Biochemical and proteomic analysis of botrytis grey mould infected susceptible and resistant chickpea lines
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Date
2019
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Punjab Agricultural University, Ludhiana
Abstract
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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