摘要
采用COⅠ、16S rRNA两种分子标记对中国浙江南部沿海岛屿岩相潮间带日本花棘石鳖(Liolophura japonica)13个群体遗传特征和亲缘关系进行了探讨。结果显示:COⅠ基因片段长度为675 bp,16S rRNA基因长度为517 bp,A+T比例分别为62.5%和64.9%。COⅠ基因总多态位点数为41个,单倍型多样性指数0.847,核苷酸多态性指数为0.006 01,平均核苷酸差异数为4.057 8;16S rRNA基因的总多态位点数为9个,单倍型多样性指数0.665,核苷酸多态性指数为0.001 69,平均核苷酸差异数为0.871 2。以Acanthopleura brevispinosa和A.granulata为外群构建了ML和BI系统发育树,其拓扑结构显示,基于COⅠ基因的日本花棘石鳖形成2个分支;基于16S rRNA基因的日本花棘石鳖群体仅聚合为一支。不同地理群体单倍型散乱分布,没有出现明显的地理结构和遗传谱系结构,表明各岛屿的日本花棘石鳖为近期分化并存在基因交流。
Liolophura japonica Lischke belonging to Liolophura( Mollusca: Polyplacophora: Chitonidae) is a common species in the southern of Zhejiang Province. Few study has been done on the population structure and genetic diversity of L. japonica. In order to discuss the population structure,genetic diversity and genetic background,we constructed the phylogenetic relationships of 13 geographical populations of L. japonica from the southern of Zhejiang Province using 675 bp length of mtDNA COⅠ and a 517 bp length of 16 S rRNA genes which can provid the basis data on its resource conservation and genetics utilization. The ratio of A + T was62. 5% and 64. 9% in COⅠ and 16 S rRNA respectively,which was in accordance with the mitochondrial composition. The polymorphic loci,the parsimony informative sites,the single variable sites,the haplotype diversity index,the nucleotide polymorphism index and the average number of nucleotide differences in the COⅠ gene were 41,22,19,0. 847,0. 006 014 and 4. 057 8,respectively. A total of 30 haplotypes in COⅠgene were detected including haplotype 6 in the major haplotype. The polymorphic loci,the parsimony informative sites,the single variable sites,the haplotype diversity index,the nucleotide polymorphism index and the average number of nucleotide differences of 16 S rRNA gene were 9,4,5,0. 665,0. 0016 9 and0. 871 2,respectively. A total of 10 haplotypes of 16 S rRNA gene were detected. The phylogenetic tree of ML and BI in L. japonica was constructed using Acanthopleura brevispinosa and A. granulata as outgroups. The topological structure of L. japonica based on COⅠ gene in ML and BI analyses formed two clades while the topological structure based on 16 S rRNA gene in ML and BI analyses formed one clade. Different geographical population haplotypes were in scattered distribution and no obvious geographical structure and genetic lineage structure was found,which suggested that the gene divergence may happen recently.
引文
[1]张永普,应雪萍,黄象栋,等.浙南岛屿岩相潮间带石鳖的种类组成与数量分布[J].动物学杂志,2001,36(3):5-9.ZHANG Y P,YING X P,HUANG X D,et al.The species composition and quantity distribution of the chitons in the rocky intertidal zone of the islands in the south Zhejiang[J].Chinese Journal of Zoology,2001,36(3):5-9.
[2]卢周,陆京明,周化斌,等.洞头南北爿山海洋特别保护区潮间带大型底栖生物群落的时空分布[J].温州大学学报(自然科学版),2016,37(3):38-48.LU Z,LU J M,ZHOU H B,et al.Spatial and temporal distribution of macro-benthos communities in the intertidal zone of special marine protection area in Nanbeipan mountain,Dongtou district[J].Journal of Wenzhou University(Natural Sciences),2016,37(3):38-48.
[3]彭茂潇,钱培力,张永普,等.洞头无居民海岛岩相潮间带夏季大型底栖动物群落格局[J].生态学杂志,2013,32(9):2469-2479.PENG M X,QIAN P L,ZHANG Y P,et al.Community structure of macrozoobenthos in rocky intertidal zone of uninhabited islands in Dongtou County,Zhejiang Province of East China in summer[J].Chinese Journal of Ecology,2013,32(9):2469-2479.
[4]余海,阙伟伟,陆京明,等.七星列岛软体动物种类组成与区系[J].温州大学学报(自然科学版),2016,37(4):33-39.YU H,QUE W W,LU J M,et al.Species composition and fauna of mollusca in Seven Star Islands[J].Journal of Wenzhou University(Natural Sciences),2016,37(4):33-39.
[5]张均龙,史振平,王承,等.基于壳板和齿舌形态对中国沿岸几种常见多板纲软体动物的分类研究[J].海洋科学,2015,39(11):96-107.ZHANG J L,SHI Z P,WANG C,et al.Taxonomic study on some common species of Polyplacophora in the coast of China based on valves and rudular morphology[J].Marine Sciences,2015,39(11):96-107.
[6]陈道海,孙世春.9种石鳖壳板的形态研究[J].中国海洋大学学报(自然科学版),2010,40(6):53-60.CHEN D H,SUN S C.Observations on the shell plates of 9 chiton species(Polyplacophora)[J].Periodical of Ocean University of China(Natural Sciences),2010,40(6):53-60.
[7]张永普,刘德庆.浙南岛屿岩相潮间带石鳖的群落结构[J].动物学杂志,2002,37(4):5-9.ZHANG Y P,LIU D Q.Community structure of chitons in the rocky intertidal zone of the islands in South Zhejiang[J].Chinese Journal of Zoology,2002,37(4):5-9.
[8]SIRENKO B I.Report on present state of our knowledge with regard to the chitons of the Magellan Strait and Falkland Islands[J].Venus,2006,65(1-2):81-89.
[9]赵青松,金珊,陈寅儿.石鳖的结构与习性[J].水利渔业,2007,27(3):46-47.ZHAO Q S,JIN S,CHEN Y E.The structure and habits of chitons[J].Reservoir Fisheries,2007,27(3):46-47.
[10]GLLER M,LIUZZI M G,ZELAYA D G.A new species of Leptochiton(Polyplacophora:Leptochitonidae)from the southwestern Atlantic[J].Malacologia,2015,58(1-2):147-155.
[11]OWADA M.A new cryptic species distinguished from Ischnochiton comptus(Gould,1859)(Polyplacophora:Ischnochitonidae)in central Honshu,Japan[J].Molluscan Research,2016,36(4):255-263.
[12]张均龙,徐凤山,张素萍.多板纲软体动物系统分类学研究进展[J].海洋科学,2013,37(4):111-117.ZHANG J L,XU F S,ZHANG S P.Progress of the systematics of Polyplacophora(Mollusca)[J].Marine Sciences,2013,37(4):111-117.
[13]CHEN D H,SUN S C.Analysis of diversity of COⅠgene and system generation of nine species of chitons[J].Wuhan University Journal of Natural Sciences,2013,18(2):171-177.
[14]BORIS S O,JAVIER S.Update of the genus Leptochiton(Mollusca:Polyplacophora)in Chilean deep waters three new reports and description of two newspecies[J].Zootaxa,2016,4173(3):259-279.
[15]钱霞,刘维,赵见高.红条毛肤石鳖齿舌牙齿内的纳米磁性矿物质[J].科学通报,2002,47(1):10-13.QIAN X,ZHAO W,ZHAO J G.Magnetic anisotropy study of the radula of chiton Acanthochton rubrolinestus Lischke[J].Chinese Science Bulletin,2002,47(1):10-13.
[16]IRISARRI I,EERNISSE D J,ZARDOYA R.Molecular phylogeny of Acanthochitonina(Mollusca:Polyplacophora:Chitonida):Three new mitochondrial genomes,rearranged gene orders and systematics[J].Journal of Natural History,2014,48(45-48):2825-2853.
[17]GARCIARIOS C I,PEREZPEREZ N M,FERNANDEZLOPEZ J.et al.Calibrating the chitons(Mollusca:Polyplacophora)molecular clock with the mitochondrial DNA cytochrome C oxidase I gene[J].Revista De Biologia Marina Y Oceanografia,2014,49(2):193-207.
[18]BONATTO S L,SALZANO F M.A simple and early migration for control region sequences in a marine mammal:Implications for conservation and biogeography of Steller sea lions(Eumetopias jubatus)[J].Journal of Mammalogy,1997,77(1):95-108.
[19]STANLEYET H F,CASEY S,CARNAHAM J M,et al.Worldwide patterns of mitochondrial DNA differentiation in the harbor seal Phocavitulina[J].Molecular Biochemical Evolution,1996,13(2):368-382.
[20]VILA C,SAVOLAINEN P,MALDONADO J E,et al.Multiple and ancient origins of the domestic dog[J].Science,1997,276(5319):1687-1689.
[21]AVISE J C.Molecular markers:Natural history and evolution[M].New York:Chapman&Hall,1993.
[22]KUMAR R,JAISWAR A K,PAVAN-KUMAR.A.A,et al.DNA barcoding of Thais species(Family:Muricidae)from west coast of India[J].Indian Journal of Fisheries,2015,62(2):92-97.
[23]田吉腾,侯丫,刘志鸿,等.基于线粒体COⅠ基因的毛蚶群体遗传多样性[J].海洋科学,2016,40(1):1-9.TIAN J T,HOU Y,LIU Z H,et al.Genetic diversity of different populations of Scapharca subcrenata based on mitochondrial COⅠgene[J].Marine Sciences,2016,40(1):1-9.
[24]LI J J,YE Y Y,WU C W,et al.Genetic diversity and population structure of Sepiella japonica(Mollusca:Cephalopoda:Decapoda)inferred by mitochondrial DNA(COⅠ)variations[J].Biochemical Systematics and Ecology,2014(56):8-15.
[25]QI P Z,GUO B Y,XIE C X,et al.Assessing the genetic diversity and population structure of Culteral burnus in China based on mitochondrial 16S rRNA and COⅠgene sequences[J].Biochemical Systematics and Ecology,2013(50):390-396.
[26]FOLMER O,BLACK M,HOEH W,et al.DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoaninvertebrates[J].Molecular Marine Biology and Biotechnology,1994,3(5):294-299.
[27]PALUMBI S R,MARTIN A P,ROMANO S,et al.The simple fool's guide to PCR version2.0[M].Honolulu:Department of Zoology,University of Hawaii,1991.
[28]TAMURA K,STECHER G,PETERSON D,et al.MEGA6:Molecular evolutionary genetics analysis version 6.0[J].Molecular Biochemical Evolution.2013,30(12):2725-2729.
[29]韦筱媚,刘凡,颜琳妙,等.基于线粒体基因的广西变色树蜥种群分子系统关系与遗传多样性[J].四川动物,2017,36(5):519-530.WEI X M,LIU F,YAN L M,et al.Phylogenetic relationship and genetic diversity of Calotes versicolor in Guangxi[J].Sichuan Journal of Zoology,2017,36(5):519-530.
[30]LIBRADO P,ROZAS J.DNASP v5:A software for comprehensive analysis of DNA polymorphism data[J].Bioinformatics,2009,25(11):1451-1452.
[31]KIMURA M.A simple method for estimating evolutionary rate of base substitutions through comparative of studies of nucleotide sequence[J].Journal of Molecular Evolution,1980(16):111-120.
[32]HUELSENBECK J P,RONDQUIST F.MRBAYES:Bayesian inference of phylogenetic trees[J].Bioinformatics Applications Note,2001,17(8):754-755.
[33]FORSTER P,TORRONI A,RENFREW C.Phylogenetic star contraction applied to Asian and Papuan mt DNA evolution[J].Molecular Biochemical Evolution,2001,18(10):1864-1881.
[34]王琳楠,闫喜武,秦艳杰,等.中国帘蛤目16种经济贝类DNA条形码及分子系统发育的研究[J].大连海洋大学学报,2013,28(5):431-437.WANG L N,YAN X W,QIN Y J,et al.DNA barcoding and molecular phylogeny of 16 species of economical shellfish in Veneroida along China Coast[J].Journal of Dalian Ocean University,2013,28(5):431-437.
[35]王超,曹奕,吴杨平,等.6个不同海域文蛤地理群体的亲缘关系分析[J].海洋渔业,2016,38(3):262-272.WANG C,CAO Y,WU Y P,et al.Genetic relationship analysis of six Meretrix meretrix populations from different sea areas[J].Marine Fisheries,2016,38(3):262-272.
[36]毛阳丽,蔡厚才,李成久,等.基于线粒体COⅠ与16S rRNA基因序列探讨贻贝属的系统发育[J].南方水产,2010,6(5):27-36.MAO Y L,CAI H C,LI C J,et al.Molecular phylogenetics of genus Mytilus based on COⅠand16S rRNA sequences[J].South China Fisheries Science,2010,6(5):27-36.
[37]孙超,刘志鸿,杨爱国,等.基于线粒体COⅠ和16S rRNA基因研究3个地理群体黑龙江河蓝蛤的遗传多样性[J].海洋科学,2015,39(1):39-45.SUN C,LIU Z H,YANG A G,et al.The genetic diversity of three populations of Potamocorbula amurensis based on mitochondrial COⅠand16S rRNA gene[J].Marine Sciences,2015,39(1):39-45.
[38]苏天凤,黄建华,吴进锋,等.两种东风螺线粒体基因序列多态性研究[J].中国水产科学,2007,14(3):369-376.SU T F,HUANG J H,WU J F,et al.Genetic polymorphism of mitochondrial DNA sequences in Babylonia areolata and Babylonia formosae[J].Journal of Fishery Sciences of China,2007,14(3):369-376.
[39]LIN X Z,ZHANG X D,XIAO S,et al.Phylogeny of the cuttlefishes(Mollusca:Cephalopoda)based on mitochondrial COⅠand 16S rRNA gene sequence data[J].Acta Oceanologica Sinica,2004,23(4):699-707.
[40]GRANT P R,GRANT B R.Hybridization of bird species[J].Science,1992,256(5054):193-197.
[41]王旭.黄口荔枝螺和疣荔枝螺遗传多样性研究[D].浙江海洋学院,2013.WANG X.Research on genetic diversity of Thais luteostoma and Thais clavigera[D].Zhejiang Ocean University,2013.
[42]李宏俊,张晶晶,袁秀堂,等.利用线粒体COⅠ和微卫星标记分析文蛤7个地理群体的遗传变异[J].生态学报,2016,36(2):499-507.LI H J,ZHANG J J,YUAN X T,et al.Genetic diversity and differentiation of seven geographical populations of hard clam(Meretrix meretrix)assessed by COⅠand microsatellite markers[J].Acta Ecologica Sinica,2016,36(2):499-507.
[43]陈灵芝.中国的生物多样性现状及其保护对策[M].北京:科学出版社,1993:99-113.CHEN L Z.Biodiversity status and protection countermeasures in China[M].Beijing:Science Press,1993:99-113.
[44]李玉龙,董婧,王彬,等.基于线粒体COⅠ基因的海蜇不同地理群体遗传特征[J].应用生态学报,2016,27(7):2340-2347.LI Y L,DONG J,WANG B,et al.Genetic characterization of different populations of Rhopilema esculentum based on the mitochondrial COⅠsequence[J].Chinese Journal of Applied Ecology,2016,27(7):2340-2347.
[45]朱叶,章群,李贵生,等.中近海角木叶鲽(Pleuronichthys cornutus)种群遗传多样性研究[J].海洋通报,2012,31(5):552-556.ZHU Y,ZHANG Q,LI G S,et al.Genetic diversity of four Pleuronichthys cornutus populations in coastal waters of China[J].Marine Science Bulletin,2012,31(5):552-556.
[46]FRANKHAM R,BALLOU J D,BRISCOE D A.Introduction to Conservation Genetics[M].Cambridge:Cambridge University Press,2002.
[47]AVISE J C,NEIGEL J E,ARNOLD J.Demographic influences on mitochondrial DNA lineage survivorship in animal population[J].Journal of Molecular Evolution,1984,20(2):99-105.
[48]CARLTON J T.The Light and Smith Manual:Intertidal Invertebrates from Central California to Oregon[M].California:University of California Press,2007:701-713.
[49]SIGWART J D.Morphological cladistic analysis as amodel for character evaluation in primitive livingchitons(Polyplacophora,Lepidopleurina)[J].American Malacological Bulletin,2009,27(1-2):95-104.