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Culture of cotyledons of Douglas‐fir on a medium for the induction of adventitious shoots induces rapid changes in polypeptide profiles and mRNA populations
Author(s) -
Campbell Michael A.,
Gaynor John J.,
Kirby Edward G.
Publication year - 1992
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.1399-3054.1992.tb04721.x
Subject(s) - rna , shoot , polyacrylamide gel electrophoresis , messenger rna , biology , population , gel electrophoresis , biochemistry , dry weight , protein biosynthesis , microbiology and biotechnology , botany , gene , enzyme , demography , sociology
Cotyledons of 3‐ to 4‐week‐old seedlings of Douglas‐fir [ Pseudotsuga menziesii (Mirb). Franco] were treated with shoot induction medium (SIM) containing 5 μ M 6N‐benzylaminopurine (BAP) and 5 n M naphthaleneacetic acid (NAA). Fresh weight, dry weight and soluble protein levels were not altered within the first 48 h of SIM treatment. SIM‐treated cotyledons were labelled in vivo with 35 SO 4 2‐ , and TCA‐insoluble proteins were analyzed electrophoretically by a 2‐dimensional system consisting of non‐equilibrium electrophoresis (NEPHGE) followed by sodium dode‐cylsulfate polyacrylamide gel electrophoresis (SDS‐PAGE). A basic polypeptide with a relative molecular weight of 14.5 kDa was detected 32 h after induction and after 48 h a number of polypeptides in the 14 to 35 kDa range were induced. Translation products of poly‐A + RNA isolated from cotyledons treated with SIM for 4, 16, 32 and 48 h were analyzed by using 2‐dimensional NEPHGE‐SDS‐PAGE. Both qualitative and quantitative differences in the translation products were observed at all time points investigated. Expression of a specific RNA coding for a 30 kDa polypeptide was demonstrated as early as 4 h after culture on SIM. This RNA was also present at 16 h, but decreased with longer SIM treatments. Thus, culture of excised cotyledons on a medium inducing the formation of adventitious shoots results in rapid quantitative and qualitative changes in the polypeptide composition and translatable RNA population prior to morphological evidence of shoot induction.