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However, vision may also be used to mediate navigation and locate preferred habitat (von Buddenbrock and Moller-Racke 1953; Hamilton and Koch 1996), as well as to recognize favorable feeding conditions (Speiser and Johnsen 2008b). We also capitalized on the first complete genome for a more distantly related bivalve lineage, the Pacific oyster () in the family Ostreidae (Zhang et al.Recently, two copies of the Gq-coupled opsin (=Gq-opsin) were identified from eye transcriptomes of two scallop species () (A. 2012), along with select molluscan opsin sequences to test the following hypotheses: (1) the timing of bivalves’ duplication of opsin was specific to the scallop lineage and (2) there was a long-term shift in selective constraints upon scallops’ opsins following this duplication.All PCR reactions contained 1× Buffer B, 2.5 m M Mg Cl, 0.2 m M of each d NTP, 0.4 M of each primer, 1.25 U of Taq DNA polymerase (Fisher Scientific, Pittsburg, Pennsylvannia, USA) in a 25 -μl reaction volume with magnesium chloride concentration and annealing temperature optimized for some taxa. These ORFs were then blasted against the NCBI non-redundant protein database using a blastx algorithm and an opsins from Genbank were also blasted against the oyster-genome assembly scaffolds to further confirm our identification of opsins and to assist in identifying opsin exons.PCR products were cloned following the manufacturer’s instructions (TOPO TA Cloning Kit with p CR2.1-TOPO) and at least five positive colonies were sequenced using M13 primers. Serb, unpublished data]) were blasted against the three molluscan genomes using the tblastx blast algorithm (translated nucleotide query against a translated nucleotide database) with an -value cutoff of E-3 in the program Geneious version 5.6.6. We ranked putative opsin candidates by the presence of three motifs ().This work thus expands on our knowledge of the evolution of opsin within bivalves, demonstrating that bivalves have a greater level of opsin diversity than previously thought.We examined eight species of scallop and a species from each of the three closely related families (Spondylidae, Limidae, and Propeamussidae) based on published work (Puslednik and Serb 2008; Alejandrino et al. Ethanol-preserved tissues from the Pectinoida were obtained from museum collections or provided by colleagues (Table 1).
Polymerase chain reaction (PCR) was performed using a combination of newly developed and published primers to amplify a region of the opsin gene that includes transmembrane (TM) 4 through TM7 (Table 2). Blast results that contained the largest open reading frames (ORFs) encompassing a region that match a scallop’s opsin were extracted first.The adult scallop also has an interesting visual system that comprises 30 to upward of 200 non-cephalic eyes found along the middle fold of the margins of the mantle that lines the shell.Each of these eyes can construct a spatially resolved representation of the animal’s environment using a reflective argentea lining the back of the eye and redirecting and focusing light on two independently functioning retinas (Land 1965, 1966; Speiser and Johnsen 2008a).Evolutionary biologists have long been interested in how expansions of the photosensory system might contribute to morphological differentiation of animals.
Comparative studies in vertebrate and arthropod lineages have provided considerable insight into how the duplication of opsin, the first gene of the phototransduction pathway, have led to functional differentiation and new ecological opportunities; however, this relationship cannot be examined in many invertebrate groups as we have yet to characterize their opsin content.We discovered that duplication of Gq-opsin is not restricted to scallops, but instead occurred earlier in the Pteriomorphia.