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根据已报道的拟南芥春化相关基因VIN3设计引物,通过RT-PCR方法首次从结球甘蓝中克隆得到两条春化相关基因BoVIN3的cDNAORF编码区序列,各1 680 bp,GenBank登录号分别为JQ394927和JQ394928,可编码由560个氨基酸组成的蛋白质;与拟南芥VIN3基因在核酸水平上同源性分别达到85.55%和79.95%;与拟南芥氨基酸序列同源性分别为80.72%和72.96%。半定量RT-PCR表达分析表明BoVIN3受春化作用的诱导,只在感应低温的茎尖生长点表达,在其它部位不表达,并且其转录产物随春化时间延长逐渐积累,在春化42 d时达到峰值;而在不同浓度GA3和KT诱导下不表达;FLC受春化作用的抑制,并且BoVIN3受诱导表达后才能使FLC的表达受到抑制,暗示在结球甘蓝中BoVIN3可能参与春化调控开花的生物学过程,属于春化特异基因,只响应低温诱导而转录。
According to the published report of vernalization-related gene VIN3 of Arabidopsis thaliana, cDNA coding region of two vernalization-related genes, BoVIN3, was cloned by RT-PCR from cabbage for the first time, with a length of 1 680 bp and a GenBank accession number Which encodes a protein of 560 amino acids for JQ394927 and JQ394928, respectively. The homology with the VIN3 gene of Arabidopsis reached 85.55% and 79.95% at the nucleic acid level, respectively. The amino acid homologies with Arabidopsis thaliana were 80.72% and 72.96%. Semi-quantitative RT-PCR analysis showed that BoVIN3 was induced by vernalization, expressed only at the induction of low temperature, but not at other sites. The transcripts of BoVIN3 accumulated gradually with the prolongation of vernalization, And peaked at different concentrations of GA3 and KT; FLC was inhibited by vernalization, and the expression of BoVIN3 was induced to inhibit the expression of FLC, suggesting that BoVIN3 may be involved in vernalization in cabbage The biological process of flowering, belonging to vernalization-specific genes, is only transcribed in response to cold induction.