根结线虫种群的线粒体DNA分析  

Analysis of root-knot nematode species and populations based on mitochondrial DNA

作　　者：孙龙华[1] 廖金铃[1] 李迅东[1] 卓侃[1] 

机构地区：[1]华南农业大学资源环境学院,广州510642

出　　处：《植物病理学报》2005年第2期134-140,共7页Acta Phytopathologica Sinica

基　　金：国家自然科学基金资助项目 (30170611);国家"十五"科技攻关资助项目(2001BA509b06)

摘　　要：在同工酶和形态学鉴定的基础上, 利用引物#C2F3和#1108对42个根结线虫种群线粒体DNA(mtDNA)中的COII和LrRNA间区域进行特异性PCR扩增, 35个种群的扩增产物约为1. 7kb,其中29个是南方根结线虫, 6个是爪哇根结线虫; 3个花生根结线虫种群的扩增产物约为1. 1kb; 1个种群的扩增产物约为0. 7kb,为根结线虫属在中国的新记录种; 3个北方根结线虫种群的扩增产物约为0. 5kb。用单条2龄幼虫提取物作模板得到的结果与大量提取DNA作模板的结果相同。为了区分产生相同大小片段的南方根结线虫和爪哇根结线虫,用限制性内切酶HinfⅠ对扩增产物进行酶切,结果表明:所有供试的南方根结线虫都可以被HinfⅠ酶切,且产生约1. 3和0. 4kb的2个限制性片段;但供试的爪哇根结线虫种群不能被酶切。由此表明,利用mtDNA PCR及酶切实验可以作为快速而准确地鉴定常见根结线虫的方法。Meloidogyne incognita, M. javanica and M. arenaria have their representative isozyme phenotypes. Primers #C2F3 and #1108 were utilized to amplify the intergenic region between COII and LrRNA genes of mtDNA of 42 Meloidogyne populations. Specific amplified fragments were about 1.7 kb for 35 Meloidogyne populations, including 29 M. incognita populations and 6 M. javanica populations; about 1.1 kb for 3 M. arenaria populations ; about 0.7 kb for 1 population of a new record root-knot nematode species; and about 0.5 kb for 3 M. hapla populations. The PCR results gained from DNA of the single juvenile and the mass juveniles were the same. The amplified products were digested with the restriction enzyme HinfⅠ, and the results showed that all populations of M. incognita can be digested into two restriction fragments of about 1.3 and 0.4 kb, but the PCR product of specific region on mtDNA of M. javanica can not be digested. It concluded that mtDNA was a rapid and reliable approach for molecular identification of common Meloidogyne species.

关 键 词：线粒体 DNA分析 种群 南方根结线虫 爪哇根结线虫 扩增产物 花生根结线虫 北方根结线虫 限制性内切酶 形态学鉴定 PCR扩增 根结线虫属 限制性片段 mtDNA 新记录种 酶切 同工酶 特异性 中29 提取物 小片段 模板 幼虫 

分 类 号：S828[农业科学—畜牧学] Q731[农业科学—畜牧兽医]