杨树ERF转录因子RAP2L15基因RNAi植物表达载体的构建及遗传转化  

Construction of an ERF transgenic factor gene RAP2L15-RNAi vector and transformation in poplar

作　　者：王星斗 黄娟娟[1] 樊艳 郭聪慧 王升级 韩有志[1] WANG Xingdou;HUANG Juanjuan;FAN Yan;GUO Conghui;WANG Shengji;HAN Youzhi(College o f Forestry,Shanxi Agricultural University,Taigu,Shanxi 030801,China)

机构地区：[1]山西农业大学林学院,山西太谷030801

出　　处：《植物生理学报》2020年第12期2705-2715,共11页Plant Physiology Journal

基　　金：国家自然科学基金(31800564);山西省高校科技创新基金(2019L0392)。

摘　　要：ERF (ethylene response factor)转录因子是植物最大的特异性转录因子基因家族之一,在植物抗逆及生长发育方面起着重要作用。本研究从‘84K杨’(Populus abla×P.glandulosa)中克隆出ERF转录因子基因RAP2L15的全长cDNA序列,对其编码的氨基酸序列进行了同源性比对分析,并利用生物信息学对RAP2L15蛋白进行理化性质分析及结构预测。结果表明, RAP2L15编码的蛋白质序列含有155个氨基酸,属于不稳定的亲水蛋白,与杨树耐盐胁迫基因ERF38具有相似结构。同源性序列比对分析表明RAP2L15与拟南芥ERF转录因子基因AtERF016具有较高同源性,推测该基因可能与植物耐盐胁迫功能有关。构建RAP2L15-RNAi抑制表达载体并利用农杆菌介导叶盘转化法进行杨树遗传转化,获得转基因杨树株系5个。本研究为进一步探索RAP2L15转录因子基因在杨树生长发育及抗盐胁迫方面发挥的功能提供理论及材料基础。The ERF(ethylene response factor) transcription factor is one of the largest plant-specific transcription factors family in plants and plays an important role in plant resistence and growth. In this study, the fulllength cDNA sequence of ERF transcription factor gene RAP2 L15 was cloned from Populus abla × P. glandulosa, and homology comparative analysis was performed on the amino acid sequence encoded by the gene. Bioinformatics was used to analyze the physicochemical properties and structure prediction of RAP2 L15 protein. The results showed that RAP2 L15 was an unstable hydrophilic protein with 155 amino acids and had a similar structure to salt-tolerant ERF38 in poplar. Homologous sequence alignments showed that RAP2 L15 had high homology with Arabidopsis ERF transcription factor AtERF016, which was speculated to be related to plant salt stress tolerance. After that, RAP2 L15-RNAi inhibitory expression vectors were constructed and agrobacterium-mediated disc transformation was used for poplar genetic transformation to obtain 5 transgenic poplar lines. This study provides theoretical and material basis for further exploring the functions of RAP2 L15 transcription factor gene in poplar growth and development and salt stress resistance.

关 键 词：ERF RAP2L15 盐胁迫 遗传转化 基因重组 

分 类 号：Q943.2[生物学—植物学] S792.11[农业科学—林木遗传育种]