不同启动子在转基因烟草中抗番茄黄化曲叶病毒的研究  

Expressing a GroEL Gene in the Different Promoters of Tobacco Plants Conferring Resistance to Tomato Yellow Leaf Curl Virus

作　　者：吴丹[1,2] 张保龙[2] 杨郁文[2] 倪万潮[2] 赵统敏[3] 张云华[1] 王荣富[1] 

机构地区：[1]安徽农业大学生命科学学院,安徽合肥230061 [2]江苏省农业科学院农业生物技术研究所,江苏南京210014 [3]江苏省农业科学院蔬菜研究所,江苏南京210014

出　　处：《江苏农业学报》2010年第1期55-60,共6页Jiangsu Journal of Agricultural Sciences

基　　金：江苏省自然科学基金项目(BK2007125)

摘　　要：为了提高植物对番茄黄化曲叶病毒的抗性,利用CaMV35S启动子、棉花韧皮部特异启动子PGHNBS、拟南芥韧皮部特异启动子SUC2分别构建了含有不同启动子控制下的烟粉虱内GroEL基因的植物表达载体P-Gro-EL、P-PGHNBS-GroEL、P-SUC2-GroEL。通过农杆菌介导转化法,获得了一批抗卡那霉素的转化再生烟草植株。RT-PCR结果表明,3个启动子皆驱动GroEL基因在转基因烟草中表达。对转化再生烟草的一次病毒注射侵染结果显示,共有19株抗性烟草植株,其中,转P-GroEL载体的有7株,转P-PGHNBS-GroEL和P-SUC2-GroEL载体的各有6株;2次病毒注射侵染的结果显示,获得7株抗性烟草植株,它们是转P-GroEL载体的1株、转P-PGHNBS-GroEL和P-SUC2-GroEL载体的各3株。2次接种的试验结果表明,PGHNBS和SUC2转基因烟草植株的抗病效果均比CaMV35S转基因植株明显。证明利用韧皮部特异表达GroEL基因可以获得抗番茄黄化曲叶病毒的转基因植物。With the purpose of enhancing the resistance against tomato yellow leaf Curl virus （TYLCV）, the GroEL gene which was driven by CaMV35S, SUC2, PGHNBS promoters was exploited from Bemisia tabaci. Three plant expression vectors （P-GruEL, P-SUC2-GroEL, P-PGHNBS-GroEL） were successfully constructed and their transgenic tobacco plants were obtained by agrobacterium-mediated transformation, respectively. The result of RT-PCR proved that GroEL gene was expressed in three regenerated kanamycin resistant tobacco plants. The detection of virus infection showed that 19 resistant tobacco plants were acquired. Seven plants were transformed with expression vector P-GroEL, and P-PGHNBS-GroEL and P-SUC2-GroEL had 6 resisitant tobacco plants respectively. Double injections showed that 7 resistant tobacco plants were acquired, only one of which was transformed with expression vector P-GroEL. However, three resisitant tobacco plants were transformed with expression vector P- PGHNBS-GroEL and P-SUC2-GroEL, respectively. Two inoculation experiments reflected that it was transgenic to- bacco with PGI-INBS and SUC2 promoters expressed higher resistance toward TYLCV. Accordingly, it is feasible to acquire anti-TYLCV tobacco by constructing the vector with phloem-specific promoters.

关 键 词：烟粉虱 番茄黄化曲叶病毒(TYLCV) 双生病毒 GroEL基因 启动子 烟草转化 

分 类 号：Q789[生物学—分子生物学]