アサガオ未熟種子中のジベレリンの機能に関する組織化学的解析

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Immature seeds of several dicotyledonous plants often contain a large amount of gibberellins (GAs). Their physiological roles, however, remain unknown. We have reported the localization of active GAs around starch grains in outer-integument of immature seeds of Pharbitis nil. This observation suggests that GAs may be concerned with the degradation of starch in the seeds via an induction of specific hydrolytic enzymes like α-amylase as in cereal grains. To verify this possibility, we cloned the genes which had a high homology to well-known α-amylase genes from cDNAs prepared from the immature seeds of P. nil., obtained two full-length cDNA (PnAmyl,2) by 5'- and 3'-RACE, and examined their responsiveness to GAs. The northern- hybridization analysis revealed that the transcription levels of these genes were up-regulated by exogeneously applied GA_3. The antibodies specific for the α-amylase homologs were prepared by using recombinant proteins, and used for the immuno-western analysis of the soluble extract from the immature seeds of P. nil. PnAmy1 was clearly detected with the anti- PnAmy1 recombinant antiserum, while PnAmy2 was not. This observation agreed with the result of the northern-hybridization that the expression level of PnAmy2 was much lower than that of PnAmyl. We performed the immunohistochemical analysis using the anti-PnAmy1 recombinant antiserum to analyze the localization of PnAmy1 in the immature seed. The specific staining due to PnAmy1 was observed at the same site where active GAs were stained by with anti-GA-antiserum.
植物化学調節学会の論文
2000-11-02

アサガオ未熟種子中のジベレリンの機能に関する組織化学的解析

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