Corresponding author: Donald Colgan (
Academic editor: Cristian Altaba
The Australian Museum
The authors declare they have no conflicts of interest
The two genera of mudflat snails of the family
The morphologies of
This study was conducted to clarify the distribution and variation of the three south-eastern Australian
Specimens were collected by hand and stored at -80°C or in 95% ethanol until use. Specimens were morphologically identified using a stereomicroscope according to the body pigmentation criteria specified in
AMS: Australian Museum Malacology collection.
MAL#: designation of Australian Museum Malacology collection locality
CASIZ: California Academy of Sciences Invertebrate Zoology collection
See Fig.
New South Wales:
N1: Port Jackson, Exile Bay, Bayview Park (MAL# 88194),
N2: Cuttagee Lake, estuary to west of the road bridge (MAL#88542),
Pittwater, Careel Bay, (MAL# 88529),
Tasmania:
T1: Tamar River, west bank, mudflats at Deviot (MAL# 72693),
T2: Henderson Lagoon, south bank at Falmouth (MAL# 76559),
T3: Snug, mudflats, estuary immediately S of town, upstream of bridge (MAL# 76557),
DNA was extracted using the QIAGEN DNeasy Blood and Tissue Extraction kit following the manufacturer’s instructions. PCR amplifications generally followed the procedures in
Data were collected from parts of the COI and 28S rRNA genes. COI was amplified using the universal primers of
Chromatograms were examined in Sequencher version 5.4.5 (Gene Codes Corporation, Ann Arbor, MI). The dataset, comprising new sequences and GenBank data, was aligned in ClustalX (
Various phylogenetic analyses were conducted, although only the Maximum Likelihood (ML) analyses are reported in detail below. These were performed on the CIPRES data portal (
Net pairwise Kimura 2-parameter genetic distances (
Statistical parsimony analyses of the COI data were conducted with TCS 1.21 (
The GenBank accession numbers for the newly-collected sequences are
Suppl. material
The alignment of the cytochrome
Suppl. material
The alignment of the 28S ribosomal sequences used here. Sequences are identified by the Australian Museum registration number or GenBank accession number.
The alignment of COI sequences (Suppl. material
There was no geographic pattern of variation within
Average pairwise distances within species were similar for
The genetic diversity measures (Table
The same sequence of the ribosomal RNA fragment (Suppl. material
The DNA sequencing results provide novel insights into the distribution of
Sympatry at the local level was observed for
There is considerable genetic diversity within each of the three species of
The authors wish to thank Tina Reutelshöfer and Pam Da Costa for technical assistance. We also thank Tina for fieldwork and collection assistance.
The authors declare they have no conflicts of interest
Map of collection localities of
Maximum Likelihood phylogeny of the relationships between
TCS analysis of the COI sequences prepared by the PopART graphical interface. The figures beside branches specify the number of mutational steps between network elements. Branches lacking figures represent one step. The provenance of sequences is colour-coded according to the legend. Abbreviations in parentheses in the legend indicate the newly-collected localities, detailed in the Materials and Methods. The numbers of occurrences of a haplotype is proportional to the size of the circle representing it. All but three haplotypes occurred only once. The specimens with a haplotype found in multiple individuals (shown by larger circles) are indicated by the accession numbers in the box nearest their symbol.
Estimates of pairwise genetic distance within species and net pairwise genetic distance between species. Analyses were conducted using the Kimura 2-parameter model. Standard error estimates are shown above the diagonal for inter-species comparisons and after the distance measure for intra-species comparisons.
|
|
|
|
|
|
|
0.007 ± 0.002 | 0.244 | 0.021 | 0.028 | 0.027 |
|
0.028 | 0.014 ± 0.004 | 0.025 | 0.027 | 0.028 |
|
0.203 | 0.225 | 0.012 ± 0.003 | 0.024 | 0.024 |
|
0.251 | 0.268 | 0.221 | 0.011 ± 0.003 | 0.025 |
|
0.224 | 0.243 | 0.206 | 0.212 | 0.055 ± 0.01 |
Measures of genetic variability in
|
n | Number of |
Hd | Nd | Tajima’s |
Fu’s Fs |
|
26 | 19 | 0.976 | 0.00757 | -1.77465 |
-15.274 |
|
11 | 7 | 0.818 | 0.01436 | 0.23394 |
0.893 |
|
5 | 5 | 1.000 | 0.01063 | -1.22485 |
-0.875 |
COI_aligned_fas.fas
DNA sequence alignment
The alignment of the cytochrome
File: oo_408505.fas
28SA_aligned_names.fas
DNA squence alignment
The alignment of the 28S ribosomal sequences used here. Sequences are identified by Australian Museum registration number or GenBank accession number.
File: oo_408506.fas