Serodiagnosis of pearl millet resistance to downy mildew by quantitating cell wall P/HRGP using polyclonal antiserum Pab-P/HRGP |
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Authors: | Shantharaj Deepak Sekhar Shailasree Neerakkal Sujeeth Ramachandra K Kini Axel Mithöfer Shekar H Shetty |
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Institution: | 1. Department of Studies in Applied Botany and Biotechnology, University of Mysore, Manasagangotri, Mysore, 570006, Karnataka, India 2. Department Bioorganic Chemistry, Max-Planck-Institute for Chemical Ecology, Hans-Kn?ll-Stra?e 8, 07745, Jena, Germany
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Abstract: | Proline/hydroxyproline-rich glycoprotein (P/HRGP) level in pearl millet genotypes resistant to downy mildew increase after
inoculation with the oomycete pathogen Sclerospora graminicola. Using purified P/HRGPs from pearl millet cell walls, polyclonal antibodies (Pab-P/HRGP) were raised in rabbit. Based on this
antiserum, an enzyme immunoassay was developed that displays a linearity detection range from 0.01 to 10 μg P/HRGP. Western
blot analysis, confirming the induction of three marker P/HRGPs in the infected resistant genotype, and immunocytochemical
studies on P/HRGP localization either in epidermal peelings or in suspension-cultured cells demonstrated the specificity of
the antiserum. Besides its characterization, Pab-P/HRGP was employed to screen various genotypes of pearl millet for fast,
sensitive and specific detection of induced P/HRGPs upon infections. The results presented are discussed with presumed importance
to downy mildew disease and the use of this new antiserum in pearl millet screening for disease resistance. |
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Keywords: | Downy mildew Enzyme immunoassay Immunological quantification Proline/hydroxyproline rich-glycoproteins |
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