Genetics and Molecular Biology, 29, 1, 174-179 (2006) Copyright by the Brazilian Society of Genetics. Printed in Brazil www.sbg.org.br Short Communication Allozymic variation and differentiation in the chilean blue mussel, Mytilus chilensis, along its natural distribution Jorge E. Toro, Grace C. Castro, Johana A. Ojeda and Ana M. Vergara Universidad Austral de Chile, Instituto de Biología Marina “Dr. Jürgen Winter”, Valdivia, Chile. Abstract Genetic differentiation in the Chilean blue mussel Mytilus chilensis (Hupé 1854) was investigated based on the varia- tion in the allozyme frequencies of Pgm, Gpi, Icd, Me, Gsr, Lap and Pep in eight samples collected along 1800 km from Arauco (VIII Region) to Punta Arenas (XII Region). Despite the large geographic separations, values of Neis un- biased genetic distance, D (0.004-0.048) and standardised genetic variation among populations, Fst (0.011-0.055) were small. The levels of gene flow (Nm = 8) found in this study prevent the effect of differentiation among popula- tions by genetic drift. This findings indicate that its long-lived planktotrophic larvae provides this species with consid- erable dispersal ability throughout its range which is favoured by the ocean currents along the chilean coast. In terms of management of the M. chilensis fishery, the results provide no evidence for discrete stocks, with the possible ex- ception of the Punta Arenas population. Considering the intensive aquaculture activities with this species the present study provide preliminary data which can be used as a baseline for further characterization and /or monitoring these mussel populations. Key words: allozyme, Mytilus chilensis, population genetics, gene flow, Chile. Received: July 21, 2004; Accepted: May 31, 2005. The Chilean blue mussel (Mytilus chilensis, Hupe in the literature. Only a few studies with limited sampling 1854) is an economically important resource in southern are reported in the scientific literature mainly focused to Chile, commonly known as chorito or quilmahue. It is dilucidate the distribution of the Mytilus edulis species widely distributed on hard substrata in the lower intertidal complex (Koehn, 1991; McDonald et al., 1991). zone to 25 m depths, along the chilean coast line (Bratts- tröm and Johanssen, 1983). The range of the species ex- An important question, from both evolutionary and tends over 40° of latitude from Arica to Cape Horn fishery amangement perspectives, is the extent of gene flow (Lancellotti and Vázquez, 2000) and it is impacted by a ma- and associated genetic differentiation among Mytilus jor oceanographic feature, the West Wind Drift (43°S) that chilensis populations at a macrogeographic (over 100 km) defines the boundary between the main current directions scale. Marine species with longer larval phases are gener- along the chilean Coast (Strub and Mesías, 1998). Winter et ally though to disperse further and have more gene flow, al. (1984), determined that there was an annual reproduc- larger geographic ranges, lower levels of genetic differenti- tive cycle, with spawning occurring during spring and sum- ation among populations, and high levels of genetic varia- mer. Toro and Sastre (1995) and Toro et al. (2004) have tion within populations (Scheltema and Williams, 1983; shown that M. chilensis possesses a planktotrophic larval Waples, 1987; Palumbi, 1995; Williams and Benzie, 1993). stage with a pelagic existence of 45 d. The species thus has However, expected patterns of gene flow and population the potential for long-distance dispersal over a scale of hun- connectivity in Mytilus chilensis could be altered by ocean- dreds of kilometers along the chilean coast. Its culture be- ographic features peculiar to its range, such as the presence gan in 1943 in the Chiloé Island, southern Chile (Osorio et of the west wind drift (WWD) which bisects the flow of al., 1979) and the aquaculture production raised from 3,864 currents along the chilean coast (Strub and Mesías, 1998). t in 1993 to 41,406 t in 2001. Despite this explosive devel- Also, human mediated dispersal may have heavely affected opment of M. chilensis aquaculture, population genetics the levels of genetic variation in several southern Chile studies in the chilean mussel (Mytilus chilensis) are scarce stocks, because most of the mussel cultures used juveniles (seed) transferred from Yaldad Bay (Winter et al., 1984) Send correspondence to Jorge E. Toro. Universidad Austral de that may caused in some locations the restocking of local Chile, Instituto de Biología Marina“Dr. Jürgen Winter”, Casilla 567, natural populations with juveniles from the Yaldad popula- Valdivia, Chile. E-mail: [email protected]. tion. Toro et al. 175 In this paper we examine the genetic structure of Mytilus chilensis collected from eight widely separated (up to 1800 km) sites along the chilean coast, using allozyme electrophoresis to establish the extend of gene flow and lev- els of genetic differentiation. Samples of mussels were collected by dredging or by diving at eight localities extending over 1800 km along the chilean coast (covering the whole range of the species natu- ral distribution) from Arauco (VIII Región) to Pta. Arenas (XII Región) (Figure 1). The mussels (N = 120 from each population) were delivered alive to the laboratory where they were immediately dissected and isolated tissues were stored in liquid nitrogen for subsequent electrophoretic analysis. Tissue samples from abductor muscle and hepato- pancreas were homogenized in 50-100 mL of 0.05 M Tris- HCl buffer, pH 8.0. Standard techniques of horizontal starch (12%) gel electrophoresis were assayed in the fol- Figure 1 - Locations of the eight natural populations of the chilean blue lowing loci: PGM (2.7.5.1), GPI (5.3.1.9), LAP (3.4.11.1), mussel (Mytilus chilensis) sampled along its whole natural distribution in EST (3.1.1.1), ODH (1.5.1.11), XDH (1.2.1.37), Alfa GPD southern Chile (•). (1.1.1.8), MDH (1.1.1.37), ME (1.1.1.40), AAT (2.6.1.1), PEP (3.4.11.13), IDH (1.1.1.42), ICD (1.1.1.2), MPI (5.3.1.8) y GSR (1.1.4.2). Starch gels were stained accord- 1978) unbiased genetic distances (D) and identities (I). A ing to (Shaw and Prassad, 1970; Harris and Hopkinson, UPGMA cluster analysis was performed using I values. 1976; Selander et al., 1983; Beaumont and Toro, 1996). Al- The Mantel test using 50,000 randomisations was carried leles were designated alphabetically according to their mi- out for association between geographic distance and the ge- gration relative to the most common electromorph. A locus netic distance, using PopTools (Hood, 2004). was considered polymorphic if the frequency of the most Only seven variable loci (40.8% range = 33.3-46.7%) common allele did not exceed 0.95. Data were analysed us- differing in gene frequencies were found across the geo- ing the computer programs POPGENE V. 1.31 (Yeh and graphic samples, Leucine aminopeptidase, glucose phos- Boyle, 1997) and GENETIX V 4.05.2. These programs phate isomerase, Malic Enzime, Peptidase, Isocitrate were used to test for Hardy-Weinberg equilibrium, to pro- Deshidrogenase, Glutatione reductase and phospho gluco- vide F-statistics with their significance and Neis (Nei, mutase. The allele frequencies for the seven polymorphic Table 1 - Allele frequencies for the seven polymorphic loci screened in eight populations of the chilean blue mussel (Mytilus chilensis). Locus allele Arauco Queule Valdivia Calbuco Ancud Yaldad Pto. M.B. Pta. Arenas PGM n 109 108 116 110 99 128 99 100 A 0.0275 0.0694 0.0603 0.0136 0.0101 0.0078 0.0051 0.2950 B 0.7936 0.7454 0.7802 0.7864 0.8283 0.7969 0.8384 0.4650 C 0.1786 0.1852 0.1595 0.2000 0.1616 0.1953 0.1566 0.2350 D 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0050 GPI n 112 80 102 110 110 111 99 107 A 0.1071 0.0312 0.0294 0.0955 0.0909 0.0721 0.1263 0.1262 B 0.7277 0.8625 0.8775 0.8000 0.6636 0.8423 0.7727 0.5794 C 0.1607 0.1062 0.0931 0.1045 0.2455 0.0856 0.1010 0.2944 D 0.0045 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 0.0000 ICD n 112 108 115 110 110 141 99 107 A 0.0268 0.0417 0.0087 0.0500 0.0227 0.0177 0.0253 0.0327 B 0.9375 0.9444 0.9565 0.8773 0.9545 0.9645 0.9646 0.9252 C 0.0357 0.0139 0.0348 0.0727 0.0227 0.0177 0.0101 0.0421 176 Allozymic variation in Mytilus chilensis Table 1 (cont.) Locus allele Arauco Queule Valdivia Calbuco Ancud Yaldad Pto. M.B. Pta. Arenas ME n 112 108 116 110 110 111 98 106 A 0.0223 0.1852 0.1078 0.0227 0.0318 0.0721 0.0816 0.0377 B 0.1652 0.2778 0.3233 0.2455 0.1864 0.2973 0.2959 0.2311 C 0.4554 0.2454 0.2974 0.4182 0.3727 0.3874 0.2959 0.3962 D 0.2857 0.1806 0.2112 0.2818 0.2364 0.2207 0.2500 0.2406 E 0.0714 0.1111 0.0603 0.0318 0.1727 0.0225 0.0765 0.0943 GSR n 110 108 116 110 110 111 97 107 A 0.0227 0.0000 0.0086 0.0364 0.0182 0.0450 0.0155 0.0514 B 0.9409 0.9306 0.9181 0.9545 0.9812 0.9505 0.8969 0.8972 C 0.0364 0.0694 0.0733 0.0091 0.0000 0.0450 0.0876 0.0514 LAP n 111 106 116 112 107 115 93 100 A 0.2252 0.2736 0.2069 0.0938 0.1963 0.1565 0.2312 0.2600 B 0.5090 0.5330 0.4784 0.4777 0.5234 0.5217 0.6720 0.4800 C 0.2432 0.1887 0.2802 0.3438 0.2523 0.2783 0.0968 0.2400 D 0.0225 0.0047 0.0345 0.0848 0.0280 0.0435 0.0000 0.0200 PEP n 111 108 107 112 80 136 96 101 A 0.1486 0.4815 0.5047 0.3705 0.2313 0.4449 0.4583 0.3119 B 0.3874 0.4398 0.3458 0.4062 0.4750 0.4338 0.4583 0.4208 C 0.3604 0.0648 0.1262 0.1786 0.2938 0.1176 0.0833 0.1881 D 0.1036 0.0139 0.0234 0.0446 0.0000 0.0037 0.0000 0.0792 Key: [PGM: phosphoglucomutase; GPI: glucose phosphate isomerase; IDH: isocitrate dehydrogenase; ME: malic enzyme; GSR: glutathione reductase; LAP: leucine aminopeptidase; PEP: peptidase ]; n = number of specimins scored.