小麦TaAAC基因克隆及功能的初步验证

为明确ATP/ADP转运蛋白在小麦籽粒粒重及淀粉合成方面的作用,以百农207 cDNA为模板,利用RT-PCR技术获得TaAAC的基因全长。生物信息学分析表明,TaAAC基因CDS长度为1 143 bp,编码380个氨基酸;TaAAC蛋白位于线粒体内膜和质体膜上。随后,以大麦条纹花叶病毒(BSMV)为载体,以GFP基因为指示基因,利用VIGS和qRT-PCR技术对TaAAC基因的功能进行研究。结果表明,在病毒诱导接种后24 d和29 d,TaAAC基因的相对表达量分别较对照下降51%和67%。BSMV病毒诱导TaAAC基因沉默后的百农207籽粒内总淀粉含量和千粒重均较对照显著降低。以上结果表明TaAAC基因正向调控小麦籽粒的淀粉合成,提高该基因表达有助于增加小麦粒重。

Cloning and Functional Analysis of TaAAC Gene in Wheat

In order to clarify the role of ATP/ADP transporter in wheat grain weight and starch synthesis,the full length of TaAAC gene was obtained by using Bainong 207 cDNA as a template,with RT-PCR technology; Bioinformatics analysis showed that the CDS of TaAAC was 1 143 bp in length,encoding 380 amino acids; the protein of TaAAC was located on the mitochondrial inner membrane and the plastid membrane. The function of TaAAC gene was studied by VIGS technology and qRT-PCR,with barley striped mosaic virus (BSMV) as the vector,and the GFP gene as the indicator gene. The results showed that the relative expression levels of TaAAC gene were decreased by 51% and 67% on the 24 and 29 days after virus-induced vaccination respectively.The total starch content and 1 000-grain weight of Bainong 207 seeds after virus-induced TaAAC silencing were significantly lower than those of the control. The above results indicated that TaAAC positively regulate starch synthesis in wheat grains,and the increased expression of this gene contribute to the increase of wheat grain weight.