5种粘孢子虫基于HSP70基因的系统分类学研究
摘要
粘孢子虫个体微小,形态相似性较大,属于国际间公认的研究难度较大的类群,仅借助于传统的形态学分类方法是很难准确的对其进行分类学研究的。本研究选用了HSP70基因序列分析,对粘孢子虫进行物种区分,并研究了一些分类阶元间的系统发生关系,同时对不同分子标记方法和不同分析方法在粘孢子虫分类和系统学研究中的应用做了较为详尽的探讨。
本研究首次完成了包含3个属、5种粘孢子虫的7条HSP70基因序列的测定,它们分别是:楚克拉虫未定种Zschokkella sp. ;瓶囊碘泡虫Myxobolus ampullicapsulatus Zhao et al., 2008;多涅茨尾孢虫Henneguya doneci Schulman, 1962 (3个种群);鳃居碘泡虫(新种) Myxobolus branchicola sp. nov.;住心碘泡虫Myxobolus hearti Chen, 1998。
本研究主要涉及两个层面的研究工作:一是基于HSP70基因,研究了多涅茨尾孢虫三个种群间的系统分类关系。利用本研究的多涅茨尾孢虫3个种群之HSP70基因序列,以及从Genbank选取得18个物种的HSP70基因序列构建分子支序树,分析了多涅茨尾孢虫3个种群的遗传多样性及多涅茨尾孢虫的系统发生。结果表明:1)遗传距离和地理距离之间呈现出一定的相关性,不同种群地理距离上的远近也直接反映于其亲缘关系上;2)HSP70较RAPD、18SrDNA更适合用来研究种群遗传距离;3)说明HSP70基因很保守,适合用于分子系统发育方面的研究;4)系统分析结果也进一步证明粘孢子虫属于后生动物,这与Smothers等(1994)依据18S rDNA所获得的研究结果是一致的;5)研究结果表明将HSP70基因应用在粘孢子虫种群多样性研究方面具有较高的可靠性。
二是基于HSP70基因,研究了五种粘孢子虫的系统分类关系。利用本研究的5种粘孢子虫之HSP70基因序列以及Genbank中选取的18个物种的HSP70基因序列构建分子支序树,研究并探讨了5种粘孢子虫的系统地位。分析研究了它们的系统发生、分子分类等问题。本研究结果表明:1)HSP70是一个可用的分子标记,在进行粘孢子虫系统分类学研究时可将海水种和淡水种分离开来;2)碘泡虫属、尾孢虫属、库道虫属可能具有共同的起源,且在进化上都呈非单系发生;3)结果支持粘孢子虫是后生动物且与水母等刺孢动物有着更近的亲缘关系的观点;4)表明以HSP70基因序列为分子标记分析高阶元粘孢子虫类群间系统发育关系的有较高的可靠性;5)研究结果将能进一步丰富粘孢子虫分子系统学数据。因此,建议物种描述时同时提供形态学数据与分子数据更为完善。
本研究的工作旨在初步研究HSP70基因应用于粘孢子虫系统分类的可行性,并利用现代技术研究手段从不同层面对多种粘孢子虫的遗传多样性、系统发生以及精细结构进行深入的探讨和研究。
Myxosporean parasites have a significant role as pathogens of freshwater and marine fishes in wild and cultured stocks with over 2,200 described species. Most species are identified on the basis of the myxospore. However, with the use of these classical zoological methods, it is very difficult to distinguish morphologically similar myxosporean species that may infect identical tissues and that develop in hosts species that are closely related taxonomically. Fortunately, molecular biological methods have become increasingly applied to parasitological studies and the taxonomic classification of myxosporeans has been expanded to incorporate phylogenetic analyses.
In this experiment, we put investigations on 5 species: Zschokkella sp., Myxobolus ampullicapsulatus Zhao et al.,2008, Henneguya doneci Schulman, 1962 (3 population), Myxobolus branchicola sp.nov., Myxobolus hearti Chen, 1998.
This work covered two main sections: one was the analysis of diversity of Henneguya doneci from the perspective of comparison of HSP70 gene. In this study, We determined the HSP70 gene sequences for 3 stocks of Henneguya doneci. The phylogeny trees were built By combining the HSP70 gene sequences of other 18 species available in the GenBank. Genetic differences among 3 stocks of Henneguya doneci and molecular phylogeny among some species were carefully compared. Through the obtained data, it’s safe to say that HSP70 gene was a considerable mark with a strong conservative character.
The other section was mainly focusing on the taxonomic and phylogenic relation of five myxosporean based on HSP70 gene. In the present study, the HSP70 gene sequences for 5 species were determined. By combining the HSP70 gene sequences of other 18 species available in the GenBank, we investigated the molecular phylogeny among some Myxosporidia species. The results of our study supported that HSP70 gene sequencing and identification of morphological characters which will enhance the taxonomy and systematics of Myxosporea.
Information delivered from both of the two aspects of this study is of great value to reaserch on different myxosporean by advanced measurements.
本研究首次完成了包含3个属、5种粘孢子虫的7条HSP70基因序列的测定,它们分别是:楚克拉虫未定种Zschokkella sp. ;瓶囊碘泡虫Myxobolus ampullicapsulatus Zhao et al., 2008;多涅茨尾孢虫Henneguya doneci Schulman, 1962 (3个种群);鳃居碘泡虫(新种) Myxobolus branchicola sp. nov.;住心碘泡虫Myxobolus hearti Chen, 1998。
本研究主要涉及两个层面的研究工作:一是基于HSP70基因,研究了多涅茨尾孢虫三个种群间的系统分类关系。利用本研究的多涅茨尾孢虫3个种群之HSP70基因序列,以及从Genbank选取得18个物种的HSP70基因序列构建分子支序树,分析了多涅茨尾孢虫3个种群的遗传多样性及多涅茨尾孢虫的系统发生。结果表明:1)遗传距离和地理距离之间呈现出一定的相关性,不同种群地理距离上的远近也直接反映于其亲缘关系上;2)HSP70较RAPD、18SrDNA更适合用来研究种群遗传距离;3)说明HSP70基因很保守,适合用于分子系统发育方面的研究;4)系统分析结果也进一步证明粘孢子虫属于后生动物,这与Smothers等(1994)依据18S rDNA所获得的研究结果是一致的;5)研究结果表明将HSP70基因应用在粘孢子虫种群多样性研究方面具有较高的可靠性。
二是基于HSP70基因,研究了五种粘孢子虫的系统分类关系。利用本研究的5种粘孢子虫之HSP70基因序列以及Genbank中选取的18个物种的HSP70基因序列构建分子支序树,研究并探讨了5种粘孢子虫的系统地位。分析研究了它们的系统发生、分子分类等问题。本研究结果表明:1)HSP70是一个可用的分子标记,在进行粘孢子虫系统分类学研究时可将海水种和淡水种分离开来;2)碘泡虫属、尾孢虫属、库道虫属可能具有共同的起源,且在进化上都呈非单系发生;3)结果支持粘孢子虫是后生动物且与水母等刺孢动物有着更近的亲缘关系的观点;4)表明以HSP70基因序列为分子标记分析高阶元粘孢子虫类群间系统发育关系的有较高的可靠性;5)研究结果将能进一步丰富粘孢子虫分子系统学数据。因此,建议物种描述时同时提供形态学数据与分子数据更为完善。
本研究的工作旨在初步研究HSP70基因应用于粘孢子虫系统分类的可行性,并利用现代技术研究手段从不同层面对多种粘孢子虫的遗传多样性、系统发生以及精细结构进行深入的探讨和研究。
Myxosporean parasites have a significant role as pathogens of freshwater and marine fishes in wild and cultured stocks with over 2,200 described species. Most species are identified on the basis of the myxospore. However, with the use of these classical zoological methods, it is very difficult to distinguish morphologically similar myxosporean species that may infect identical tissues and that develop in hosts species that are closely related taxonomically. Fortunately, molecular biological methods have become increasingly applied to parasitological studies and the taxonomic classification of myxosporeans has been expanded to incorporate phylogenetic analyses.
In this experiment, we put investigations on 5 species: Zschokkella sp., Myxobolus ampullicapsulatus Zhao et al.,2008, Henneguya doneci Schulman, 1962 (3 population), Myxobolus branchicola sp.nov., Myxobolus hearti Chen, 1998.
This work covered two main sections: one was the analysis of diversity of Henneguya doneci from the perspective of comparison of HSP70 gene. In this study, We determined the HSP70 gene sequences for 3 stocks of Henneguya doneci. The phylogeny trees were built By combining the HSP70 gene sequences of other 18 species available in the GenBank. Genetic differences among 3 stocks of Henneguya doneci and molecular phylogeny among some species were carefully compared. Through the obtained data, it’s safe to say that HSP70 gene was a considerable mark with a strong conservative character.
The other section was mainly focusing on the taxonomic and phylogenic relation of five myxosporean based on HSP70 gene. In the present study, the HSP70 gene sequences for 5 species were determined. By combining the HSP70 gene sequences of other 18 species available in the GenBank, we investigated the molecular phylogeny among some Myxosporidia species. The results of our study supported that HSP70 gene sequencing and identification of morphological characters which will enhance the taxonomy and systematics of Myxosporea.
Information delivered from both of the two aspects of this study is of great value to reaserch on different myxosporean by advanced measurements.
引文
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