辣椒CaERF70的表达特征和转录自激活活性分析  

Expression patterns and transcriptional autoactivation analysis of CaERF70 in chili pepper

作　　者：张余 金明伟 任丽 章毅颖 赵洪 刘昆[1,2] 邓姗 褚云霞[1,2,3] 李寿国 张靖立[1,2] 黄静艳 陈海荣[1,2,3] ZHANG Yu;JIN Mingwei;REN Li;ZHANG Yiying;ZHAO Hong;LIU Kun;DENG Shan;CHU Yunxia;LI Shouguo;ZHANG Jingli;HUANG Jingyan;CHEN Hairong(Institute for Agri-Food Standards and Testing Technology,Shanghai Academy of Agricultural Sciences,Shanghai 201403,China;Shanghai Sub-Center for New Plant Variety Tests,Ministry of Agriculture and Rural Affairs,Shanghai 201415,China;Shanghai Key Lab of Protected Horticultural Technology,Horticultural Research Institute,Shanghai Academy of Agricultural Sciences,Shanghai 201403,China;Agricultural College,AnShun University,AnShun 561000,Guizhou,China)

机构地区：[1]上海市农业科学院农产品质量标准与检测技术研究所,上海201403 [2]农业农村部植物新品种测试(上海)分中心,上海201415 [3]上海市设施园艺技术重点实验室·上海市农业科学院园艺研究所,上海201403 [4]安顺学院农学院,贵州安顺561000

出　　处：《浙江农业学报》2024年第10期2247-2256,共10页Acta Agriculturae Zhejiangensis

基　　金：上海市农业科学院农业科技创新支撑领域研究专项(JCYJ231101)。

摘　　要：AP2/ERF转录因子家族广泛参与植物生长发育以及逆境应答等生物学过程,对于AP2/ERF转录因子的研究有助于提高作物抗逆性。该研究以辣椒耐盐品种SHA2022119为材料,克隆得到CaERF70,该基因cDNA全长为819 bp,编码272个氨基酸,分子量约为30.59 ku,等电点pI为7.67。生物信息学分析表明,该蛋白带负电荷,属于不稳定蛋白,存在34个潜在发生磷酸化修饰的位点,为非跨膜蛋白且在细胞核中表达。系统发育树结果显示,辣椒CaERF70与番茄、烟草、马铃薯的ERF聚为一类,暗示着CaERF70在茄科进化过程中非常保守。定量PCR结果显示,CaERF70基因受低温诱导、高温抑制表达。原核表达实验表明,CaERF70编码蛋白大小约为30 ku,与预测大小一致。转录激活活性分析表明,CaERF70具有转录激活活性。综合以上结果表明,CaERF70为AP2/ERF2转录因子,属于ERF亚组第Ⅶ亚组成员,具有转录激活活性,高温或者低温胁迫导致CaERF70表达发生显著变化,暗示着CaERF70可能在辣椒应对高温或低温胁迫过程中发挥作用。The AP2/ERF transcription factor family is widely involved in biological processes such as plant growth and development and stress response,so it is of great significance for the study of AP2/ERF transcription factors.In this study,we cloned a ERF gene,CaERF70 from salt-tolerant variety SHA2022119 of pepper,which contains 819 bp,encoding 272 amino acids,molecular weight of about 30.59 ku,and isoelectric point pI of 7.67.Bioinformatics analysis showed that the protein had a character with negatively charged,unstable,34 potential phosphorylation sites,non-transmembrane and expressed in the nucleus.The phylogenetic tree results indicated that CaERF70 was clustered with the ERFs of tomato,tobacco and potato,suggesting that CaERF70 was very conserved in the evolution of Solanaceae.qRT-PCR showed that the expression of CaERF70 gene was induced by low temperature and suppressed by high temperature.Prokaryotic expression experiments showed that the size of the protein encoded by CaERF70 was about 30 ku consistent with the predicted size.Transcriptional activation activity analysis showed that CaERF70 had transcriptional activation activity.Taken together,the above results indicated that CaERF70 was an AP2/ERF2 transcription factor,belonging to the ERFⅦgroup,with transcriptional activating activity,and the expression of CaERF70 was significantly changed due to high or low temperature stress,suggesting that CaERF70 was involved in the response of pepper to high or low temperature stress.

关 键 词：辣椒 基因克隆 转录活性 原核表达 非生物胁迫 

分 类 号：S641.3[农业科学—蔬菜学]