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Theses

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2020 - 20227
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Subject: Biotechnology and molecular Biology × End date: 2022 × Start date: 2020 × Thesis Type: M.Sc ×
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Now showing 1 - 7 of 7
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    ThesisItemOpen Access
    Physiological and biochemical changes in selected Brassica juncea varieties under drought stress (Brassica juncea L. Czern. & Coss.)
    (CCS HAU, Hisar, 2022-08) Sharma, Reena; Lakra, Nita
    Mustard is the major oilseed crop that can assist meet India's demand for edible oil, and it ranks third after soybean and oil palm as a source of edible oil. As a result of the drought, its development and seed yield output have been seriously hindered. Therefore, the present study aimed to screen for drought tolerance at different stages among six (RH1566, RH725, RH749, RH725, RH761 and RH30) varieties of Indian mustard. These six varieties were grown as hydroponic in green house and for further analysis four varieties (RH30, RH 8812, RH1566 and RH725) were selected. Screening was done using PEG-6000 treatment to maintain different drought level. Various morpho-physiological (chlorophyll content, root and shoot length, RWC, osmotic potential, Electrolyte leakage), Biochemical, Metabolites were analysed at seedling and pot level. By investigating this study, we found the increasing trend of protective osmolyte (sugar, proline, glycine betaine) and increased activity of oxygen scavenging enzymes (Catalase, ascorbic peroxidase, superoxide dismutase) in severe stress (50 Field capacity %) as compared to its control. Plants under drought stress indicated a significant increase in antioxidative enzyme but this increase was more pronounced in drought tolerant variety. The content of ascorbic acid, carotenoids, total glutathione content were also recorded higher in tolerant variety than in sensitive variety under drought stress. We also studied PAL enzyme activity and reported increased activity under drought stress. This study helps Plant breeders and plant physiologists to design particular ways to alleviate the negative impacts of drought and maximise Indian mustard crop output by better understanding the processes of drought tolerance in Indian mustard.
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    ThesisItemOpen Access
    In-silico characterization and expression analysis of genes responsible for Zn uptake in wheat-Aegilops lines
    (CCS HAU, Hisar, 2022-07) Raja Kumar; Upendra Kumar
    Zinc deficiency is the most ubiquitous micronutrient deficiency in the world affecting nearly 2.6 billion peoples resulting in disease such as growth retardation, hypogonadism, immune dysfunction. It is a particular problem in developing countries, where most people rely on cereals as their staple food. One-third of the world’s cereal growing area is deficient in Zn. Wheat (Triticum aestivum L., 2n = 6x = 42) belongs to family Gramineae is a major cereal crop grown all over the world constitutes approximately 30% of the total cereals consumed by world’s population making it a major source of minerals. Therefore, it is imperative that wheat cultivars with improved concentrations of Zn is necessary to be developed to alleviate malnutrition among peoples. Lack of genetic variation in wheat is a major drawback in mineral biofortification and bioinformatics tools can be used as a potential resource in investigation of genes related to Zn homeostasis. In the present investigation 7 Zn regulatory gene families namely NAS, NAAT, DMAS, ZIP, ZIFL, YSL and TOM had been identified and analyzed for their importance in biosynthesis of phytosiderophores, their uptake and translocation under Zn deficiency. Chinese spring, an elite wheat landrace and UPHAU-3, a wheat-Aegilops addition/substitution line were grown hydroponically under Zn sufficient and deficient conditions. The relative expression of NAS, NAAT, DMAS, ZIP, ZIFL, YSL and TOM measured in UP HAU-3 and CS revealed that DMAS, ZIP and TOM genes are the most potential genes majorly responsible for Zn uptake and transport by release of phytosiderophores under Zn deficiency. The content of phytosiderophores released in the root exudates induced under Zn deficiency was found 17.41% more higher for UPHAU-3 as compared to CS. Therefore, this comparative study represents that under Zn deficiency UPHAU-3 was more potential genotype for more Zn uptake and transport in comparison with Chinese spring, which can be used as a promising wheat genotype for further studies to make Zn biofortified wheat crop for mitigating Zn malnutrition in human population.
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    ThesisItemOpen Access
    Morpho-physiological, biochemical and protein profiling under drought stress in selected chickpea (Cicer arietinum L.) varieties
    (CCSHAU Hisar, 2022-07) Yama Keerthi Sree; Lakra, Nita
    Chickpea (Cicer arieitnum L.) is an important food legume crop of the family Fabaceae with high protein levels that is widely grown in rainfed areas prone to drought stress. It is a self-pollinated cool season crop with true diploid (2n=16) nature. They are relatively cheaper and higher source of protein. About 90% chickpea crop is grown in rainfed conditions without depending on irrigation. In the present study, two varieties of chickpea namely ICC 4958 and HC-6 were grown under 3 field capacities (100% FC, 50% FC and 25% FC). Sampling was taken 3 times i.e., 15, 30 and 45 days after sowing (DAS). Parameters like morphological, physiological, biochemical, metabolite and protein profiling of the two varieties was done. Under morphological parameters, shoot length, number of branches number of leaves, number of flowers declined as the drought level increased. Under physiological parameters, relative water content (RWC), cell membrane stability (CMS), chlorophyll content decreased in both the varieties as field capacity decreased. Under biochemical parameters, proline, sugar, glycine betaine, superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR) increased as drought stress increased. Under metabolite analysis, MDA (Malondialdehyde) content, total anthocyanin, flavonoid content, phenolic content increased as drought progressed. We also performed protein profiling of the two varieties using SDS-PAGE (Sodium dodecyl-sulfate polyacrylamide gel electrophoresis) to differentiate the expression analysis of the two varieties.
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    ThesisItemOpen Access
    Development of an efficient protocol for micropropagation of strawberry (Fragaria ananassa Duch.)
    (CCSHAU, Hisar, 2020-07) Swati Rani; Upendra Kumar
    Strawberry (Fragaria × ananassa Duch.) is the most important fruit worldwide. Micropropagation is mainly for the clonal multiplication.Strawberry cvs. Grenada and Petaluma were micropropagated for rapid shoot and root multiplication. Meristematic part and leaf primordia is source of explants. Foe the sterilization process, the explants were dipped in two to three drops of Tween 20 per 100 ml for 10 minutes with distilled water, 0.5% Bavisitin & Streptocycline for 30 mintues with antimicrobial supplement and 0.1% HgCL2 for 2 minutes gave the maximum aseptic cultures. After the surface sterilization of meristemetic part of plants 3-5 mm long was used as a explant. Multiplication stage results indicate that highest auxiliary buds were observed when MS medium supplemented with 1 mg/l and 1.5 mg/l BAP in Petaluma and Grenada. At rooting stage, it was clear visually that MS medium supplemented with 0.5 mg/l of BAP with 1.5 mg/l of IBA in both the cultivars gave the best results of enhanced number roots with higher length and number shoots with higher length per explants. The best result for root multiplication indicating the use of IBA with ( 1.0, 1.5 mg/l) concentration as compared to other treatments.The highest response for the shoot multiplication was obtained with MS containing 1.5 mg/l and 1.0 mg/l BAP respectively. The present research is very useful in commercializing the new cultivars of strawberry into north Indian conditions which gives higher yield of fruits.
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    ThesisItemOpen Access
    In silico identification and physical mapping of gene(s) responsible for β-glucan in Bread Wheat (Triticum aestivum. L)
    (CCSHAU, Hisar, 2020-07) Dhamija, Aryan; Upendra Kumar
    Dietary fibers from plant cell wall are an essential component of healthy foods. Higher intake of dietary fiber reduces the risk of diet related chronic disease like type 2 diabetes, obesity and also improves gastrointestinal health. In human diets, cereal fiber is the largest contributor to total dietary fiber consumption. The soluble dietary fiber (1-3) (1-4) mixed linked β-D-glucan from cereal grains is a valuable component of a healthy diet. In the present investigation, the gene responsible for β-glucan in hexploid wheat was identified using both in silico and molecular methods.The in silico analysis confirmed that CslF6 gene in wheat was located on the chromosome 7A. Sequence similarity search was conducted between HvCslF6 and wheat survey sequences displayed a similarity of 96% with chromosome 7A. The structure of this gene had 3 exons and codes for a protein of 945 amino acids. Homology modeling of putative CslF6 protein was described as Probable cellulose synthase A catalytic subunit 8.Different cytogenetic stocks of Chinese spring revealed that CslF6 gene was located on centromeric region of 7AL (FL=0.29). Identified CslF6 in wheat can be utilized to make β-glucan efficient wheat and can be used in biofortification program.
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    ThesisItemOpen Access
    Screening of wheat genotypes for multiple fungal resistance using functional markers
    (CCSHAU, Hisar, 2021-09) Garg, Bharat; Yashveer, Shikha
    In the present study, 58 wheat genotypes were characterized for grain yield, biochemical 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, 2019-2020 for the evaluation of ten yield and its component characters viz. plant height, number of tiller per meter, spike length, number of grain per spike, number of spikelet per spike, grain yield per meter, biological yield per meter, harvest index and 1000-grain weight. In this study 58 wheat genotypes were screened for morphological and biochemical traits, showed varied mean range signifying ample amount of variability present in the wheat genotypes Genetic variability of genotypes was assessed by PCV, GCV, heritability and genetic advance over mean. 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 maximum value of the GCV & PCV was observed for the grain yield per meter (11.87) and number of spikelet per spike (16.65). Highest broad sense heritability was recorded for biological yield per meter (80.92) The positive and significant genotypic correlation for grain yield per meter was found with number of tillers per meter (0.503), spike length (0.217), number of grains per spike (0.332), biological yield per meter (0.864), harvest index (0.616). 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. total phenolic content, total flavonoids, proline content, glycine betaine, phenylalanine ammonia lyase (PAL) and tyrosine ammonia lyase (TAL) at vegetative and maturity stages. Total phenolic content range for vegetative stage is 0.41 to 4.74 mg CE/g and 1.07 to 6.17 mg CE/g in the maturity stage. Total flavonoids range for the vegetative stage, is 0.30 to 0.97 mg QE/g and 1.07 to 9.18 mg QE/g in the maturity stage. Proline content range for vegetative stage is 0.07 to 0.45 mg /g and 0.08 to 0.55 mg /g in maturity stage. Glycine betaine range for vegetative stage is 0.59 to 9.59 mg /g DW and 2.67 to 4.24 mg /g DW in the maturity stage. PAL activity range for vegetative stage is 4.15 to 6.23 μmole/h/g and 4.99 to 6.51 μmole/h/g in the maturity stage. TAL activity range for vegetative stage is 1.45 to 12.43 μmole/h/g and 1.74 to 12.10 μmole/h/g in the maturity stage. Total flavonoids content had high GCV and PCV in biochemical traits. On heritability analysis, Tyrosine ammonia lyase (TAL) had higher heritability. High heritability coupled with high genetic advance was found in characters such as total phenolic content, total flavonoids content, proline content, glycine betaine and TAL. These characters are recommended in selecting genotypes for screening for biotic and abiotic stress. For functional marker analysis, a total of 24 functional markers were used showed amplification. These selected primers are linked with specific fungal disease resistance gene and amplification of specific primer on wheat genotypes gave the idea of presence of that allele.
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    ThesisItemOpen Access
    Estimation of phytosiderophores in wheat-Aegilops addition/substitution lines under iron deficiency
    (CCSHAU, Hisar, 2021-07) Bisla, Bindu; Upendra Kumar
    Wheat is a major cereal crop all over the world in terms of cultivated area and food source. Most elite wheat cultivars, on the other hand, have low grain iron (Fe) content. Over three billion people, mostly women and children, suffer from micronutrient deficiency, mainly Fe deficiency. However, plants have developed systems to ensure that Fe homeostasis is kept precisely regulated at all levels. The roots improve the nutrient availability in the rhizosphere by removing proton and phenolic compounds from their roots into the rhizosphere which is intended to improve the solubility of ferric ions (Strategy I in dicots) or the solubilization of Fe3+ and other metal micronutrients by phytosiderophores (PS) of the mugineic acid (MA) family secreted from roots (Strategy II, in monocots). In the present investigation, two wheat-Aegilops addition/substitution lines (UP HAU-3 and CS) were grown hydroponically under Fe sufficient and deficient condition. Phytosiderophore release in the root exudates was induced under Fe deficiency and was higher for UP HAU-3 as compared to CS. Significantly higher expression of NAS, NAAT, DMAS and Ids3 was measured in UP HAU-3 than CS under Fe deficiency. We can use biotechnological tools and techniques to introduce the gene responsible for phytosiderophores secretion into field crops. It will enhance the crop uptake in nutrient deficient conditions and indirectly crop yield.