葡萄Alfin-like转录因子家族的鉴定和表达分析

Identification and Expression Analysis of Alfin-like Transcription Factor Family in Grape

Alfin-like(AL)转录因子家族对高盐、低温、干旱等非生物胁迫反应具有重要的调控作用。该研究采用同源比对的方法检索鉴定葡萄AL转录因子家族基因、分析其生物信息学特性,并采用qRT-PCR方法分析非生物胁迫下AL基因的表达特征,以探究葡萄AL基因在非生物逆境胁迫中的功能。结果表明:(1)在葡萄中共鉴定出6个AL基因家族成员,分别命名为VvAL1~VvAL6,且6个成员分别分布在6条染色体上。(2)葡萄中AL转录因子具有高度保守的DUF3594结构域和PHD结构域,各家族成员均含有5个外显子和4个内含子;上游启动子区域分析发现大量植物激素与非生物胁迫响应相关的顺式作用元件。(3)基因芯片表达模式分析显示,盐、干旱、ABA胁迫以及低温(5℃)处理均显著影响葡萄AL家族基因(VvAL1~VvAL6)的表达。(4)qRT-PCR检测显示,不同胁迫处理下AL基因在葡萄叶片中的表达水平不同;ABA处理下葡萄AL转录因子家族基因表达量较对照均显著下调,但在PEG处理下差异不显著;在盐胁迫处理下,VvAL2、VvAL4、VvAL5基因的表达量均显著上调,分别是对照的23倍、8.5倍和10.5倍,而VvAL1和VvAL6基因的表达量均显著下调,分别是对照的33倍和25倍。研究发现,葡萄AL转录因子家族与植物激素和非生物胁迫密切相关,尤其是该家族基因强烈响应高盐胁迫。

The Alfin-like(AL)transcription factor family plays an important role in abiotic stress such as high salt,low temperature,and drought.In this study,the homology ratio method was used to retrieve and identify the genes of grape AL transcription factor family,analyze their bioinformatics characteristics and the expression characteristics of AL gene under abiotic stress by qRT-PCR,so as to explore the function of grape AL gene under abiotic stress.The results showed that:(1)we identified 6 members of the AL gene family in grape,they were named VvAL1-VvAL6,and the 6 members were distributed on 6 chromosomes.(2)The AL transcription factors in grape had highly conserved DUF3594 domain and PHD domain and each gene contained 5 exons and 4 introns.The upstream promoter region analysis showed that there was a lot of cis-elements associated with various stress and hormone responses.(3)Gene chip expression pattern analysis showed that salt,drought,ABA stress and low temperature(5℃)treatments all significantly affected the expression of AL family genes(VvAL1-VvAL6).(4)qRT-PCR showed that the expression level of AL gene in grape leaves was different under different stress treatments;under ABA treatment,the gene expression of grape AL transcription factor family was significantly down-regulated compared with CK,while PEG treatment not significant.Under salt stress,the expression levels of VvAL2,VvAL4 and VvAL5 genes were significantly up-regulated,which were 23 times,8.5 times and 10.5 times of the control,respectively.The expression levels of VvAL1 and VvAL6 genes were significantly down-regulated,which were 33 times and 25 times of the control,respectively.It was found that grape AL transcription factor family is closely related to plant hormone and abiotic stress,especially the genes of this gene family strongly respond to high salt stress.