枸杞WRKY3基因克隆及组织表达分析  

Cloning and Tissue Expression Analysis of WRKY_3 Gene in Lycium barbarum L.

作　　者：徐惠娟[1] 郑蕊[1] 陈任[1] 王彦才[1] 王丽娟[1] 

机构地区：[1]宁夏大学生命科学学院,西部特色生物资源保护与利用教育部重点实验室,银川750021

出　　处：《西北植物学报》2016年第9期1721-1727,共7页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家自然科学基金(31260065;31360363);2013年中国科学院西部之光项目

摘　　要：以枸杞为材料,采用PCR及RACE方法,克隆了枸杞WRKY转录因子基因cDNA序列,命名为Lb WRKY3,GenBank登录号为KX196192。在生物信息学分析的基础上,进行亚细胞定位、基因表达分析。结果显示:(1)Lb WRKY3开放阅读框ORF长度为1 068bp,编码356个氨基酸。(2)生物信息学分析显示,Lb WRKY3编码蛋白具有一个WRKY结构域,二级结构中不规则卷曲结构所占比例最大(58.67%),延伸链结构次之(18.88%),α螺旋比例为15.82%,β转角最少,仅为6.63%;Lb WRKY3蛋白与案头菊WRKY蛋白、黄花蒿WRKY蛋白相似性较高。(3)亚细胞定位显示,Lb WRKY3蛋白定位于细胞核。(4)实时定量PCR分析表明,Lb WRKY3在根中表达量最高,在花中表达量最低;在枸杞果实发育过程中Lb WRKY3均有表达,表达量随果实成熟逐渐升高,并于35d达到峰值;Lb WRKY3基因在果实中的表达具有组织特异性表达特性(果肉>果皮>种子)。研究表明,Lb WRKY3基因参与了枸杞果实生长发育调控。With wolfberry（Lycium barbarum L.）as material,polymerase chain reaction（PCR）combined with RACE technology were used to clone a cDNA of WRKY from wolfberry,named as Lb WRKY3.（GenBank accession No.KX196192）.Meanwhile,on the basis of bioinformatic analysis,we performed the subcellular localization assays and tissue-specific expression analysis.The results indicated that：（1）Lb WRKY3 has an open reading frame（ORF）of 1 068 bp,which encoded a protein of 356 amino acid residues.（2）Bioinformatic analysis indicated that Lb WRKY3 protein contains the one conserved WRKY motifs,the predictive secondary structure showed that Lb WRKY3 protein was made up of 15.82%alpha-helix,6.63% beta-turn,18.88% extended strand and 58.67%random coil.Lb WRKY3 protein showed the highest homology identity with WRKY protein fromChrysanthemum x morifolium and Artemisia annua.（3）Subcellular localization assays showed that the Lb WRKY3 protein was located in the nucleus.（4）Realtime PCR analysis indicated that Lb WRKY3 was expressed in high transcript level in roots,low levels in flower.The Lb WRKY3 gene expression could be detected during the whole period of fruit development.Interestingly,Lb WRKY3 gene showed a high transcription level in 35 days.The expression level in thepulp was higher than that in the seed and peel.Lb WRKY3 gene takes part in fruit development and sugar accumulation in wolfberry.

关 键 词：枸杞 WRKY转录因子 果实发育 表达模式 

分 类 号：Q785[生物学—分子生物学] Q789