Molecular resolution of the family Dreissenidae (Mollusca: Bivalvia) with emphasis on Ponto-Caspian species, including first report of Mytilopsis leucophaeata in the Black Sea basin
Introduction
Dreissenid molluscs are an important group of biofouling bivalves that are rapidly invading habitats around the world (e.g., Hebert et al., 1989; May and Marsden, 1992; Nuttall, 1990). Dreissenids have been reclassified many times, at many levels (i.e., genus, subgenus, species, subspecies, and variety). These analyses have resulted in confusion regarding phylogenetic relationships within the family. For example, Russian systematists have been unable to develop a uniform taxonomic history for the genus Dreissena. Andrusov (1897) outlined the first major taxonomic classification scheme for dreissenids and included Dreissena polymorpha (Pallas), D. rostriformis (Deshayes), and D. bugensis (Andrusov), amongst others, as legitimate species. Zhadin (1952) did not recognize D. rostriformis as a species but subsequent reclassifications by Logvinenko (1965), Logvinenko and Starobogatov (1968), and Starobogatov (1994) did. Zhadin (1952) recognized D. bugensis as a species, while Mordukhai-Boltovskoi (1960) classified D. bugensis as a subspecies of D. rostriformis. Starobogatov (1994) re-elevated D. bugensis to species level within the subgenus Pontodreissena and maintained species level classification for both D. polymorpha (subgenus Dreissena) and D. rostriformis (subgenus Pontodreissena) and included the newly identified D. stankovici (subgenus Carinodreissena). Rosenberg and Ludyanskiy’s (1994) comparative review of dreissenid taxonomy included additional species, subspecies, and varieties based primarily on Russian accounts (e.g., Andrusov, 1897; Babak, 1983; Logvinenko and Starobogatov, 1968; Nevesskaya, 1963; Starobogatov, 1970; Starobogatov, 1994; Taktakishvili, 1973).
Some workers have suggested that Dreissena and Mytilopsis evolved from extinct branches of the genus Congeria (Andrusov, 1897; Babak, 1983; Mackie et al., 1989; Starobogatov, 1994), while others maintain that Dreissena and Congeria arose from Mytilopsis (Marelli, 1994). Mytilopsis was considered a subgenus of Congeria by Russian taxonomists (Andrusov, 1897; Babak, 1983; Starobogatov, 1970), but was elevated to genus level by Nuttall (1990), a classification scheme later supported by Rosenberg and Ludyanskiy (1994). Marelli and Gray (1985) suggested that there exist only five extant species of Mytilopsis, including M. leucophaeata. Unfortunately, as with Dreissena, traditional taxonomic classification of the genus Mytilopsis is complex, discordant and variable through time. We argue that the use of molecular techniques can help clarify the phylogenetics of this group.
Two of four Dreissena species used herein, D. rostriformis and D. stankovici, have never been reported outside their historical ranges (Table 1). Dreissena rostriformis occurs in the Middle and South Caspian Sea at salinities between 12 and 13.5‰, while the closely related D. bugensis is typical of freshwater or oligohaline habitats both within its historical range in the Black Sea basin, and in its introduced range in the Volga River. D. polymorpha occurs in similar habitats as those reported for D. bugensis, but is capable of inhabiting mesohaline waters typical of the northern Caspian Sea (Table 1).
Human activities are rapidly changing aquatic ecosystems. Most notable are activities related to transoceanic shipping and canal creation, both of which link water bodies and allow transfer of nonindigenous species between previously isolated aquatic ecosystems. The Dreissenidae have undergone considerable global redistribution as a result of shipping activities (Nuttall, 1990). Typically considered Ponto-Caspian “endemics” (Geary et al., 2000), two dreissenids have recently invaded the Laurentian Great Lakes. Dreissena polymorpha, the zebra mussel, was first discovered in Lake St. Clair in 1988 (Hebert et al., 1989), while D. bugensis, the quagga mussel, was first reported from Lake Ontario in 1991 (May and Marsden, 1992). A “profundal” variety was reported from deep-water habitats in Lake Erie in 1992 (Dermott and Munawar, 1993) and later identified as D. bugensis using allozymes (Marsden et al., 1996; Spidle et al., 1994). Another Dreissenidae, the dark false mussel Mytilopsis leucophaeata, is native to the Gulf of Mexico, but invaded the Hudson River, New York in the 1930s. The species also has recently been identified in the Upper Mississippi River (Koch, 1989) and at several locations in southern New England (Smith and Boss, 1996). Mytilopsis leucophaeata also has been reported from European waters as early as 1835 (Wolff, 1999) and is found along North Sea coasts from Germany to France (Marelli and Gray, 1983; Oliver et al., 1998) and the River Thames estuary, England (Bamber and Taylor, 2002). European populations occupy both freshwater and brackish estuary habitats (Reise et al., 1999).
Dreissena polymorpha has an extensive distribution in both European and North American freshwaters (Nalepa and Schloesser, 1993). In contrast, D. bugensis has a more restricted distribution in European and North American freshwaters, but is currently undergoing range expansion in the Volga River, Russia, and is replacing D. polymorpha in the lower Great Lakes (Berkman et al., 2000; Mills et al., 1999). Dreissenids are nuisance species in many invaded habitats owing to biofouling (Kharchenko, 1995; Marelli and Gray, 1983), but are considered beneficial in some habitats where they improve water quality (Reeders et al., 1993). Owing largely to human-induced range expansion, co-occurrence of dreissenid species is increasing globally and the ability to discern morphologically similar species has become increasingly important. Dreissena polymorpha, D. bugensis, D. rostriformis, D. stankovici, and M. leucophaeata share many life-history characteristics (e.g., use of byssal threads for attachment, and possession of a free-swimming veliger larva) and exhibit strong morphological and shell colour similarities (Biochino, 1994; Lukashev, 2000; May and Marsden, 1992; O’Neill, 1990; Pathy and Mackie, 1993; Protasov and Gorpinchuk, 2000). Moreover, each species may exhibit pronounced intraspecific variability. Genetic markers may prove particularly useful for discrimination of species, such as dreissenids, with high intraspecific variability or small larval or juvenile size (Claxton et al., 1997, Claxton et al., 1998; Skurikhina et al., 2001). Identification of a few individuals early on in an incipient invasion may allow for implementation of rapid control measures. For example, shortly after M. sallei arrived in Darwin, Australia, a comprehensive eradication campaign was undertaken before the species could become established (Pyne, 1999).
In this study, we use mitochondrial gene sequencing to assess the phylogenetic relationships among members of the family Dreissenidae identified from Ponto-Caspian and Mediterranean regions including the genera Dreissena, Mytilopsis, and Congeria. This is the first study to use molecular techniques to resolve the placement of D. stankovici and D. rostriformis within the family Dreissenidae. In addition, we attempt to resolve the relationship between D. rostriformis and D. bugensis using sequence data. The relationship between these taxa has been highly discordant over time based on traditional taxonomical accounts as some authors consider each a species while others consider D. bugensis a subspecies of D. rostriformis (e.g., Andrusov, 1897; Mordukhai-Boltovskoi, 1960; Starobogatov, 1994; Zhadin, 1952). Furthermore, we use nuclear and mitochondrial DNA restriction digests to identify species that are difficult to distinguish based on morphological characteristics alone.
Section snippets
DNA isolation and PCR amplification
Specimens used in sequencing were collected from their current European ranges (Table 2). Two cryptic dreissenid specimens were collected from the Dniester Liman, Black Sea, and included in our analyses. External shell morphology of these individuals was similar but not identical to those of D. bugensis and D. polymorpha. Total DNA was extracted from mantle muscle tissues of specimens preserved in 95% ethanol or frozen using either a standard phenol–chloroform method or a DNA purification kit
Phylogenetic analysis
The NJ tree based on 16S showed that D. rostriformis and D. bugensis differed by a single nucleotide (Fig. 1). Bootstrap support for distinct nodes in this part of the tree was weak, providing the first molecular evidence that these individuals might represent a common species. However, both NJ and maximum parsimony analyses maintained each taxon as monophyletic. Considering D. rostriformis and D. bugensis as a single species with two possible races (see below), intraspecific differences ranged
Dreissenid phylogenetics
In contrast to traditional dreissenid taxonomy based principally on morphological attributes, our molecular analyses, in combination with environmental tolerances, suggest that D. bugensis and D. rostriformis may represent a single species with two distinct races. This view is supported by only a single base pair difference (0.23%) between D. bugensis and D. rostriformis in the 16S gene, and by 2–3 bp differences (0.36–0.54%) in the COI gene. Consequently, we suggest the ancestral name of D.
Acknowledgements
Drs. I. Grigorovich and C. Lee collected samples of D. bugensis and D. stankovici, without which this work would not have been possible. Financial support was provided by NSERC (T.W.T.) and GLIER (M.F.D.) postdoctoral fellowships, Canada Research Chair (DDH), Premier’s Research Excellence Award (H.J.M.), and by NSERC research grants to D.D.H. and H.J.M.
References (85)
- et al.
Molecular phylogeny and biogeography of the marine shrimp Penaeus
Mol. Phylogen. Evol.
(1998) - et al.
Ancient Lake Pannon and its endemic molluscan fauna (Central Europe; Mio-Pliocene)
Adv. Ecol. Res.
(2000) - et al.
Taxonomic status and phylogenetic relationships of some species of the genus Gammarus (Crustacea, Amphipoda) deduced from mitochondrial DNA sequence
Mol. Phylogenet. Evol.
(1997) - et al.
Changes in the dreissenid community in the Lower Great Lakes with emphasis on southern Lake Ontario
J. Great Lakes Res.
(1999) - et al.
Sphaeriid and Corbiculid clams represent separate heterodont bivalve radiations into freshwater environments
Mol. Phylogen. Evol.
(2000) - et al.
Recent mass invasion of the North American Great Lakes by Ponto-Caspian species
Tr. Ecol. Evol.
(2000) - et al.
Diagnostic genetic markers and evolutionary relationships among invasive dreissenoid and corbiculoid bivalves in North America: phylogenetic signal from mitochondrial 16S rDNA
Mol. Phylogen. Evol.
(1999) Fossile und lebende Dreissensidae Eurasiens
Trav. Soc. Nat. St.-Pétersbourg (Sect. Géol. Minérol.)
(1897)- et al.
Population structure and genetic variability of pearl oyster Pinctada mazatlanica along Pacific coasts from Mexico to Panama
Cons. Gen.
(2000) The Pliocene and Quaternary Dreissenidae of the Evsinsk Basin
Akad. Nauk SSSR Tr. Paleontol. Inst.
(1983)
A diagnostic molecular marker for zebra mussels (Dreissena polymorpha) and potentially co-occurring bivalves: mitochondrial COI
Mol. Mar. Biol. Biotechnol.
The brackish water mussel Mytilopsis leucophaeata (Conrad, 1831) (Bivalvia: Dreissenidae) in the River Thames
J. Conchol.
Habitat shift in invading species: zebra and quagga mussel population characteristics on shallow soft substrates
Biol. Invas.
Polymorphism and geographical variability
A biological survey of ballast water in container ships entering Hong Kong
Hydrobiologia
Discrimination of field-collected juveniles of two introduced dreissenids (Dreissena polymorpha and Dreissena bugensis) using mitochondrial DNA and shell morphology
Can. J. Fish. Aquat. Sci.
A genetic and morphological comparison of shallow- and deep-water populations of the introduced dreissenid bivalve Dreissena bugensis
Can. J. Zool.
An invasion history for Cercopagis pengoi based on mitochondrial gene sequences
Limnol. Oceanogr.
Invasion of Lake Erie offshore sediments by Dreissena, and its ecological implications
Can. J. Fish. Aquat. Sci.
The Caspian lake: history, biota, structure, and function
Limnol. Oceanogr.
DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates
Mol. Mar. Biol. Biotechnol.
Size-dependent, spatial and temporal genetic variation at a leucine aminopeptidase (LAP) locus among blue mussel (Mytilus galloprovincialis) populations along a salinity gradient
Mar. Biol.
Ecological and genetic studies on Dreissena polymorpha (Pallas): a new mollusc in the Great Lakes
Can. J. Fish. Aquat. Sci.
Genetics of physiological differentiation within the marine mussel genus Mytilus
Evolution
Distribution of Mytilus edulis, M. galloprovincialis, and their hybrids in open-coast populations of mussels in southwestern England
Mar. Biol.
Phylogenetic evidence for role-reversals of gender-associated mitochondrial DNA in Mytilus (Bivalvia: Mytilidae)
Mol. Biol. Evol.
Congeria leucophaeata (Conrad) in the Hudson River
Nautilus
Physical factors that limit the distribution and abundance of Dreissena polymorpha (Pall.)
J. Shellfish Res.
Some data on morphological evolution in bivalve molluscs
Zool. Z.
Dreissena: range, ecology, biofouling
Gidrobiol. Z.
Mytilopsis leucophaeata (Conrad, 1831) from the upper Mississippi River (Bivalvia: Dreissenidae)
Malacol. Data Net
Maintenance of an aminopeptidase allele frequency cline by natural selection
Proc. Natl. Acad. Sci. USA
MEGA: Molecular Evolutionary Genetics Analysis, version 1.01
Changes in the fauna of Caspian molluscs Dreissena after intrusion of Mytilaster lineatus (Gmelin)
Nauchn. Dokl. Vyssh. Shk. Biol. Nauki
Phillum Mollusca
Morphological variability of Dreissena bugensis Andrusov under conditions of the regulated Dnieper outflow
Hydrobiol. J.
Molecular systematics and evolution of reproductive traits of North American freshwater unionacean mussels (Mollusca: Bivalvia) as inferred from 16S rRNA gene sequences
Philos. Trans. R. Soc. Lond. B
Comments on the status of recent members of the genus Mytilopsis (Bivalvia: Dreissenidae)
Malacol. Rev.
Cited by (99)
An impact of non-native species invasions on the Caspian Sea biota
2023, Advances in Marine BiologyThe Late Pleistocene mollusk fauna of Selitrennoye (Astrakhan province, Russia): A natural baseline for endemic Caspian Sea faunas
2020, Journal of Great Lakes ResearchAn integrated reconstruction of the early Pleistocene palaeoenvironment of Homo erectus in the Denizli Basin (SW Turkey)
2019, GeobiosCitation Excerpt :The species was found in Quaternary deposits of the Konya Basin, located east of the lake (Schütt, 1991). The bivalve Dreissena is an epifaunal filter feeder dwelling on hard substrates in rivers, estuaries and lakes from freshwater to mesohaline conditions down to depths of over 130 m (Therriault et al., 2004; Orlova et al., 2005; Welter-Schultes, 2012; Cummings and Graf, 2015). Lymnocardiinae occur in a wide range of well-oxygenated habitats in lagoons, coastal lakes or river mouths, and are typical of oligohaline to mesohaline conditions; some species occasionally also extend into freshwater (Kijashko in Bogutskaya et al., 2013; Albrecht et al., 2014).
Alien Species Invasion: Case Study of the Black Sea
2019, Coasts and Estuaries: The FutureStrategies and consequences of indigenous and invasive freshwater mussels living in cyanobacterial and anthropogenic impacted waters
2019, Ecotoxicology: New Challenges and New Approaches