| Summary: | 碩士 === 國立臺灣海洋大學 === 水產養殖學系 === 107 === Potato grouper (Epinephelus tukula) is a new aquaculture species and fast growth which is a high economic potential species, however, it still has different degrees of variation during rearing. The aquaculture sector in Taiwan is significantly behind foreign countries in applying selective breeding program, thus, aquaculture sector whom in southern Taiwan hoped to raise their provenance’s quality and management through molecular markers-assisted selection and breeding platform in potato grouper species. The purpose of this study was to develop a scheme which manage of broodstock and identify paternity. We injected RFID in the broodstock of potato grouper in two years for tracking and analyzed the genetic diversity. We constructed and sequenced cDNA libraries from potato grouper with seven tissues of brain, gill, head kidney, heart, spleen, liver and muscle in the transcriptome. Of these, approximately 51.3411G nt raw sequencing reads were generated and assembled 111,490 unigene, total length 172,770,477 bp and average length 1,549 bp. Through five protein databases and Venny analysis have annotated 22,031 unigenes. Furthermore, a total of 44,565 putative SSRs and 122,220 putative SNPs were identified. We also screened the molecular markers involved in growth-related genes from the transcriptomic database of potato grouper established by the next-generation sequencing. The experience not only screened the transcriptome database in growth-related genes which established out of 65 functional SSR and 46 SNP candidate markers but also contained two SSR markers studied before in our Lab.
These molecular markers were genotyped in potato grouper’s offspring which subjected to growing test less than ninety days. We utilized mtDNA COI to identify
the offspring of maternal potato grouper. The PCR- RFLP technology could help us to distinguish between three maternal potato grouper with restriction enzyme Hha I and
Hinc II in mtDNA D-loop region. The offspring that we got from the aquaculture sector had been confirmed to breed from the M6 maternal grouper. The results of 10
functional SSRs showed polymorphism in the offspring by using capillary electrophoresis technology, especially; the offspring had more than three genotypes in five functional SSR markers. MassARRAY platform showed 46 SNPs average call rate were 76.73%. After statistical analyze the association between growth-related with molecular markers which had 2 SNP makers (Unigene 7626、CL8791.Contig3) showed significant association with the wieght (p<0.05). The purpose in our experiment not only provide a genetic management of broodstock but also assist the seedling industry in growth breeding, accelerated aquaculture traceability systems and improvement of quality-based potato grouper in the future.
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