漾濞大泡核桃JsWRKY1过表达增强转基因烟草对胶孢炭疽菌的抗性  

Overexpression of a WRKY Transcription Factor Gene from Juglans sigillata Dode Confers Resistance to Colletotrichum gloeosporioides in Transgenic Tobacco Plants

作　　者：王国东[1] 陈朝银[1] 李金晶[1] 普丽梅 关瑞攀 葛锋[1] 刘迪秋[1] WANG Guodong;CHEN Chaoyin;LI Jinjing;PU Limei;GUAN Ruipan;GE Feng;LIU Diqiu(Faculty of Life Science and Technology,Kunming University of Science and Technology,Kunming 650500,China)

机构地区：[1]昆明理工大学生命科学与技术学院,云南昆明650500

出　　处：《华北农学报》2016年第3期155-162,共8页Acta Agriculturae Boreali-Sinica

基　　金：科技部支撑计划项目(2011BAD46B00)

摘　　要：为进一步研究JsWRKY1的生物学功能,构建JsWRKY1的植物超表达载体并转入烟草中过表达。再生的JsWRKY1转基因烟草与野生型烟草在表型上无明显差异。JsWRKY1过表达上调了转基因烟草体内几个防卫相关基因MnSOD、Cu/Zn SOD、CN、NADPH oxidase和PR1的转录水平。与野生型烟草相比,JsWRKY1转基因烟草体内的超氧化物歧化酶、过氧化物酶和抗坏血酸过氧化物酶活性在正常生长以及胶孢炭疽菌接种后均显著增强。平板抑菌试验结果显示,JsWRKY1转基因烟草叶片总蛋白对病原真菌葡萄座腔菌、串珠状赤霉菌、胶孢炭疽菌和尖孢镰刀菌的菌丝生长具有不同程度的抑制作用。此外,用胶孢炭疽菌接种烟草叶片,转基因烟草对胶孢炭疽菌的抗性明显增强。显然,JsWRKY1是漾濞大泡核桃中正调控病害防卫反应的转录因子。To investigate the function of JsWRKY1,a constitutive expression vector of JsWRKY1 was constructed and transferred into Nicotiana tabacum L. cv Xanthi. There were no visible differences between the positive transgenic plants and WT. The expression levels of several defense-related genes Mn SOD,Cu / Zn SOD,CN,NADPH oxidase,and PR1 resistance gene were up-regulated in the transgenic tobacco lines,and the SOD,APX and POD showed significantly higher activities in the transgenic lines than in wild type under normal conditions or after inoculation with C. gloeosporioides. The crude protein extract of transgenic tobacco lines inhibited the hyphal growth of the following four fungi,Botrosphaeria dothidea,Gibberella moniliformis,C. gloeosporioides and Fusarium oxysporum at different degrees. The antifungal activity in vitro plates demonstrated that the activities of SOD in the transgenic tobacco lines were significantly higher than those in WT. Moreover,the transgenic tobacco plants showed strong resistance after inoculation with C. gloeosporioides in the leaves. In conclusion,the JsWRKY1 is a positive transcription factor of J. sigiuata regulating the defense response to pathogens.

关 键 词：漾濞大泡核桃 WRKY转录因子 防卫相关基因 抗氧化酶类 胶孢炭疽菌 

分 类 号：Q78[生物学—分子生物学] S436.62[农业科学—农业昆虫与害虫防治]