基因工程微生物生态学研究进展

基因工程微生物(genctically enginccred microorganism,GEM)生态学的研究已成为微生物分子生态学的一项主要研究内容之一．随着分子标记和分子生物学检测手段的引入,传统的微生物生态学研究被注入了新的活力,在分子水平上探讨基因工程微生物与环境及环境中土著生物之间的关系已成为可能．基因工程微生物生态学是一门内容涉及分子生物学、微生物学、生态学等诸多学科的新型交叉边缘学科．本文提出加紧进行转基因生物生态学和转基因生物的风险评价的研究工作,建立适合中国国情的检测手段和评价标准,有助于我国基因工程微生物生态学的健康发展．     

环境中基因工程微生物生态学研究

详细阐述了基因工程微生物(GEM)在环境释放中需考虑的主要问题：GEM的构建、基因转移、适合度、扩散、转运和潜在的生态影响等,并针对GEM特定的生态学特征和生态影响。提出了基因工程微生物生态学研究宜采取“具体问题具体分析”的策略。制定相应的研究方案,为安全有效地在非受控条件下应用GEM铺平道路．     

转基因的逃逸及生态风险

转基因技术的发展为提高农作物产量和解决全球人口不断增长而引发的粮食问题带来了无限的机遇,但生物技术的应用和转基因作物的环境释放也带来了一系列生物安全问题．转基因产品是否会对植物、动物、人类健康、遗传资源和环境带来危害已成为公众关注的焦点．诸多生物安全问题中最引人注目的问题之一就是转基因的逃逸及其可能导致的生态风险．文中就转基因逃逸的可能性和逃逸的不同途径、转基因逃逸后可能导致的各种生态风险、转基因逃逸的不同控制方法以及转基因作物安全距离设立应该考虑的因素等问题进行了讨论,旨在了解转基因作物的环境释放和外源基因的逃逸可能导致的生物安全问题,以及如何控制和避免转基因逃逸．     

转基因植物的环境生物安全：转基因逃逸及其潜在生态风险的研究和评价

转基因作物的商品化生产和大规模环境释放在带来巨大利益的同时,也引起了全球对其生物安全问题的广泛关注和争议,其中转基因通过花粉介导的基因漂移逃逸到非转基因作物及其野生近缘种,进而导致的潜在环境和生态风险就是备受争议的生物安全问题之一。转基因植物的环境生物安全涉及两方面关键问题：如何科学评价转基因植物商品化种植以后带来的环境和生态影响;如何利用环境生物安全的研究成果来制定科学有效的风险监测和管理措施。对转基因逃逸及其潜在生态风险的科学评价应包括三个重要环节：（1）检测转基因的逃逸的频率;（2）检测转基因逃逸后的表达和遗传规律;（3）确定逃逸后的转基因对野生近缘种群体适合度的影响及其进化潜力,本文将围绕对转基因逃逸及其潜在环境风险的科学评价,以转基因水稻为案例来对转基因逃逸带来生态影响的研究好评价的进展进行简要介绍,并对目前依据风险评价研究成果制定的各种管理策略进行了讨论。只有提高对转基因生物环境安全研究和评价的水平,并制定有效的风险监测和管理措施,才能为我国转基因技术的发展和转基因产品的商品化应用保驾护航。     

我国转基因水稻商品化应用的潜在环境生物安全问题

转基因水稻的研发和商品化应用将为提高我国水稻的生产力提供新的机遇,并缓解我国的粮食安全问题.转基凶水稻的人规模环境释放和商品化生产可能会带来一定的环境生物安全问题,处理不好会影响转基因水稻的进一步研究和发展.通常所指的环境生物安全问题主要包括以下几个方面:(1)抗生物胁迫转基因对非靶标生物的影响及效应;(2)外源基因向非转基因作物和野生近缘种逃逸及其可能带来的生态后果;(3)转基因作物对农业生态系统、土壤微生物以及生物多样性的潜在影响;(4)抗生物胁迫转基因的长期使用导致靶标生物对转基因产生抗性等.为了安全有效和持续利用转基因生物技术及其产品,有必要对转基因水稻的环境生物安全性进行科学评价.基于风险评价的原则,本文对转基因水稻在我国商品化生产和大规模种植可能带来的环境生物安全问题进行了理性分析,希望为我国转基因水稻商品化应用的决策和生物安全评价提供科学依据.     

转基因植物环境安全评价策略

构建完善的转基因植物环境安全评价技术体系是保障转基因生物产业健康发展的重要组成部分。本文综述了转基因植物环境安全评价技术发展历程与趋势,归纳了转基因植物环境安全评价的思路与内容。转基因植物环境安全评价应分为潜在风险分析、风险假设验证、风险特征描述等3个步骤,并采用逐层评价模式;安全评价应贯穿转基因植物新品种研发与产业化全程,包括应用前预测、研发中筛选、推广前评价、推广后监测。此外,基于科学性和个案分析原则,本文对复合性状、非生物胁迫抗性等新型转基因植物环境安全评价策略进行了探讨。     

杂交-渐渗进化理论在转基因逃逸及其环境风险评价和研究中的意义

转基因作物的商品化生产和大规模环境释放, 引起了全球对生物安全问题的广泛关注和争议, 其中转基因通过花粉介导的基因漂移逃逸到非转基因作物及其野生近缘种, 进而带来不同类型的环境风险就是备受争议的生物安全问题之一。有效的生物安全评价和研究能够为转基因作物的安全持久利用保驾护航。按照风险评价的原则, 对于转基因逃逸及其潜在环境风险的评价应包括两个重要步骤: (1)检测转基因向野生近缘种(包括杂草类型)群体逃逸的频率; (2)确定逃逸后的转基因能否通过遗传渐渗在野生近缘种群体中存留和扩散。杂交－渐渗是进化生物学中非常重要的科学命题和普遍的自然现象, 杂交－渐渗的进化理论与转基因逃逸及其潜在环境风险的研究和评价有密切的关系。杂交－渐渗过程往往导致物种形成、适应性进化和自然群体的濒危与灭绝, 这是因为在杂交－渐渗过程中, 不同的机制如遗传同化作用、群体湮没效应以及群体的选择性剔除效应等都会在很大程度上影响群体的进化过程。转基因通过杂交－渐渗进入野生群体, 使这一过程更加复杂化。如果转基因能提高群体的适合度, 则更有利于其渐渗速率, 从而在群体中迅速扩散并带来一定的生态后果。杂交－渐渗的进化理论和思想将有益于指导转基因逃逸及其潜在环境风险的研究和评价。     

土壤环境中转基因植物重组DNA持留与水平转移研究进展

基因水平转移(horizontal gene transfer,HGT)是转基因植物环境风险评估的重要内容之一。转基因植物重组DNA通过根系分泌、花粉、残体等方式向土壤环境释放。已有研究表明,外源重组DNA很可能被土壤微生物通过同源重组的方式整合到基因组中,直接或间接地造成微生物群落结构和功能的改变,这将造成土壤生态环境系统的改变。本文论述了转基因植物重组DNA在土壤环境中的持留、水平转移及其影响因素和相关检测方法,讨论了转基因植物重组DNA在土壤环境中持留和水平转移的研究重点,并对其研究方法进行比较分析,提出今后的重点研究方向和方法,以期为转基因植物风险评估和安全管理提供技术支撑。     

转基因作物对土壤生态系统的影响

综述了转基因作物对土壤生态系统影响的研究进展,包括转基因作物中的外源基因在土壤中的活性,转基因作物对土壤微生物区系有土壤酶活性的影响以及转基因作物对土壤动物区系的影响,转基因作物对土壤生态系统的影响与导入的外源基因特性和土壤类型相关,转基因产物进入土壤后引起的土壤生物变化的程度依赖于许多因素,最重要的决定因素是生态系统的复杂性和稳定性,评价不同转基因作物对土壤生态系统的影响具有重要的生态学意义,急需发展和完善以分子生物学为主的风险评价方法。     

转基因植物的生态风险评价

自从１９８３年第一株转基因植物诞生以来,至今各种类型的转基因植物进入大田试验的已不计其数,近１０种转基因作物的产物已经商品化。与此同时,转基因植物向环境释放后可能带来的生态风险问题也越来越受到人们的重视。关于转基因植物的生态风险或对环境的危害,科学家提出了不同的概念和测试方法。生态毒理学的经验以及８０年代发展起来的,为作环境决策用的生态风险评价的经验可以借鉴以作转基因植物生态风险的评价。本文介绍了转基因植物对农田生态系统和自然生态系统可能带来的危害以及从基因、基因组、个体、种群以至生态系统等各级水平上危害测试的方法。对风险的判断作了详细的论述,对风险的管理也作了概略的介绍,并对生态风险评价当前发展的水平进行了讨论。     

Use of genetically engineered microorganisms (GEMs) for the bioremediation of contaminants

This paper presents a critical review of the literature on the application of genetically engineered microorganisms (GEMs) in bioremediation. The important aspects of using GEMs in bioremediation, such as development of novel strains with desirable properties through pathway construction and the modification of enzyme specificity and affinity, are discussed in detail. Particular attention is given to the genetic engineering of bacteria using bacterial hemoglobin (VHb) for the treatment of aromatic organic compounds under hypoxic conditions. The application of VHb technology may advance treatment of contaminated sites, where oxygen availability limits the growth of aerobic bioremediating bacteria, as well as the functioning of oxygenases required for mineralization of many organic pollutants. Despite the many advantages of GEMs, there are still concerns that their introduction into polluted sites to enhance bioremediation may have adverse environmental effects, such as gene transfer. The extent of horizontal gene transfer from GEMs in the environment, compared to that of native organisms including benefits regarding bacterial bioremediation that may occur as a result of such transfer, is discussed. Recent advances in tracking methods and containment strategies for GEMs, including several biological systems that have been developed to detect the fate of GEMs in the environment, are also summarized in this review. Critical research questions pertaining to the development and implementation of GEMs for enhanced bioremediation have been identified and posed for possible future research.     

Use of Genetically Engineered Microorganisms (GEMs) for the Bioremediation of Contaminants

ABSTRACT

This paper presents a critical review of the literature on the application of genetically engineered microorganisms (GEMs) in bioremediation. The important aspects of using GEMs in bioremediation, such as development of novel strains with desirable properties through pathway construction and the modification of enzyme specificity and affinity, are discussed in detail. Particular attention is given to the genetic engineering of bacteria using bacterial hemoglobin (VHb) for the treatment of aromatic organic compounds under hypoxic conditions. The application of VHb technology may advance treatment of contaminated sites, where oxygen availability limits the growth of aerobic bioremediating bacteria, as well as the functioning of oxygenases required for mineralization of many organic pollutants. Despite the many advantages of GEMs, there are still concerns that their introduction into polluted sites to enhance bioremediation may have adverse environmental effects, such as gene transfer. The extent of horizontal gene transfer from GEMs in the environment, compared to that of native organisms including benefits regarding bacterial bioremediation that may occur as a result of such transfer, is discussed. Recent advances in tracking methods and containment strategies for GEMs, including several biological systems that have been developed to detect the fate of GEMs in the environment, are also summarized in this review. Critical research questions pertaining to the development and implementation of GEMs for enhanced bioremediation have been identified and posed for possible future research.     

基因工程微生物的环境监测及生物防御体系研究进展

随着生物技术的发展, 研究人员构建出了大量具有特定功能的基因工程微生物, 这些基因工程微生物在实际应用时常受到限制, 因为它们释放到环境中有可能带来新的污染。为了减少或消除其对环境的潜在危害, 有必要采取措施对这些基因工程微生物进行监测和安全控制。通常要求这类基因工程微生物带有便于监测的检测标记以及能进行自消亡的主动生物防御体系。对基因工程微生物的检测标记以及主动生物防御体系的研究现状进行了综述。     

Models to examine containment and spread of genetically engineered microbes

Genetically engineered microbes (GEMs) have the potential to revolutionize agricultural techniques by facilitating crop protection and increased productivity. However, there has been widespread concern regarding the potential impact these microbes may have on the environment. Here we mathematically model the dynamics of GEMs in an agricultural setting, focusing on parameters that can be used to summarize the potential of modified microbes for persistence and spread. First developing a comprehensive model for the dynamics of GEMs which includes mobile and stationary classes of GEMs as well as competition from indigenous microflora, we then analyse a sequence of simplified mathematical models with a view to answering two fundamental questions: (1) will the GEMs spread (or invade), and if so how quickly? and (2) what are the best strategies for containing the spread of GEMs in a spatially varying environment?     

基因工程微生物的环境监测及生物防御体系研究进展

随着生物技术的发展, 研究人员构建出了大量具有特定功能的基因工程微生物, 这些基因工程微生物在实际应用时常受到限制, 因为它们释放到环境中有可能带来新的污染。为了减少或消除其对环境的潜在危害, 有必要采取措施对这些基因工程微生物进行监测和安全控制。通常要求这类基因工程微生物带有便于监测的检测标记以及能进行自消亡的主动生物防御体系。对基因工程微生物的检测标记以及主动生物防御体系的研究现状进行了综述。     

同源重组法构建多功能农药降解基因工程菌研究

构建遗传稳定的多功能农药降解基因工程菌可以为农药污染的生物修复提供良好的菌种资源,然而,构建遗传稳定且不带入外源抗性的基因工程菌是一个难点。通过以受体菌的16S rDNA为同源重组指导序列、sacB基因为双交换正筛选标记构建同源重组载体,二亲结合的方法将甲基对硫磷水解酶基因（mpd）整合到呋喃丹降解菌Sphingomonas sp.CDS1染色体的16S rDNA位点,分别成功构建了含1个和2个mpd基因插入到rDNA位点且不带入外源抗性的基因工程菌株CDSmpd和CDS-2mpd。同源重组单交换的效率为3.7×10^－7～6.8×10^－7。通过PCR和Southern杂交的方法验证了同源重组事件。基因工程菌遗传稳定,能同时降解甲基对硫磷和呋喃丹。甲基对硫磷水解酶（MPH）的比活在各生长时期均高于原始出发菌株,比活最高达6.22 mu/μg。     

Platelet glycoprotein Ib beta/IX mediates glycoprotein Ib alpha localization to membrane lipid domain critical for von Willebrand factor interaction at high shear

The localization of the platelet glycoprotein GP Ib-IX complex (GP Ibα, GP Ibβ, and GP IX) to membrane lipid domain, also known as glycosphingolipid-enriched membranes (GEMs or raft) lipid domain, is essential for the GP Ib-IX complex mediated platelet adhesion to von Willebrand factor (vWf) and subsequent platelet activation. To date, the mechanism for the complex association with the GEMs remains unclear. Although the palmitate modifications of GP Ibβ and GP IX were thought to be critical for the complex presence in the GEMs, we found that the removal of the putative palmitoylation sites of GP Ibβ and GP IX had no effects on the localization of the GP Ib-IX complex to the GEMs. Instead, the disruption of GP Ibα disulfide linkage with GP Ibβ markedly decreased the amount of the GEM-associated GP Ibα without altering the GEM association of GP Ibβ and GP IX. Furthermore, partial dissociation with the GEMs greatly inhibited GP Ibα interaction with vWf at high shear instead of in static condition or under low shear stress. Thus, for the first time, we demonstrated that GP Ibβ/GP IX mediates the disulfide-linked GP Ibα localization to the GEMs, which is critical for vWf interaction at high shear.     

Effects of Genetically Engineered Microorganisms on Nitrogen Transformations and Nitrogen-Transforming Microbial Populations in Soil

The principal concern about releasing genetically engineered microorganisms (GEMs) into the environment is their potential adverse effects on the environment, whether caused directly or indirectly by the GEMs. The effects of five GEMs on ammonification, nitrification, and denitrification in soil were studied. With the possible exception of a strain of Enterobacter cloacae carrying a plasmid, no consistent statistically or ecologically significant differences in effects on these processes or on the population dynamics of the microorganisms responsible for the processes were observed between soils inoculated with the GEMs or their homologous plasmidless hosts and those that were not inoculated. Increasing the concentration of montmorillonite in the soil enhanced the rate of nitrification, regardless of the inoculum, indicating that the perfusion technique used was sensitive enough to detect changes in nitrification rates when they occurred.     

Use of green fluorescent protein to monitor survival of genetically engineered bacteria in aquatic environments.

Many methods for detecting model genetically engineered microorganisms (GEMs) in experimental ecosystems rely on cultivation of introduced cells. In this study, survival of Escherichia coli was monitored with the green fluorescent protein (GFP) gene. This approach allowed enumeration of GEMs by both plating and microscopy. Use of the GFP-marked GEMs revealed that E. coli persisted in stream water at higher densities as determined microscopically than as determined by CFU enumeration. The GFP gene did not negatively impact the fitness of the host strain.