转木霉菌激发子基因的链霉工程菌株构建及检测  

Construction and Detection of Streptomyces lydicus A01 Transformants with Elicitor Genes from Trichoderma

作　　者：吴琼[1] 陈捷[1] 李莹莹[1] 高士刚[1] 余传金[1] 李雅乾[1] 

机构地区：[1]上海交通大学农业与生物学院/农业部都市农业南方重点开放实验室,上海200240

出　　处：《中国生物防治学报》2013年第1期157-162,共6页Chinese Journal of Biological Control

基　　金：上海市科委重点项目(09391910900);国家"863"计划项目(2011AA10A205);国家"948"项目(2011-G4)

摘　　要：里氏木霉菌Trichoderma reesei QM 6a蛋白激发子基因xyn2及绿木霉菌蛋白激发子基因sm1通过调节植物乙烯、茉莉酸等信号途径激发其诱导抗性,提高其免疫力。本研究根据天蓝色链霉菌偏爱密码子人工合成了这两个基因,将其单价连接到链霉菌表达载体pIB139中。通过酶切鉴定的阳性质粒pIB139-xyn2(sm1)转化入ET12567(pUZ8002)中间菌株,利用接合转移技术定点整合到利迪链霉菌A01基因组中,采用PCR技术鉴定工程菌株A01-xyn2(sm1)。随后构建A01-xyn2+sm1双价链霉菌工程菌株并验证。实验结果表明:利用接合转移技术已成功获得含xyn2,sm1,xyn2+sm1的链霉菌工程菌株,将实现抗生与诱抗的协同作用,提高防治植物病害的效果。Trichoderma reesei QM 6a is characterized with a high level of XYN2 elicitor, which induces ethylene signaling pathway in plants, while sml elicitor produced by Trichoderma virens Gv29-8 is proved to induce jasmonic acid and ethylene signaling pathway. Both the elicitors could enhance the resistance of pathogens in plants. In this study, in order to improve the control effect of fungal pathogens by synergy of natamycin and elicitors, xyn2 and sml genes were synthetized according to the codon preference ofStreptomyces coelicolor, linked into plB139 expression vector and identified via enzyme digestion. The positive plasmid pIB139-xyn2 （sml） was transformed to ET12567 （pUZ8002） intermediate strain and further into the genome of S. lydicus A01 by conjugal-transformed method, respectively. Then, for higher induce resistanc, S. lydicus A01 transformant with both xyn2 and sml was constructed. The results showed that： The three conjugal-transformants （A01-xyn2, A01-sm1, A01-xyn2＋sml ） were constructed successfully, and they will have the synergistic effects of antagonism and induced resistance on plant disease.

关 键 词：利迪链霉菌A01 木霉菌xyn2基因 木霉菌sm1基因 工程菌株 

分 类 号：S482.292[农业科学—农药学] S476[农业科学—植物保护]