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DETECTION AND QUANTITATION OF THREE PHYTOCHROMES IN UNIMBIBED SEEDS OF: Avena sativa L.
Author(s) -
Wang YuChie,
CordonnierPratt MarieMichèle,
Pratt Lee H.
Publication year - 1992
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/j.1751-1097.1992.tb02225.x
Subject(s) - phytochrome , endosperm , avena , biology , etiolation , sodium dodecyl sulfate , phytochrome a , embryo , biochemistry , botany , microbiology and biotechnology , arabidopsis , enzyme , gene , red light , mutant
— Three phytochrome apoproteins in unimbibed seeds of Avena saliva L. were identified with monoclonal antibodies directed to, and specific for, three oat phytochromes with monomeric molecular masses of 125, 124 and 123 kDa [Wang et al ., 1991 , Planta 184, 96–104]. All three phytochromes were readily detected in embryo‐containing portions. Only trace amounts were found in endosperm tissue. Phytochrome photoreversibility was detected after concentration and partial purification of embryo extracts by fractionation with ammonium sulfate, indicating that at least one of these seed phytochromes had its chromophore prosthetic group bound to it. Immunoblot analyses were performed to quantitate each of the three phytochromes in unimbibed seeds. Quantitation of phytochromes in detergent‐free extracts led to serious underestimates of phytochrome contents in the unimbibed seeds. In contrast, more than 93% of each of the three phytochromes in the unimbibed seeds was extracted when a modified sodium dodecyl sulfate sample buffer was used as the extraction medium. In such extracts, we measured per embryo 1.40 ± 0.12. 1.60 ± 0.05 and 6.13 ± 0.31 ng of 125–, 124– and 123‐kDa phytochrome, respectively.