›› 2012, Vol. 55 ›› Issue (11): 1306-1314.doi:

• 研究论文 • 上一篇    下一篇

萧氏松茎象线粒体基因组全序列测定与分析

李国宏,尚娜,魏建荣   

  • 收稿日期:2012-09-24 修回日期:2012-11-19 出版日期:2012-11-20 发布日期:2012-11-20
  • 通讯作者: 魏建荣 E-mail:weijr@hbu.edu.cn
  • 作者简介:lmz9827@yahoo.com.cn

Sequencing and analysis of the complete mitochondrial genome of Hylobitelus xiaoi (Coleptera: Curculionidae)

LI Guo-Hong, SHANG Na, WEI Jian-Rong   

  • Received:2012-09-24 Revised:2012-11-19 Online:2012-11-20 Published:2012-11-20
  • Contact: WEI Jian-Rong E-mail:weijr@hbu.edu.cn
  • About author:lmz9827@yahoo.com.cn

摘要: 象甲是鞘翅目中物种最丰富的类群, 目前关于其线粒体基因组全序列的研究还未见报道。本研究利用长距PCR和引物步移法对萧氏松茎象Hylobitelus xiaoi Zhang线粒体基因组全序列进行了测定。结果显示: 萧氏松茎象线粒体基因组序列全长16 123 bp(GenBank登录号为JX847496), 共编码37个基因和1个非编码的控制区, 基因次序与典型的六足动物线粒体基因排列一致, 未发现基因重排现象。在基因组中两个值得注意的发现分别是: 1)N链上存在1个额外的trnV-like序列, 反密码子为GAC, 长度为69 bp, 其中65 bp与J链上的trnD重叠; 2)trnSUCNnad1之间存在1个长度为232 bp的基因间隔区。全部13个蛋白质编码基因的起始密码子均为ATN, 9个蛋白质编码基因的终止密码子为TAA, 其余4个蛋白质编码基因中, nad1cox2的终止密码子为TAG, nad4nad5则以不完整的终止密码子T作为终止信号。除trnSAGN外, 其余的tRNAs均可形成典型的三叶草结构。而trnSAGN的反密码子由TCT替代GCT, 反密码子臂延长形成9 bp(中间含1个碱基突起), TΨC臂由正常的5 bp变为6 bp, DHU臂缩短仅1 bp, 各个臂之间没有连接碱基。线粒体控制区中包括10处长度不少于5 bp的poly-T(最长poly-T长度为14 bp)和2处微卫星样重复序列 (TA)6和(TA)9。本研究结果为探讨象甲总科在鞘翅目中的系统学地位及其与其他总科间的系统发生关系等问题提供了重要的分子生物学数据。

关键词: 鞘翅目, 象甲总科, 萧氏松茎象, 线粒体基因组, 长距PCR, 引物步移法

Abstract: The Curculionoidea is one of the richest groups in Coleptera, but the research about its complete mitochondrial genome has not been reported. In this study, the complete mitochondrial genome (mitogenome) of Hylobitelus xiaoi Zhang was determined using long PCR (L-PCR) and primer walking technology. The entire mitochondrial DNA (mtDNA) molecule is 16 123 bp in length and its overall nucleotide composition is heavily biased towards As and Ts, which account for 75.66% of the whole genome. The H. xiaoi genes are arranged in the same order and orientation as the other hexapod species, except for the occurrence of one 69 bp trnV-like region with anticodon GAC that overlaps 65 bp with the reversely encoded trnD and the presence of 232 bp long intergenic spacer between trnSUCN and nad1. All protein-coding genes (PCGs) start with a typical ATN codon. Nine of 13 PCGs have a complete termination codon TAA, while the remaining nad1 and cox2 are terminated with TAG, nad4 and nad5 with the incomplete T. All tRNAs show the typical clover-leaf structure, except for trnSAGN. It habors anticodon TCT instead of GCT, a lengthened 9 bp anticodon stem with a bulged nuleotide in the middle, an unusual 6 bp TΨC-stem, and a mini 1 bp DHU-stem and no connector nucleotides. The control region (CR) contains one 14 bp Poly-T sequence (ten Poly-T sequences more than 5 bp and two microsatellite-like repeating sequences (TA)6 and (TA)9. The results of this study will provide some important molecular data for discovering phylogenetic relationships among the superfamilies of Coleptera.

Key words: Coleoptera, Curculionoidea, Hylobitelus xiaoi, mitochondrial genome, long PCR (LPCR), primer walking