The Molecular Identification Technology Of Anti-Counterfeiting And Protecting Ling Etc.Marine Species

With the rapid development of our country’s marine product international trade, some merchants in pursuit of profit adulterated some inexpensive marine products in costly producst, who sold shoddy and masqueraded, which seriously effected the normal order in the international trade. However, there is not a standard or method of detecting original components of marine product in domestic and foreign country. We study ling, callop and horseshoe crab, set up the PCR method specific for ling identification, the species-specifc microsatellite markers method specific for scallop identification, and the real-time PCR method specific for horseshoe crab identification, which effectually probe into the molecular identification for anti-counterfeiting and protecting the marine species.1. Anti-counterfeiting of ling. Avoiding ling and its product was imitated with cheap catfish etc. in the marine product trade, we set up a PCR method specifically amplifying DNA of ling species, mtDNA COXII sequences of7ling subspecies and5other fish species were download from Genbank, and compared by DNASTAR7.0software, designed PCR primers,.The applied results indicated that this method was specific to identification of ling species, which offered the powerful technical guarantee in the ling trade.From March2010to March2011, the PCR method of identifying ling species has been popularized and applied to Yantai, Weihai, Qingdao, Rongcheng, Penglai, Longkou, Laizhou and so on,10port cities of Shandong province, fishery corporations, every city’s entry exit inspection and quarantine bureaus, and scientific research centers in9provinces, Guangdong, Liaoning, Beijing, Shanghai, Jiangsu, Fujian, Sichuan, Guangxi and so on.3,256samples have been inspected in one year, of which345shipments of fake and dubious ling in total have been detected at the end of march2011. The total value was 49,060,000RMB.2. Molecular identification of scallop species. Scallops,which comprise hundreds of species, have very different price, and then the cheap one is usually imitated the rare one for sale in the market. Identification of commercial scallops to the species level is an important issue owing to the difficulty of distinguishing the species by morphological characters. In the present study, we established the species-specific microsatellite markers for Zhikong scallop, Yesso scallop, Noble scallop and Bay scallop to identify commercial scallop..This method well demonstrated high precision and high sensitivity, and more competence in the identification of heavily processed product, which provides a technical support for anti-fake commercial scallops in the trade.From2009, this method has been popularized and applied to identify the importing scallop larvas in Shandong Dongfang Ocean Ltd, Co., Beijing Huadu Yukou Aquatic Products Ltd, Co..and so on, the main aquatic products corporations of our country. The polyculture of different scallop larva which should be in different clutured conditions may cause profit losses. This method well demontrated more competence in the identification of scallop larva species. Separately culturing scallop larvas can reduce the losses caused by polyculture to minimum. The total benefit has been31.53million RMB in this3years.In the1990s, we have found that the importing Noble scallop larvas have been adulterated the other species, but we couldn’t counterclaim for lack of scientific proof. With our method, the low purity of Noble scallop larvas imported from an American company have been detected in the early2010, which urged the company to provide the purest Noble scallop larvas for the Chinese imported company, and maintained the rights and interests of our country’s aquatic product company in the international trade.The method of species-specific microsatellite markers for scallop species identifi-cation has been formed a standard of inspection and quarantine industry.3. Molecular identification of horseshoe crab. We set up a SYBR Green I real-time PCR method specific for horseshoe crab identification for protecting the rare aquatic animal existed only in our country. mtDNA COXIII sequences of horseshoe crab and17other arfhropoda species were download from Genbank, and compared by DNASTAR7.0software. The established method probed into the protection of our country’s rare animals. In2010, a passel of bio-reagent which may comprise some dubious horseshoe crab elements was captured in a city of Shandong province. It was proved that the bio-reagent contained horseshoe crab elements with our method. When confronted the result, the manufacturer confessed.