Polyphyly across oceans: a molecular phylogeny
of the Chromodorididae (Mollusca,Nudibranchia)
Lucy Turner
School of Biological Sciences, University of Bristol, Woodland Road,
Bristol, BS8 1UG, UK. Email: lucy...@bristol.ac.uk
Having done my first degree in marine biology (Marine Biology BSc,
University of Wales, Swansea, 2003) where I completed my dissertation
on opisthobranch behavioural ecology and being a keen diver I had
for some time been interested in opisthobranchs. Upon the start of
my MRes studies at the University of Plymouth (where I was based at
the Marine Biological Association) I started researching topics for
my nine month research project and came upon a particularly intriguing
phylogenetic problem, one that the more I researched it, the more
fascinating it became; that of the phylogenetics of one of the largest
and most spectacularly coloured groups of opisthobranch molluscs,
the Chromodorididae.
History of the problem
Representatives of the Chromodorididae are found in all ocean basins,
at all latitudes, with the highest diversity occurring in tropical
regions (Figure. 1 will
open in a separate window when you click). The family is known
to comprise over 300 described species although it is thought that
there are many more members yet to be discovered, and many more await
description. The classification and phylogenetic analysis of the Chromodorididae
have an extremely confused history. Early attempts at the classification
of the group were based primarily on the examination of morphological
characteristics and external colouration, but there has remained a
poor knowledge of the comparative anatomy of the many species within
the family. Rudman (1984) carried out a comprehensive revision of
the family, using previously untapped reproductive characters, together
with external colouration and radular morphology to produce a detailed
discussion about potential evolutionary trends. Rudman proposed that
the Chromodorididae comprised 16 genera which could be split into
three subgroups: the Cadlina subgroup, which he considered
to contain the three genera exhibiting the most primitive characters
(Cadlina, Cadlinella and Tyrinna); the Chromodoris
subgroup, containing six genera (Chromodoris, Ardeadoris, Glossodoris,
Noumea, Pectenodoris and Verconia); and the Hypselodoris
subgroup, containing seven genera (Hypselodoris, Ceratosoma,
Digidentis, Durvilledoris, Mexichromis, Risbecia and Thorunna)
which he considered to possess the most advanced characters, having
characters derived from those found within the Chromodoris subgroup.
Before I started this project Rudman's work remained the best revision
of the family to date, although it only addressed representatives
from the Indo-west Pacific, despite the fact that the Chromodorididae
have a worldwide distribution. Since Rudman's 1984 paper a few studies
have been conducted examining some of the 17 genera of the family
in phylogenetic detail (Ceratosoma, Hypselodoris and
Chromodoris (Wilson & Lee, 2005)). Wilson and Lee's paper
suggested that Chromodoris was paraphyletic, although broad
geographic sampling was not undertaken. However, in general, the comparison
of congeners from different ocean basins had not yet been addressed
with appropriate outgroups, and the monophyly of broadly distributed
genera remained largely untested. Also, despite recent studies suggesting
new sources of taxonomically informative characters for the Chromodorididae,
such as histological structures, sperm ultrastructure and egg mass
shape there has been no further work on the phylogenetics of the Chromodorididae
as a complete family and the phylogeny of the group has remained unresolved.
Master's work
For my master's project I decided to use molecular methods to investigate
the phylogenetics of the Chromodorididae, specifically by sequencing
two genes from members of this family: the mitochondrial 16S gene
and the nuclear 18S gene and then use these results to: (a) reconstruct
chromodorid phylogeny; (b) to establish the position of the basal
genera Cadlina, Cadlinella and Tyrinna either within the
Chromodorididae or outside the family; (c) to test the monophyly of
the more widely distributed genera of the Chromodorididae (i.e. Chromodoris,
Glossodoris and Hypselodoris).
However, a stronger phylogenetic signal was observed from the 16S
gene compared to that of the 18S gene and it was concluded that the
phylogenetic information obtained from the 18S gene was not enough
to produce a fully resolved hypothesis for the phylogeny of the Chromodorididae.
I was therefore keen to examine another gene for phylogenetic information
and after further research decided that the mitochondrial COI gene
was a good candidate. It was here that the money awarded by the Malacalogical
Society was invaluable as it was sufficient to sequence the mitochondrial
COI gene from the specimens I had already sequenced 16S and 18S for,
as well as enabling me to sequence 16S and COI for further species
that I previously did not have access to material for.
Results and Discussion
In total sequences were used from 61 species of chromodorids, including
at least one representative from each genus. Where possible the type
species were used. Some additional sequences, where they already existed
were taken from GenBank. An additional 7 species (from the Actinocyclidae,
Dorididae and Discodorididae) were used as outgroups to root the trees
produced.
Our most resolved tree is shown in Figure 2 (click
here to open this in a separate window). This tree was produced
using a combination of the 16S and COI data. Some key findings included:
This work has been presented at the 2nd International Opisthobranch
Workshop (Bonn, Germany, September 2006) and The Society for Integrative
and Comparative Biology Annual Meeting (Phoenix, Arizona, USA, January
2007). The results have been submitted to Zoologica Scripta where
the paper is currently in review: Turner LM and Wilson N "Paraphyly
across oceans: a molecular phylogeny of the family Chromodorididae
(Gastropoda: Nudibranchia) using three molecular markers".
I would like to thank again the Malacological Society of London for their grant which has enabled this project to be seen through to completion. I would also like to thank all the people who so generously donated material, without which this project would not be so wide ranging.
Literature Cited
Rudman, W.B., 1984. The Chromodorididae (Opisthobranchia: Mollusca)
of the Indo-West Pacific: a review of the genera. Zool. J. Linn.
Soc. 81, 115-273.
Wilson, N.G., Lee, M.S.Y., 2005. Molecular phylogeny of Chromodoris
(Mollusca, Nudibranchia) and the identification of a planar spawning
clade. Mol. Phylogenet. Evol. 36, 722-727.