青花菜锌指蛋白基因BoCCCH1的克隆和表达分析  

Isolation and Expression Analysis of a Zinc Finger Protein Gene Bo CCCH1 From Broccoli

作　　者：蒋明[1] 刘青娥[2] 管铭[1] 张雪[1] 陈雅[1] 左诗璇 

机构地区：[1]台州学院生命科学学院/生态学浙江省重点学科,浙江椒江317000 [2]丽水学院生态学院,浙江丽水323000

出　　处：《核农学报》2016年第3期444-450,共7页Journal of Nuclear Agricultural Sciences

基　　金：浙江省本科院校中青年学科带头人学术攀登项目(pd2013420);浙江省自然科学基金项目(LY13C150003);浙江省重点学科(浙江省台州学院生态学)开放课题(EKD2013-03)

摘　　要：CCCH锌指蛋白广泛存在于真核生物中,在植物的生长、发育和逆境胁迫响应中起着重要作用。为获知青花菜CCCH转录因子基因的序列特征和表达特性,从青花菜叶片中克隆到1个CCCH基因,命名为Bo CCCH1,同时,利用RT-PCR方法研究了其在不同器官及霜霉菌侵染叶片中的表达模式。序列分析结果表明,Bo CCCH1的基因组全长为1 568 bp,包含1个长度为599 bp的内含子,2个长度分别为627 bp、342 bp的外显子,编码322个氨基酸,蛋白质的分子量与等电点分别为35 296.42 Da和8.47;Bo CCCH1有2个锌指结构,类型均为C-X8-C-X5-C-X3-H。系统发育分析结果表明,Bo CCCH1与其它植物的21条同源序列在进化树上分为6组,来自十字花科的CCCH与Bo CCCH1处于同一分支。基因表达结果表明,Bo CCCH1在叶、花茎、嫩角果、花蕾和花中表达,其中花茎和嫩角果的表达量最高,但在根中未检测到表达;在霜霉菌侵染下,叶片Bo CCCH1的表达量升高,到24 h和36 h时达到最大,推测Bo CCCH1与霜霉病抗性相关。本研究为进一步探明Bo CCCH1在霜霉病抗性反应中的功能奠定了基础。CCCH zinc finger proteins are widely distributed in eukaryotic organisms,and they play important roles in plant growth,development and stress responses. To learn about sequence features and expression characters of broccoli CCCH transcription factor gene,a CCCH gene designated Bo CCCH1,was isolated from broccoli（ Brassica oleracea var.italica）,and expression patterns of Bo CCCH1 in different organs as well as leaves invaded by Hyaloperonospora parasiticawere performed by means of RT-PCR. Sequence analysis results indicated that the full length genome of Bo CCCH1 was 1 568 bp with a sole intron of 599 bp. The two exons were 627 and 342 bp in length encode 322 amino acids,the molecular weight and isoelectric point were 35296. 42 Da and 8. 47,respectively. Phylogenetic analysis results indicated that Bo CCCH1 and its homogeneous sequences were divided into 6 groups,and it was grouped with sequences from other Cruciferae plants. Expression analysis results revealed that Bo CCCH1 expressed in leaves,stalks,young siliques,flower buds and flower,with the highest expression levels in stalks and siliques,however,no expression was detected in roots. The expression were increased in leaves when infected by H. parasitica,and the highest expression levels were identified on 24 h and 36 h,indicating Bo CCCH1 with resistance responses against H. parasitica.Isolation and expression analysis of Bo CCCH1 will provide evidences for further studies on gene functions during downy mildew resistance responses.

关 键 词：CCCH锌指蛋白 霜霉病 基因表达 青花菜 

分 类 号：S635.3[农业科学—蔬菜学]