中国科学院水生生物研究所机构知识库
Advanced  
IHB OpenIR  > 中科院水生所知识产出(2009年前)  > 学位论文
题名: 浮游生物群落DNA指纹分析及其在生态学研究中的应用
作者: 颜庆云
答辩日期: 2009-06-02
导师: 余育和
授予单位: 中国科学院水生生物研究所
授予地点: 水生生物研究所
学位: 博士
关键词: 浮游生物群落 ; 宏基因组 ; DNA指纹分析 ; RAPD ; PCR-DGGE
其他题名: DNA fingerprinting analysis of plankton community and its applications in ecological study
摘要: 本研究从生态系统功能角度将包括生产者(浮游植物)、消费者(浮游动物)和分解者(浮游细菌)等不同功能类群在内的浮游生物群落视为一个整体,将广泛应用于个体、种群层次的DNA指纹分析拓展至以真核生物为主的浮游生物群落研究。浮游生物因其丰富的多样性和特殊的生态位而在水体生态系统中发挥着极其重要的作用,其结构和功能是水体生态学重要的研究对象。很早以来人们就开始从不同层面和视角来研究浮游生物的分布、时空变化规律及其与环境的关系。以期揭示浮游生物群落在维持水体生态系统平衡中的作用,并探讨持续其生态系统服务功能的机制。不过,由于一般的浮游生物网通常难以采集到浮游细菌(bacterioplankton),且它们的鉴定和计数也非常困难。因此,浮游细菌一般被排除在常规的浮游生物生态调查范围之外,通常所说的浮游生物只包括浮游植物和浮游动物两大类。但浮游细菌作为水体生态系统物质循环过程不可缺少的环节,其生物量大、多样性高,是浮游生物生态学研究不容忽视的层面。本研究以浮游生物群落DNA(宏基因组)为媒介来研究浮游生物多样性及其与环境的相互关系。 首先选择生态背景资料(如水体理化特征、浮游生物组成及时空变化规律、滤食性鱼类放养密度等)比较丰富的武汉东湖为对象,以两种比较常用的指纹分析方法(RAPD和PCR-DGGE)为例,探索了DNA指纹分析在以真核生物为主的浮游生物群落应用的可行性。在此基础上,针对不同水体生态系统(湖泊、河流、水库等)的浮游生物群落,首先利用RAPD指纹分析进行整体扫描,然后针对16S rRNA和18S rRNA基因,通过PCR-DGGE分别对其中的细菌(包括蓝藻)和真核类群进行指纹分析,从不同层面来探讨浮游生物群落DNA多态性。同时,对不同时空范围采集的水体样品平行进行浮游生物的形态分类鉴定、定量计数,以及常规的理化分析。并对不同研究方法所得结果进行比较和综合分析,以期揭示浮游生物群落结构及其动态变化与环境的相互关系,进而探讨浮游生物对环境变化的响应机制及其在水环境预警中的作用。 在武汉东湖、洞庭湖、三峡库区的研究结果表明,浮游生物群落DNA指纹结构与物种组成及环境理化因子存在较好的相关性。武汉东湖浮游生物群落时空异质性研究发现,对浮游生物时空分布起主要作用的环境因子存在明显的季节差异,其中总氮(TN)、总磷(TP)与浮游生物群落结构的相关性较强,且TP在营养水平较高的环境与浮游生物群落的相关性更明显;研究结果还表明当营养水平相近时,重金属含量对浮游生物群落PCR-DGGE指纹结构具有较明显影响。人工模拟实验研究表明,TP与浮游生物群落PCR-DGGE指纹结构最相关,且壳聚糖改性土壤除藻过程对蓝藻以外的其它浮游生物演替没有明显影响。此外,本研究还对鲢、鳙肠含物、特定功能基因(nifH)和极端环境特定类群(水华蓝藻)进行了探索研究,结果显示这些都能为探讨、揭示水体生态学的规律提供分子生物学依据。通过群落DNA指纹结构来表征浮游生物群落组成,这样不同研究者在不同时空范围所开展的研究就能进行有效的比较,进而实现对浮游生物三大功能类群(浮游细菌、浮游植物和浮游动物)的综合分析研究,而这正是探究及阐明水体生态系统浮游生物生态学机理真谛的重要前提。
英文摘要: The present study extended DNA fingerprinting analysis, which has been successfully applied in individual- and population-level research, to ecological investigation of plankton community. As we all know, the planktonic organisms play important roles in functioning of aquatic ecosystems and also in biogeochemical cycles. Therefore, researchers started to investigate their distribution, spatiotemporal dynamics and their potential relationships with environments since the very start of marine research expeditions. Traditional plankton investigations primarily focus on the composition and dynamics based on the morphospecies. However, morphological identification has historically been a difficult task due to the paucity of readily observable characters of taxonomic value. Additionally, only the planktonic organisms with certain size can be collected by the commonly used plankton nets, and many small organisms (e.g. bacterioplankton) were generally excluded in traditional plankton investigations. But these ignored microorganisms are usually very important in maintaining the balance of aquatic ecosystem. Here, the whole plankton community (including bacterioplankton, phytoplankton and zooplankton) were studied with metagenome-based fingerprinting methods. This study characterized the biodiversity and dynamics of plankton community in target aquatic ecosystems, from species composition to metagenomic DNA fingerprints, to study the relationships between planktonic organisms and also between plankton community and environment. Firstly, the feasibility of applying DNA fingerprinting analysis to plankton community was explored and proved targeted the plankton communities from Donghu Lake. Then, RAPD and PCR-DGGE fingerprinting were applied to study the relationship between DNA polymorphisms and species composition of plankton communities in different ecosystems. Because community composition is dictated to a large degree by environmental factors, it is also a natural extension of this relationship between DNA fingerprints of plankton community and environmental conditions. Studies in the Donghu Lake, Dongting Lake and the Three-Gorges Reservoir suggested that DNA fingerprints of plankton communities were generally correlated with species composition, as well as with physiochemical factors. Spatiotemporal heterogeneity of plankton communities in Donghu Lake indicated that factors strongly correlated with the distribution of plankton community were seasonally different. But the concentrations of total nitrogen (TN) and total phosphorus (TP) were generally the main factors related to the PCR-DGGE fingerprints of plankton communities. Additionally, environments that exhibit similar trophic level but different concentrations of heavy metals could be differentiated by the PCR-DGGE fingerprinting patterns. During ecological restoration in artificial simulation systems, plankton community succession (except the target cyanobacterial cells) was not significantly affected by the cyanobacteria removal process using chitosan-modified soils. TP was the most important environmental predictor of PCR-DGGE fingerprints of plankton community in the particular artificial niche. Additionally, the present study also explored DNA fingerprinting and sequencing for analyzing intestinal contents of filter-feeding fishes (silver carp and bighead carp), particular functional gene (nifH) and particular planktonic group (cyanobacteria). All these results suggested that metagenome-based fingerprinting analysis was promising tool for ecological investigation of plankton community. In fingerprinting analysis, multiple samples can be analyzed simultaneously with a standardized set of procedures, which making it practical to perform comparative analyses. It is reasonable to believe that metagenome-based study of plankton community will be an exciting direction for ecological study.
语种: 中文
内容类型: 学位论文
URI标识: http://ir.ihb.ac.cn/handle/342005/12382
Appears in Collections:中科院水生所知识产出(2009年前)_学位论文

Files in This Item:
File Name/ File Size Content Type Version Access License
10001_200618011915013颜庆云_paper.pdf(6716KB)----暂不开放-- 联系获取全文

Recommended Citation:
浮游生物群落DNA指纹分析及其在生态学研究中的应用.颜庆云[d].中国科学院水生生物研究所,2009.20-25
Service
Recommend this item
Sava as my favorate item
Show this item's statistics
Export Endnote File
Google Scholar
Similar articles in Google Scholar
[颜庆云]'s Articles
CSDL cross search
Similar articles in CSDL Cross Search
[颜庆云]‘s Articles
Related Copyright Policies
Null
Social Bookmarking
Add to CiteULike Add to Connotea Add to Del.icio.us Add to Digg Add to Reddit
所有评论 (0)
暂无评论
 
评注功能仅针对注册用户开放,请您登录
您对该条目有什么异议,请填写以下表单,管理员会尽快联系您。
内 容:
Email:  *
单位:
验证码:   刷新
您在IR的使用过程中有什么好的想法或者建议可以反馈给我们。
标 题:
 *
内 容:
Email:  *
验证码:   刷新

Items in IR are protected by copyright, with all rights reserved, unless otherwise indicated.

 

 

Valid XHTML 1.0!
Copyright © 2007-2016  中国科学院水生生物研究所 - Feedback
Powered by CSpace