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Sodium Dodecyl sulphate Polyacrylamide gel Electrophoresis Pattern of Horse Gram Seed Storage Proteins during Germination

Author Affiliations

  • 1Faculty of Agro Based Industry, University Malaysia Kelantan (Jeli campus), Beg Berkunci No. 100, 17600 Jeli, Kelantan, MALAYSIA
  • 2Faculty of Earth science, University Malaysia Kelantan (Jeli campus), Beg Berkunci No. 100, 17600 Jeli, Kelantan, MALAYSIA
  • 3Faculty of Agro Based Industry, University Malaysia Kelantan (Jeli campus), Beg Berkunci No. 100, 17600 Jeli, Kelantan, MALAYSIA

Int. Res. J. Biological Sci., Volume 1, Issue (4), Pages 39-50, August,10 (2012)

Abstract

The aim of the experiment was to study the effect of germination on the three varieties of horse gram seed storage proteins. Seeds were germinated for 24, 48, 72, 96 and 110h in a dark place. Germinated seeds were frozen at -18C for 12h to stop the germination process. Spouted seeds were freeze-dried and ground to pass through a 40 mesh sieve for analysis. Total proteins, water soluble and water insoluble proteins were estimated by standard methods. Seed storage protein profiles of three varieties germinated horse gram were analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Extracted protein fractions from germinated horse gram seeds in different solvents were studied by HPLC. The highest amount of storage protein degradation was observed in all varieties after 72-80h of germination. Characterization of protein fractions by HPLC showed that albumins/globulins, prolamins and glutelins increases slightly during germination time, as germinating seeds usually accompany by interconversion and production of new compounds. Electrophoregrams for each variety were shown and the high molecular weight proteins band intensity decreases during different germination period were noted. Genetic diversity of germinated horse gram was evaluated by constructing the dendrogram for high molecular weight (HMW) and low molecular weight (LMW) gluten subunit bands. In conclusion, SDS-PAGE of germinated seed storage proteins can be economically used to assess genetic variation and relation in germplasm. The specific bands of germinated seed storage protein profiles may be used as markers for identification of the mutants/genotypes

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