Non-Specific Association of Phytochrome to Nuclei during Isolation from Dark-Grown Pea (Pisum sativum cv. Alaska) Plumules

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Although phytochrome is known to regulate gene expression in plants, the location of the phytochrome molecules involved in the response remains unclear. Thus the aim of this investigation was to test the possibility that nuclei contain phytochrome. Nuclei were isolated from dark-grown pea (Pisum sativum cv. Alaska) plumules in the dark and the nuclear proteins were resolved on SDS polyacrylamide gels and transferred to nitrocellulose by western blotting. A significant amount of phytochrome was detected in the nuclear proteins by immunostaining using monoclonal and polyclonal anti-phytochrome antibodies. Most of the phytochrome associated with the nuclei could not be removed by treatment with high salt and low magnesium, indicating that the binding was rather stable. However, the isolation of nuclei in the presence of exogenously added [^<125>I]phytochrome of P_R form in the dark suggested that significant amount of the phytochrome detected in the nuclei was derived from contamination by the soluble fraction during isolation. It is an open question as to whether, besides this contaminated phytochrome, isolated nuclei endogenously contain very small amount of phytochrome.
日本植物生理学会の論文