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20203
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Subject: Biotechnology and molecular Biology × End date: 2020 × Start date: 2020 × Type: Thesis ×
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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
    Molecular approaches for detection and forecasting of wheat yellow rust
    (CCSHAU, Hisar, 2020-03) Rizwana Rehsawla; Yadav, Neelam R.
    Yellow or stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is a devastating airborne disease that affects bread wheat in the major wheat growing regions of India. The understanding of the origin, evolution, pathogenicity, avirulence/virulence behaviour of Pst is very important for the development of more effective management strategies to combat the disease spread. To understand inter and intraspecific phylogenetic relationship among Indian Pst pathotypes, multigene sequence analysis was done. Molecular marker study along with sequencing technology was used to collect information, which was more effective than virulence characterization. The molecular diversity analysis among 13 different Pst pathotypes showed two major cluster formations at similarity coefficient of 0.78. Sometimes, all three wheat rust or two rusts in combination occur simultaneously in the field. Under such conditions identification and differentiation of the yellow rust is needed for precise identification and high throughput DNA based detection protocols. To address this problem PCR based markers were developed which can specifically detect and differentiate Pst from two other rust species of Puccinia and other wheat pathogens. DNA-based methods such as conventional PCR have revolutionized plant disease detection; they are not very reliable at asymptomatic stage. Therefore, a simple and reproducible LAMP assay was developed which could detect the pathogen at an early stage i.e. 3rd day of post infection without any visible sign of pathogen attack on the leaf sample using LAMP primers available in public domain. Four novel sets of LAMP primers from ketopantotate reductase gene were also designed for Pst detection which worked successfully. Biosensing by electrochemical and SPR for the detection of yellow rust was also undertaken. Electrochemical based sensing was done using different sequences of Pst specific genes as probes. Linear response over wide DNA concentration range from 10 pg/μl to 115ng/μl was obtained with a high sensitivity, accuracy and reproducibility. The lowest detection limit was observed for microRNA like RNA 1 gene i.e. 10 pg/μl. Electrochemical DNA based biosensing was developed to distinguish between the yellow rust susceptible and resistant wheat genotypes using TaATG8j gene sequence. Linear response over wide DNA concentration range from 1 ng/μl to 50 ng/μl was obtained with detection limit of 4 pg/μl. The SPR biosensor demonstrated high specificity and long shelf life thus promising its application in Pst diagnosis. The developed biosensor exhibited a high sensitivity (0.18°/ (ng/μl)), good linearity, low detection limit (1 ng/μl) and high specificity over a wide concentration range of DNA (1–150 ng/μl).