MpCYS4基因对苹果褐斑病病原菌侵染的响应表达及其转化株系抗性鉴定  

Response of MpCYS4 Gene to Diplocarpon mali Infection and Pathogen Resistance Assay in Transgenic Apple Plants

作　　者：谭延肖 郑现和 石莹 赵文超 马锋旺[2] 刘长海[2] TAN Yanxiao;ZHENG Xianhe;SHI Ying;ZHAO Wenchao;MA Fengwang;LIU Changhai(Dezhou Academy of Agricultural Sciences,Dezhou,Shandong 253015;College of Horticulture,Northwest A&F University,Yangling,Shaanxi 712100)

机构地区：[1]德州市农业科学研究院,山东德州253015 [2]西北农林科技大学园艺学院,陕西杨凌712100

出　　处：《核农学报》2020年第10期2161-2167,共7页Journal of Nuclear Agricultural Sciences

基　　金：国家自然科学基金资助项目(31572108)。

摘　　要：为探究半胱氨酸蛋白酶抑制剂(cystatin)基因MpCYS4在苹果响应苹果褐斑病病原菌侵染过程中的功能,以苹果矮化砧木M26野生型和MpCYS4过表达株系#1、#3、#4为材料,利用实时荧光定量PCR分析苹果褐斑病病菌侵染后MpCYS4基因在苹果叶片中的表达变化;同时,对转MpCYS4基因苹果叶片的褐斑病抗性进行鉴定,并对MpCYS4融合蛋白进行体外诱导纯化和抑菌活性分析。结果表明,MpCYS4基因的表达受褐斑病侵染诱导,其过量表达提高了苹果叶片对褐斑病的抗性,且经体外纯化的MpCYS4融合蛋白能够有效抑制褐斑病病原菌孢子的萌发和菌丝生长,表现出显著的抑菌活性。本研究结果为进一步探索MPCYS4的生物学功能及其抗病作用机制奠定了基础。In order to explore the function of apple cystatin gene MpCYS4 in response to Diplocarpon mali infection, we used apple dwarf rootstock M26 wild type and MpCYS4 transgenic lines #1, #3 and #4 as the experimental materials, and monitored MpCYS4 expression in apple leaves after the infection of Diplocarpon mali by quantitative real-time PCR. Meanwhile, the disease resistance of MpCYS4 transgenic apple leaves was tested, the MpCYS4 fusion protein was induced and purified, and the in vitro antibacterial activity was analyzed. The results showed that the expression of MpCYS4 gene was induced by Diplocarpon mali, and overexpression of MpCYS4 significantly improved the resistance of apple leaves to Diplocarpon mali infection. Moreover, MpCYS4 protein purified in vitro can effectively inhibit the spore germination and mycelial growth of Diplocarpon mali, showing significant antibacterial activity. These results provide a theoretical reference for further research on the biological function of apple cystatin gene MpCYS4 and the relationship between cystatin and plant disease resistance.

关 键 词：苹果 褐斑病 MpCYS4基因 转基因株系抗性鉴定 

分 类 号：S436.611.1[农业科学—农业昆虫与害虫防治]