红地球葡萄生长素响应因子VvARF7基因克隆及其光响应分析

Cloning of Red Earth grape auxin response factor VvARF7 and its light response analysis

生长素是一类重要的植物内源激素,在植物的生长发育过程中具有重要作用,而生长素响应因子(ARF)是一类受生长素调节的转录因子,在生长素信号调控途径中起重要作用。因此,本研究以设施栽培的红地球葡萄VvARF7基因为对象,对其进行生物信息学及在红蓝光处理下的表达响应分析,为进一步研究其作用机制提供参考。结果显示,VvARF7基因全长3468 bp,编码1155个氨基酸,相对分子质量为12958,蛋白质等电点为6.66。VvARF7与河岸葡萄VrARF7-like亲缘关系最近,为亲水蛋白质;二级结构中β-转角占8.14%,延伸链占14.63%,α-螺旋占27.27%,无规则卷曲占49.96%,属于核定位蛋白质。启动子序列分析结果表明,VvARF7主要包括激素应答和光调控顺式作用元件。qRT-PCR结果表明,在花芽分化过程中处理组VvARF7相对表达量在5月30日-8月15日低于对照组,花芽分化后期(6月30日-7月30日)在处理组中的相对表达量显著低于对照组。由此推测VvARF7响应红蓝光(4∶1)参与葡萄花芽分化过程的调控。

Auxin is a kind of important plant endogenous hormones which plays important roles in the growth and development of plants.Auxin reaction factor(ARF)is a kind of transcription factors regulated by auxin and plays an important role in the auxin signaling pathway.Therefore,VvARF7 gene in facility cultivated Red Earth grape was used as the object in this study,bioinformatics analysis and expression response analysis under red and blue light treatment were made to provide reference for further study of the action mechanism.The results showed that,the VvARF7 gene was 3468 bp in full-length,encoded 1155 amino acids,the relative molecular mass was 12958,the protein isoelectric point was 6.66.VvARF7 showed the closest relationship with VrARF7-like of riparian grape,both of them were hydrophilic proteins.In the secondary structure,theβ-angle accounted for 8.14%,the extension chain accounted for 14.63%,the α-spiral accounted for 27.27% and the irregular curl accounted for 49.96%,which indicated that the VvARF7 was nuclear located protein.The analysis results of promoter sequence showed that,VvARF7 mainly included hormone response and cis-acting elements of light regulation.The qRT-PCR results showed that,during the stage of grape bud differentiation,the relative expression amount of VvARF7 gene in the treatment group was lower than that in the control group between May 30 th and August 15 th.Between June 30 th and July 30 th,which was the end stage of grape bud differentiation,the relative expression amount of VvARF7 gene in the treatment group was significantly lower than that in the control group.It is speculated that VvARF7 responded to the participation of red and blue light(4∶1)into the regulation of grape bud differentiation.