Multi-Gene Analysis Reveals a Lack of Genetic Divergence between Calanus agulhensis and C. sinicus (Copepoda; Calanoida)

The discrimination and taxonomic identification of marine species continues to pose a challenge despite the growing number of diagnostic metrics and approaches. This study examined the genetic relationship between two sibling species of the genus Calanus (Crustacea; Copepoda; Calanidae), C. agulhensis and C. sinicus, using a multi-gene analysis. DNA sequences were determined for portions of the mitochondrial cytochrome c oxidase I (mtCOI); nuclear citrate synthase (CS), and large subunit (28S) rRNA genes for specimens collected from the Sea of Japan and North East (NE) Pacific Ocean for C. sinicus and from the Benguela Current and Agulhas Bank, off South Africa, for C. agulhensis. For mtCOI, C. sinicus and C. agulhensis showed similar levels of haplotype diversity (Hd = 0.695 and 0.660, respectively) and nucleotide diversity (π = 0.003 and 0.002, respectively). Pairwise FST distances for mtCOI were significant only between C. agulhensis collected from the Agulhas and two C. sinicus populations: the Sea of Japan (FST = 0.152, p<0.01) and NE Pacific (FST = 0.228, p<0.005). Between the species, FST distances were low for both mtCOI (FST = 0.083, p = 0.003) and CS (FST = 0.050, p = 0.021). Large subunit (28S) rRNA showed no variation between the species. Our results provide evidence of the lack of genetic distinction of C. sinicus and C. agulhensis, raise questions of whether C. agulhensis warrants status as a distinct species, and indicate the clear need for more intensive and extensive ecological and genetic analysis.

[1]  M. Nei,et al.  MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. , 2011, Molecular biology and evolution.

[2]  A. Bucklin,et al.  Comparative phylogeography and connectivity of sibling species of the marine copepod Clausocalanus (Calanoida) , 2011 .

[3]  R. Harris,et al.  Barriers in the pelagic: population structuring of Calanus helgolandicus and C. euxinus in European waters , 2011 .

[4]  P. Ng,et al.  Recent vicariant and dispersal events affecting the phylogeny and biogeography of East Asian freshwater crab genus Nanhaipotamon (Decapoda: Potamidae). , 2011, Molecular phylogenetics and evolution.

[5]  J. Lutjeharms,et al.  Cyclonic eddies identified in the Cape Basin of the South Atlantic Ocean , 2011 .

[6]  D. Steinke,et al.  DNA barcoding of marine metazoa. , 2011, Annual review of marine science.

[7]  F. Dong,et al.  Molecular systematics and diversification of the Asian scimitar babblers (Timaliidae, Aves) based on mitochondrial and nuclear DNA sequences. , 2010, Molecular phylogenetics and evolution.

[8]  A. Bucklin,et al.  Basin-scale population genetic structure of the planktonic copepod Calanus finmarchicus in the North Atlantic Ocean , 2010 .

[9]  V. V. Kasyan Holoplankton of ship ballast water in the Port of Vladivostok , 2010, Russian Journal of Marine Biology.

[10]  L. Excoffier,et al.  Arlequin suite ver 3.5: a new series of programs to perform population genetics analyses under Linux and Windows , 2010, Molecular ecology resources.

[11]  M. Mundo‐Ocampo,et al.  Diversity of free-living marine nematodes (Enoplida) from Baja California assessed by integrative taxonomy , 2010, Marine biology.

[12]  S. Ahyong,et al.  Phylogenetic relationships among genera in the Calanidae (Crustacea: Copepoda) based on morphology , 2010 .

[13]  C. Schubart,et al.  Phylogenetic and morphometric differentiation reveal geographic radiation and pseudo-cryptic speciation in a mangrove crab from the Indo-West Pacific. , 2009, Molecular phylogenetics and evolution.

[14]  K. Holsinger,et al.  Genetics in geographically structured populations: defining, estimating and interpreting FST , 2009, Nature Reviews Genetics.

[15]  Pablo Librado,et al.  DnaSP v5: a software for comprehensive analysis of DNA polymorphism data , 2009, Bioinform..

[16]  R. Harrison,et al.  Deep genetic divergences among morphologically similar and parapatric Skistodiaptomus (Copepoda: Calanoida: Diaptomidae) challenge the hypothesis of Pleistocene speciation , 2008 .

[17]  E. Virginia Armbrust,et al.  Population structure of the planktonic copepod Calanus pacificus in the North Pacific Ocean , 2008 .

[18]  D. Posada jModelTest: phylogenetic model averaging. , 2008, Molecular biology and evolution.

[19]  P. Hebert,et al.  Comparative phylogeography of marine cladocerans , 2008 .

[20]  Alexandros Stamatakis,et al.  Exploring New Search Algorithms and Hardware for Phylogenetics: RAxML Meets the IBM Cell , 2007, J. VLSI Signal Process..

[21]  F. Cipriano,et al.  Genetic diversity of the pelagic harpacticoid copepod Macrosetella gracilis on colonies of the cyanobacterium Trichodesmium spp. , 2007 .

[22]  David J. Lohman,et al.  Cryptic species as a window on diversity and conservation. , 2007, Trends in ecology & evolution.

[23]  S. Adamowicz,et al.  Molecular systematics and patterns of morphological evolution in the Centropagidae (Copepoda: Calanoida) of Argentina , 2007 .

[24]  S. Rice,et al.  An analysis of species boundaries and biogeographic patterns in a cryptic species complex: the rotifer--Brachionus plicatilis. , 2006, Molecular phylogenetics and evolution.

[25]  Alexandros Stamatakis,et al.  RAxML-VI-HPC: maximum likelihood-based phylogenetic analyses with thousands of taxa and mixed models , 2006, Bioinform..

[26]  B. Frost,et al.  Phylogeography of Calanus helgolandicus and the Black Sea copepod Calanus euxinus, with notes on Pseudocalanus elongatus (Copepoda, Calanoida) , 2006 .

[27]  M. Miya,et al.  Molecular phylogeny and evolution of the pelagic copepod genus Neocalanus (Crustacea: Copepoda) , 2006 .

[28]  C. Meyer,et al.  DNA Barcoding: Error Rates Based on Comprehensive Sampling , 2005, PLoS biology.

[29]  E. Goetze GLOBAL POPULATION GENETIC STRUCTURE AND BIOGEOGRAPHY OF THE OCEANIC COPEPODS EUCALANUS HYALINUS AND E. SPINIFER , 2005, Evolution; international journal of organic evolution.

[30]  T. Robinson,et al.  Marine alien species of South Africa — status and impacts , 2005 .

[31]  T. Sawabe,et al.  Molecular phylogeny of Neocalanus copepods in the subarctic Pacific Ocean, with notes on non-geographical genetic variations for Neocalanus cristatus , 2004 .

[32]  E. Goetze Cryptic speciation on the high seas; global phylogenetics of the copepod family Eucalanidae , 2003, Proceedings of the Royal Society of London. Series B: Biological Sciences.

[33]  Peter Donnelly,et al.  A comparison of bayesian methods for haplotype reconstruction from population genotype data. , 2003, American journal of human genetics.

[34]  J. G. Field,et al.  Seasonal and event-scale variation in growth of Calanus agulhensis (Copepoda) in the Benguela upwelling system and implications for spawning of sardine Sardinops sagax , 2003 .

[35]  Christian Mullon,et al.  Modelling the transport success of anchovy Engraulis encrasicolus eggs and larvae in the southern Benguela: the effect of spatio-temporal spawning patterns , 2003 .

[36]  Jeremy R. deWaard,et al.  Biological identifications through DNA barcodes , 2003, Proceedings of the Royal Society of London. Series B: Biological Sciences.

[37]  N. Copley,et al.  Molecular systematic and phylogenetic assessment of 34 calanoid copepod species of the Calanidae and Clausocalanidae , 2003 .

[38]  R. Hill,et al.  Multiplexed species-specific PCR protocol to discriminate four N. Atlantic Calanus species, with an mtCOI gene tree for ten Calanus species , 2001 .

[39]  P. Donnelly,et al.  A new statistical method for haplotype reconstruction from population data. , 2001, American journal of human genetics.

[40]  S. Uye Why does Calanus sinicus prosper in the shelf ecosystem of the Northwest Pacific Ocean , 2000 .

[41]  A. Richardson,et al.  A review of the biology and ecology of Calanus agulhensis off South Africa , 2000 .

[42]  K. Crandall,et al.  TCS: a computer program to estimate gene genealogies , 2000, Molecular ecology.

[43]  N. Knowlton Molecular genetic analyses of species boundaries in the sea , 2000, Hydrobiologia.

[44]  J. Pawlowski,et al.  Molecular evidence of cryptic speciation in planktonic foraminifers and their relation to oceanic provinces. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[45]  Malcolm B. Jones,et al.  Simple molecular method to distinguish the identity of Calanus species (Copepoda: Calanoida) at any developmental stage , 1999 .

[46]  A. Meyer,et al.  Mitochondrial and nuclear rRNA based copepod phylogeny with emphasis on the Euchaetidae (Calanoida) , 1999 .

[47]  A. Bucklin,et al.  Distribution and relative abundance of Pseudocalanus moultoni and P. newmani (Copepoda: Calanoida) on Georges Bank using molecular identification of sibling species , 1998 .

[48]  R. Petit,et al.  Identifying Populations for Conservation on the Basis of Genetic Markers , 1998 .

[49]  W. Peterson Life cycle strategies of copepods in coastal upwelling zones , 1998 .

[50]  Y. Fu,et al.  Statistical tests of neutrality of mutations against population growth, hitchhiking and background selection. , 1997, Genetics.

[51]  A. Murase,et al.  Relationship of egg production rates of the planktonic copepod Calanus sinicus to phytoplankton availability in the Inland Sea of Japan , 1997 .

[52]  T. Kocher,et al.  Molecular systematics of six Calanus and three Metridia species (Calanoida: Copepoda) , 1995 .

[53]  R. Vrijenhoek,et al.  DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. , 1994, Molecular marine biology and biotechnology.

[54]  S. Uye,et al.  Ontogenetic diel vertical migration of the planktonic copepod Calanus sinicus in the Inland Sea of Japan , 1993 .

[55]  L. Excoffier,et al.  Analysis of molecular variance inferred from metric distances among DNA haplotypes: application to human mitochondrial DNA restriction data. , 1992, Genetics.

[56]  F. Tajima Statistical method for testing the neutral mutation hypothesis by DNA polymorphism. , 1989, Genetics.

[57]  J. Bradford Review of the taxonomy of the Calanidae (Copepoda) and the limits to the genus Calanus , 1988, Hydrobiologia.

[58]  E. V. Wal,et al.  Cargo vessel ballast water as a vector for the transport of non-indigenous marine species , 1988 .

[59]  Charles R. McClain,et al.  Observations of upwelling around the Izu Peninsula, Japan: May 1982 , 1987 .

[60]  Arnold L. Gordon,et al.  Interocean Exchange of Thermocline Water , 1986 .

[61]  M. Kimura A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences , 1980, Journal of Molecular Evolution.

[62]  Y. Fujita,et al.  Upwelling plumes in sagami bay and adjacent water around the Izu Islands, Japan , 1980 .

[63]  B. Frost Calanus marshallae, a new species of calanoid copepod closely allied to the sibling species C. finmarchicus and C. glacialis , 1974 .

[64]  Mikelle L. Nuwer Genetic structure and speciation in planktonic copepods: Global phylogeography of the Calanus helgolandicus clade , 2008 .

[65]  Brian D. Ortman DNA barcoding the Medusozoa and Ctenophora , 2008 .

[66]  李少菁,et al.  Mitochodrial DNA COI Sequence Analysis of Calanus sinicus (Copepod) in Qingdao Waters , 2005 .

[67]  T. Kocher,et al.  Source regions for recruitment of Calanus finmarchicus to Georges Bank: evidence from molecular population genetic analysis of mtDNA , 1996 .

[68]  K. Hulsemann Calanus sinicus Brodsky and C. jashmovi, nom. nov. (Copepoda : Calanoida) of the north-western Pacific Ocean: a comparison, with notes on the integumental pore pattern in Calanus s. str. , 1994 .

[69]  S. Uye,et al.  Geographic distribution, seasonal life cycle, biomass and production of a planktonic copepod Calarms sinicus in the Inland Sea of Japan and its neighboring Pacific Ocean , 1993 .

[70]  N. Knowlton Sibling species in the sea , 1993 .

[71]  A. Decker,et al.  A new species of Calanus (Copepoda, Calanoida) from South African waters , 1991 .

[72]  A. Fleminger,et al.  Geographical variation in Calanus helgolandicus s. l. (Copepoda, Calanoida) and evidence of recent speciation of the Black Sea population , 1987 .

[73]  B. Frost Taxonomic Status of Calanus finmarchicus and C. glacialis (Copepoda), with Special Reference to Adult Males , 1971 .