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1.
抗除草剂转基因作物面临的机遇与挑战及其发展策略   总被引：11，自引：0，他引：11  
2008年全球抗除草剂转基因作物已占转基因作物总种植面积的80%以上,抗除草剂成为最主要的转基因应用的性状.该技术的成功应用极大地降低了杂草防除成本、增加了安全性和减少了除草剂的残留药害.但是,抗除草剂转基因作物的大规模产业化将导致除草剂市场单一化,从而冲击除草剂产业,还会产生杂草抗药性和基因逃逸等环境安全问题,因此抗除草剂转基因作物面临着挑战.在我国转基因抗除草剂作物还没有实际的商业化,不过由于市场巨大、政策支持等面临良好发展机遇.此外,从抗除草剂基因利用、转基因抗除草剂作物的研究与开发策略等方面进行综述,以期为我国转基因抗除草剂作物的研究与开发提供可能有益的信息.  相似文献   

2.
为建立耐除草剂转基因作物的高通量检测方法,本试验以目前生产上广泛应用的5种除草剂抗性基因dmo、pat、CP4EPSPS、bar和aad1为靶标进行多重PCR(MPCR)研究。通过引物适用性测试、反应体系中的不同引物浓度和反应程序中的退火温度测试、灵敏度和特异性验证等,建立了能同时检测5种除草剂抗性基因的MPCR检测方法。结果表明,当dmo、pat、CP4EPSPS、bar、aad1基因的检测引物终浓度分别为0.2、0.2、0.3、0.4、0.2μmol·L-1,退火温度为63℃时5种靶标扩增效果较好,且特异性条带清晰且均一。此外,MPCR检测方法具有较好的特异性,对每种靶标的检测灵敏度均可达到0.1%。适用性测试结果显示,MPCR检测方法可对含有5种除草剂抗性基因的多种转基因作物的单个品系或多个品系混合物进行筛选检测。无假阳性和假阴性结果表明,MPCR检测方法对实际样品具有很好的适用性。本试验结果为筛选高效耐除草剂转基因作物检测技术提供了一定的理论依据。  相似文献   

3.
全球抗除草剂转基因作物转化事件分析   总被引：2，自引：0，他引：2  
本文根据国际农业生物技术应用服务组织(International Agricultural Biotechnology Application Service Organization,ISAAA)的相关数据,归纳总结了棉花(Gossypium hirsutum)、大豆(Glycine max)、油菜(Brassica napus)和玉米(Zea mays)这4种作物的抗除草剂转基因转化事件,以便为我国抗除草剂转基因作物的培育提供参考。经统计发现截止2017-05-21,全球抗除草剂的转基因棉花、大豆、油菜和玉米转化事件分别为39、28、32和201个。在这4种作物中,抗除草剂基因有19种,来源于16种生物,涉及到的除草剂共有9种;分别是草甘膦(glyphosate)、草铵膦(glufosinate)、咪唑啉酮类(imidazolinone)、2,4-D(2,4-dichlorophenoxy)、异噁唑草酮(isoxaflutole)、麦草畏(dicamba)、磺酰脲类(sulfonylurea)、硝磺草酮(mesotrione)和溴苯腈(bromoxynil)。单抗事件、多抗事件和复合抗性事件分别为25个、18个和257个,分别占抗除草剂总转化事件的8.3%、6%和85.7%。抗除草剂转化事件涉及的公司有8个,分别是先正达、孟山都公司、杜邦、拜耳作物科学、陶氏益农有限公司、巴斯夫、Genective S.A.和美国斯泰恩种子农场股份有限公司。本文能为我国抗除草剂转基因作物的培育提供重要参考。  相似文献   

4.
快速检测抗除草剂拿捕净基因流的方法初探   总被引：2，自引：0，他引：2  
实验探讨了快速检测抗除草剂拿捕净（Sethoxydim)基因流的方法，该方法核心是用双氧水和赤霉素（GA3）打破青狗尾草种子的休眠，让其在含适宜剂量的除草剂基质上发芽，在暗处生长4d后仅通过测量种子发芽后的幼芽生长、形态差异即鉴定出抗、感除草剂种子，从而达到快速检测的目的。  相似文献   

5.
具有多选择标记的植物基因表达载体有利于转基因植物研究中的转基因植株的筛选。本研究对植物基因表达载体pCAMBIA1301进行了改造,产生了1个具有可溶性的红移绿色荧光蛋白基因（smRS-GFP）、抗除草剂Basta、葡萄糖苷酸酶（GUS）及潮霉素（Hpt）的多选择标记的新的植物基因表达载体。运用这一表达载体的多选择标记可以有效降低检测和筛选转化植株时的假阳性率。此外,如此的基因表达载体也能满足实验室的不同筛选方法的需求。  相似文献   

6.
抗除草剂转基因水稻的安全性评价   总被引：11，自引：0，他引：11  
以转基因抗除草剂水稻（Oryza sativa ssp.japonica）为例,对转基因作物进行安全性评价.对转bar基因水稻进行了基因流动实验、小鼠急性毒性实验、致突变实验和Ames实验.结果表明,转bar基因水稻花粉在5 m内有少量的漂移到普通野生稻（O.rufipogon）,但没有向其它野生稻（O.officinalis和O.meyeriana）和杂草漂移.小鼠经口急性毒性半数致死量LD50＞20 g/kg,属实际无急性毒性物质;经微核实验、精子畸变实验及Ames实验均未发现有致突变作用,初步表明转bar基因水稻对小鼠是安全的.  相似文献   

7.
我国抗草甘膦基因的发掘现状   总被引：1，自引：0，他引：1  
草甘膦是世界上用途最广的除草剂之一。抗除草剂特性是目前转基因作物中最主要的农艺性状。本文简要陈述了草甘膦及其抗性基因的作用机理,并对我国抗草甘膦基因的发掘现状及抗草甘膦专利的授予情况予以介绍。截止到2012年底国内单位及个人已有15项抗草甘膦的专利申请获得中国专利授权,另有22项申请正在审查中。可以看到自2007年以后,国内单位在发掘新的抗草甘膦基因并在获得自主知识产权方面已经有了长足的进步。通过在转基因植物中对草甘膦抗性的验证,其中部分抗性基因已经具有潜在的商业应用价值。  相似文献   

8.
直接产生抗除草剂转基因水稻纯系的新方法   总被引：3，自引：0，他引：3  
提出了一种提高转基因杂交后代育种选择效率的新途径，即在花药培养过程中添加抗性筛选物质，直接筛选含有目的基因的纯合体。以转bar基因水稻植株和非转基因植株杂交F1为试格进行花药培养，并对分化的绿苗进行PCR检测和田间抗性检测。结果表明，在愈伤组织诱导培养基和分化培养基中分别添加PPT0.05mg/L与未添加PPT的对照，均产生较多的愈伤组织和绿苗，并有较高比例不抗除草剂的假阳性株。当PPT浓度增至0.1mg/L时，所得花培苗全部抗除草剂，显示很好的筛选作用。PPT浓度继续增加，愈伤组织诱导率和绿苗分化率急剧下降。当PPT的浓度增至5mg/L时，愈伤组织的诱导和幼苗分化完全被抑制。对30个加倍单倍体系作连续两代农艺性状考察，约73％以上的株系在遗传率较高的性状中，变异系数很小（＜10％），说明这些株系的基因型已经纯合，证实了该方法的有效性。  相似文献   

9.
美国转基因作物田间试验频次分析   总被引：1，自引：0，他引：1  
本研究通过对美国转基因作物田间试验频次分析揭示转基因作物研发的概貌和规律.采用EXCEL"数据透视表与数据透视图"软件,对美国转基因作物田间试验数据库和世界经济合作组织转基因作物田间试验数据库近20年数据进行统计分析.结果表明,至2008年9月止,美国转基因作物田间试验累计达13782次,涉及158种作物,10大类性状和98家公司或研究机构.1987-2008年间,审批的田间试验次数先呈直线上升,其后稳定在每年1000次左右.试验物种以玉米、大豆、棉花为主,分别占45.8%、9.7%和6.4%.涉及的主要性状有抗除草剂、抗虫和品质性状.62.7%的转基因作物田间试验由孟山都和国际先锋等五家私人公司完成.分析认为,只有经济价值高、安全性好、技术成熟、消费者容易接受的转基因作物才拥有更广阔的发展前景.建议我国加强非粮食作物转基因技术的研究.  相似文献   

10.
我国在转基因水稻研究方面处于国际领先地位,但由于研究材料扩散到商品化生产水稻中而引起的贸易纠纷时有发生。加强转基因水稻检测与监管,从源头控制转基因水稻基因扩散对于保障生物安全、促进米制品贸易有重要意义。本研究针对我国当前转基因水稻监管重点,结合已有相关检测方法标准,针对三种不同转基因水稻：抗虫水稻TT51-1、抗病水稻M12、抗除草剂水稻LLRICE62,以水稻内源基因RBE4为参照基因,建立转基因水稻四重复合PCR检测方法,可在同一反应体系中同时检测鉴定三种转基因水稻品系,检测限可达250个拷贝。利用此方法可快速地定性检测和鉴定转基因水稻TT51、M12、LL RICE62。  相似文献   

11.
Glyphosate-tolerant, Roundup Ready (RR) soybeans account for about 57% of all genetically modified (GM) crops grown worldwide. The entry of recombinant DNA into soil from GM crops has been identified as an environmental concern due to the possibility of their horizontal transfer to soil microorganisms. RR soybeans contain recombinant gene sequences that can be differentiated from wild-type plant and microbial genes in soil by using a sequence-specific molecular beacon and real-time polymerase chain reaction (PCR). A molecular beacon-based real-time PCR system to quantify a wild-type soybean lectin ( le1) gene was designed to compare amounts of endogenous soybean genes to recombinant DNA in soil. Microcosm studies were carried out to develop methodologies for the detection of recombinant DNA from RR soybeans in soil. RR soybean leaf litterbags were imbedded in the soil under controlled environmental conditions (60% water holding capacity, 10/15 degrees C, and 8/16 h day/night) for 30 days. The soybean biomass decomposition was described using a single-phase exponential equation, and the DNA concentration in planta and in soil was quantified using real-time PCR using sequence-specific molecular beacons for the recombinant cp4 epsps and endogenous soybean lectin ( le1) genes. The biomass of RR soybean leaves was 8.6% less than nontransgenic (NT) soybean leaves after 30 days. The pooled half-disappearance time for cp4 epsps and le1 in RR and of le1 in NT soybean leaves was 1.4 days. All genes from leaves were detected in soil after 30 days. This study provides a methodology for monitoring the entry of RR and NT soybean DNA into soil from decomposing plant residues.  相似文献   

12.
The presence of DNA in foodstuffs derived from or containing genetically modified organisms (GMO) is the basic requirement for labeling of GMO foods in Council Directive 2001/18/CE (Off. J. Eur. Communities 2001, L1 06/2). In this work, four different methods for DNA extraction were evaluated and compared. To rank the different methods, the quality and quantity of DNA extracted from standards, containing known percentages of GMO material and from different food products, were considered. The food products analyzed derived from both soybean and maize and were chosen on the basis of the mechanical, technological, and chemical treatment they had been subjected to during processing. Degree of DNA degradation at various stages of food production was evaluated through the amplification of different DNA fragments belonging to the endogenous genes of both maize and soybean. Genomic DNA was extracted from Roundup Ready soybean and maize MON810 standard flours, according to four different methods, and quantified by real-time Polymerase Chain Reaction (PCR), with the aim of determining the influence of the extraction methods on the DNA quantification through real-time PCR.  相似文献   

13.
Milling fractions from conventional and transgenic corn were prepared at laboratory scale and used to study the influence of sample composition and heat-induced DNA degradation on the relative quantification of genetically modified organisms (GMO) in food products. Particle size distributions of the obtained fractions (coarse grits, regular grits, meal, and flour) were characterized using a laser diffraction system. The application of two DNA isolation protocols revealed a strong correlation between the degree of comminution of the milling fractions and the DNA yield in the extracts. Mixtures of milling fractions from conventional and transgenic material (1%) were prepared and analyzed via real-time polymerase chain reaction. Accurate quantification of the adjusted GMO content was only possible in mixtures containing conventional and transgenic material in the form of analogous milling fractions, whereas mixtures of fractions exhibiting different particle size distributions delivered significantly over- and underestimated GMO contents depending on their compositions. The process of heat-induced nucleic acid degradation was followed by applying two established quantitative assays showing differences between the lengths of the recombinant and reference target sequences (A, deltal(A) = -25 bp; B, deltal(B) = +16 bp; values related to the amplicon length of the reference gene). Data obtained by the application of method A resulted in underestimated recoveries of GMO contents in the samples of heat-treated products, reflecting the favored degradation of the longer target sequence used for the detection of the transgene. In contrast, data yielded by the application of method B resulted in increasingly overestimated recoveries of GMO contents. The results show how commonly used food technological processes may lead to distortions in the results of quantitative GMO analyses.  相似文献   

14.
The presence of genetically modified organisms (GMOs) in food and feed products is subject to regulation in the European Union (EU) and elsewhere. As part of the EU authorization procedure for GMOs intended for food and feed use, reference materials must be produced for the quality control of measurements to quantify the GMOs. Certified reference materials (CRMs) are available for a range of herbicide- and insect-resistant genetically modified crops such as corn, soybean, and cotton. Here the development of the first CRM for a GMO that differs from its non-GMO counterpart in a major compositional constituent, that is, starch, is described. It is shown that the modification of the starch composition of potato (Solanum tuberosum L.) tubers, together with other characteristics of the delivered materials, have important consequences for the certification strategy. Moreover, the processing and characterization of the EH92-527-1 potato material required both new and modified procedures, different from those used routinely for CRMs produced from genetically modified seeds.  相似文献   

15.
The labeling of products containing genetically modified organisms (GMO) is linked to their quantification since a threshold for the presence of fortuitous GMOs in food has been established. This threshold is calculated from a combination of two absolute quantification values: one for the specific GMO target and the second for an endogenous reference gene specific to the taxon. Thus, the development of reliable methods to quantify GMOs using endogenous reference genes in complex matrixes such as food and feed is needed. Plant identification can be difficult in the case of closely related taxa, which moreover are subject to introgression events. Based on the homology of beta-fructosidase sequences obtained from public databases, two couples of consensus primers were designed for the detection, quantification, and differentiation of four Solanaceae: potato (Solanum tuberosum), tomato (Solanum lycopersicum), pepper (Capsicum annuum), and eggplant (Solanum melongena). Sequence variability was studied first using lines and cultivars (intraspecies sequence variability), then using taxa involved in gene introgressions, and finally, using taxonomically close taxa (interspecies sequence variability). This study allowed us to design four highly specific TaqMan-MGB probes. A duplex real time PCR assay was developed for simultaneous quantification of tomato and potato. For eggplant and pepper, only simplex real time PCR tests were developed. The results demonstrated the high specificity and sensitivity of the assays. We therefore conclude that beta-fructosidase can be used as an endogenous reference gene for GMO analysis.  相似文献   

16.
In many countries, including the European Union member states, Switzerland, Australia, New Zealand, and Japan, legislation has been set up for labeling of genetically modified organisms (GMOs) in food and feed products. To comply with these regulations, reliable detection methods are necessary. If the detection is based on DNA, a GMO analysis may contain several steps where qualitative and quantitative species-specific, GMO screening, GMO construct, and GMO line-specific polymerase chain reactions (PCRs) are used. A limit of detection (LOD) thereby defines to what extent a target molecule may be detected in a sample. In this study, cookies were made with variable levels of a soy sample containing 2 wt% Roundup Ready soy. For all PCRs described, detection limits based on dilution series and practical LODs were determined. The practical LODs are used to determine to what extent a GMO ingredient may be detected in a real food product. Results reveal that, due to the baking process, the overall DNA fragment length is reduced, rendering GMO analyses more difficult. Furthermore, Roundup Ready soy line-specific and real-time quantitative PCR are less sensitive than GMO screening PCRs, whereas just these PCRs are crucial in the decision-making process regarding the presence of GMOs in a food product. Moreover, high standard deviations and errors render the precise quantification of GMOs difficult.  相似文献   

17.
The genetic modification in fruit and vegetables could lead to changes in metabolic pathways and, therefore, to the variation of the molecular pattern, with particular attention to antioxidant compounds not well-described in the literature. The aim of the present study was to compare the quality composition of transgenic wheat ( Triticum durum L.), corn ( Zea mays L.), and tomato ( Lycopersicum esculentum Mill.) to the nontransgenic control with a similar genetic background. In the first experiment, Ofanto wheat cultivar containing the tobacco rab1 gene and nontransgenic Ofanto were used. The second experiment compared two transgenic lines of corn containing Bacillus thuringiensis "Cry toxin" gene (PR33P67 and Pegaso Bt) to their nontransgenic forms. The third experiment was conducted on transgenic tomato ( Lycopersicum esculentum Mill.) containing the Agrobacterium rhizogenes rolD gene and its nontransgenic control (cv. Tondino). Conventional and genetically modified crops were compared in terms of fatty acids content, unsaponifiable fraction of antioxidants, total phenols, polyphenols, carotenoids, vitamin C, total antioxidant activity, and mineral composition. No significant differences were observed for qualitative traits analyzed in wheat and corn samples. In tomato samples, the total antioxidant activity (TAA), measured by FRAP assay, and the naringenin content showed a lower value in genetically modified organism (GMO) samples (0.35 mmol of Fe (2+) 100 g (-1) and 2.82 mg 100 g (-1), respectively), in comparison to its nontransgenic control (0.41 mmol of Fe (2+) 100 g (-1) and 4.17 mg 100 g (-1), respectively). On the basis of the principle of substantial equivalence, as articulated by the World Health Organization, the Organization for Economic Cooperation and Development, and the United Nations Food and Agriculture Organization, these data support the conclusion that GM events are nutritionally similar to conventional varieties of wheat, corn, and tomato on the market today.  相似文献   

18.
Plants derived through agricultural biotechnology, or genetically modified organisms (GMOs), may affect human health and ecological environment. A living GMO is also called a living modified organism (LMO). Biotech cotton is a GMO in food or feed and also an LMO in the environment. Recently, two varieties of biotech cotton, MON 15985 and MON 88913, were developed by Monsanto Co. The detection method is an essential element for the GMO labeling system or LMO management of biotech plants. In this paper, two primer pairs and probes were designed for specific amplification of 116 and 120 bp PCR products from MON 15985 and MON 88913, respectively, with no amplification from any other biotech cotton. Limits of detection of the qualitative method were all 0.05% for MON 15985 and MON 88913. The quantitative method was developed using a TaqMan real-time PCR. A synthetic plasmid, as a reference molecule, was constructed from a taxon-specific DNA sequence of cotton and two construct-specific DNA sequences of MON 15985 and MON 88913. The quantitative method was validated using six samples that contained levels of biotech cotton mixed with conventional cotton ranging from 0.1 to 10.0%. As a result, the biases from the true value and the relative deviations were all within the range of +/-20%. Limits of quantitation of the quantitative method were all 0.1%. Consequently, it is reported that the proposed detection methods were applicable for qualitative and quantitative analyses for biotech cotton MON 15985 and MON 88913.  相似文献   

19.
With the increasing development of genetically modified organism (GMO) detection techniques, the polymerase chain reaction (PCR) technique has been the mainstay for GMO detection. An oligonucleotide microarray is a glass chip to the surface of which an array of oligonucleotides was fixed as spots, each containing numerous copies of a sequence-specific probe that is complementary to a gene of interest. So it is used to detect ten or more targets synchronously. In this research, an event-specific detection strategy based on the unique and specific integration junction sequences between the host plant genome DNA and the integrated gene is being developed for its high specificity using multiplex-PCR together with oligonucleotide microarray. A commercial GM soybean (GTS 40-3-2) and six GM maize events (MON810, MON863, Bt176, Bt11, GA21, and T25) were detected by this method. The results indicate that it is a suitable method for the identification of these GM soybean and maizes.  相似文献   

20.
In many countries, the labeling of grains and feed- and foodstuffs is mandatory if the genetically modified organism (GMO) content exceeds a certain level of approved GM varieties. The GMO content in a maize sample containing the combined-trait (stacked) GM maize as determined by the currently available methodology is likely to be overestimated. However, there has been little information in the literature on the mixing level and varieties of stacked GM maize in real sample grains. For the first time, the GMO content of non-identity-preserved (non-IP) maize samples imported from the United States has been successfully determined by using a previously developed individual kernel detection system coupled to a multiplex qualitative PCR method followed by multichannel capillary gel electrophoresis system analysis. To clarify the GMO content in the maize samples imported from the United States, determine how many stacked GM traits are contained therein, and which GM trait varieties frequently appeared in 2005, the GMO content (percent) on a kernel basis and the varieties of the GM kernels in the non-IP maize samples imported from the United States were investigated using the individual kernel analysis system. The average (+/-standard deviation) of the GMO contents on a kernel basis in five non-IP sample lots was determined to be 51.0+/-21.6%, the percentage of a single GM trait grains was 39%, and the percentage of the stacked GM trait grains was 12%. The MON810 grains and NK603 grains were the most frequent varieties in the single GM traits. The most frequent stacked GM traits were the MON810xNK603 grains. In addition, the present study would provide the answer and impact for the quantification of GM maize content in the GM maize kernels on labeling regulation.  相似文献   

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