野生黄毛草莓NAC转录因子基因FnNAC7克隆与表达分析  

Cloning and Expressing Analysis of a NAC Transcription Gene FnNAC7 from Wild Fragaria nilgerrensis Schidl.

作　　者：何淑敏 姚立萍 张童 胡玉慈 陈清西[1] 文志丰[1] He Shumin;Yao Liping;Zhang Tong;Hu Yuci;Chen Qingxi;Wen Zhifeng(College of Horticulture,Fujian Agriculture and Forestry University,Fuzhou,350002;Fuzhou Institute of Agricultural Science,Fuzhou,350018)

机构地区：[1]福建农林大学园艺学院,福州350002 [2]福州市农业科学研究所,福州350018

出　　处：《分子植物育种》2020年第11期3461-3469,共9页Molecular Plant Breeding

基　　金：国家自然科学基金(青年项目)(31701907);福建省自然科学基金(高校联合)项目(2018J01703);福建省现代农业产业技术体系建设专项(闽财指[2017]625号);中央引导地方科技发展专项(NO.2017L3001)共同资助。

摘　　要：NAC转录因子基因为植物所特有,在植物的生长发育、非生物和生物胁迫的抗逆反应中起着重要作用。本研究以野生黄毛草莓(Fragaria nilgerrensis Schidl.)为材料,利用RT-PCR技术,从中克隆到1个NAC转录因子基因FnNAC7(GenBank登录号:MK685675)。序列分析结果表明,FnNAC7开放阅读框为1137 bp,编码378个氨基酸,包含一个保守的NAM结构域。同源性分析结果表明,FnNAC7基因编码的氨基酸序列与森林草莓编码的氨基酸相似性为94.87%。蛋白结果预测显示,FnNAC7蛋白为不稳定的酸性疏水蛋白,不存在跨膜结构域,且无信号肽存在;亚细胞定位试验结果显示,该基因编码的蛋白质在细胞核中表达;实时定量PCR结果显示,接种草莓尖孢炭疽菌(Colletotrichum acutatum)后,草莓叶片FnNAC7基因表达上调,接种24 h后该基因表达量最高,是对照组的17.0倍。研究结果为进一步深入开展NAC基因在草莓炭疽病抗性中功能分析和分子育种提供了科学的依据,也为进一步研究FnNAC7基因在野生黄毛草莓抗草莓炭疽病中所起的调控作用提供了参考。NAC proteins are plant-specific transcription factors,they have been suggested to play important regulatory roles in plant development,abiotic and biotic stress responses.In this study,we cloned a NAC transcription factor gene by RT-PCR from wild Fragaria nilgerrensis Schidl.,and the gene designed as FnNAC7(GenBank No.:MK685675).The result of sequence analysis shows that the length of FnANC7 was 1137 bp and encoded 378 amino acids,the predicted protein contained a conserved NAM domain.Homogeneous analysis indicated that the deduced sequence of FnANC7 shares 94.87%with the predicted protein from Fragaria vesca.The FnNAC7 protein structure predicted showed that should be an unstable,acidic and hydrophobic protein,lack of predicted transmembrane region and without signal peptides.The results of subcellular localization experiments showed that the FnNAC7 protein is localized to the nucleus of the Arabidopsis.Quantitative real-time PCR showed that the expression of FnANC7 was upregulated after inoculated Colletotrichum acutatum,expression levels reached a maximum at 24 h,over 17.0-fold than controls.The results provide a scientific basis for further analysis of function and molecular breeding of NAC gene in resistance to strawberry anthracnose.It also provides a reference for further study on the regulatory role of FnNAC7 gene in resistance to strawberry anthracnose in wild Fragaria nilgerrensis.

关 键 词：野生黄毛草莓(Fragaria nilgerrensis Schidl.) 草莓炭疽病 FnNAC7 生物信息学分析 表达分析 

分 类 号：S436.68[农业科学—农业昆虫与害虫防治] Q943.2[农业科学—植物保护]