›› 2009, Vol. 52 ›› Issue (10): 1083-1089.

• 研究论文 • 上一篇    下一篇

中华卵索线虫线粒体基因组多态性分析

何芳, 姜爱兰, 李神斌, 吴运梅, 王国秀   

  • 出版日期:2009-10-20 发布日期:2009-10-20
  • 通讯作者: 王国秀

Sequencing and polymorphism analysis of the complete mitochondrial genome of the entomopathogenic nematode Ovomermis sinensis (Nematoda: Mermithidae)

  • Online:2009-10-20 Published:2009-10-20

摘要: 为完善昆虫病原索科线虫线粒体基因组全序列数据库, 更系统地研究其基因组特征和系统演化规律, 进而为发挥该线虫生防潜力打下基础, 我们开展了中华卵索线虫Ovomermis sinensis线粒体全基因组的研究。该研究通过线粒体基因组滚环复制及酶切图谱, 揭示了中华卵索线虫线粒体基因组具有种内遗传多态性, 即群体中单体线虫具有独特的酶切条带, 且条带累加之和变化范围较大, 为16.5~24.5 kb。为进一步了解线粒体基因组多态性特征及产生的分子机制, 采用两步长PCR方法对2条代表性成虫线粒体基因组进行了测序及拼接, 得其全长分别为18 864和16 777 bp。对这2条序列的比对表明, 线粒体基因组中位于ND2ND4之间的可变区域, 不仅基因排列顺序不同, 且存在ND3基因重复现象, 这是导致中华卵索线虫线粒体基因组呈现多态性的主要原因。通过对以上研究结果的分析及与GenBank中已有的6种索科线虫线粒体基因组序列进行比对, 概括出其线粒体基因组基本特点: ①线粒体基因排列顺序各不相同;②部分线虫线粒体基因存在重复现象, 且重复次数不同;③线粒体基因组大小存在很大差异。

关键词: 中华卵索线虫, 昆虫病原线虫, 线粒体基因组, 多态性

Abstract: To augment the databases of Mermithidae mitochondrial genome, to study the mitochondrial genome characteristics and phylogenies of the nematodes, and to provide the theoretical information for the biological control, we carried out the research on the entomopathogenic nematode Ovomermis sinensis. The characterization of mitochondrial genomes from individual O. sinensis nematodes showed that numerous mtDNA haplotypes, ranging in size from 16.5 to 24.5 kb. To understand the molecular basis of this hypervariation, complete nucleotide sequences of two O. sinensis mtDNA haplotypes were determined by two-step long-PCR, with the length of 18 864 and 16 777 bp, respectively. Our data revealed that O. sinensis mtDNA can be divided into a constant region and a hypervariable region. The hypervariable segment, residing between ND2 and ND4, contributes exclusively to O. sinensis mtDNA size variation. Within this region, mtDNA coding genes and putative nonfunctional sequences are duplicated and rearranged to varying extents. Based on the above analysis and the comparison of the mitochondrial genomes of O. sinensis and other six species of Mermithidae, the following characteristics of the mitochondrial genomes of Mermithidae were summarized: firstly, the gene orders of mitochondrial genomes in Mermithidae are different; secondly, the frequency of gene duplication is variable among some species; and thirdly, the sizes of complete mitochondrial genomes are different.

Key words: Ovomermis sinensis, entomogenous nematodes, mitochondrial genome, polymorphism