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本研究运用反转录PCR(reverse transcription PCR,RT-PCR)技术,从油桐(Vernicia fordii)近成熟种子中克隆了油桐乙醛脱氢酶基因家族两个成员的全长编码序列(CDS),命名为VfALDH2B4和VfALDH2B7,NCBI登记号分别为MF179125和MF179126,CDS长度分别为1 617和1 626 bp,分别编码538和541个氨基酸。通过蛋白结构分析发现这两个蛋白均含有乙醛脱氢酶基因家族特有的保守结构域,且两个蛋白都与麻疯树(Jatropha curcas)的ALDH2B4和ALDH2B7亲缘关系最近。实时荧光定量PCR分析表明,这两个基因在油桐各组织器官中均有表达,其中VfALDH2B4在嫩叶的表达量最高,是其他组织的21倍左右,而f ALDH2B7在花瓣的表达量最高,是其他组织的62倍左右;在油桐种子不同发育时期,两个基因的表达也存在明显差异。此外,两个基因表达还受高盐、干旱和脱落酸调控,它们可能在油桐逆境胁迫应答和脱落酸信号途径中发挥作用。
In this study, we cloned the full-length coding sequence of two members of the gene family of aldahydrate aldehyde dehydrogenase from the mature seeds of Vernicia fordii by reverse transcription PCR (RT-PCR) ), Named as VfALDH2B4 and VfALDH2B7. The NCBI accessions were MF179125 and MF179126, respectively. The CDS length was 1 617 and 1 626 bp, encoding 538 and 541 amino acids respectively. Both proteins were found to contain a conserved domain specific to the aldehyde dehydrogenase gene family by protein structural analysis, and both proteins are most closely related to ALDH2B4 and ALDH2B7 from Jatropha curcas. Real-time PCR analysis showed that these two genes were expressed in all tissues and tissues of Tung tree, in which the expression level of VfALDH2B4 was the highest in young leaves about 21 times that of other tissues, while the expression level of f ALDH2B7 was the highest in petals, Which is about 62 times higher than that of other tissues. There is also a significant difference in the expression of the two genes during the different developmental stages of the tung oil seed. In addition, the expression of both genes was also regulated by high salt, drought and abscisic acid, which may play roles in the stress response and abscisic acid signaling pathway in Tung oil tree.