6个相似穿孔线虫种群的形态特征及rDNA-ITS序列分析
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摘要
运用光学显微镜对相似穿孔线虫进行观察及测量其主要特征;采用线虫通用引物18S和28S对6个相似穿孔线虫种群进行rDNA-ITS区进行了序列扩增,获得片段长度大约850 bp,克隆测序后使用UPGMA法进行聚类分析和构建系统发育树。结果表明:6个相似穿孔线虫的相似度高达99.3%,其中1RS、2RS、3RS和6RS 4个种群与来自台湾的KJ845638.1、广州的KF234224.1和马来西亚半岛的FJ455830.1在一个小类群中,表明它们之间的亲缘关系较近;而4RS和5RS种群在另一个小类群中。本研究为深入相似穿孔线虫不同种群间的形态差异与系统发育关系、及两者之间的关系等研究提供科学的理论依据。
In this research the main features of Radopholus similis were observed and measured by using the optical microscope. The r DNA-ITS sequences from six R. similis populations were amplified by using nematode universal primers 18 S and 28 S, the length of obtained fragment was about 850 bp, and then sequences were cloned to have a cluster analysis by using UPGMA method and construct a phylogenetic tree. The results showed that the similarity of six R. similis was as high as 99.3%, of which four populations(1 RS, 2 RS, 3 RS, 6 RS), KJ845638.1 from Taiwan, KF234224.1 form Guangzhou, and FJ455830.1 from Malaysia Peninsular were contained in the same branch, which indicated the genetic relationship between them was close, and 4 RS, 5 RS were in another branch. This research might provide the scientific theoretical basis for further analyzing the morphological differences and phylogenetic relationship among different species of R. similis, as well as the relationship between morphological and phylogenetic relationship.
In this research the main features of Radopholus similis were observed and measured by using the optical microscope. The r DNA-ITS sequences from six R. similis populations were amplified by using nematode universal primers 18 S and 28 S, the length of obtained fragment was about 850 bp, and then sequences were cloned to have a cluster analysis by using UPGMA method and construct a phylogenetic tree. The results showed that the similarity of six R. similis was as high as 99.3%, of which four populations(1 RS, 2 RS, 3 RS, 6 RS), KJ845638.1 from Taiwan, KF234224.1 form Guangzhou, and FJ455830.1 from Malaysia Peninsular were contained in the same branch, which indicated the genetic relationship between them was close, and 4 RS, 5 RS were in another branch. This research might provide the scientific theoretical basis for further analyzing the morphological differences and phylogenetic relationship among different species of R. similis, as well as the relationship between morphological and phylogenetic relationship.
引文
Elbadri G.A.A.,Geraert E.,and Moens M.,1999,Morphological differences among Radopholus similis(Cobb,1893)thorne,1949 populations,Russian Journal of Nematology,50(7):139-153
Li J.,Peng D.L.,and Huang W.K.,2008,Phylogenticanalysis of Radopholus similis from D2 and D3 fragments of the 28Sr RNA gene sequences,Huzhong Nongye Daxue Xuebao(Journal of Huazhong Agricultural University),27(5):585-589(李佳,彭德良,黄文坤,2008,香蕉穿孔线虫28S r RNA基因的D2/D3区序列分析,华中农业大学学报,27(5):585-589)
Peng D.L.,and Xie B.Y.,2004,Invasive biological Radopholus similis and their distribution,Zhiwu Baohu(Plant Protection),30(5):83-87(彭德良,谢丙炎,2004,外来入侵生物香蕉相似穿孔线虫及其发生分布,植物保护,30(5):83-87)
Subbotin S.A.,Vierstraete A.,Ley P.D.,Rowe J.,Waeyenberge L.,Moens M.,and Vanfleteren J.R.,2001,Phylogentic relationships within the cyst-forming nematodes(Nemagoda,Heteroderidae)based on analysis of sequences from the ITSregions of ribosomal DNA,Molecular Phylogenetics and Evolution,21(1):1-16
Wang Q.,Geng L.F.,Zhang D.S.,and Peng D.L.,2011,Species specific molecular diagnosis of burrowing nematode(Radopholus similis),Zhiwu Bingli Xuebao(Acta Phytopathologica Sinica),41(2):171-177(王琼,耿丽凤,张东升,彭德良,2011,香蕉穿孔线虫(Radopholus similis)特异性分子检测技术研究,植物病理学报,41(2):171-177)
Xie H.,2006,Radopholus similis and their detection and prevention controls,Zhiwu Jianyi(Plant Quarantine),20(5):321-324(谢辉,2006,香蕉穿孔线虫及其检测和防疫控制,植物检疫,20(5):321-324)
Zhao H.,Peng D.L.,and Zhu J.L.,2004,Using the r DNAITS-PCR technology to molecular diagnosis in the plant parasitic nematodes,Zhiwu Jianyi(Plant Quarantine),18(2):100-105(赵鸿,彭德良,朱建兰,2004,r DNA-ITS-PCR技术在植物寄生线虫分子诊断中的应用,植物检疫,18(2):100-105)
Zhou G.L.,Chen C.,Qi L.J.,Hu B.S.,and Bao Y.X.,2005,The classification and identification of Radopholus similis and research progress,Zhiwu Jianyi(Plant Quarantine),19(6):366-369(周国梁,陈晨,戚龙君,胡白石,包云轩,2005,相似穿孔线虫的分类鉴定及其研究进展,植物检疫,19(6):366-369)
Li J.,Peng D.L.,and Huang W.K.,2008,Phylogenticanalysis of Radopholus similis from D2 and D3 fragments of the 28Sr RNA gene sequences,Huzhong Nongye Daxue Xuebao(Journal of Huazhong Agricultural University),27(5):585-589(李佳,彭德良,黄文坤,2008,香蕉穿孔线虫28S r RNA基因的D2/D3区序列分析,华中农业大学学报,27(5):585-589)
Peng D.L.,and Xie B.Y.,2004,Invasive biological Radopholus similis and their distribution,Zhiwu Baohu(Plant Protection),30(5):83-87(彭德良,谢丙炎,2004,外来入侵生物香蕉相似穿孔线虫及其发生分布,植物保护,30(5):83-87)
Subbotin S.A.,Vierstraete A.,Ley P.D.,Rowe J.,Waeyenberge L.,Moens M.,and Vanfleteren J.R.,2001,Phylogentic relationships within the cyst-forming nematodes(Nemagoda,Heteroderidae)based on analysis of sequences from the ITSregions of ribosomal DNA,Molecular Phylogenetics and Evolution,21(1):1-16
Wang Q.,Geng L.F.,Zhang D.S.,and Peng D.L.,2011,Species specific molecular diagnosis of burrowing nematode(Radopholus similis),Zhiwu Bingli Xuebao(Acta Phytopathologica Sinica),41(2):171-177(王琼,耿丽凤,张东升,彭德良,2011,香蕉穿孔线虫(Radopholus similis)特异性分子检测技术研究,植物病理学报,41(2):171-177)
Xie H.,2006,Radopholus similis and their detection and prevention controls,Zhiwu Jianyi(Plant Quarantine),20(5):321-324(谢辉,2006,香蕉穿孔线虫及其检测和防疫控制,植物检疫,20(5):321-324)
Zhao H.,Peng D.L.,and Zhu J.L.,2004,Using the r DNAITS-PCR technology to molecular diagnosis in the plant parasitic nematodes,Zhiwu Jianyi(Plant Quarantine),18(2):100-105(赵鸿,彭德良,朱建兰,2004,r DNA-ITS-PCR技术在植物寄生线虫分子诊断中的应用,植物检疫,18(2):100-105)
Zhou G.L.,Chen C.,Qi L.J.,Hu B.S.,and Bao Y.X.,2005,The classification and identification of Radopholus similis and research progress,Zhiwu Jianyi(Plant Quarantine),19(6):366-369(周国梁,陈晨,戚龙君,胡白石,包云轩,2005,相似穿孔线虫的分类鉴定及其研究进展,植物检疫,19(6):366-369)