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Theses

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2018 - 20195
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Subject: Biotechnology and Molecular Biology × End date: 2019 × Start date: 2018 × Type: Thesis × Thesis Type: M.Sc ×
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Now showing 1 - 5 of 5
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    ThesisItemOpen Access
    Molecular characterization of cluster bean (Cyamopsis tetragonoloba L. Taub.) genotypes
    (CCSHAU, 2019) Alka Rani; Yadav, Neelam R.
    Cluster bean (Cyamopsis tetragonoloba L. Taub.) is an annual, diploid legume crop with 14 number of chromosomes and highly valued for presence of galactomannan in its seeds. For commercial use of seeds, their genetic purity determination is very important. The present study was carried out for development of molecular profile of commercial varieties of cluster bean using SSRs and to study molecular diversity in selected genotypes of cluster bean using EST-SSRs. For this study, seeds of 22 commercial varieties and 41 germplasm lines of guar were grown in net house of the department and Farm area of Forage Section, Department of Genetics and Plant Breeding. DNA was extracted using CTAB method. Molecular analysis was carried out using 79 SSRs which included 40 EST-SSRs and 39 miSSRs from Medicago truncatula and cluster bean (developed in our lab). Out of 79 SSRs, 55 SSRs were found polymorphic and employed for varietal identification. Among these polymorphic SSRs, eight SSRs identified selected varieties. Ct-miSSR-17, Ct-miSSR-19, Mt-miRNA-SSR45, Mt-miRNA-SSR47 and Mt-miRNA-SSR164, EST-SSR2, EST-SSR17 and EST-SSR40 identified selected varieties. Among 55 polymorphic SSRs, size of amplified products ranged from 100-1160 bp and PIC value ranged from 0.28 to 0.91. The molecular analysis divided the 22 cluster bean varieties into two major clusters at similarity coefficient of 0.06 and HG 6 existed as an isolated genotype. Cross-transferability of Medicago truncatula miRNA-SSRs showed 31% cross-transferability among cluster bean varieties. Molecular analysis showed that IC 402295-2, IC 522486, IC 113452, IC 415161 and IC 113489 were extreme and diverse group with around 30 % gum content while IC 113472 and IC 370725 were low gum (<20%) genotypes. The molecular analysis of 63 genotypes together showed that all the 22 varieties are in different genetic pool than 41 germplasm lines and can be exploited in cluster bean breeding programme.
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    ThesisItemOpen Access
    Development and characterization of transgenic pigeon pea (Cajanus cajan L. Millspaugh) plants with Psp68 for improvement against salinity stress
    (CCSHAU, 2019) Chhabra, Neha; 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 salt stress is the most deleterious accounting for upto 100% yield losses. In the present study, transgenic pigeon pea plants (var. Manak) carrying Psp68 gene were developed for salt stress tolerance using Agrobacterium tumefaciens strain LBA4404 with pCAMBIA1300-Psp68 (Kharb et al., 2018). Putative transformants were screened at early stage through PCR amplification using gene specific primers and a transformation frequency of 16% was observed. Southern blot and real time PCR analysis revealed stable and single copy insertion of the transgene in pigeon pea genome. Seeds from each of the PCR positive T0 pigeon pea plants were sown to raise T1 generation and a total of 42 plants (out of 100) showed the presence of Psp68 gene. Physio-biochemical analysis of selected T1 transgenic plants subjected to 75 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 reduced as compared to the wild type plants under 75 mM stress. Among all transgenic lines, line 53 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.
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    ThesisItemOpen Access
    Genetic diversity analysis in Kiwifruit (Actinidia spp.) using RAPD/SSR markers
    (CCSHAU, 2019) Kamboj, Aarti; Kharb, Pushpa
    Kiwifruit (Actinidia spp.) is a very important plantation crop, generally dioecious, deciduous and scrambling vines with chromosome number 2n=58, belonging to family Actinidiaceae. Dioecy in kiwifruit lays an important constraint for breeding programs. The present study was undertaken to study genetic diversity in kiwifruit genotypes using RAPD markers. Molecular polymorphism in 7 kiwifruit genotypes (2 males viz. Tomuri & Allision and 5 females viz. Bruno, Hayward, Monty, Allision & Abbott) was evaluated using 94 RAPD primers, out of which 23 were found to amplify the genomic DNA in all the seven genotypes. Analysis of RAPD data was done using NTSYS-pc software and dendrogram was constructed using UPGMA method. The 7 kiwifruit genotypes grouped in two main clusters comprising of male and female genotypes separately. Similarity matrix indices ranged from 0.521 to 0.719 showing maximum similarity between Allision (M) and Tomuri (M) with a similarity coefficient of 0.719 while Allision (M) and Abbott (F) were found least similar with a similarity coefficient of 0.521. Primers OPA-01, OPB-02, OPB-20 and OPC-05 amplified unique amplicons in Bruno, Hayward, Abbott, Monty and Allision (M), and primers OPA-02 & OPN-02 amplified unique amplicons in Tomuri (M) & Allision (M) and male and female plants of Allision genotype respectively. Thus, these primers can be used to distinguish the kiwifruit genotypes and can also be validated as putative sex linked markers which have the prospective to be used for further characterization in specific kiwifruit genotypes.
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    ThesisItemOpen Access
    Phenotypic and molecular characterization of F5 and backcross generations for salt tolerance in wheat (Triticum aestivum L. em. Thell)
    (CCSHAU, 2019) Tiwari, Prakash Narayan; Shikha Yashveer
    Soil salinity is one of the major abiotic stress having multifarious adverse effects on all growth stages of wheat. In Haryana, WH 1105 is widely cultivated wheat variety with many agronomically superior characteristics but is affected by soil salinity. In the present investigationNax1 and Nax2 genes were introgressed from Kharchia 65 into WH 1105 for salinity tolerance through marker assisted backcross breeding. F5, BC1F4, BC2F3 and BC3F1 generations of WH 1105 x Kharchia 65, were evaluated under initial salt stress condition for various morphological traits. On the basis of genotypic and phenotypic variations 57 high yielding plants of WH 1105 x Kharchia 65, were selected. Out of total 210 screened SSRs 30 markers were found polymorphic for WH 1105 and Kharchia 65. These polymorphic SSRs were utilized to produce molecular diversity among selected plants. Cluster tree analysis of parents and all the four generations of the cross, portrayed that all the selected plants were inclined toward recurrent parent WH1105 indicating higher similarity with the recurrent parent. Seven best plants were selected from the cross as high grain yielding and salt tolerant plants. These plants could be further backcrossed with the recurrent parent to develop salt tolerant wheat lines.
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    ThesisItemOpen Access
    Phenotyping and molecular marker analysis in backcross and F4 generations of wheat for salt tolerance
    (CCSHAU, 2018) Varsha; Shikha Yashveer
    Wheat is a major staple food of world population and occupies about 21.8% of total cultivated area accounting for 35.5 percent of total food grain production at global level. In India, approximately 8.5 million hectare of land area is affected by high salinity (EC ≥ 5 dS m-1). In Haryana alone, 2Mha of soil is affected by salinity which contributes to 20% of the total yield loss. Development of salinity tolerant varieties through marker assisted breeding is most efficient and effective strategy for management of soil salinity. In Haryana, HD 2967 and WH 1105 are widely cultivated wheat varieties with many agronomically superior qualities but are affected by soil salinity. In the present study, two genes (Nax1 and Nax2) for salinity tolerance were introgressed from Kharchia 65 into the genetic background of wheat varieties, HD 2967 and WH 1105 through marker assisted backcross breeding and BC1F3, BC2F2 and F4 generations of the two crosses i.e. HD 2967 x Kharchia 65 and WH 1105 x Kharchia 65, were evaluated for various morphological traits under initial salt stress condition. On the basis of phenotypic and genotypic variations 68 and 44 plants of the cross HD 2967 x Kharchia 65 and WH 1105 x Kharchia 65, respectively, high yielding plants were selected. A total of 178 parental polymorphic markers were used for background selection of the foreground selected plants. Out of total 178 SSR markers screened, 31 markers were found polymorphic for the parents HD 2967 and Kharchia 65 and 30 markers were polymorphic for WH 1105 and Kharchia 65. These polymorphic SSR markers were used to produce molecular diversity among selected plants. Cluster tree analysis of parents and all the three generations of both the crosses, showed that all the selected plants were inclined toward recurrent parent HD 2967 or WH1105 indicating higher similarity with the recurrent parent. Six and four best plants were selected from the respective crosses as high grain yielding and salt tolerant plants. These plants could be further backcrossed with the recurrent parent to develop salt tolerant wheat lines.