Phylogenetic Studies of Plant Parasitical Nematodes Based on 18S rRNA Gene
LI Xu-Qing1,2, ZHENG Jing-Wu1*
1 College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, China; 2 Hangzhou Academy of Agricultural Sciences, Hangzhou 310024, China
Abstract Traditional methods for identification and classification of plant nematodes are mainly based on morphological characters and morphometrics. These methods are subjective and cumbersome, and are difficult to make reliably taxonomy and identification. The molecular method for taxonomy and identification based on DNA not only provides objective information, but also can reveal the evolution of different groups of nematodes. In this study, 18S rRNA gene fragments of 94 plant parasitic nematode populations, which belonging to Rhabditida, Dorylaimida and Triplonchida, were amplified with primer sets NEMF1/S3 and A/S3, respectively. The sequences were compared and edited by DNAstar software, and were used to construct the phylogenetic trees by Mega 6.0 software. The results showed that the phylogenetic tree constructed based on the sequences, which PCR amplified with primers A/S3, can reflect the phylogenetic relations of nematode species within different groups and the same group. The results provide reference to study phylogeny of plant nematodes in the future.
[1] 李戌清. 2010. 毛刺线虫科(Trichodoridae)一些种群形态及分子系统学研究[D]. 博士毕业论文, 浙江大学, 导师: 郑经武, pp. 135. (Li X Q.2010. Morphological and molecular phylogeny on some Trichodoridae species and populations[D]. Thesis of Ph.D., Zhejiang University, Supervisor: Zheng J W, pp. 135.) [2] 王宝庆, 葛菁萍, 平文祥. 2005. 18S rRNA在系统进化研究中的应用[J]. 黑龙江医药, 18(4): 251-252. (Wang B Q, Ge J P, Ping W X.2005. Application of 18srDNA in systematic evolution[J]. Heilongjiang Medicine Journal, 18(4): 251-252.) [3] Blaxter M L, De Ley P, Garey J R, et al.1998. A molecular evolutionary framework for the phylum nematode[J]. Nature, 392: 71-75. [4] Boutsika K, Blok V C, Phillips M S, et al.2004. Confirmation of the synonymy of Paratrichodorus christiei (Allen, 1957) Siddiqi, 1974 with P. minor (Colbran, 1956) Siddiqi, 1974 (Nematoda: Triplonchida) based on sequence data obtained for the ribosomal DNA 18S gene[J]. Nematology, 6: 145-151. [5] De Ley P, Blaxter M L.2004. A new system for Nematoda: combining morphological characters with molecular trees, and translation clades into ranks and taxa[C]. In: Cook R C, Hunt D J, eds., Nematology Monographs and Perspectives, 2: 633-653. [6] Duarte I M, De Almeida M T M, Brown D J F, et al.2010. Phylogenentic relationships, based on SSU rDNA sequence, among the didelphic genera of the family Trichodoridae from Portugal[J]. Nematology, 12(2): 171-180. [7] He Y, Subbotin S A, Rubtsova T V, et al.2005. A molecular phylogenetic approach to Longidoridae (Nematoda: Dorylaimida)[J]. Nematology, 7(1): 111-124. [8] Mailwald M, Kappe R, Sonntag H G, 1994. Rapid presumptive identification of medically relevant yeasts to the species level by polymerase chain reaction and restriction enzyme analysis[J]. Journal of Medical and Veterinary Mycology, 32: 115-122. [9] Perry R N, Moens M主编, 简恒主译. 2011. 植物线虫学[M]. 中国农业大学出版社: 22-28. (Perry R N, Moens M Edited, Jian H Translated.2011. Plant Nematology[M]. China Agricultural University Press: 22-28.) [10] Subbotin S A, Madani M, Krall E, et al.2005. Molecular diagnostics, taxonomy, and phylogeny of the stem Nematode Ditylenchus dipsaci species complex based on the sequences of the internal transcribed spacer-rDNA[J]. Phytopathology, 95: 1308-1315. [11] Subbotin S A, Sturhan D, Chizhov V N, et al.2006. Phylogenetic analysis of Tylenchida Thorne, 1949 as inferred from D2 and D3 expansion fragments of the 28S rRNA gene sequences[J]. Nematology, 8(3): 455-474. [12] Waite I S, O'Donnell A G, Harrison A, et al.2003. Design and evaluation of nematode 18S rDNA primers for PCR and denaturing gradient gel electrophoresis (DGGE) of soil community DNA[J]. Soil Biology & Biochemistry, 35: 1165-1173. [13] Ye W M, Giblin-Davis R M, Braasch H, et al.2007. Phylogenetic relationships among Bursaphelenchus species (Nematoda: Parasitaphelenchidae) inferred from nuclear ribosomal and mitochondrial DNA sequence data[J]. Molecular Phylogenetics and Evolution, 43: 1185-1197.
