胭脂萝卜RsUFGTs的克隆与表达分析

Cloning and expression of RsUFGTs in red radish

为确定参与胭脂萝卜花青苷生物合成的类黄酮糖基转移酶基因(RsUFGTs),从萝卜基因组中筛选出3个候选的RsUFGTs基因,以胭脂萝卜为材料,克隆了3个RsUFGTs的开放阅读框(ORF)并进行序列及表达分析。结果表明:RsUFGT1和RsUFGT2的ORF长度分别为1383和1374bp,分别编码460和457个氨基酸,而RsUFGT3提前终止;氨基酸序列比对和系统进化树分析表明RsUFGT1和RsUFGT2均含有PSPG保守特征结构域,且属于ClusterⅠ亚族成员;利用实时荧光定量PCR分析它们在不同着色萝卜品种中的表达发现,RsUFGT1在‘红心1号’胭脂萝卜皮和肉组织中表达量较高,在‘满堂红’萝卜皮和‘春不老’的萝卜皮与肉中均不表达,其表达量与花青苷含量呈正相关,而RsUFGT2的表达则无明显规律。因此,RsUFGT1可能是胭脂萝卜花青苷生物合成的重要结构基因之一。

To clarify the flavonoid glycosyltransferase gene( RsUFGTs ) involved in the biosynthesis of anthocyanin in red radish,three candidate RsUFGTs genes were screened from the radish genome,and the open reading frames (ORFs) of three RsUFGTs were cloned and sequenced and expressed. Results showed that the ORF length of RsUFGT1 and RsUFGT2 was 1 383 bp and 1 374 bp,encoding 460 and 457 amino acids,respectively. However, RsUFGT3 was mutated with early termination. Protein sequence alignment and phylogenetic tree analyses indicated that RsUFGT1 and RsUFGT2 both contain PSPG conserved domain and belong to the same UFGT class. The results of real-time quantitative PCR analysis showed that the expression of RsUFGT1 in ‘Hongxin No.1’ was relatively high and no expression was detected in skin of ‘Mantanghong’ and in skin and flesh of ‘Chunbulao’. The expression of RsUFGT1 was positively correlated with anthocyanins content in tissues of different radish cultivars. However,no obvious rules of RsUFGT2 expression were detected. It is indicated that RsUFGT1 may be one of the important structural genes of anthocyanin biosynthesis in red radish.