10. タバコ培養細胞BY-2におけるGA応答系の解析

概要

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Tobacco cultured cell BY-2 is widely-used, because its synchronous culture and transformation systems have been established. The cells haven't been used for the research of GAs very often, however, because GA has no obvious effect on their growth. According to Amador et al. (2001), the expression level of a gene for GA-catabolism, GA 2-oxidase increased responding to GA, which suggested that the system required for GA-response still remained in BY-2 cells. Thus, we planed to elucidate the mechanism for the GA-response in the cells. We had obtained a GA 2-oxidase gene (NtGA2ox1) expressed in the cells by degenerate-PCR and RACEs, examined its expression patterns to applied GA_4 by using RNA-blottings, and confirmed its GA-repsonsiveness reported by Amador et al. Then we further studied its expression patterns in various culture conditions. Reaching the stationary phase of the cell-growth, the expression level of NtGA2ox1 increased. A halt of shaking elevated the expression level of NtGA2ox1, which made it difficult to see the response of the gene to GA. Incomplete aeration to the cells might have activated the expression of the gene. We examined the response of the gene to various GAs, and showed that some GAs with less or no biological activities showed lower effect on its expression than GA_4. We are going to evaluate the affinity of each GA to NtGA2ox1, using its recombinant.
植物化学調節学会の論文
2004-10-13