| Summary: | 碩士 === 國立臺灣海洋大學 === 水產養殖學系 === 105 === Asian seabass (Lates calcarifer) is important commercial aquaculture species in Taiwan and Southeast Asia. Asian seabass has advantages of fast-growing, resistance to disease and tender meat, so is popular in farmers and consumer. In early days, Asian seabass been known as good for recover after surgery. Due to it’s price is higher than normal aquaculture species, only in the restaurant have meal made by Asian seabass. In recent years, production has gradually increased, price reduced to the general public have the ability to afford. Global warming caused the trend of climate change, the incidence of extreme climate increased significantly in recent years. Because of the cold spell from December to February every year cause sudden drops in water temperatures, important economic species such as milkfish, bass, grouper and white shrimp freeze to death, cause a great losses in economic. The purpose of this study is to develop putative molecular markers of cold tolerance-related genes in Asian seabass. Afier Asian seabass were subjected to cold stress, we sampled five tissues: brain, gill, brain, liver, head kidney and muscle from two groups, the cold- tolerance group (SPT), which survived under the cold stress; the cold-sensitive (SPS) group, which couldn’t survived at the low temperature. Five tissues sample were performed using the Illumina HiSeq 2000 platform for transcriptome sequencing. Transcriptome data base produce 44,183,427,900 nt, we obtained 142,824 unigenes with an average length of approximately 1320 base pairs. Total 19,370 unigenes were co-annotated in public protein databases, including the NCBI non-redundant (NR), nucleotide (NT), SwissPort, Kyoto Encyclopedia of Genes and Genome (KEGG), Clusters of Orthologous Groups (COG) and Gene Ontology (GO) database. In addition, 62,034 SSRs were identified using molecular marker loci detection software MicroSAtellite (MISA). We use Venny(2.1) to analysis Up-regulated and down-regulate genes between tissues, founds 308 co-differential expressed genes. Cold related gene sequences were established out of 20 functionality SSR markers. The result shown that 14 of 20 SSR markers were polymorphism . Using discriminant analysis to discriminate 14 SSR markers, founds the correct rate of CL12362_4 marker in SPT and SPS group were 75% & 69%, respectively.Genotype EE frequency in SPT and SPS were 50%& 18.75%, respectively. These polymorphism results among and within the strains can be further use to identify the strain-specific markers for identification of cold-tolerance and improve the genetic stability of breeding programs for L. calcarifer.
|
|---|
