葡萄VvSUC27基因的克隆及病原响应表达分析  

Gene cloning and pathogenic response expression analysis of VvSUC27 in grapevine

作　　者：吴佳鸿 黄金宝[1] 刘梅[1] 彭军波 乔广行[1] 邢启凯 WU Jiahong;HUANG Jinbao;LIU Mei;PENG Junbo;QIAO Guanghang;XING Qikai(Beijing Key Laboratory of Environment-Friendly Management of Fruit Diseases and Pests in North China,Institute of Plant and Environmental Protection,Beijing Academy of Agricultural and Forestry Sciences,Beijing 100097,China)

机构地区：[1]北京市农林科学院植物保护环境保护研究所,北方果树病虫害绿色防控北京市重点实验室,北京100097

出　　处：《植物保护》2021年第4期20-27,共8页Plant Protection

基　　金：国家自然科学基金(31801686);北京市农林科学院科技创新能力建设专项(KJCX20190406);北京市农林科学院青年科研基金(QNJJ201904)。

摘　　要：由葡萄座腔菌科Botryosphaeriaceae真菌引起的葡萄溃疡病(Botryosphaeria dieback)是葡萄上的主要枝干病害之一,严重影响葡萄产业的健康发展。其中,致病力最强的为可可毛色二孢Lasiodiplodia theobromae。糖转运蛋白在调节植物对病原菌的免疫过程中起着至关重要的作用,关于葡萄中糖转运蛋白参与植物免疫机制的研究尚未见报道。本试验从‘无核白’葡萄叶片中克隆得到蔗糖转运蛋白VvSUC 27基因,其开放阅读框全长1515 bp,预测蛋白质分子量为54 kD,理论等电点(pI)为9.58。生物信息学预测该蛋白含有12个跨膜结构。实时荧光定量PCR(qRT-PCR)分析表明:氨基环丙烷羧酸和脱落酸处理后VvSUC 27的表达水平持续上调;水杨酸和茉莉酸甲酯处理后,VvSUC 27的表达呈现上调表现但无规律的变化特征;激发子chitin和flg22处理后,VvSUC 27基因信号响应快速且明显;感病品种‘夏黑’接种可可毛色二孢后,VvSUC 27被迅速诱导,3 h时表达量达到最高,而后逐渐减弱。本研究将为今后阐明可可毛色二孢与葡萄互作的分子机制提供参考依据。Botryosphaeria dieback seriously affects the healthy development of the grape industry,which is one of the main stem diseases on grapevines caused by Botryosphaeriaceae.Lasiodiplodia theobromae is the most pathogenic among them in China.Sugar transporters play a vital role in regulating plant immunity to the pathogens.However,the role of sugar transporters in the immune mechanisms of grapes has not been studied.In this study,the sucrose transporter VvSUC 27 gene was cloned from the cDNAs of grapevine leaves,with an open reading frame length of 1515 bp.The predicted molecular weight of the protein was 54 kD and the theoretical isoelectric point(pI)was 9.58.Besides,the protein contained 12 transmembrane structures.Real-time quantitative PCR analysis was conducted in different treatments.The results showed that the expression level of VvSUC 27 was continuously up-regulated after treated with 1-aminocyclopropane-1-carboxylic acid(ACC)and abscisic acid(ABA).After treated with salicylic acid(SA)and methyl jasmonate(MeJA),VvSUC 27 showed an up-regulation without regular changes.The response of VvSUC 27 gene signal was rapid and obvious after chitin or flg22 treatment.In addition,the gene in the susceptible cultivar‘summer black’was rapidly induced by inoculation with L.theobromae,and the expression level reached the highest 3 h after inoculation and then gradually weakened.This study provided a reference for elucidating the molecular mechanism of the interaction between L.theobromae and grapevine.

关 键 词：葡萄溃疡病 可可毛色二孢 蔗糖转运蛋白基因 基因克隆 表达分析 

分 类 号：S663.1[农业科学—果树学]