Genetic variability of Chilean and Peruvian surf clams ... % sequence divergence D. hanleyanus 12 D....
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10 % sequence divergence D. hanleyanus 12 D. hanleyanus 12 D. hanleyanus 12 D. hanleyanus 12 D. hanleyanus 12 D. hanleyanus 12 D. hanleyanus 12 D. hanleyanus 12 D. asper 10 D. asper 10 D. asper 10 D. asper 10 D. asper 10 D. asper 10 D. asper 10 D. marincovichi 1 D. obesulus 11 D. obesulus 11 D. obesulus 11 D. obesulus 11 D. marincovichi 7 D. marincovichi 2 D. marincovichi 9 D. marincovichi 8 D. marincovichi 5 D. marincovichi 7 D. marincovichi 2 D. marincovichi 7 D. marincovichi 7 D. marincovichi 6 D. marincovichi 3 D. marincovichi 3 D. marincovichi 1 D. marincovichi 2 D. marincovichi 9 D. marincovichi 1 D. marincovichi 6 D. marincovichi 1 D. marincovichi 5 D. marincovichi 9 D. asper 10 D. marincovichi 2 D. marincovichi 4 D. marincovichi 8 D. marincovichi 9 D. marincovichi 3 D. marincovichi 1 D. marincovichi 3 D. obesulus 11 D. marincovichi 5 D. marincovichi 5 D. marincovichi 6 D. marincovichi 5 D. obesulus 11 D. marincovichi 1 D. marincovichi 3 D. marincovichi 3 D. marincovichi 5 D. marincovichi 6 D. marincovichi 6 D. marincovichi 7 D. marincovichi 7 D. marincovichi 8 D. marincovichi 9 D. marincovichi 9 D. marincovichi 9 D. obesulus 11 D. obesulus 11 D. obesulus 11 D. marincovichi 1 D. marincovichi 3 D. marincovichi 1 D. marincovichi 9 D. marincovichi 8 D. marincovichi 5 D. marincovichi 6 cytochrome oxidase I, 606 bp Neighbour-Joining Tree ti + tv, uncorrected p-distances 15.2 - 16.3 % 17.7 - 18.2 % 16.2 - 17.2 % 0 - 0.7 % 0 - 0.5 % 0 - 1.2 % Genetic variability of Chilean and Peruvian surf clams (Donax marincovichi and Donax obesulus) Daniel Carstensen: Alfred Wegener Institute for Polar and Marine Research Am Alten Hafen 26 27568 Bremerhaven tel: +49-471-48311315 fax: +49-471-48311918 e-mail: [email protected] References Ansell, A.D. (1983). The biology of the genus Donax. In: McLachlan, A., Erasmus, T. & Junk, W.E. (eds). Developments in Hydrobiology Vol. 19. Sandy Beaches as Ecosystems. Dr. W. Junk, 19: 607-635. Coan, E. (1983). The Eastern Pacific Donacidae. 25 (4): 273-298. Guzmán, N., Saá, S., Ortlieb, L. (1998). Descriptive catalogue of nearshore mulluscs (gastropoda and pelecypoda) from Antofogasta area, 23°S (Chile). 17 (17-86). Folmer, O., Black, M., Hoeh, W., Lutz, R., Vrijenhoek, R. (1994): DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates, Molecular Marine Biology and Biotechnologie, 3(5), 294-299. Held, C.(2000). Phylogeny and biogeography of serolid isopods (Crustacea, Isopoda, Serolidae) and the use of ribosomal expansion segments in molecular systematics, Molecular phylogenetics and evolution,15(2),165-178. D. Carstensen, M. Herrmann, J. Laudien, S. Schiel, W. Arntz, F. Leese, C. Held Acknowledgments Thanks are due to Jaime Mendo (UNA-La Molina), Matthias Wolff and Marc Taylor (ZMT), the people of the University San Marco of Lima. For helpful discussions and technical support we would like to thank Uwe John and Annergret Müller (AWI). This study was financed and conducted in the frame of the EU-project CENSOR (Climate variability and El Nino Southern Oscillation: Impacts for natural resources and management, contract 511071) and is CENSOR publication 0053. Introduction Exposed intertidal sandy beaches are commonly dominated by surf clams of the genus Donax (Ansell 1983). In Peru and Chile these bivalves play an important role for artisanal fisheries. Within this genus, the taxonomic status of Donax marincovichi and D. obesulus, distributed along the Peruvian coastline, is controversially discussed (Coan 1983, Guzmán et al. 1998). As morphometric comparisons reveal no significant differences we possibly deal with a single, rather than with two species. The aim of this study is twofold: First, we want to establish a molecular marker suitable for barcoding and providing evidence, based on sequence data, concerning this taxonomic controversy. Second, we want to estimate the genetic relatedness among geographically distant populations along the Peruvian coastline. Material and Methods Specimens of the two putative species D. marincovichi and D. obesulus were sampled at nine beaches along their distribution (S18°27’53.8"/W70°18’24.3" to S3°33'57"/W80°27'5"). Additionally, specimens of D. asper and D. hanleyanus were sampled as outgroups from one beach each. Opened and closed shells were conserved in >80% ethanol. DNA was extracted with the Qiagen Dneasy kit according to the manufacturer’s recommendations. 10-100 ng DNA were used for PCR-amplification of a fragment of the cytochrome oxidase I (COI) using the primers HCO (5'-TAAACTTCAGGTGACCAAAAAATCA-3') and LCO (5'- GGTCAACAAATCATAAAGATATTGG-3') (Folmer et al. 1994) in 25 μl reactions. PCR products were purified using the Qiagen DNA purification kit according to the manufacturer’s recommendations. Sequencing was conducted on an ABI 3730xl automated sequencer. Sequence data were processed and aligned using the ClustalW algorithm. Phylogenetic analysis was performed using PAUP4beta10. Results and Discussion Sequence data from the COI proved to be useful for species discrimination within the genus Donax: The taxonomic status of both species, D. asper and D. hanleyanus is well supported. However, there is no indication of reproductive isolation between D. marincovichi and D. obesulus in the COI data (Fig. 2a). With 0 – 1.2% sequence divergence, the divergence between D. marincovichi and D. obesulus is on the order of known intraspecific variability in the COI gene (Held 2000; and Fig. 2b). Therefore, the taxonomic status of the two species must be questioned. No genetic differentiation between the geographically separated D. marincovichi populations could be observed from the sequence data. In the future, molecular markers with higher resolving power (e.g. AFLP, Microsatellites) must be analysed for these purposes. D. marincovichi (23 mm) D. asper (30 mm) Fig. 1: Geographical distribution (-------) and sampling localities (1-11) of D. marincovichi and D. obesulus along the Chilean and Peruvian coast. Locality 12 showes the beach where D. hanleyanus was sampled. Fig. 2: (a) Relatedness between the Donax species investigated in this study based on the investigation of a 606 bp fragment of the mitochondrial cytochrome oxidase I. (b) interspecific and intraspecific (in circles) genetic distances between D. asper, D. hanleyanus, D. obesulus, and D. marincovichi. D. obesulus and D. marincovichhi cannot be distinguished based on the COI data. Fig. 2b D. obesulus (17 mm) D. hanlyanus (30 mm) 0 - 0,7 % 0 - 1,2 % 0 - 0,5 %
Transcript of Genetic variability of Chilean and Peruvian surf clams ... % sequence divergence D. hanleyanus 12 D....
10 % sequence divergence
D. hanleyanus 12D. hanleyanus 12D. hanleyanus 12D. hanleyanus 12D. hanleyanus 12
D. hanleyanus 12D. hanleyanus 12D. hanleyanus 12
D. asper 10D. asper 10
D. asper 10D. asper 10
D. asper 10D. asper 10D. asper 10
D. marincovichi 1D. obesulus 11D. obesulus 11D. obesulus 11
D. obesulus 11D. marincovichi 7
D. marincovichi 2D. marincovichi 9D. marincovichi 8
D. marincovichi 5D. marincovichi 7D. marincovichi 2
D. marincovichi 7D. marincovichi 7D. marincovichi 6D. marincovichi 3D. marincovichi 3D. marincovichi 1D. marincovichi 2
D. marincovichi 9D. marincovichi 1D. marincovichi 6
D. marincovichi 1D. marincovichi 5
D. marincovichi 9D. asper 10D. marincovichi 2
D. marincovichi 4D. marincovichi 8D. marincovichi 9
D. marincovichi 3D. marincovichi 1
D. marincovichi 3D. obesulus 11
D. marincovichi 5D. marincovichi 5D. marincovichi 6D. marincovichi 5D. obesulus 11
D. marincovichi 1D. marincovichi 3D. marincovichi 3D. marincovichi 5D. marincovichi 6D. marincovichi 6D. marincovichi 7D. marincovichi 7D. marincovichi 8D. marincovichi 9D. marincovichi 9D. marincovichi 9D. obesulus 11D. obesulus 11D. obesulus 11
D. marincovichi 1D. marincovichi 3D. marincovichi 1
D. marincovichi 9D. marincovichi 8D. marincovichi 5D. marincovichi 6
cytochrome oxidase I, 606 bpNeighbour-Joining Treeti + tv, uncorrected p-distances
15.2 - 16.3 % 17.7 - 18.2 %
16.2 - 17.2 %0 - 0.7 % 0 - 0.5 %
0 - 1.2 %
Genetic variability of Chilean andPeruvian surf clams (Donax marincovichi and Donax obesulus)
Daniel Carstensen:
Alfred Wegener Institute forPolar and Marine ResearchAm Alten Hafen 2627568 Bremerhaven tel: +49-471-48311315fax: +49-471-48311918
e-mail: [email protected]
ReferencesAnsell, A.D. (1983). The biology of the genus Donax. In: McLachlan, A., Erasmus, T. & Junk, W.E. (eds). Developments in
Hydrobiology Vol. 19. Sandy Beaches as Ecosystems. Dr. W. Junk, 19: 607-635.Coan, E. (1983). The Eastern Pacific Donacidae. 25 (4): 273-298.Guzmán, N., Saá, S., Ortlieb, L. (1998). Descriptive catalogue of nearshore mulluscs (gastropoda and pelecypoda) from
Antofogasta area, 23°S (Chile). 17 (17-86).Folmer, O., Black, M., Hoeh, W., Lutz, R., Vrijenhoek, R. (1994): DNA primers for amplification of mitochondrial cytochrome c
oxidase subunit I from diverse metazoan invertebrates, Molecular Marine Biology and Biotechnologie, 3(5), 294-299.Held, C.(2000). Phylogeny and biogeography of serolid isopods (Crustacea, Isopoda, Serolidae) and the use of ribosomal
expansion segments in molecular systematics, Molecular phylogenetics and evolution,15(2),165-178.
D. Carstensen, M. Herrmann, J. Laudien, S. Schiel, W. Arntz, F. Leese, C. Held
AcknowledgmentsThanks are due to Jaime Mendo (UNA-La Molina), Matthias Wolff and Marc Taylor (ZMT), the people of theUniversity San Marco of Lima. For helpful discussionsand technical support we would like to thank Uwe John and Annergret Müller (AWI). This study was financedand conducted in the frame of the EU-project CENSOR (Climate variability and El Nino Southern Oscillation: Impacts for natural resources and management, contract 511071) and is CENSOR publication 0053.
Introduction
Exposed intertidal sandy beaches are commonly dominated by surf clams of the genus Donax (Ansell 1983). In Peru and Chile these bivalves play an important role for artisanal fisheries. Within this genus, the taxonomic status of Donaxmarincovichi and D. obesulus, distributed along the Peruvian coastline, is controversially discussed (Coan 1983, Guzmán et al. 1998). As morphometriccomparisons reveal no significant differences we possibly deal with a single, rather than with two species.
The aim of this study is twofold: First, we want to establish a molecular marker suitable for barcoding and providing evidence, based on sequence data, concerning this taxonomic controversy. Second, we want to estimate the genetic relatedness among geographically distant populations along the Peruvian coastline.
Material and Methods
Specimens of the two putative species D. marincovichi and D. obesulus were sampled at nine beaches along their distribution (S18°27’53.8"/W70°18’24.3" to S3°33'57"/W80°27'5"). Additionally, specimens of D. asper and D. hanleyanuswere sampled as outgroups from one beach each. Opened and closed shells were conserved in >80% ethanol. DNA was extracted with the Qiagen Dneasy kit according to the manufacturer’s recommendations. 10-100 ng DNA were used for PCR-amplification of a fragment of the cytochrome oxidase I (COI) using the primers HCO (5'-TAAACTTCAGGTGACCAAAAAATCA-3') and LCO (5'-GGTCAACAAATCATAAAGATATTGG-3') (Folmer et al. 1994) in 25 µl reactions. PCR products were purified using the Qiagen DNA purification kit according to the manufacturer’s recommendations. Sequencing was conducted on an ABI 3730xl automated sequencer. Sequence data were processed and aligned using the ClustalW algorithm. Phylogenetic analysis was performed using PAUP4beta10.
Results and Discussion
Sequence data from the COI proved to be useful for species discrimination within the genus Donax: The taxonomic status of both species, D. asper and D. hanleyanus is well supported. However, there is no indication of reproductiveisolation between D. marincovichi and D. obesulus in the COI data (Fig. 2a). With0 – 1.2% sequence divergence, the divergence between D. marincovichi and D. obesulus is on the order of known intraspecific variability in the COI gene (Held 2000; and Fig. 2b). Therefore, the taxonomic status of the two species must be questioned.
No genetic differentiation between the geographically separated D. marincovichipopulations could be observed from the sequence data. In the future, molecularmarkers with higher resolving power (e.g. AFLP, Microsatellites) must be analysedfor these purposes.
D. marincovichi (23 mm)
D. asper (30 mm)
Fig. 1: Geographical distribution (-------) and sampling localities (1-11) of D. marincovichi and D. obesulus along theChilean and Peruvian coast. Locality 12 showes the beach where D. hanleyanus was sampled.
Fig. 2: (a) Relatedness between the Donax species investigated in this study based on the investigation of a 606 bp fragment of the mitochondrial cytochrome oxidase I. (b) interspecific and intraspecific (in circles) geneticdistances between D. asper, D. hanleyanus, D. obesulus, and D. marincovichi. D. obesulus and D. marincovichhicannot be distinguished based on the COI data.
Fig. 2b
D. obesulus (17 mm)
D. hanlyanus (30 mm)
0 - 0,7 %
0 - 1,2 %
0 - 0,5 %
