Bioscience Methods 2025, Vol.16, No.3, 137-153 http://bioscipublisher.com/index.php/bm 146 metabarcoding, which can be used for environmental DNA monitoring. By extracting environmental DNA from aquaculture water or sediments and amplifying barcode sequences such as COI, it is possible to detect whether there is recruitment of wild juvenile abalone or invasion of alien abalone in the aquaculture area (Li et al., 2021). This non-invasive monitoring method is very beneficial for aquaculture ecological management and wild resource protection. In addition, mini-barcoding technology can solve the problem of DNA fragmentation in processed products by selecting shorter barcode sequences to achieve species identification, which has been proven to be feasible in dried abalone and canned products. In general, the application prospects of DNA barcoding technology in the identification of abalone species are very broad, and it can play a unique role in scientific research, industrial supervision and resource protection. Of course, to fully realize its potential, it is necessary to establish a complete reference database and standard operating procedures. At present, a large number of species barcode sequences including abalone have been entered into GenBank and BOLD systems around the world, but it is still necessary to ensure that the sequences are accurately annotated and cover all species. With the advancement of these works, DNA barcoding will surely become a powerful tool for the study and management of abalone species diversity. 5.3 Framework for rapid identification and traceability systems In order to better apply molecular identification technology to abalone species monitoring and aquatic product traceability, it is necessary to build a set of rapid, accurate and economical comprehensive detection systems. Based on the current research progress, we put forward the following suggestions: First, establish a reference database and detection standards for DNA barcodes of abalone species. At the national or industry level, mitochondrial COI and other sequences of various abalone species should be collected, an authoritative reference sequence library should be constructed, and standard PCR amplification and sequencing protocols should be formulated. In this way, when an unknown sample is sent for inspection, each laboratory can operate according to a unified process, and the identification conclusion can be obtained by comparing the sequencing results with the database. Second, develop on-site rapid detection tools for abalone species. In law enforcement or market supervision, in order to improve the detection efficiency, a rapid method that does not require sequencing can be used. For example, ARMS-PCR primers are designed based on species-specific SNP sites to achieve species specificity of the amplified product. Real-time fluorescence PCR technology can also be applied to complete the simultaneous detection of multiple species within a few hours using specific probes. For example, a patented method has developed fluorescent quantitative PCR using the mitochondrial sequence specific to the wrinkled abalone, which can determine whether the sample contains wrinkled abalone components with one click. Third, use high-throughput genotyping technology to strengthen the traceability of abalone products. For high-value abalone species (such as Australian abalone and Japanese abalone), it is possible to consider developing a typing detection platform based on SNP chips or microsatellites to achieve species identification and origin certification for individual or batch products. Peng et al. (2021) have constructed an abalone SNP chip containing 60K markers, which can be simplified for species identification in the future. Chip testing can type thousands of loci simultaneously within a few hours, which improves accuracy. Fourth, combine molecular identification technology with information technologies such as blockchain to form a full traceability system for abalone products. Specifically, DNA identity tags are assigned to each batch of abalone during the breeding or fishing stage (through typing records), and the test verification results are uploaded at each link of the product circulation. Consumers can obtain authenticity information such as species and origin by scanning the product QR code. Finally, to improve the rapid identification system in practice, cost and convenience must also be considered. For example, for grassroots supervision, portable PCR equipment and kits can be promoted, which can be operated without complex laboratory conditions. At present, some companies have launched on-site DNA detection kits for aquatic products, which contain pre-freeze-dried PCR reagents. Only sample lysis solution needs to be added to amplify and the results can be read through test strips, which is very suitable for rapid screening. We believe that with the continuous maturity of molecular detection technology, it is feasible and necessary to establish a comprehensive system covering abalone species identification and product traceability. This will help combat seafood trade fraud, protect the reputation of geographical indication products, and safeguard the sustainable use of abalone resources.
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