园艺学报 ›› 2019, Vol. 46 ›› Issue (5): 939-954.doi: 10.16420/j.issn.0513-353x.2018-0671

• 研究报告 • 上一篇    下一篇



  1. 浙江省农业科学院园艺研究所,杭州 310021
  • 出版日期:2019-05-25 发布日期:2019-05-25
  • 基金资助:

Identification and Expression Analysis of WRKY Transcription Factors in Eriobotrya japonica

LI Xiaoying,XU Hongxia,and CHEN Junwei*   

  1. Institute of Horticulture,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,China
  • Online:2019-05-25 Published:2019-05-25

摘要: 以枇杷(Eriobotrya japonica)转录组数据为材料,利用生物信息学手段对枇杷WRKY 转录因子进行鉴定与特征分析。分离鉴定出33条具典型WRKY 结构域的序列,CDS长度分布在492 ~ 2 040 bp,命名为EjWRKY01 ~ EjWRKY33。分组鉴定和进化树分析结果显示,枇杷WRKY 转录因子可分为3 大类,其中Ⅰ类数量为6个,Ⅱ类和Ⅲ类的数量分别为22和5个,其中Ⅱ类又进一步分为a ~ e等5个亚类。WRKY 结构域分析显示,WRKY 结构域高度保守,绝大多数都含有WRKYGQK七肽和锌指结构。枇杷WRKY转录因子基因组DNA 全长895 ~ 3 873 bp,编码的蛋白在163 ~ 679 个氨基酸范围内,具有1 ~ 5个内含子。WRKY启动子顺式作用元件预测结果表明,大部分WRKY启动子序列具有典型的 TATA 和 CAAT 元件。将获得的所有WRKY基因与已报道的拟南芥WRKY进行进化比对分析发现,所有EjWRKY转录因子并没有位于独立的分支上,表明枇杷的基因序列虽与拟南芥物种有差别,但枇杷WRKY 蛋白在调控植物生物和非生物逆境反应、信号传导、生长发育等方面的功能相似。此外,荧光定量PCR结果显示,大部分EjWRKY均在枇杷的根、茎、叶、花和果实中表达,但相对表达水平不同,说明WRKY家族基因在枇杷的生长发育中可能具有不同的功能。

关键词: 枇杷, WRKY 转录因子, 特征分析, 系统进化, 组织表达

Abstract: In this study,the data of loquat transcriptome were used to identify and characterize loquat WRKY transcription factors. The results show that this study isolated 33 sequences with typical WRKY structure domains,and the CDS length was distributed in 492–2 040 bp,named EjWRKY01–EjWRKY33. The analysis results of group identification and evolutionary tree show that the loquat WRKY TF can be divided into three categories,including classⅠ,Ⅱ and Ⅲ,the number of each class was 6,22 and 5,respectively. Class Ⅱ is further divided into five subgroups(a–e). The genomic DNA sequence of EjWRKY TF is from 895 to 3 873 bp with 1–5 introns,whose encoded protein was in the range of 163–679 amino acids. The prediction results of cis-acting elements of WRKY promoter show that most WRKY promoter sequences have typical TATA and CAAT elements. The phylogenetic analysis of WRKY TF among Arabidopsis and loquat showed that many WRKY proteins clustered together,displaying close phylogenetic relationships,which indicated that although there were differences among the gene sequences of different species,the functional features of the WRKY proteins in regulating biotic and abiotic stress responses,metabolic regulation,signal transduction,growth and development and morphogenesis were similar. Furthermore,the RT-PCR results showed that most of EjWRKY genes were expressed in root,stem,leaf,flower and fruit of loquat,but the relative expression levels were different,which suggested that WRKY family genes may have different functions in the growth and development of loquat.

Key words: Eriobotrya japonica, WRKY transcription factor, characteristic analysis, phylogenetic analysis, tissue expression