平榛ChWRKY1转录因子的克隆及表达模式分析  

CLONING AND EXPRESSION PROFILE ANALYSIS OF ChWRKY1 FROM Corylus heterophylla Fisch.

作　　者：赵天田[1] 王贵禧[1] 梁丽松[1] 马庆华[1] 陈新[1] 

机构地区：[1]中国林业科学研究院林业研究所/林木遗传育种国家重点实验室,北京100091

出　　处：《核农学报》2012年第2期238-244,共7页Journal of Nuclear Agricultural Sciences

基　　金：国家"十一五"科技支撑计划(2006BAD01A1701);国家林业局(2008204208)

摘　　要：根据平榛花芽转录本高通量测序的结果,采用RACE-PCR法克隆到1个平榛WRKY基因,全长1273bp,推断其编码317个氨基酸,命名为ChWRKY1。序列分析表明,该蛋白属于WRKY家族第2类成员,只含有1个WRKY结构域,锌指结构为C-X5-C-X23-H-X1-H,构建的系统发育树结果表明,它与杨树PtWRKY5、葡萄VvWRKY4的亲缘关系较近,相似性分别为69%和64%。采用qRT-PCR,以Actin为内参对ChWRKY1基因在自然条件下花芽部位的表达模式进行分析,结果显示在自然条件下12月份达到最高的表达量,随后表达量呈现下降的趋势;对平榛根蘖苗进行4℃低温胁迫处理,叶片中ChWRKY1基因快速上调表达,4h达到最大表达量,表明该基因可能参与平榛对低温胁迫的早期应答反应;冬季(1月份)ChWRKY1基因在韧皮部中的表达高于花芽和雄花序,具有组织表达特异性。According to the high-throughput sequencing transcripts of hazelnut buds, a 1273bp fragment including the whole coding region was obtained through RACE-PCR. It encoded a polypeptide of 317 amino acids, and designated as ChWRKY1. The deduced protein sequence showed that this protein belonged to the second group of WRKY family and the zinc-finger structure is C-X5-C-X23-H-X1-H. Phylogeny tree results showed that the ChWRKY1 was much closer to PtWRKY5 from poplar and VvWRKY4 from grape, with 69% and 64% amino acids similarity, respectively. The ChWRKY1 expression in hazelnut buds was analyzed with Actinas an internal control gene under natural conditions and the result displayed that the highest expression level was in December and then decreased. ChWRKY1 in leaves was induced rapidly and highly when suckers were exposed to low temperatures（4℃ ） , and reached a peak at 4h after cold treatment, suggesting that it was possibly involved in early-stage responses to low temperature. Spatial expression analyses revealed that the transcription level of ChWRKY1 was higher in phloem than in bud and male inflorescence which indicated a tissue-specific expression pattern.

关 键 词：平榛 WRKY 基因克隆 低温 表达模式 

分 类 号：S664.4[农业科学—果树学]