南方根结线虫Col基因分子克隆及其VIGS效应分析  

Molecular clone of Col gene in Meloidogyne incognita and analysis of its VIGS effect

作　　者：黄永红[1,2] 梅眉[1,3] 张吉祥[1] 马金慧[1] 茆振川[1] 

机构地区：[1]中国农业科学院蔬菜花卉研究所,北京100081 [2]广东省农业科学院果树研究所,农业部南亚热带果树生物学与遗传资源利用重点实验室,广州510640 [3]湖南农业大学园林园艺学院,长沙410128

出　　处：《植物保护学报》2013年第4期333-338,共6页Journal of Plant Protection

基　　金：国家重点基础发展计划项目(2009CB119000);公益性行业(农业)科研专项(201103018);国家自然科学基金重点项目(31030057);国家大宗蔬菜产业技术体系(CARS-25)

摘　　要：为了明确线虫角质层胶原蛋白基因(Col)与根结线虫病害的关系,利用从线虫基因组中预测的Col设计特异引物,克隆了南方根结线虫Meloidogyne incognitacol基因MiCol。该基因完整编码区长903bp,编码300个氨基酸。克隆的MiCol与预测的MiCol基因序列一致性高达99.67%,氨基酸序列一致性高达100%。利用病毒介导的基因沉默技术,将其导入番茄植株并接种南方根结线虫,60 d后,沉默载体pTV-MiColi处理番茄植株的根结数比空载体对照及清水对照分别减少49.4%和49.2%,表明MiCol基因沉默显著降低了南方根结线虫的侵染数量。In order to elucidate the relationship between nematode cuticle collagen gene （Col） and dis- ease caused by root-knot nematode, the MiCol gene in Meloidogyne incognita was cloned with special primers designed based on the predicted Col sequence in nematode genomes. The complete coding se- quence of the cloned MiCol was 903 bp encoding 300 amino acids. The identity between the cloned MiCol gene and the predicted MiCol gene was as high as 99.67% in nucleotide sequence and 100% in the de- duced amino acid sequence, respectively. Using virus-induced gene silence （VIGS） technique, the Mi- Col gene was transferred into tomato plantlets followed by inoculating with M. incognita. Sixty days later, the amounts of root knot on the tomato plantlets treated with the gene silencing vector pTV-MiColi de- creased by 49.4% or 49.2% compared with the empty vector control or water control, respectively. The study revealed that the MiCol gene silencing significantly reduced the infection of M. incognita

关 键 词：根结线虫 RNA干扰 角质层胶原蛋白 病毒介导的基因沉默 

分 类 号：S432.45[农业科学—植物病理学]