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淡水魚類翹嘴紅鮊之種原培育及遺傳資源分析研究

  • 日期:98-04-17
  • 年度:2008
  • 領域:漁業領域
  • 主持人:黃家富
  • 研究人員:黃家富、黃德威、劉富光

近年來由於生態保育與環境監測日益受到重視,種原之多樣性研究與保種,日趨重要。翹嘴紅鮊(Erythroculter ilishaeformis)分佈於日月潭、曾文水庫、烏山頭水庫等地區,為當地觀光發展重要的特產品。人工繁殖部分:本中心進行了不同激素對翹嘴紅鮊產卵效果影響試驗,結果顯示以HCG混合LHRH-A2 (HCG 600 IU+LHRH-A2 10微克)注射催熟效果較佳;於水溫29℃下,效應時間約9-10小時;受精卵約15-19小時孵化。本中心本研究之RAPD引子係利用400組商業化的引子組中,篩選寡核苷酸序列G+C含量分析均高於80﹪適當引子,其計有50組作為分析研究。結果有34組RAPD引子在選定標準魚DNA上之隨機擴增DNA片段具重複性、專一性的模式,其中有25組引子組對翹嘴紅鮊或紅鰭鮊產生專一性的(unique)標記,可用於鑑別分析。此外應用微衛星DNA作為核苷酸指紋鑑定研究;運用內切性限制酶將翹嘴紅鮊染色體DNA剪切,並與質體接合,再轉形到大腸桿菌(E. coli)中培養,並運用點墨雜合檢定,結果僅篩選284株可能具有微衛星DNA片段之菌株。再經南方氏雜合檢定分析,結果僅有14株具明顯的呈色反應,進行DNA序列分析;結果其轉殖成功率偏低。檢送DNA序列定序,獲得6組明顯的微衛星DNA序列。其中microsatellite DNA有(GT)n等形式;目前完成6組之最適條件之探討,建立種源專一性特定序列位點之開發,但由於樣品與引子組無法作為族群群分析之用。目前仍積極探討偏低原因與繼續篩選適用的引子中。

研究報告摘要(英)


Research of different hormone combination on the spawning effect of Erythroculter ilishaeformis was evaluated. The results showed that hormone administration of HCG., LHRH-a2 and HCG+LHRH-a2 can induce the E. ilishaeformis to spawn.We selectrd 50 RAPD primers that G/C contant is 80 % from UBC (University of British Columbia) for their utility in identifying genetic polymorphism in E. ilishaeformis. Of the 50 primers, 34 were good and medium quality RAPD profiles and 25 RAPD primer are unique marker, its different between E. ilishaeformis and Culter erythropterus. Our study present RAPD markers are useful genetic markers in E. ilishaeformis. Genetic marker are molecular labels which identify genes and are valuable to the animal industry as tools of animal selection. In this study, Microsatellite DNA techniques will chose for identify the genetic diversity within and among different populations for Culter spp. Microsatellite loci were characterized in the E. ilishaeformis random clones from small genomic library using (GT)15 and (CT)15 prob. Primers for DNA amplification using PCR were designed and synthesized for 6 loci in this research. Developed microsatellite primers should prove useful for population studies of the E. ilishaeformis.