球茎甘蓝MYB62转录因子的克隆与表达特征分析  

Cloning and Expression Analysis of MYB62 Transcription Factors from Kohlrabi

作　　者：王鑫淼 赵孟良[1,2] 邵登魁[1] 尕桑 任延靖 WANG Xinmiao;ZHAO Mengliang;SHAO Dengkui;GA Sang;REN Yanjing(Qinghai Key Laboratory of Vegetable Genetics and Physiology,Qinghai University,Xining 810016,China;State Key Laboratory of Plateau Ecology and Agriculture,Qinghai University,Xining 810016,China;Comprehensive Agriculture and Animal Husbandry Service Center of Yushu Tibetan Autonomous Prefecture,Yushu 815000,China)

机构地区：[1]青海大学青海省蔬菜遗传与生理重点实验室,青海西宁810016 [2]青海大学省部共建三江源生态和高原农牧业国家重点实验室,青海西宁810016 [3]玉树藏族自治州农牧业综合服务中心,青海玉树815000

出　　处：《华北农学报》2024年第2期62-70,共9页Acta Agriculturae Boreali-Sinica

基　　金：国家大宗蔬菜产业技术体系项目(CARS-23-G26);科技成果转化专项(2023-NK-144)

摘　　要：为了明晰球茎甘蓝MYB62转录因子的序列特征及其在逆境胁迫后的表达特征,进一步探索球茎甘蓝MYB62转录因子的生物学功能,为后续MYB62转录因子功能鉴定提供理论依据,以球茎甘蓝为研究对象,采用同源克隆的方法获得球茎甘蓝MYB62转录因子基因,并对其进行生物信息学分析,通过实时荧光定量PCR的方法分析MYB62在球茎甘蓝不同组织间以及逆境胁迫后的表达特征。基因克隆结果表明,BocMYB62基因gDNA全长1353 bp,CDS为837 bp,包括4个外显子和3个内含子,编码278个氨基酸。序列结构分析结果显示,BocMYB62为亲水性蛋白,具有2个SANT-MYB结构域,属于R2R3-MYB型MYB转录因子;空间结构预测显示其具有典型的α-螺旋结构;系统发育分析表明,BocMYB62与甘蓝型油菜MYB62亲缘关系最近。时空表达结果显示,BocMYB62在绿色球茎甘蓝中的表达量始终高于在紫色球茎甘蓝中,且存在明显的组织特异性,在干旱胁迫后BocMYB62表达量显著升高,胁迫12 h表达量最高,低温胁迫后BocMYB62表达量显著低于对照,在4℃低温胁迫时表达量最低,推断BocMYB62可能参与花青素生物合成调控,并可能参与逆境胁迫的调控作用。In order to clarify the sequence characteristics of the MYB62 transcription factor,expression changes after stress,and further explore the biological function of the MYB62 transcription factor,kohlrabis were used as plant materials.MYB62 transcription factors were obtained by homologous cloning method and bioinformatics was analyzed.Spatiotemporal expression analysis of MYB62 and its expression after stress were performed by Real-time PCR.Results of gene cloning showed that gDNA length of BocMYB62 gene was 1353 bp,the length of CDS was 837 bp,which contained four exons and three introns and encoded 278 amino acids.The sequence structure analysis showed that BocMYB62 was a hydrophilic protein with two SANT-MYB domains,belonging to the R2R3-MYB type in the MYB transcription factor family.The spatial structure prediction revealed a typicalα-helix structure.Phylogenetic analysis indicated that BocMYB62 was closely related to MYB62 in Brassica napus.The spatiotemporal expression results showed that BocMYB62 was consistently higher in green kohlrabi than that in purple with clear tissue specificity.The expression of BocMYB62 was significantly increased during drought stress with the highest expression under 12 h.The BocMYB62 expression was significantly lower than control and the lowest at 4℃cold stress.These results inferred that BocMYB62 may be involved in the regulation of anthocyanin biosynthesis and the regulation of stress.It provides a theoretical basis for the subsequent functional identification of MYB62 transcription factors.

关 键 词：球茎甘蓝 MYB62 序列分析 保守基序 亲缘关系 荧光定量PCR 

分 类 号：S635.03[农业科学—蔬菜学] Q78[农业科学—园艺学]