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Characterization of embryogenesis‐related proteins in alfalfa ( Medicago sativa )
Author(s) -
Giroux Randal W.,
Pauls K. Peter
Publication year - 1996
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.1996.tb00230.x
Subject(s) - medicago sativa , biology , somatic embryogenesis , gel electrophoresis , in vitro , embryo , biochemistry , genotype , medicago , polyacrylamide gel electrophoresis , molecular mass , microbiology and biotechnology , botany , tissue culture , gene , enzyme
Large samples of embryogenic cells can be obtained from tetraploid alfalfa ( Medicago sativa L.) suspension cultures following induction by auxin and culture on growth regulator‐free medium. A one–dimensional sodium dodecyl sulphate‐polyacrylarnide gel electrophoresis (SDS‐PAGE) comparison of the polypeptides synthesized by embryogenic and nonembryogenic alfalfa genotypes during inductive conditions identified a 50‐kDa polypeptide band that was specific to embryogenic genotypes. Two‐dimensional (iso electric focusing SDS‐PAGE) electrophoretic separations resolved the 50–kDa band into three 50‐kDa polypeptides with different isolectric points. A comparison of the polypeptides in the soluble and microsomal membrane fractions from embryogenic cell masses of an embryogenic genotype suggested that the 50‐kDa pep‐tides are membrane associated. The in vitro translation products from RNAs of an embryogenic genotype had a 50‐kDa band that was absent from the in vitro translation products obtained from RNAs of a nonernbryogenic genotype. Therefore, we concluded that the membrane‐localized 50‐kDa proteins are involved in somatic embryo–genesis in alfalfa and that their expression is transcriptionally determined.