辣椒Me3基因介导抗根结线虫相关基因的SSH分析  

Analysis of Pepper Me3 Gene Resistance to Root Knot Nematodes by SSH

作　　者：茆振川[1] 谢丙炎[2] 杨宇红[2] 冯东昕[2] 冯兰香[2] 杨之为[1] 

机构地区：[1]西北农林科技大学植物保护学院,陕西杨凌712100 [2]中国农业科学院蔬菜花卉研究所,北京100081

出　　处：《园艺学报》2007年第6期1453-1458,共6页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(30671412;30270236);科技部‘973’重大基础研究前期研究专项(2004CCA05300);‘十一五’国家科技支撑计划项目(2006BAD08A08);农业公益性行业科研专项(nyhyzx07-050)

摘　　要：以辣椒(Capsicum annuum L.)双单倍体HDA149为试验材料,接种南方根结线虫(Meloido-gyne incognita)12、24、36h的根尖材料作为测验方(Tester),相应的未接种根尖材料作为驱动方(Driv-er),构建一个南方根结线虫诱导Me3基因表达早期的抑制消减杂交cDNA文库,通过高密度点阵膜杂交差异筛选,获得了309条表达序列标签(EST)。通过测序,除去重复序列共得到259条EST序列,在Gen-Bank上进行BLAST分析,30个EST片段未发现同源性基因,229个为具有同源性的已知基因,其中71个为功能未知的基因。在显微观察和抗性相关EST表达谱分析的基础上,推测在Me3基因介导的不亲和互作中,编码NBS-LRR结构的抗病基因参与了寄主对根结线虫的识别,激发了过敏性坏死反应的信号基因,同时多种抗性相关的转录因子基因表达,使以代谢为基础的防御反应和保护机制发挥抗线虫作用。To investigate the early up-regulated expression gene profile in pepper line HAD149 induced by root-knot nematode （RKN） Meloidogyne incognita, a forward subtracted cDNA library of pepper seedling root tips was constructed using suppression subtractive hybridization （ SSH ）. The cDNA of pepper seedling root tips inoculated with J2 RKN on 12, 24 and 36 h were used as tester, and that from untreated root tips as driver. Totally 309 SSH cDNA fragments were selected with DIG Nonradioactive Nucleic Acid Labeling and Detection System. Through the DNA sequencing, we got 259 ESTs. With the Blast, 30 EST fragments didn＇t find homologous gene, 229 had homologous genes, and among of them, 71 fragments were function unknown. Base on the observing with microscope and the ESTs function analyzing, we putative that disease resistant genes encode nucleotide binding/leucine-rich repeat （ NBS - LRR） have a key rule to recognizes root knot nematode, result in the signal transduction-related genes expressed, activate a suitable defensive response and protection mechanism, it often including hypersensitive response （HR） and Systemic Acquired Resistance （SAR）. In addition, all of these responses are base on the first and secondary metabolites.

关 键 词：辣椒 Me3基因 根结线虫 抑制性消减杂交 

分 类 号：S641.3[农业科学—蔬菜学]