The byssal-producing glands and proteins of the silverlip pearl oyster Pinctada maxima (Jameson, 1901).

Pinctada maxima are most well known for their production of high-quality natural pearls. They also generate another natural material, the byssus, an adhesive thread critical for steadfast attachment underwater. Herein, P. maxima byssal threads were analysed via proteotranscriptomics to reveal 49 proteins. Further characterisation was undertaken on five highly expressed genes: glycine-rich thread protein (GRT; also known as PUF3), apfp1/perlucin-like protein (Pmfp1); peroxidase; thrombospondin 1, and Balbiani ring 3 (BR3), which showed localised tissue expression. The spatial distribution of GRT and Pmfp1 via immunodetection combined with histology helped to identify glandular regions of the foot that contribute to byssal thread production: the byssal gland, the duct gland, and two thread-forming glands of basophilic and acidophilic serous-like cells. This work advanced primary knowledge on the glands involved in the creation of byssal threads and the protein composition of the byssus for P. maxima, providing a platform for the design of marine biopolymers.

Structure, function and parallel evolution of the bivalve byssus, with insights from proteomes and the zebra mussel genome.

The byssus is a structure unique to bivalves. Byssal threads composed of many proteins extend like tendons from muscle cells, ending in adhesive pads that attach underwater. Crucial to settlement and metamorphosis, larvae of virtually all species are byssate. By contrast, in adults, the byssus is scattered throughout bivalves, where it has had profound effects on morphological evolution and been key to adaptive radiations of epifaunal species. I compare byssus structure and proteins in blue mussels (Mytilus), by far the best characterized, to zebra mussels (Dreissena polymorpha), in which several byssal proteins have been isolated and sequenced. By mapping the adult byssus onto a recent phylogenomic tree, I confirm its independent evolution in these and other lineages, likely parallelisms with common origins in development. While the byssus is superficially similar in Dreissena and Mytilus, in finer detail it is not, and byssal proteins are dramatically different. I used the chromosome-scale D. polymorpha genome we recently assembled to search for byssal genes and found 37 byssal loci on 10 of the 16 chromosomes. Most byssal genes are in small families, with several amino acid substitutions between paralogs. Byssal proteins of zebra mussels and related quagga mussels (D. rostriformis) are divergent, suggesting rapid evolution typical of proteins with repetitive low complexity domains. Opportunities abound for proteomic and genomic work to further our understanding of this textbook example of a marine natural material. A priority should be invasive bivalves, given the role of byssal attachment in the spread of, and ecological and economic damage caused by zebra mussels, quagga mussels and others. This article is part of the Theo Murphy meeting issue 'Molluscan genomics: broad insights and future directions for a neglected phylum'.

Experimental study of transgenerational effects, pH and predation risk on byssus production in a swiftly spreading invasive fouling Asian mussel, Musculista senhousia (Benson).

Marine biofouling by the highly invasive Asian date mussel, Musculista senhousia (Benson), has caused devastating ecological and economic consequences in most coastal seas. Acute and short-term exposure experiments have demonstrated the susceptibility of mussel byssus - a holdfast structure by which mussels strongly adhere to underwater substrates, to pH. Yet, the influence of long-term exposures, especially across multiple generations, is largely unknown. Here, we evaluated transgenerational effects of pH on byssal threads secreted by M. senhousia, and compared byssus performance in absence versus presence of predators. If no predation occurred, neither pH nor transgenerational exposure significantly affected the number, length and diameter of byssal threads. Under predation risk, mussels, even exposed to low pH, significantly enhanced byssus production. In particular, individuals originating from parents grown under low conditions produced significantly more, longer and stronger byssal threads compared with those spawn from parents exposed to high pH, demonstrating positive transgenerational effects which can confer mussel byssus resilience at low pH. Given the energetically expensive nature of byssus production, these observations can be in line with previously documented plasticity of energy metabolism arose following transgenerational exposure to low pH, which allows mussels to allocate more energy to fulfill the synthesis and secretion of byssal proteins. Our findings demonstrate the remarkable ability of highly invasive fouling mussel species to respond plastically and adapt behaviorally to low pH and hence provide important implications for linking marine biofouling, biological invasion, and coastal acidification.

Near-future levels of ocean temperature weaken the byssus production and performance of the mussel Mytilus coruscus.

Marine mussels are key ecological engineers that form dense aggregations to maintain the vital habitat in benthic systems. It is essential to understand the consequences of mussel byssus attachment in elevated temperatures associated with ocean warming. We evaluated byssus production and the mechanical performance of threads in the mussel Mytilus coruscus at 21° (control), 27 °C (average temperature in the M. coruscus habitat during the summer season) and 31 °C (4 °C raised) for 72 h. We quantified byssus secretion and shedding number, measured byssal breaking force, byssal polyphenol oxidase (PPO) activity, byssal thread length and diameter. Expression of byssus foot protein genes was analyzed by quantitative real-time PCR in foot tissue. High seawater temperature decreased the number of newly secreted byssus and the diameter of byssal threads, leading to the reduction of byssal breaking force and the alteration of the weakest part of the thread. Increased breakpoints in the upper part of the thread (proximal region) were higher at 27 °C than at 21 °C. High-temperature stress significantly reduced the PPO activity in byssus at 31 °C in comparison to 21 °C. The expression of mussel foot protein genes was affected by elevated temperature. The increased gene expression of byssus collagen-like protein 2 (Mccol2) at 31 °C conflicted with the number of byssuses produced. Suggesting the reduction of mussel foot protein abundance is not the cause of decreased byssus production at 31 °C. These results show that byssus, as an extracellular structure of mussels, may be highly susceptible to the adverse effects of ocean warming.