Preliminary Assessment Of Genus-specific Genes As Molecular Markers To Study Early Process Of Marine Biofouling

Marine creatures adhering to the bottom of hulls, water pipes and other surfaces of artificial facilities have caused huge losses to the development of marine industry. Marine antifouling has become an important issues to be solved. Prevention of marine biofouling could start from the adhesion of marine organisms at pioneer period of biofouling, so the study on fouling organisms in the early attachment process is a significant basis for the development of antifouling technology. Currently, most of the study on fouling organisms focuses on the aspects of fouling biological ecology and antifouling technology. Little study was conducted on dynamic process of biofouling, especially in the molecular level. So this topic is committed to using molecular markers to explore the early dynamic process of biofouling.On the one hand, possible corresponding relationship was investigated between specific gene expression of marine fouling prokaryotic microbes adhering to different surfaces of marine engineering coating and different types of antifouling coating surfaces. Through consulting related references and literatures,78kinds of different marine fouling organisms were collected. Names of marine fouling organisms which have whole genome sequence were retrieved and determined in the EBI database. Genera-specific and species-specific genes were determined through consulting the whole genome specific gene database constructed by Harvard Biostatistics Laboratory. Finally21kinds of genera-specific genes and10kinds of species-specific genes were obtained. According to principles for primer design, for each of the genus-specific genes, primers were designed and verified one by one until suitable primer pairs were obtained. A total of31primer pairs were acquired (21primer pairs for specific genes).Through marine adhesion test lasting for one year,6kinds of promotion and5kinds of inhibition of the growth of oyster different kinds of antifouling coatings were selected. The nano-sized additives are silicon nanopowder, carbon nanopowder, graphite powder and MWNT (<8nm). Fouling samples were collected through short-term marine adhesion test at different sampling time (15d). Mixed genomic DNA extracted from early fouling samples were scanned and verified by31kinds of specific primers and six primer have been successfully identified as different genera-specific primers, namelyAcinetobacter, Flavobacterium, Glaciecola, Jannaschia, Rhodococcus and Sulfitobacter. The identified six genus-specific primers were applied to scan genome DNA extracted from samples collected from different coating surfaces at different periods.The results showed that there are certain differences between the expression of Jannaschia and Rhodococcus and Sulfitobacter in the two types of surfaces. Acinetobacter only appeared in surface of coating containing MWNT additives.On the other hand, preliminary endeavor was provided to clone marine eukaryotic genes with the most richness in the whole cDNA library of early-stage (about14days at15℃). Main strategies adopted are using the restriction enzymes to cut cDNA fragments with adapter containing enzyme loci provided by kit. A variety of and combinations included HindIII and BamHI, PstI and XbaI. Then cDNA were ligated into vector at the same restriction sites. Over300white colonies were collected and subjected to DNA sequencing. The experimental result revealed that though it is difficult to combine the whole cDNA sequence with the vector, our study provides a new strategy and insights into marine fouling eukaryotic gene cloning, which provided theoretical basis to delve into marine fouling early signaling pathway and promoted the development and utilization to gene expression suppressor gene research.