| 不同引物扩增对东海陆架沉积物真核微型生物多样性的影响 |
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| 引用本文： | 俞凯成,王健鑫,陶 诗,刘明华,蒋 然,刘雪珠,黄 备.不同引物扩增对东海陆架沉积物真核微型生物多样性的影响[J].海洋与湖沼,2014,45(4):806-816. |
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| 作者姓名： | 俞凯成 王健鑫 陶 诗 刘明华 蒋 然 刘雪珠 黄 备 |
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| 作者单位： | 浙江海洋学院海洋微生物分子生态与应用实验室 舟山,浙江海洋学院海洋微生物分子生态与应用实验室 舟山,浙江海洋学院海洋微生物分子生态与应用实验室 舟山,浙江海洋学院海洋微生物分子生态与应用实验室 舟山,浙江海洋学院海洋微生物分子生态与应用实验室 舟山,浙江海洋学院海洋微生物分子生态与应用实验室 舟山,浙江省舟山海洋生态环境监测站 舟山 |
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| 基金项目： | 国家自然科学基金面上项目, 31270160号 |
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| 摘 要： | 采用三对通用引物(ITS1/ITS4,EF3/EF4,NS1/NS4)检测其对东海陆架DH-13站点表层沉积物真核微型生物的扩增特异性和多样性差异。对323个有效克隆子进行OTU分型和多样性分析,结果表明,NS1/NS4引物扩增所获得的文库多样性最高,EF3/EF4其次,ITS1/ITS4最后。通过NCBI比对和分类学研究,发现优势种群主要为原生生物界,包括丝足虫门(Cercozoa,占所有OTU的36.4%)、横裂甲藻纲(Dinophyceae,占14.2%)、不等鞭毛门(Stramenopiles,占7.4%)和纤毛虫门(Ciliophora,占6.3%);另含部分真菌界如子囊菌门(Ascomycota,占7.4%)和壶菌门(Chytridiomycota,占6.3%)。原生生物方面,NS1/NS4引物表现的多样性最高,适合于扩增丝足虫门、横裂甲藻纲和不等鞭毛门,并扩增到独有的眼虫门(Euglenozoa)、隐藻门(Cryptophyta)和顶复门(Apicomplexa);ITS1/ITS4适合于扩增甲藻和纤毛虫门;EF3/EF4则适合于丝足虫门。真菌方面,ITS1/ITS4适合于扩增子囊菌门;EF3/EF4适合于子囊菌门和壶菌门,而NS1/NS4适合于壶菌门。系统发育分析表明,三对引物的原生生物系统树中的相似序列多来自海洋环境样品,真菌序列则分别来自陆地土壤真菌和海洋真菌,说明三对引物更适合于海洋沉积环境微型原生生物的分子生态分析。
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| 关 键 词： | 真核微型生物 多样性 引物 S rDNA ITS |
| 收稿时间： | 2013/11/6 0:00:00 |
| 修稿时间： | 6/2/2014 12:00:00 AM |
EFFECTS OF AMPLIFICATION WITH DIFFERENT PRIMERS ON MICROEUKARYOTIC DIVERSITY IN THE SURFACE LAYER SEDIMENTS OF THE EAST CHINA SEA |
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| YU Kai-Cheng,WANG Jian-Xin,TAO Shi,LIU Ming-Hu,JIANG Ran,LIU Xue-Zhu and HUANG Bei.EFFECTS OF AMPLIFICATION WITH DIFFERENT PRIMERS ON MICROEUKARYOTIC DIVERSITY IN THE SURFACE LAYER SEDIMENTS OF THE EAST CHINA SEA[J].Oceanologia Et Limnologia Sinica,2014,45(4):806-816. |
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| Authors: | YU Kai-Cheng WANG Jian-Xin TAO Shi LIU Ming-Hu JIANG Ran LIU Xue-Zhu and HUANG Bei |
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| Institution: | Zhejiang Ocean University, Laboratory for Marine Microbial Molecular Ecology and Application, Zhoushan, 316000,(Marine Science & T echnology College,Zhejiang Ocean University,Zhoushan,316000),(Marine Science & T echnology College,Zhejiang Ocean University,Zhoushan,316000),(Marine Science & T echnology College,Zhejiang Ocean University,Zhoushan,316000),(Marine Science & T echnology College,Zhejiang Ocean University,Zhoushan,316000),(Marine Science & T echnology College,Zhejiang Ocean University,Zhoushan,316000),(Marine Science & T echnology College,Zhejiang Ocean University,Zhoushan,316000) |
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| Abstract: | Three pairs of universal PCR primer which are specific to 18S rDNA and ITS sequence were used for testing specificity of amplification for microeukaryotic from sediments ,and comparing their ability of evaluation for diversity about microeukaryotic from ocean surface sedimentary .Three pairs of PCR primer acquired 323 valid clones which will used for taxonomy comparison and phylogenetic analysis after classify OUT. The results suggest that NS1/NS4 show the highest diversity in the protest taxon from the East China Sea continental shelf surface sediments. ITS1/ITS4 was followed .EF3/EF4 fit for diversity analysis about species within genus Cercozoa.For fungus, the main groups of ITS1/ITS4 were Ascomycota and Basidiomycota, the main groups of EF3/EF4 were Ascomycota and Chytridiomycota,and the main groups of NS1/NS4 were Chytridiomycota. With the various amplification results, we should integrate apply base on intent and object. By comparing the effects to microeukaryotic diversity from the surface layer sediments amplified with different primers, this paper illustrates the integrated use of varied primers to assess the importance of eukaryotic microorganism biodiversity in complex environment. |
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| Keywords: | microeukaryotic diversity amplification primer 18S rDNA ITS |
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