应用分子系统发育学的方法,以蟹类18S rDNA和线粒体细胞色素C氧化酶亚单位Ⅰ(COⅠ)基因序列片段为分子标记,结合形态学特征对10种蟹类的分类地位进行探讨。实验共获得10 条18S rDNA序列,长度为1780~1787 bp,其中A、T, G,C平均含量分别为23.72%, 24.58%,24.52%和27.17%;通过序列对比,发现18S rDNA序列相对保守,只含有1个从88 bp 到130 bp 约 50 bp的高可变区;物种间碱基距离比较小,从0.001到0.017。18S rDNA系统发育树为方蟹总科、沙蟹总科和梭子蟹总科起源于同-海洋蟹类祖先提供了分于生物q证惦,开上火亚N内尔量分别为26.88%.弓蟹科。获得的5条CO Ⅰ基因序列,长度均为709 bp,A、T,G、C平均含量分别为26.88%,37.62%,17.50%和18.00%;COⅠ基因片段变异性高,物种间碱基距离从0.016到0.138。COⅠ基因系统发育树真实地反映了住属各物种之间的进化关系,和传统分类非常吻合,为形态特征相似的姆类鉴定提供了-种快速准确的方法。
Abstract
Using molecular phylogenetic methodology with gene sequence fragments of 18S rDNA and subunitl of C enzyme oxide of mitochondrial dell pigment(CO I) as the molecular mark and combined withmorphological character,evolutionary history of selected brachyuran crabs were discussed. Totally 10 18SrDNA sequnces and 5 CO I gene fragments were obtained by Polymerase Chain Reaction (PCR),andsummited to the Genebank. The length of 18S rDNA sequences from 10 crabs,ranges from 1 780 to l787 bp,in which the shortest one is Hemigrapsus sinensis, the longest one is Parthenope validus. In the 18S rDNAsequnces, the average content of base A,T,G and C is 23.72%,24.58%,24.52%,and 27.17% separately.Sequence alignment shows that 18S rDNA sequences are relatively conservative,containing only onehypervariable region of about 50 bp between 88 bp and 130 bp. The base distance of the species is very small,from 0.001 to 0.017.The phylogenetic trees of 18S rDNA give molecular biological proof for the same originof Grapsioidea,Ocypodioidea and Portunoidea, and also support moving the Helice crab from Sesarminae toVaruninae.The length of mitochondrial DNA sequence from 5 charybdis crabs is 709 bp,and the averagecontent of the base A,T,G and C is 26.88%,37.62%,17.50% and 18.00%separately.The fragment ofco I gene has high species variability,more suitable for the study of the interspecific evolutionary history.The base distance of the charybdis is long,from 0.016 to 0.138.The phylogenetic trees of CO I gene truereflect the evolutionary relations of different charybdis species and are concordant with traditional taxology.This provides a molecular method for the identification of charybdis species of similar morphologicalcharact ers.
关键词
蟹类 /
18S rDNA /
COl /
分子系统发育
Key words
crab /
18S rDNA /
COl /
phylogenesis
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参考文献
[1] BOWMAN T E,ABELE L G. Classification of recent Crustacea[M]//ABELE L G. Biology of Crustacea. New York: Academic Press,1982: 1-92.
[2] DAI Ai-yu,YNAG Si-qiong,SONG Yu-zhi,et al. China sea crabs[M]. Beijing: OceanPress,1986.
戴爱云,杨思琼,宋玉枝,等.中国海洋蟹类[M].北京:海洋出版社, 986.
[3] HARRISON M K, CRESPI B J. Phylogenetics of cancer crabs (Crustacea: Decapoda: Brachyura) [J]. Molecular Phylogenetics and Evolution, 1999, 2(2): 186-199.
[4] GAO Tian-xiang,WANG Yu-jiang,LIU Jin-xian,et al. Study on phylogenetic relationships of four scylla species and taxonomic sta-tus of mud crab in China based on mitochondrial 12S rRNA sequences. Periodical of Ocean University of China, 2007,37 (1):57-60.
高天翔,王玉江,刘进贤,等.基于线粒体12S rRNA序列探讨4种青蟹系统发育关系及中国沿海青蟹的分类地位[J].中国海洋大学学报, 007,37(1):57-60.
[5] SPEARS T,ABELE L G,KIM W. The monophyly of brachyuran crabs: A phylogenetic study based on 18S rRNA[J]. System-atic Biology, 1992,41(4): 446-461.
[6] AHYONG S T,LAI J C Y,SHARKEY D. Phylogenetics of the brachyuran crabs (Crustacea: Decapoda):The status of Podotremata based on small subunit nuclear ribosomal RNA[J]. Molecu-lar Phylogenetics andEvolution,2007,45(2):576-586.
[7] MARTIN J W. Crabs of the family Homolodromiidae. First record of the larvae[J]. Journal of Crustacean Biology, 1991, 11(I), 156-161.
[8] JAMIESON B G M. Ultrastructural comparison of the spermato-zoa of Ranina ranina (Oxystomata) and of other crabs exemplified by Portunus pelagicus (Brachygnatha) (Crustacea, Brachyura)[J]. Zoomorphology,989,109(2): 103-111.
[9] JAMIESON BG M. The ultrastructure of the spermatozoa of Petalomera lateralis (Gray) (Crustacea,Brachyura, Dromiacea) and its phylogenetic significance. Invertebrate Reproduction q De-velopment, 1990,7(1);39-45.
[10] SCHRAM FR. The fossil record and evolution of Crustacea[M]//ABELE L G. Biology of Crustacea. New York, Academic Press,1982: 94-147.
[11] IGUCHI A,ITO H,UENO M,et al Molecular phylogeny of the deep-sea Buccinum species (Gastropoda: Buccinidae) around Japan: In.ter-and intraspecific relationships inferred from mito-chondrial 16S rRNA sequences[J]. Molecular Phylogenetics and Evolution,2007,44(3):1 342-1 345.
[12] LIAO Xiu-zhen, LIN Rong-cheng. Phylogenetic analysis of polychaetes based on sequences of 18S rDNA and CO I segments[J]. Journal of Oceanography in Taiwan Strait, 2006,25(4): 490-497.
廖秀珍,林荣澄.多毛类18S rDNA和CO I基因序列片段及其分子系统发育研究[J].台湾海峡, 006,25(4):490-497.
[13] WEI Guo-qing, DAI Jun-jun, LIU Chao-liang,et al. Sequence of the 18S Ribosomal RNA Gene of of silkwormCBombyx mori) and molecular systematic analysis[J]. Journal of Economic Animal, 2006,10(3):151-155.
魏国清,代君君,刘朝良,等.家蚕核糖体18S RNA基因的序列分析及分子系统学研究[J].经济动物学报,2006,10(3):151-155)
[14] WEINBERG J R,DAHLGREN T G,TROWBRIDGE N,et al. Genetic differences within and between species of deep-sea crabs (Chaceon)from the North Atlantic Ocean[J]. Biological Bulletin,2003,204(3):318-326.
[15] GAO Tian-xing,WANG Yu-jiang,LIU Jin-xian,et al Sequence analysis of mitochondrial 12S rRNA gene in three species of Scylla[J]. Journal of Fishery Science of China,2005,29(3);313-317.
高天翔,王玉江,刘进贤,等.3种青蟹线粒体12S rRNA基因序列分析[J].中国水产学报, 005,29(3): 313-317.
[16] SUN Hong-ying. The mitochondrial genome of the mitten crab, Eriocheir japonica sinensis and genetic marker from mitochondri-al 16S rDNA[D]. Nanjing: Nanjing Normal University,002.
孙红英.中华绒螯蟹线粒体基因组与16SrDNA遗传标记研究 [D].南京:南京师范大学, 002.
[17] GUO Tian-hui,XIAO Yu, CHEN Si-qing,et al. A study of the sequences of mitochondrial 16S rRNA and CO I gene fragment of portunus trituberculatus[J]. Journal of Ocean University of China,2004,34(1):022-028.
郭天慧,晓瑜,陈四清,等.三疣梭子蟹线粒体DNA 16S rRNA和CO I基因片段序列的比较研究[J].中国海洋大学报,2004,34 (1)022-028.
[18] LUNT D H,ZHANG D X,SZYMURA J M. The insect cyto-chrome oxidase I gene: evolutionary patterns and conserved primers for phylogenetic studies [J]. Insect Mol Biol,1996,5(3):153-165.
[19] HU Peng-fei,Wang-qian,DAI Wei,et al. Study on mitochon-drial DNA diversity of Eriocheir sinensis in Changjiang River[J]. Chinese Journal of Fisheries,2006,9(1): 1-5.
胡鹏飞,王茜,戴伟,等.长江水系中华绒螯蟹线粒体DNA的遗传多样性研究[J].水产学杂志, 006,19(1):1-5.
基金
中国大洋协会国际海底区域研究开发“十一五”资助项目(DYXM-115-02-4-05)
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