Identification and characterization of endosymbiosis-related immune genes in deep-sea mussels Gigantidas platifrons

Deep-sea mussels of the subfamily Bathymodiolinae are common and numerically dominant species widely distributed in cold seeps and hydrothermal vents.During long-time evolution,deep-sea mussels have evolved to be well adapted to the local environment of cold seeps and hydrothermal vents by various ways,especially by establishing endosymbiosis with chemotrophic bacteria.However,biological processes underlying the establishment and maintenance of symbiosis between host mussels and symbionts are largely unclear.In the present study,Gigantidas platifrons genes possibly involved in the symbiosis with methane oxidation symbionts were identified and characterized by Lipopolysaccharide(LPS)pull-down and in situ hybridization.Five immune related proteins including Toll-like receptor 2(TLR2),integrin,vacuolar sorting protein(VSP),matrix metalloproteinase 1(MMP1),and leucine-rich repeat(LRR-1)were identified by LPS pull-down assay.These five proteins were all conserved in either molecular sequences or functional domains and known to be key molecules in host immune recognition,phagocytosis,and lysosome-mediated digestion.Furthermore,in situ hybridization of LRR-1,TLR2 and VSP genes was conducted to investigate their expression patterns in gill tissues of G.platifrons.Consequently,LRR-1,TLR2,and VSP genes were found expressed exclusively in the bacteriocytes of G.platifrons.Therefore,it was suggested that TLR2,integrin,VSP,MMP1,and LRR-1 might be crucial molecules in the symbiosis between G.platifrons and methane oxidation bacteria by participating in symbiosis-related immune processes.

Mg/Ca,Ba/Ca,and S/Ca ratios as environmental and growth proxies for bivalve shells from the Haima cold seep,South China Sea

Bivalve shell fossils,cemented by authigenic carbonates,are widely spread in the Haima cold seep,Qiongdongnan Basin of the South China Sea(SCS).In this study,we examined an element profile of Gigantidas platifrons in the Haima cold seeps at a water depth of 1381 m.Based on the scanning electron microscope(SEM)analyses,the prismatic layer and nacreous layer were identified,which are characterized by prismatic structure and stratified structure,respectively.In addition,the profile can be subdivided into two parts:altered and unaltered zones.Laser inductively coupled plasma mass spectrometry(LA-ICP-MS)mapping shows that the element concentrations of the altered zones were influenced by the authigenic carbonate rocks,whereas the element concentrations of unaltered zones remain stable.In-situ X-ray diffraction(XRD)analyses show that the mineral constituent of the prismatic layer is mainly composed of aragonite.Along with the growth profile,Mg/Ca ratios of unaltered zones have minor variations,ranging 0.72-0.97 mmol/mol(mean=0.87 mmol/mol),with estimated temperatures of 3.8-4.1℃,indicating that the temperature of the surrounding seawater remains constant and agree with the measured data of 3.9℃which was conducted by a conductivity-temperature-depth system(CTD).The minor variations of Ba/Ca ratios(0.01-0.06 mmol/mol;mean=0.04 mmol/mol)indicate a relatively stabilized salinity of the surrounding seawater.S/Ca ratios show large variations of 0.04-4.15 mmol/mol(mean=1.37 mmol/mol).S/Ca ratios have regular variations which generally correspond to the variations of the Mg/Ca ratios,highlighting that the S/Ca ratios of bivalve shells show the potential to reflect the growth rate of the Gigantides.However,further studies should be carried out on the understanding of the links between the S/Ca ratios and seepage intensity of cold-seep fluids.