基于5S核糖体间隔序列的丝虫寄生虫分子系统发育研究。

Molecular phylogenetic studies on filarial parasites based on 5S ribosomal spacer sequences.

作者信息

Xie H, Bain O, Williams S A

机构信息

Program in Molecular and Cellular Biology, University of Massachusetts at Amherst, MA 01003, USA.

出版信息

Parasite. 1994 Jun;1(2):141-51. doi: 10.1051/parasite/1994012141.

DOI:10.1051/parasite/1994012141

PMID:9140481

Abstract

This paper is the first large-scale molecular phylogenetic study on filarial parasites (family Onchocercidae) which includes 16 species of 6 genera: Brugia beaveri Ash et Little, 1962, B. buckleyi Dissanaike et Paramananthan, 1961; B. malayi (Brug, 1927) Buckley, 1960; B. pahangi (Buckley et Edeson, 1956) Buckley, 1960; B. patei (Buckley, Nelson et Heisch, 1958) Buckley, 1960; B. timori Partono et al, 1977; Wuchereria bancrofti (Cobbold, 1877) Seurat, 1921: W. kalimantani Palmieri. Purnomo, Dennis and Marwoto, 1980: Mansonella perstans (Manson, 1891) Eberhard et Orihel, 1984; loa loc, Stiles, 1905; Onchocerca volvulus (Leuckart, 1983) Railliet er Henry, 1910; O. ochengi Bwangamoi, 1969; O. gutturosa Neumann, 1910; Dirofilaria immitis (Leidy, 1856) Railliet e Henry, 1911; Acanthocheilonema viteae (Krepkogorskaya, 1933) Bain, Baker et Chabaud, 1982 and Litomosoides sigmodontis Chandler, 1931. 5S rRNA gene spacer region sequence data were collected by PCR, cloning and dideoxy sequencing. The 5S rRNA gene spacer region sequences were aligned and analyzed by maximum parsimony algorithms, distance methods and maximum likelihood methods to construct phylogenetic trees. Bootstrap analysis was used to test the robustness of the different phylogenetic reconstructions. The data indicated that 5S spacer region sequences are highly conserved within species yet differ significantly between species. Spliced leader sequences were observed in all of the 5S rDNA spacers with no sequence variation, although flanking region sequence and length heterogeneity was observed even within species. All of the various tree-building methods gave very similar results. This study identified four clades which are strongly supported by bootstrap analysis the Brugia clade; the Wuchereria clade; the Brugia-Wuchereria clade and the Onchocerca clade. The analyses indicated that L. sigmodontis and A. viteae may be the most primitive among the 16 species studied. The data did not show any close relationship between Loa loa and D. immitis presently classified in the same subfamily, and the constitution of the Dirofilariinae subfamily is questionable.

摘要

本文是关于丝虫寄生虫（盘尾丝虫科）的首次大规模分子系统发育研究，涵盖6个属的16个物种：1962年的比氏布鲁线虫（Ash et Little）、1961年的巴克利布鲁线虫（Dissanaike et Paramananthan）；马来布鲁线虫（Brug, 1927）Buckley, 1960；彭亨布鲁线虫（Buckley et Edeson, 1956）Buckley, 1960；派氏布鲁线虫（Buckley, Nelson et Heisch, 1958）Buckley, 1960；帝汶布鲁线虫（Partono et al, 1977）；班氏吴策线虫（Cobbold, 1877）Seurat, 1921：卡里曼丹吴策线虫（Palmieri. Purnomo, Dennis and Marwoto, 1980）；常现曼森线虫（Manson, 1891）Eberhard et Orihel, 1984；罗阿罗阿丝虫（Stiles, 1905）；旋盘尾丝虫（Leuckart, 1983）Railliet er Henry, 1910；奥氏盘尾丝虫（Bwangamoi, 1969）；喉盘尾丝虫（Neumann, 1910）；犬恶丝虫（Leidy, 1856）Railliet e Henry, 1911；链尾唇棘线虫（Krepkogorskaya, 1933）Bain, Baker et Chabaud, 1982和猴丝虫（Chandler, 1931）。通过聚合酶链反应（PCR）、克隆和双脱氧测序收集5S核糖体RNA（rRNA）基因间隔区序列数据。采用最大简约算法、距离法和最大似然法对5S rRNA基因间隔区序列进行比对和分析，以构建系统发育树。使用自展分析来检验不同系统发育重建的稳健性。数据表明，5S间隔区序列在物种内高度保守，但在物种间差异显著。在所有5S rDNA间隔区均观察到拼接前导序列，且无序列变异，尽管即使在物种内侧翼区序列和长度也存在异质性。所有不同的建树方法都得到了非常相似的结果。本研究确定了四个分支，自展分析有力支持了这些分支：布鲁线虫分支；吴策线虫分支；布鲁线虫 - 吴策线虫分支和盘尾丝虫分支。分析表明，在研究的16个物种中，猴丝虫和链尾唇棘线虫可能是最原始的。数据未显示目前分类在同一亚科的罗阿罗阿丝虫和犬恶丝虫之间有任何密切关系，恶丝虫亚科的构成值得怀疑。