台灣及其鄰近地區養殖牡蠣（Crassostrea spp.）族群遺傳變異之研究
國立中山大學海洋生物研究所碩士論文摘要

研 究 生：余正平
指導教授：鄭金華 博士

本研究以台灣、金門以及福建沿海牡蠣為材料，利用分子生物方法分析其族群間的差異性。首先以粒腺體16S rRNA以及COI兩個基因的DNA片段序列，得到各種序列間的差異以及限制脢切位等資料，再利用限制脢切割片段形式分析各族群間的差異性。
在16S rRNA 530 bp DNA片段序列的分析結果顯示：葡萄牙牡蠣與台灣養殖的C. angulata差異很小（0-1 bp），與C. gigas的差異較大（3-4 bp）。在COI 700 bp DNA片段序列的分析中也得到類似的結果。葡萄牙牡蠣與台灣養殖的C. angulata無法利用這兩個片段的差異加以區分。利用COI的PCR RFLP分析，本研究首次發現金門、福建沿海也有C. angulata的分布。以COI PCR RFLP分析則發現從葡萄牙採集的C. angulata與金門、福建沿海的牡蠣族群遺傳距離最為相近。

Population genetic variation of cultured oyster Crassostrea spp. in Taiwan and neighborhood

Cheng-Pin Yu

（Advisor：Dr. Jin-Hua Cheng）
Institute of Marine Biology, National Sun Yat-sen University, Kaohsuang 804 Taiwan R.O.C.

Thesis Abstract
The genetic variations among oyster populations of Taiwan, Ginmen and Fu-kein coast are analyzed using molecular method. Firstly, DNA sequences of mitochondrial 16S rRNA and COI genes are analyzed according to nucleotide differences and restriction enzyme recognition site. Then, population variations were analyzed by the lane pattern of restriction enzyme fragment（RFLP）.
The sequences of 530 bp DNA fragment of 16S rRNA gene showed little difference（0-1 bp）between Portuguese oyster and oyster cultured in Taiwan, both have larger differences（3-4 bp）compared with C. gigas. The sequences of 700 bp DNA fragment of COI gene showed similar results. C. angulata is found in Ginmen and Fu-kein coast for the first time which is identified by the PCR-RFLP of COI gene fragment. PCR-RFLP analysis of COI gene fragment showed C. angulata from Portugal are genetically closest to the oysters collected from Ginmen and Fu-kein.

目 錄

章次 頁數
中文摘要 ............................................ III
英文摘要 ............................................. IV
謝辭 .................................................. V
目錄 ................................................. VI
表目錄 ............................................. VIII
圖目錄 ............................................... IX

章次 頁數
前言 .............................................. 1
材料及方法 ........................................ 3
I. 材料收集及處理 .............................. 3
II. Genomic DNA萃取 ............................ 3
III. 16S rRNA與COI DNA之聚合脢連鎖反應 ........ 3
IV. 洋菜膠電泳分析 ............................. 4
V. 16S及COI之定序 ............................. 5
VI. 16S與COI定序結果分析 ...................... 5
VII. DNA序列比對及限制脢切位分析 ............... 6
VIII. 16S與COI之限制脢片段多型性分析 .......... 6
IX. COI　RFLP在各族群間之比較 .................. 7
結果 .............................................. 9
討論 ............................................. 12
參考文獻 ......................................... 17

表目錄
表次 頁數
表一：牡蠣粒腺體16S rRNA基因中530 bp DNA片段序列之差異數量及遺傳距離 ............................. 22
表二：牡蠣粒腺體16S rRNA基因中530 bp DNA片段之限制脢切割資料 ..................................... 23
表三：牡蠣粒腺體COI基因中700 bp DNA片段序列差異數及遺傳距離 ..................................... 24
表四：牡蠣粒腺體COI基因中700 bp DNA片段之限制脢切割資料 ......................................... 25
表五：十一種COI RFLP type與其對應的binary code ... 26
表六：各族群中不同COI RFLP type的分布情形 ........ 27
表七：各COI RFLP type相互間之遺傳距離 ............ 28
表八：各族群間之遺傳距離 .......................... 29

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