Molecular Phylogeny of the Calappidae (Crustacea: Decapoda) by K4X04V74

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									Systematics of Calappidae
     (Decapoda, Brachyura)

   S. L. Boyce, T. Spears, & L. G. Abele
          Florida State University
          Tallahassee, FL U.S.A.
          Outline

• The family Calappidae
• Molecular phylogenetic study
• Morphological phylogenetic study
  based on larval characters
• Tests of alternative hypotheses
 The Calappidae
     (sensu lato)

Calappinae    (9 genera)
Hepatinae     (3 genera)
Matutinae     (4 genera)
Orithyiinae   (1 genus)
         Calappid Monophyly?

•   Guinot (1979) morphological characters
•   Rice (1980) larval evidence
•   Stevcic (1983) physiological evidence
•   Bellwood (1996) morphological cladistic
    analysis
           Morphological Hypothesis
              (Bellwood, 1996)
                               Portunids
                               Matutinae
                               Leucosiids
                               Orithyiinae
                               Dorippids
                               Xanthids
                               Calappinae
                               Hepatinae

Based on 78 adult characters
 To further investigate calappid
          systematics

• Molecular phylogenetic study using
  mt16S rDNA sequence data
• Morphological phylogenetic study
  using larval characters
• Tests of alternative hypotheses
  using adult morphology, larval
  morphology, and molecular data
  sets
        Molecular Methods

• Extract genomic DNA
• Amplify, purify, and sequence a 550
  bp region of the mt16S rRNA gene
• Align sequences, adjust by
  secondary structure, and remove
  ambiguous regions
• Perform maximum-parsimony (MP)
  and maximum-likelihood (ML)
  analyses with PAUP
                       Maximum-Parsimony Tree
                                                                      Calappa hepatica
                                                    98
169 p. i. characters                                      100     Calappa flammea
                                          58
                                                                  Calappa sulcata
TL= 670 steps
                                                                   Cryptosoma baguerri
CI= 0.503                            52             100
                                                                Cryptosoma cristata
                                                                                              Calappids
                                84                     Acanthocarpus alexandri
                                          100        Mursia armata
                                     100             Mursia cristiata
                                                     Platymera guadachardii
                                          80              Callinectes sapidus
                                  99                    Charybdis helleri                     Portunids
                                                       Portunus spinicarpus
                                                             100
                                                                  Hepatus epheliticus
                                                            100   Hepatus pudibundus          Hepatids
                                               52                 Osachila kaiserae
                                                              Ashtoret lunaris
                                                        100
                                                              Ashtoret planipes
                                                                                              Matutids
                                                               Leucosia ocellata
                                               93
                                                             Persephona mediterranea
                                                                                              Leucosiids
                                          67          Eurytium limosum
                                     100              Neopanope sp.                           Xanthids
                                                     Panopeus sp.
                                              Ethusina sp.                                    Dorippid
                                     Orityia sinica                                           Orithyia
                                     Lyreidus bairdii
                        89
                                                                                              Raninids
                                                Ranilia muricata
                             10 changes
                                                                                    Bootstrap values based on 1000 reps
                             Maximum-Likelihood Tree
                                                                                Calappa hepatica
TVM + I +                                                 100
                                                                 99    Calappa flammea
 = 0.69942
                                                                       Calappa sulcata
I = 0.3302
                                                                              Cryptosoma baguerri
                                                                                                     Calappids
                                                                 100
-ln L= 3669.29119
                                                                        Cryptosoma cristata
Bootstrap values based on 100 reps        81                             Acanthocarpus alexandri
                                                              100    Mursia armata
                                                           98        Mursia cristiata
                                                                     Platymera guadachardii
                                                      75                      Callinectes sapidus
                                                99                    Charybdis helleri
                                                                        Portunus spinicarpus
                                                                                                     Portunids/Dorippid
                                                                       Ethusina sp.
                                                                               Hepatus epheliticus
                                                                  99
                                                                          100
                                                                                Hepatus pudibundus   Hepatids
                                        54                               Osachila kaiserae
                                                                           Ashtoret lunaris
                                                                    100
                                                                           Ashtoret planipes
                                                                                                     Matutids
                             55
                                                     65      Eurytium limosum
                                                100            Neopanope sp.                         Xanthids
                        97                                       Panopeus sp.
                                                                         Leucosia ocellata
                                               97
                                                                                                     Leucosiids
                                                              Persephona mediterranea
                                                Orithyia sinica                                      Orithyia
                                                Lyreidus bairdii
                                                          Ranilia muricata                           Raninids
                                     0.05 substitutions/site
                                 Larval Characters




     antennule                                  Calappa japonica                 maxilliped 1 & 2
                                           (Taishaku and Konishi, 1995)




    antenna                           maxillule                      maxilla   abdomen & telson
Appendages from Portunus pelagicus (Shinkarenko, 1979)
                       Maximum-Parsimony Tree
47 p. i. characters               Raninoides benedicti                                         Raninid
TL= 155 steps                                             Calappa japonica
                                                  99
CI= 0.574                                                 Calappa gallus                       Calappids
Bootstrap based on 1000 reps
                                             Charybdis bimaculata
                                                 Callinectes similis
                                                                                               Portunids
                                                  Portunus rubromarginatus
                                                 Portunus pelagicus
                                             Hepatus epheliticus
                                                                                               Hepatids
                                                        Hepatus pudibundus
                                                           Eurytium limosum
                                                    74                                         Xanthids
                                                           Panopeus herbstii

                                            82
                                                   Ashtoret lunaris                            Matutids
                                                   Ashtoret planipes
                                  62
                                                               Persephona mediterranea
                                                    88
                                                                     Philya globosa
                                                                                               Leucosiids
                                                               94

                                                                          Leucosia pubescens

                                                   85           Meodorippe lanata              Dorippids
                                       74                       Ethusa microphthalma
                                                        Orithyia sinica                        Orithyia
                               5 changes
             Calappidae Monophyly
                 Calappidae                                         MP or ML
                                                                     Tree
                                       vs
                 All Other Crabs




                                  Parsimony                   Likelihood
Data set                 KH              Templeton            KH          SH
Adult Morphology         0.0001*         0.0003*              N/A        N/A
Larval Morphology        0.009-0.001* 0.01-0.02*              N/A         N/A
mt16S rDNA               0.06-0.08       0.06-0.08            0.05*      0.049*


     KH = Kishino-Hasegawa Test    SH = Shimodaira-Hasegawa Test    * = P < 0.05
Calappid Sister Relationships
 Calappinae                          Matutinae
 Hepatinae                           Leucosiids

 All Other Crabs                     All Other Crabs


                   Orithyia
                   Dorippids


                   All Other Crabs


                                     Matutinae
Calappinae                           Hepatinae
Portunids

                                     All Other Crabs
 All Other Crabs
       Calappid Sister Relationships


                       Adult Morphology    Larval Morphology      mt16S rDNA
 Hypothesis              KH Test                KH Test            SH Test


Calappinae-Hepatinae        --                   0.16-0.29            0.07

Matutinae-Leucosiid         --                   --                   0.17

Orithyia-Dorippid           --                   --                   0.18

Calappinae-Portunid         0.003*               --                   --

Matutinae-Hepatinae         0.06                 0.29-0.30            --


   KH = Kishino-Hasegawa Test    SH = Shimodaira-Hasegawa Test   * = P < 0.05
              Maximum-Likelihood Tree
                                                                                          Calappa hepatica
                                                                     100
                                                                           99    Calappa flammea
                                                                                 Calappa sulcata
                                                                           100           Cryptosoma baguerri
Short basal branches                                81
                                                                                Cryptosoma cristata
                                                                                   Acanthocarpus alexandri
                                                                        100    Mursia armata
                                                                     98        Mursia cristiata
                                                                               Platymera guadachardii
                                                                75                      Callinectes sapidus
                                                          99                    Charybdis helleri
In an attempt to resolve                                                          Portunus spinicarpus
this problem I’ve sequenced                                                      Ethusina sp.
2 regions of the nuclear 28S rRNA                                                   100  Hepatus epheliticus
gene:                                                                       99            Hepatus pudibundus
     ~40 p.i. chars out of 1300 bp                54                               Osachila kaiserae
                                                                              100    Ashtoret lunaris
                                          55                                         Ashtoret planipes
                                                               65      Eurytium limosum
                                                          100            Neopanope sp.
                                     97                                    Panopeus sp.
                                                         97                                Leucosia ocellata
                                                                        Persephona mediterranea
                                                          Orithyia sinica
                                                          Lyreidus bairdii
                                                                    Ranilia muricata
                                               0.05 substitutions/site
             Conclusions
• The subfamilies Calappinae, Hepatinae,
  and Matutinae are monophyletic

• The family Calappidae is not monophyletic

• Relationships among the subfamilies and
  potential eubrachyuran sister taxa need
  further investigation

• New nuclear markers are needed
               Acknowledgements
For providing specimens:       For providing support:
•Orpha Bellwood               • Kasia Chodyla
•Savel Daniels                • Congress of Graduate
                                Students, FSU
•Won Kim
                              • Department of Biological
•Timothy Stebbins               Sciences, FSU
•Regina Wetzer                • FSU Sequencing Facility
•Peter Wirtz                  • NSF
•Natural History Museum of
Los Angeles County
•National Museum of Natural
History

								
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