Pooja; Yashveer, Shikha
    Soil salinity is emerging environmental stress that reduces rate of germination, delays emergence and affects almost all stages of growth and development of wheat plants which ultimately leads to reduction in grain yield of crop.The present investigation focused on phenotypic and molecular characterization of selected wheat (Triticum aestivum L. em. Thell)genotypes of F5 and backcross (BC1F4, BC2F3, BC3F1) generations derived from HD 2967 X Kharicha 65 for salinity tolerance on the basis of presence of both Nax loci and their agronomic performance at salt stress level 8 dS/m. A total of 92 plants confirmed the presence of Nax1 locus at a band size of 210 bp consisting of 16 F5, 41 BC1F3, 9 BC2F2 and 27 BC3F1, a total of 53 plants confirmed the presence of Nax2 locus at a band size of 225 bp comprising of 6 F5, 35 BC1F4, 8 BC2F3 and 4 BC3F1.Overall, plants comprising of 6 F5,26 BC1F4, and 4 BC3F1 of HD 2967 x Kharchia 65were found to have both the Nax loci. On the basis of phenotypic and genotypic variations, 56 high yielding plants were selected for genotyping using polymorphic SSR markers. A total of 180 SSR primers were used to study the polymorphism between parents HD 2967 and Kharchia 65, out of which 27 SSR primers showed polymorphism. Polymorphism information content (PIC) value in the studied genotyped ranged from 0.191 (xgwm480) to 0.425 (wmc3) with an average value of 0.339. Cluster tree analysis using NTSYS-pc software, F5 and backcross (BC1F4, BC2F3, BC3F1) generations showed that all the selected plants were inclined toward recurrent parent HD 2967 representing higher similarity with HD 2967. From above analysis, high grain yielding and salt tolerant plants were selected such as plant number 259Q3c, 271Q3c (F5); 202P9b, 303P2b and 202P3b (BC1F4); 241R2c and 243R1a (BC2F3); 25(3)1b and 24(1)1c (BC3F1) and these plants could be further backcrossed with the parent HD 2967 to develop salt tolerant wheat lines.
  • Thumbnail Image
    ThesisItemOpen Access
    Characterization of wheat germplasm for grain yield, quality traits and functional markers
    (CCSHAU, Hisar, 2019-08) Verma, Priyanka; Yashveer, Shikha
    In the present study, 98 wheat genotypes were characterized for grain yield, quality traits and functional markers. The wheat genotypes were sown in RBD (Randomized Block Design) design in two replications in the field of Wheat and Barley Section, Department of Genetics and Plant Breeding, CCS Haryana Agricultural University, Hisar, Haryana (India) during Rabi, 2018-2019 for the evaluation of nine yield and its component characters viz. plant height, number of effective tiller per meter, spike length, number of grain per spike, number of spikelet per spike, 1000-grain weight, grain yield per plot, biological yield per plot. The values of GCV were lower than their corresponding PCV for all the characters studied, indicated the negligible effect of environment on the development of plants. The highest value for GCV and PCV was observed on biological yield per plot (18.09 & 19.18 %) followed by harvest index (15.76 & 16.04 %) and number of grains per spike (14.29 & 14.60 %), respectively. Heritability for broad sense (h2) showed highest for harvest index (96.50 %) followed by number of grains per spike (95.90 %) and biological yield per plot (89%). Genetic advance as 5% of mean was observed highest for biological yield (35.17%) and harvest index (31.89%), indicating a good scope for improvement in grain yield and its component characters. Grain yield per plot was highly and significantly positive correlated with number of effective tillers per plot (0.448), spike length (0.551), number of grains per spike (0.611), number of spikelets per spike (0.588), 1000 grain weight (0.585), biological yield per plot (0.805) and harvest index (0.113). Path analysis revealed that biological yield per plot had the maximum direct positive effect on grain yield per plot (0.600) followed by harvest index (0.573), whereas, all traits contributed indirectly towards grain yield per plot via biological yield per plot except plant height (-0202). The grain yield component traits viz. biological yield per plot, harvest index and number of grains per spike were showing high GCV, PCV, heritability and genetic advance as 5% of mean. Thus, these component traits may be used in wheat breeding programme for further improvement in grain yield. For characterization of quality traits, various biochemical attributes were analyzed viz. protein content, mineral (Fe, Zn), phytates, gluten content, amylose content, amylopectin content, hectolitre weight and sedimentation value. Protein content ranged from 9.4 to 15.2 per cent with mean of 12.2 per cent, iron content from 21.3 to 39.3 mg/kg with mean of 29.3 mg/kg, zinc content from.15.7 to 40.70 mg/kg with mean of 29.60 mg/kg, phytates (μg/g) from 4816.0 to 8757.3 (μg/g) with mean of 7008.5 μg/g, wet gluten content from 8.00 to 37.90 per cent with mean of 24.20 per cent, dry gluten content ranged from 3.00 to 14.11 per cent with mean of 10.96 per cent, total soluble sugar content from 0.86 to 6.16 per cent with mean of 3.05 per cent, starch content from 49.8 to 69.4 per cent with mean of 56.7 per cent, amylose content from 3.02 to 8.45 per cent with mean of 5.48 per cent, amylopectin content from 49.36 to 68.97 per cent with mean of 56.17 per cent, hectolitre weight from 72.0 to 87.30 kg/hl with mean of 80.10 kg/hl, sedimentation value from 19 to 41 ml with mean of 31.2 ml. For functional marker analysis, a total of 6 functional markers were used showed amplification
  • Thumbnail Image
    ThesisItemOpen Access
    ISSR Markers Based Genetic Diversity Analysis in Kiwifruit (Actinidia deliciosa (A.Chev.) CF Liang et AR Ferguson) Genotypes
    (CCSHAU, Hisar, 2019-09) Jhilta, Agrim; Kharb, Pushpa
    Kiwifruit (Actinidia deliciosa (A. Chev.) CF Liang et AR Ferguson) (2n=6x=174) also known as Chinese goose berry and the horticultural wonder of New Zealand, is a commercially important crop and a very good source of Vitamin C. The present study was undertaken for assessing genetic diversity in Kiwifruit genotypes using ISSR primers and for identification of any probable ISSR marker linked to sex in Kiwifruit genotypes. A total of seven genotypes of Kiwifruit were used that included two male (Tomuri and Allison (M)) genotypes and five female genotypes (Allison, Abbott, Bruno, Hayward, Monty). Genomic DNA was isolated from young leaves of Kiwifruit genotypes. For molecular analysis 104 ISSR primers were screened, out of which 24 ISSR primers amplified and were polymorphic. These were employed for genetic diversity analysis and identification of genotype. Among these polymorphic ISSRs, eight ISSRs were found to identify selected genotypes on the basis of unique amplicons. IS7, IS11, IS93, IS88, IS114, IS107, IS101 and IS79 could identify genotypes . One SSR marker, A 003 reported earlier was also validated. It could identify Allison (M). Similarity matrices of seven kiwifruit genotypes revealed that similarity value among different genotypes ranged from 0.5298 to 0.7094. A maximum similarity value of 0.7094 was observed between Allison (M) and Tomuri whereas Abott and Allison (F) were found to be most diverse. Molecular analysis showed that Allison (M) and Tomuri were much similar. Abbott was found most diverse when compared to other kiwifruit genotypes. The results of present study can be utilized for sex and genotype identification at seedling stage and in kiwifruit improvement programme.
  • Thumbnail Image
    ThesisItemOpen Access
    Development of micro-RNA and EST-SSR markers for diversity analysis in clusterbean [Cyamopsis tetragonoloba (L.) Taub]
    (CCSHAU, Hisar, 2019-09) Chaudhary, Vrantika; Yadav, Neelam R
    Cluster bean [Cyamopsis tetragonoloba (L.) Taub] is an important industrial crop due to the presence of soluble dietary fibre, high protein content and high value gum. MicroRNA are tiny non-coding small RNAs master regulators that play important role in multiple biological processes by degrading targeted mRNAs or repressing mRNA translation. In silico computational analysis based on Cluster bean ESTs led to identification of 57 miRNA families along with 37 targets. The length of most of the mature miRNAs was found to be 21nt long and average MFE value of 25.4 (-kcal/mol). The gene ontology study revealed the genes encoding transcription factors or enzymes involved in the regulation of development, growth, metabolism, physiological processes and stress response. Identification of transcription factors and miRNA resulted into model prediction. Most abundant miRNA families predicted in this study were miR156, miR172 and miR2606. The miRNA related to gum metabolism were also identified i.e. miR5658, miR1533 and miR414. A total of 40 unigene SSRs were screened among hundred germplasm lines including selected cultivars. The molecular analysis divided the 100 cluster bean genotypes into two major clusters at similarity coefficient of 0.49. Genic SSRs related to galactomannan metabolism were also used for diversity analysis of low and high gum clusterbean genotypes. A new class of functional and regulatory markers i.e., miRNA-SSRs have been identified in Cluster bean with mononucleotides repeats in highest proportion (76%) followed by di- (10%), tri- (10%) and tetranucleotides (4%) in the pre-miRNA sequences. Cross transferability of Medicago truncatula miRNA SSRs showed 34% cross transferability among Cluster bean genotypes. A key enzyme in galactomannan metabolism, galactosyltransferase was used for structure prediction using in silico approaches. In silico molecular dynamics revealed its stability and structure was dominated by random coil and α-helix followed by β sheet region. Cluster bean galactosyltransferase sequence showed highest similarity with fenugreek and least similarity with Medicago truncatula
  • Thumbnail Image
    ThesisItemOpen Access
    Development of transgenic pigeon pea (Cajanus cajan L.) plants containing Lectine receptor like kinase gene for improving abiotic stress tolerance
    (CCSHAU, Hisar, 2019-05) Pratibha; Kharb, Pushpa
    Pigeon pea (Cajnus cajan L.) is a nutrient rich, sixth most important and second most important legume pulse crop of the world and India respectively after chickpea. Production and productivity of pigeon pea is affected by various abiotic stresses but salinity is the most deleterious stress which causes considerable loss in crop’s productivity. In the present study, transgenic pigeon pea plants (cv.. Manak) carrying OsLecRLK gene, were developed for salinity tolerance using Agrobacterium tumefaciens strain LBA4404 with pCAMBIA1300- OsLecRLK gene (Kharb et al., 2018 Patent Application No. 201811012099). Putative transformants were screened through PCR amplification using gene specific primers and out of 86 plants 16 plants were found positive giving a transformation efficiency of 18.6%. Six higher yielding lines were selected and checked for stable transgene integration. Southern hybridization and Real-time PCR analysis was done to find out the copy no. of transgene in selected transgenic lines. Seeds from T0 pigeon pea plants were sown to raise T1 generation and a total of 52 plants (out of 120) showed the presence of OsLecRLK gene. Physiobiochemical analysis was done to assess the efficacy of transgene via subjecting wild type and selected T1 transgenic plants to 75 mM salt stress. The results showed that transgenic line performed better in terms of maintaining higher chlorophyll content, relative water content, proline content, total soluble sugar content, peroxidase and catalase activity in comparison to the wild type plants. Moreover, MDA content and membrane injury index were significantly reduced in transgenic lines when compared to the wild type plants indicating that the transgenic lines were less affected by salt stress. Among all transgenic lines, L-19 performed well with respect to all the parameters studied and can be taken further for the development of transgenic pigeon pea plants for salt stress tolerance.