应用DGGE技术分析扇贝养殖海区真核浮游生物种群多样性  

Diversity of Eukaryotic Plankton Assemblages at Culture Area for Scallop by Denaturing Gradient Gel Electrophoresis(DGGE)

作　　者：王娜[1] 蔡玉勇[1,2] 李赟[1] 王崇明[2] 崔红 

机构地区：[1]中国海洋大学海水养殖教育部重点实验室,山东青岛266003 [2]中国水产科学研究院黄海水产研究所,山东青岛266071 [3]山东省潍坊市畜牧检测中心,山东潍坊261061

出　　处：《中国海洋大学学报（自然科学版）》2010年第12期51-56,共6页Periodical of Ocean University of China

基　　金：国家高技术研究发展计划项目(2006AA100307);现代农业产业技术体系建设专项(nycytx-4)资助

摘　　要：为分析扇贝养殖海区真核浮游生物种类和丰度,本研究首先以实验室培养的9种饵料微藻总DNA为模板PCR扩增18S rDNA V1~V3区的高变区序列,筛选用于DGGE分析该扩增序列的变性剂浓度,结果表明,扩增序列长560 bp,DGGE分析可将不同藻种分离开。随后对青岛沙子口扇贝养殖海区9个月份真核浮游生物多样性进行了DGGE分析,结果从9个月份2 m深的海水中总共扩增出38条谱带,其中共有谱带3条,占总数的7.69%,不同月份的特征性谱带11个,占总谱带数的28.2%。不同月份水样中的谱带数显著不同,香农多样性指数为0.4632。相似性分析表明,2009年3和4月养殖海区真核微生物种类相似性最高,为89.7%。基于不同月份水样微生物种类多样性进行UPGMA聚类分析,结果表明,2008年5,6,8月和2009年1,2,3,4月分别聚为一支,显示取样时间相近的月份微生物种类相似度最高。The 18S rRNA genes（V1 to V3 region） was amplified by two universal primers（Euk516r-GC and Euk1A） from 9 cultured microalgae species.Then,the denaturing gradient gel electrophoresis（DGGE） with denaturing gradient of 25% to 65% urea were optimized.The result indicated that the primer sets gave a single band with the length of amplified fragment approximately 560 bp,and the DGGE with denaturing gradient of 35% to 48% urea could separated nine cultured microalgae species in different position.The diversity of eukaryotic planktons from scallop cultivation area was analyzed by DGGE,and total 38 electrophoric bands were showed from the 9 months water collected from 2 meters deep,of which 3 bands existed in all nine months,taking 7.69 percent of total bands number,and there were 11 characteristic bands in different month,taking 28.2 percent.The result indicated that the diversity of eukaryotic planktons was various significantly among different months,Shannon index is 0.4632.Similarity analysis showed that the highest similarity was 89.2% between March and April 2009.UPGMA clustering analysis based on different eukaryotic plankton assemblages showed that May,June and August of 2008 were grouped into a clad,January,February,March,April of 2009 into the other clad,which implied that similarity of the eukaryotic plankton diversity among close month was high.

关 键 词：PCR-DGGE 18S RDNA序列 真核浮游生物多样性 

分 类 号：S917.1[农业科学—水产科学]