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

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 ／ZnSOD、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 trans-genic plants and WT.The expression levels of several defense-related genes MnSOD,Cu /ZnSOD,CN,NADPH oxi-dase,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 inocu-lation 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 to-bacco lines were significantly higher than those in WT.Moreover,the transgenic tobacco plants showed strong resist-ance 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.