结球甘蓝春化相关基因BoVIN3的克隆及其表达分析

Cloning and Expression Analysis of BoVIN3 from Cabbage

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。

The mid harvest season cabbage variety ＇Guangrong 1＇ was used in this experiment. Fourteen leaves cabbage plants were vernalized in the incubator at 8℃. Two vernalization-related genes were isolated by RT-PCR. Sequencing analysis results showed that their ORF were both 1 680 bp in length, which encoded 560 amino acids, and GenBank accession number are JQ394927 and JQ394928 respectively. Their nucleotide sequences shared 85.55% and 79.95% identity with .4t VIN3 respectively, and the deduced amino acid sequence showed homology to .4tVIN3 with 80.72% and 72.96%. The expression patterns of the BoVIN3 genes were investigated by semi-quantity RT-PCR, results showed that BoVIN3 gene could express induced by low temperature treatment, and expressed in the stem apex specifically, transcript levels increased with increasing treatment time. The transcription reach the peak at 42 d of vernalization. At the same time the expression of FLC was suppressed after Bo VIN3 was induced by low temperature treatment, results suggest that both genes maybe involved in the biological process of floweringregulation in vernalization. However both BoVIN3 could not be induced by GA3 and KT treatment. It indicated that cabbage BoVIN3 was specific vernalization genes, whose transcripts could specially response to vernalization.