绿色荧光蛋白标记检测鸦胆子素D抗TMV活性  

Detection of Anti-TMV Activity of Bruceine D by Green Fluorescent Protein Labeling

作　　者：陈启建[1] 董雷[1] 欧阳明安[1] 谢联辉[1] 林奇英[1] 

机构地区：[1]福建农林大学植物病毒研究所生物农药与化学生物学教育部重点实验室,福建福州350002

出　　处：《激光生物学报》2010年第6期742-747,共6页Acta Laser Biology Sinica

基　　金：国家自然科学基金项目(30770089);福建省自然科学基金项目(2009J01069)

摘　　要：以绿色荧光蛋白(green fluorescent protein,GFP)为报告基因,将含TMV表达载体的质粒p35S-30B:GFP转化农杆菌EHA 105,通过渗透法把经MMA诱导后的农杆菌悬浮液注射到本氏烟叶片内,测定了鸦胆子素D(Bruceine D)对烟草植株内TMV的增殖和运动的抑制作用;通过PEG介导法把p35S-30B:GFP转化到本氏烟叶肉细胞原生质体内,测定了Bruceine D对烟草原生质体中TMV增殖的抑制效果。结果表明,在10μg/mL浓度下,Bruceine D不仅可抑制烟草叶肉细胞原生质体中TMV的增殖,还可以抑制烟草接种叶中TMV向茎部及植株上部叶片移动,且对寄主植物不造成明显的毒害。To investigate the effects of Bruceine D on multiplication and movement of TMV in Nicotiana benthamiana, the green fluorescent protein （GFP） gene was used as a reporter gene, and agrobacterium strain EHA 105 harbouring TMV based expression vector p35S-30B：GFP was infiltrated into Nicotiaaa benthamiana leaves after induction by MMA. For assessment of Bruceine D inhibitting replication of TMV in tobacco protoplasts, p35S-30B：GFP after PEG-mediated was delivered into protoplasts derived from Nicotiana benthamiana leaves. The results showed that Bruceine D could not only inhibit the multiplication of TMV in the protoplasts, but also restrict systemic movement of TMV in tobacco at 10 μg/mL, while no significant negative effects on the normal physiology of the tested plants were observed.

关 键 词：绿色荧光蛋白 烟草花叶病毒 鸦胆子素D 

分 类 号：S432.41[农业科学—植物病理学]