浙江海洋学院学报(自然科学版)
浙江海洋學院學報(自然科學版)
절강해양학원학보(자연과학판)
JOURNAL OF ZHEJIANG OCEAN UNIVERSITY(NATURAL SCIENCE)
2013年
6期
484-487,493
,共5页
李鹏飞%朱文斌%贺舟挺%王跃斌
李鵬飛%硃文斌%賀舟挺%王躍斌
리붕비%주문빈%하주정%왕약빈
浙江近海%三疣梭子蟹%控制区%遗传多样性
浙江近海%三疣梭子蟹%控製區%遺傳多樣性
절강근해%삼우사자해%공제구%유전다양성
offing of Zhejaing%Portunus trituberculatus%D-loop%genetic diversity
以浙江近海三疣梭子蟹大螯肌肉的线粒体DNA作为模板,对4群体共67个个体的控制区(D-loop)序列进行了PCR扩增。通过对PCR产物进行双向测序,最终得到了控制区3’端的rRNA部分序列(64 bp)、D-loop全序列(1179 bp)及控制区5’端tRNA-Ile部分序列(73 bp)。经DNASP软件分析得知,67个D-loop序列定义了65个单倍型,在65个单倍型中,除去插入/缺失位点,所有单倍型共检测到344个多态位点,主要发生在D-loop序列的中间区域;D-loop序列检测到26个插入或缺失位点,碱基插入主要发生在中央后端区域,以13 bp重复片断插入的形式进行。每个个体含有0~3个连续重复片断。通过对三疣梭子蟹的遗传多样性分析发现,D-loop序列的单倍型多样性为0.982~1.000,核苷酸多样性指数为0.02770~0.03633,平均核苷酸差异数为31.790~43.304。综合分析,三个成蟹群体的遗传多样性相近,都低于仔蟹的遗传多样性,说明经过遗传育种,可以提高三疣梭子蟹的种质情况,真正实现对三疣梭子蟹的科学利用。
以浙江近海三疣梭子蟹大螯肌肉的線粒體DNA作為模闆,對4群體共67箇箇體的控製區(D-loop)序列進行瞭PCR擴增。通過對PCR產物進行雙嚮測序,最終得到瞭控製區3’耑的rRNA部分序列(64 bp)、D-loop全序列(1179 bp)及控製區5’耑tRNA-Ile部分序列(73 bp)。經DNASP軟件分析得知,67箇D-loop序列定義瞭65箇單倍型,在65箇單倍型中,除去插入/缺失位點,所有單倍型共檢測到344箇多態位點,主要髮生在D-loop序列的中間區域;D-loop序列檢測到26箇插入或缺失位點,堿基插入主要髮生在中央後耑區域,以13 bp重複片斷插入的形式進行。每箇箇體含有0~3箇連續重複片斷。通過對三疣梭子蟹的遺傳多樣性分析髮現,D-loop序列的單倍型多樣性為0.982~1.000,覈苷痠多樣性指數為0.02770~0.03633,平均覈苷痠差異數為31.790~43.304。綜閤分析,三箇成蟹群體的遺傳多樣性相近,都低于仔蟹的遺傳多樣性,說明經過遺傳育種,可以提高三疣梭子蟹的種質情況,真正實現對三疣梭子蟹的科學利用。
이절강근해삼우사자해대오기육적선립체DNA작위모판,대4군체공67개개체적공제구(D-loop)서렬진행료PCR확증。통과대PCR산물진행쌍향측서,최종득도료공제구3’단적rRNA부분서렬(64 bp)、D-loop전서렬(1179 bp)급공제구5’단tRNA-Ile부분서렬(73 bp)。경DNASP연건분석득지,67개D-loop서렬정의료65개단배형,재65개단배형중,제거삽입/결실위점,소유단배형공검측도344개다태위점,주요발생재D-loop서렬적중간구역;D-loop서렬검측도26개삽입혹결실위점,감기삽입주요발생재중앙후단구역,이13 bp중복편단삽입적형식진행。매개개체함유0~3개련속중복편단。통과대삼우사자해적유전다양성분석발현,D-loop서렬적단배형다양성위0.982~1.000,핵감산다양성지수위0.02770~0.03633,평균핵감산차이수위31.790~43.304。종합분석,삼개성해군체적유전다양성상근,도저우자해적유전다양성,설명경과유전육충,가이제고삼우사자해적충질정황,진정실현대삼우사자해적과학이용。
Mitochondrial DNA (mt DNA) of Portunus trituberculatus was prepared as templates from their muscle. The PCR technique was used to amplify the D-loop sequence in 67 individuals of 4 population. The PCR products were sequenced and 64 bp rRNA sequence of 3’end and 1 179 bp of complete D-loop sequences and 73 bp tRNA-Ile of 5’end were obtained (Remove partial sequences of the two ends). By using DNASP soft-ware to analyze the genetic diversity, the result showed that there were 65 haplotypes and 334 variation sites were observed (no inserting/missing sites). The variation sites occurred in the middle of the D-loop sequences. 26 inserting/missing sites were observed in the end of the D-loop sequences too. Results revealed a 13 bp length repetitive sequence in the sequences and individuals differed from each other in frequency and length of the repetitive sequences. As estimated by DNASP, the haplotypic diversity region was 0.982-1.000, the nu-cleotide diversity and the average number of nucleotide defferences were 0.027 70-0.036 33 in D-loop se-quences. It can be concluded that the genetic diversity of P. trituberculatus in the offing of Zhejiang was rich.