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20192
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Subject: Biochemistry × End date: 2019 × Start date: 2019 × Type: Thesis × Thesis Type: Ph.D ×
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    Biochemical and proteomic analysis of botrytis grey mould infected susceptible and resistant chickpea lines
    (Punjab Agricultural University, Ludhiana, 2019) Thakur, Richa; Sharma, Sucheta
    Present investigation reports the morphological, ultrastructural and biochemical analysis of botrytis grey mould infected susceptible (PBG5) and resistant (C. pinnatifidum) chickpea genotypes. Seedlings of both the genotypes were inoculated with pregrown culture of B. cinerea (1 x 104 spores mL-1), using growth chamber and cut twig techniques. Inoculated leaves from both the genotypes were collected at 1, 3, 5 and 7 days post inoculation and used for further analysis. Visual disease symptoms were observed only in susceptible genotype PBG5 in terms of conidial colonies and water soaked lesions on leaves and stems. Scanning electron microscopic analysis revealed differences in cell surface topography between two genotypes with more number of stomata in PBG5 which were further validated by stomatal frequency analysis. Fourier transform infrared spectra of proteins were almost similar for both the genotypes. B. cinerea inoculation resulted in the induction of biochemical constituents viz. total phenols, malondialdehyde content and activities of CAT, APX, SOD, POX, PPO and PAL in both the genotypes with higher activities in resistant genotype than susceptible one at different days post inoculation. Isoenzymic analysis revealed the presence of one isoform of catalase, two in superoxide dismutase & three each in guaiacol peroxidase and polyphenol oxidase at 1 to 7 days postinoculation. SDS-PAGE analysis revealed absence of polypeptide bands with Rf (0.5, 0.21, 0.49 and 0.64) in PBG5 and (0.44, 0.60, 0.64 and 0.34) in C. pinnatifidum leaves at 3-7 days post B. cinerea inoculation in comparison to respective controls. At 5 and 7 days postinoculation, bands of molecular weight (97.4 and 66 kDa) were missing in the C. pinnatifidum leaves. Uninoculated chickpea genotypes, screened for botrytis responsive genes using PCR approach, indicated the presence of DNA band (~300 bp) in resistant genotype (C. pinnatifidum) suggesting its possible role for resistance against B. cinerea.
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    ThesisItemOpen Access
    Biochemical basis of micronutrient deficiency tolerance in triticale x wheat derivative lines
    (Punjab Agricultural University, Ludhiana, 2019) Ravneet Kaur; Bhatia, Surekha
    The present study was aimed to investigate the biochemical basis and molecular marker profile of genes associated with micronutrient uptake in parental lines viz rye, triticale (TL 2908, TL 2942), bread wheat lines (PBW 343, PBW550, HD 2967, HD 3086) and durum wheat lines (PDW 233, PDW 291, WHD 943). Five days old seedlings of these parental lines were transferred to three set of treatments (T1-control, T2- micronutrient deficient (stress), T3-with micronutrient supply (alkaline stress pH-9) for 30 days. Rye and triticale lines have longer, hairy and thick roots with high number as compared to bread wheat and durum wheat lines. Roots and shoots of rye and triticale lines accumulated higher fresh/dry weight than bread wheat and durum wheat lines irrespective of treatments. Both stressed conditions caused reduction in roots/shoots fresh weight and dry weight of all parental lines however, rye and triticale lines accumulated greater biomass under alkaline stress conditions on 21 DAT. Rye and triticale lines had higher phytosiderophore content (118 to 498 µmole of Cu equivalents/gFW) than bread wheat (19 to 112 µmole of Cu equivalents/gFW) and durum wheat (6 to 68 µmole of Cu equivalents/gFW) under different treatments during whole period of experiment. In rye and triticale, it was increased from 7 DAT to 21 DAT and decreased afterward on 30 DAT. However, in bread wheat and durum wheat lines, this increase was found only up to 14 DAT. Increase or decrease in release of PSs from roots of parental lines was positively correlated with the higher micronutrient content in roots (r=>0.9) and shoots (r=>0.8) of these lines. Highest micronutrient efficiency was found in rye (73.62-86.08%) followed by triticale lines (70.61-81.06%), bread wheat lines (36.82-66.14%) and durum wheat lines (29.42-62.38%). Further higher activities of SOD, CAT and CA enzymes in rye and triticale lines than bread wheat and durum wheat lines were found which was probably linked with higher micronutrient efficiency corresponding to efficient utilization of micronutrients in rye and triticale lines. Rye and triticale also exhibited higher activities of NAAT and MAS enzymes. Parental lines and triticale X wheat derivative lines were tested for presence of 5R chromosomes using seven 5R chromosome specific markers. Only three markers (SCM 138, KU 140, KU 640) showed expected polymorphism in parental lines were further used to evaluate triticale X wheat derivative lines. Fifteen rye specific primers of Ids3 gene were designed after in-silico sequence analysis. Five primers showed polymorphism in parental lines as well as triticale X wheat derivative lines. Higher release of PSs was estimated from the roots of triticale X wheat derivative lines which were positive for Ids3 and 5R chromosome specific markers. Fourteen triticale X wheat derivative lines with higher PSs under micronutrient deficient conditions, were selected for their further utilization in wheat breeding program for improvement and development of efficient varieties with higher nutritional value.