‘红地球’葡萄花芽分化过程中VvARF18基因克隆与表达分析  

Cloning and expression analysis of VvARF18 gene during flower bud differentiation in'Red Globe'grape

作　　者：袁苗 徐伟荣[1] 刘鑫 党仕卓 周娟 张亚红[1] YUAN Miao;XU Weirong;LIU Xin;DANG Shizhuo;ZHOU Juan;ZHANG Yahong(College of Agriculture,Ningxia University,Yinchuan 750021,China)

机构地区：[1]宁夏大学农学院,银川750021

出　　处：《植物生理学报》2023年第6期1184-1194,共11页Plant Physiology Journal

基　　金：宁夏回族自治区重点研发计划(2021BEF02016)。

摘　　要：ARF是植物中一类重要的转录因子,广泛参与植物生长发育及各种生理过程。本研究以葡萄(Vitis vinifera) VvARF18基因为研究对象,进行生物信息学分析,并探究设施栽培条件下‘红地球’葡萄花芽分化进程中其表达丰度与光环境的关系,为设施栽培葡萄花芽分化的分子调控机理研究提供理论基础与参考。利用PCR技术从‘红地球’葡萄中同源克隆出VvARF18基因,该基因全长2 040 bp,编码679氨基酸,分子质量为60.399 kDa,理论等电点为8.81;VvARF18蛋白具有ARF家族蛋白特有的B3 DNA结合结构域和Auxin_resp结构域。系统进化分析表明VvARF18和茶树CsARF18亲缘关系最近,亚细胞定位与转录自激活结果表明VvARF18是一个定位在细胞核中且具有转录激活活性的转录因子。实时荧光定量PCR分析结果表明,在花芽分化进程中处理组VvARF18表达量均高于对照组,推测VvARF18响应红/蓝光4:1诱导表达参与葡萄花芽分化进程,其具体调控机制有待进一步研究。ARF genes are an important class of transcription factor genes in plants.Their encoded products are widely involved in plant growth and development and various physiological processes.In this paper,grape(Vitis vinifera)VvARF18 gene was used as the research object for bioinformatics analysis.We investigated the relationship between the expression abundance and light environment in the process of flower bud differentiation of'Red Globe'grape,to provide a basis for the regulation mechanism during the differentiation process.VvARF18 gene was homologously cloned from'Red Globe'grape by PCR,which is 2040 bp in length,encoding 679 amino acids,with a molecular mass of 60.399 kDa and a theoretical isoelectric point of 8.81.VvARF18 protein has a B3 DNA binding domain and Auxin_resp domain that is unique to ARF family proteins.Phylogenetic tree shows that VvARF18 and tea tree CsARF18 are most closely related.The results of subcellular localization confirm that VvARF18 protein belongs to the nuclear localization protein and has yeast self-activation activity,which is consistent with the characteristics of a transcription factor.The results of quantitative real-time PCR(qRT-PCR)show that the expression level of VvARF18 in the treatment group was higher than that in the control group during the process of flower bud differentiation,which led to the speculation that VvARF18 might be involved in grape flower bud differentiation in response to 4:1 red/blue light,but the specific mechanism needs to be further investigated.

关 键 词：花芽分化 ARF基因 基因克隆 表达分析 

分 类 号：S663.1[农业科学—果树学]