海洋微微型真核生物分子多样性的研究方法

摘要

Picoeukaryotes are widely distributed in marine ecosystems. They play crucial roles in primary productivity and constitute one of the essential components of the microbial food webs. Because of their small size and the lack of morphological characteristics to be identified, traditional approaches such as electron microscopy can hardly be used in the study of microeukaryotic diversity. However, molecular biological approach has shed light in this area and has become an essential research method in the study of microeukaryotic diversity in the past decade. We generalized the present results of the molecular diversity study of marine picoeukaryotes and especially pointed out the study results in China and reviewed the target genes commonly used in the area, including conserved genes such as 18S rDNA, ITS regions and functional genes such as psbA, rbcL;the molecular biological methods such as clone library, DGGE, TGGE, PCR-RFLP, T-RFLP, FISH and metagenomics;the problems in the study of marine microeukaryotic diversity;and our study of microeukaryotic diversity in the North Yellow Sea.%微微型真核生物在海洋中广泛存在,是初级生产力的重要贡献者,也是微食物环的重要组成部分.由于微微型真核生物形体微小且不同种类缺乏明显形态学特点,故而难以利用电镜等传统方法精确研究其系统发生关系和种类组成.运用分子生物学技术直接对微微型真核生物种类组成进行研究已成为微微型真核生物多样性研究的重要方法.本文概述了目前世界范围内的微微型真核生物分子多样性研究进展以及我国在该领域中的研究状况.综述了微微型真核生物分子多样性研究中常用的靶标基因,包括18S rDNA和ITS序列等真核生物保守基因及psbA和rbcL等功能基因;常用的分子生物学方法,如克隆文库构建、DGGE/TGGE、PCR-RFLP/T-RFLP、FISH、宏基因组等;以及目前研究方法中存在的一些问题.