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DNA Barcoding of the Endangered Freshwater Mussel Cristaria plicata Using Shell-derived DNA KCI 등재
- 언어ENG
- URLhttps://db.koreascholar.com/Article/Detail/451781
pp.121-131
Freshwater unionid bivalves are difficult to monitor due to their cryptic, burrowing behavior and their residence in soft substrates. DNA barcode reference sequences are essential for molecular identification and conservation studies of endangered freshwater unionids. For endangered species, the collection of live individuals is often restricted, which limits the construction of such reference databases. In this study, we evaluated the feasibility of using shell-derived DNA from Cristaria plicata as a source for mitochondrial DNA barcoding. Naturally deposited shells were collected from a freshwater environment, and DNA was extracted from five distinct shell regions under different preservation conditions. Mitochondrial 16S rDNA was targeted using both short (180 bp) and long (450 bp) barcode primers. PCR products were sequenced and compared with GenBank reference sequences, and phylogenetic analyses were conducted to confirm taxonomic identity. DNA yield differed substantially among shell regions and preservation states. Phylogenetic analyses demonstrated that sequences derived from shells clustered with established C. plicata references from South Korea and other East Asian populations. However, degraded shell materials occasionally produced divergent or nonspecific sequences, particularly when longer barcode regions were amplified. These results indicate that shell-derived DNA can serve as a viable, noninvasive source for generating DNA barcode references of endangered unionid bivalves. This approach may facilitate the expansion of reference databases and support molecular monitoring and conservation efforts without requiring the collection of live specimens.
INTRODUCTION
MATERIALS AND METHODS
1. Collection of Cristaria plicata shells
2. DNA extraction from shells
3. DNA barcode amplification
4. Sequencing of DNA barcodes andphylogenetic analysis
5. Data resources
RESULTS
1. Variation in shell-derived DNA concentrationbased on shell condition
2. Mitochondrial 16S rRNA gene amplification
3. Phylogenetic analysis of mitochondrial16S rDNA sequences
DISCUSSION
REFERENCES
Supplementary data 1. Estimates of pairwise distance between short sequences (Unio 16S F/R primer) based on Unionidae 16S rDNA.
Supplementary data 2. Estimates of pairwise distance between long sequences (16S-FW1F/FW1R, FW2R primer) based on Unionidae 16S rDNA.
