To detect genetic diversity of seafood by single strand conformation polymorphism method

• Main Authors: Ching-Wen Kuo, 郭晴文
• Other Authors: Tsai-Hsin Chiu
• Format: Others
• Language: zh-TW
• Published: 2015
• Online Access: http://ndltd.ncl.edu.tw/handle/31860178993082235796

碩士 === 國立澎湖科技大學 === 食品科學研究所 === 103 === Ivory shell（Babylonia areolata) and Coral trout (Plectropomus leopardus) are high-valued seafood in Taiwan,and this study use ISSR (Inter-simple sequence repeat)、DGGE (Denaturing gradient gel electrophoresis) and SSCP (Single strand conformation polymorphism) of molecular marker technique to analyze genetic diversity of B. areolata and P. leopardus .The samples were collected from Penghu Marine Biology Research Center.All samples were confirmed to the species and extracted DNAs were analyzed for COXI gene、16S rRNA gene and ITS gene sequences,and than 59 primers of ISSR,and use DGGE and SSCP were used to identify the genetic diversity for all samples. Results showed that COXI and 16S rRNA gene sequences be undifferentiated of B. areolata ; in ISSR analysis, 59 primers were selected in this study. A total of 3 primers, ISSR3, ISSR7 and ISSR13, yielded polymorphic bands and had a good distinguishing ability; DGGE analysis had banding patterns being different in each sample, this results also showed which patterns and clusters were similar to COXI sequencing analysis, thus DGGE method could discriminate Babylonia samples between and within species; SSCP result this method could discriminate Babylonia samples between. In this study, results showed that COXI could be amplified the target fragment 710 bp,and ITS gene could be amplified the target fragment 543 bp,but this sequences be undifferentiated of Plectropomus leopardus ;and than used DGGE and SSCP method, SSCP result can't analysis Plectropomus leopardus but DGGE method could discriminate Plectropomus leopardus and Serranidae fish samples between and within species.In this study,the DNA moleculer marker were developed and could be provide information for variety identification, clonal propagation, and genetic improvement of species-specific in aqultues.