Delaying evolution of insect resistance to transgenic crops by decreasing dominance and heritability

The refuge strategy is used widely for delaying evolution of insect resistance to transgenic crops that produce Bacillus thuringiensis (Bt) toxins. Farmers grow refuges of host plants that do not produce Bt toxins to promote survival of susceptible pests. Many modelling studies predict that refuges will delay resistance longest if alleles conferring resistance are rare, most resistant adults mate with susceptible adults, and Bt plants have sufficiently high toxin concentration to kill heterozygous progeny from such matings. In contrast, based on their model of the cotton pest Heliothis virescens, Vacher et al. (Journal of Evolutionary Biology, 16, 2003, 378) concluded that low rather than high toxin doses would delay resistance most effectively. We demonstrate here that their conclusion arises from invalid assumptions about larval concentration-mortality responses and dominance of resistance. Incorporation of bioassay data from H. virescens and another key cotton pest (Pectinophora gossypiella) into a population genetic model shows that toxin concentrations high enough to kill all or nearly all heterozygotes should delay resistance longer than lower concentrations.     

Advances in development of transgenic pulse crops

It is three decades since the first transgenic pulse crop has been developed. Todate, genetic transformation has been reported in all the major pulse crops like Vigna species, Cicer arietinum, Cajanus cajan, Phaseolus spp, Lupinus spp, Vicia spp and Pisum sativum, but transgenic pulse crops have not yet been commercially released. Despite the crucial role played by pulse crops in tropical agriculture, transgenic pulse crops have not moved out from laboratories to large farm lands compared to their counterparts - 'cereals' and the closely related leguminous oil crop - 'soybean'. The reason for lack of commercialization of transgenic pulse crops can be attributed to the difficulty in developing transgenics with reproducibility, which in turn is due to lack of competent totipotent cells for transformation, long periods required for developing transgenics and lack of coordinated research efforts by the scientific community and long term funding. With optimization of various factors which influence genetic transformation of pulse crops, it will be possible to develop transgenic plants in this important group of crop species with more precision and reproducibility. A translation of knowledge from information available in genomics and functional genomics in model legumes like Medicago truncatula and Lotus japonicus relating to factors which contribute to enhancing crop yield and ameliorate the negative consequences of biotic and abiotic stress factors may provide novel insights for genetic manipulation to improve the productivity of pulse crops.     

生命伦理学视野下的转基因作物产业化问题研究

转基因作物的产业化作为当前生命伦理学的重要问题之一日益受到我国学术界、政府和广大公众的关注。转基因作物产业化不仅关涉到我国13亿人的吃饭问题,也与我国公众的身心健康、基本权利密切相关。转基因作物产业化的生命伦理意蕴主要体现在：转基因作物产业化的基础是确保公众健康与生命安全,关键是尊重公众权利,核心是促进社会公正。     

转基因作物与非转基因作物的共存立法动态研究——以美、日、欧应对基因污染事件为视角

随着基因工程技术的运用与发展,越来越多的转基因作物基因污染事件的发生也引起了世界各国对转基因作物与非转基因作物之间的和谐发展问题的高度重视。以美、日、欧为代表,世界各国对转基因作物与非转基因作物的共存问题都进行了积极地探索。借鉴美、日、欧转基因与非转基因作物共存法律制度,以期为我国转基因与非转基因作物共存法律制度的构建提供参考。     

转基因抗虫作物对捕食性瓢虫的安全性研究进展

转基因抗虫作物的商业化种植带来了显著的经济、生态和社会效益,对转基因抗虫作物的安全性评价也一直是国内外学者研究的热点。对生物非靶标效应的研究是转基因抗虫作物安全性评价的重要组成部分,农田天敌类生物是其中的重点内容之一。瓢虫是农田生态系统重要的捕食性天敌,评价其在转基因抗虫作物田间是否能通过捕食猎物或取食花粉而接触到杀虫蛋白并富集于体内,进而对自身产生一定的非靶标效应,这对捕食性瓢虫的安全性研究具有重要的意义。本文从捕食性天敌瓢虫的生命表参数、行为功能参数、田间群落参数及体内微环境指标等方面综述了转基因抗虫作物对瓢虫的安全性研究进展,并对后期转基因抗虫作物对田间捕食性天敌的研究方向提出了建议,以期为转基因抗虫作物的环境安全性研究提供理论指导和为进一步完善转基因抗虫作物对瓢虫安全性评价的技术体系提供系统的数据资料。     

转基因抗虫作物对非靶标昆虫的影响

转基因抗虫作物自 1996年被批准商业化种植以来 ,它的抗虫性和经济效益已得到了普遍肯定 ,同时 ,转基因抗虫作物对非靶标生物的影响 ,如转基因抗虫作物的长期种植 ,是否会导致次要害虫上升为主要害虫 ,是否会影响有益昆虫 ,包括重要经济昆虫、捕食性和寄生性天敌以及重要蝶类的种类及种群数量 ,已成为转基因抗虫作物生态风险评估的重要内容。一些研究结果表明 ,转基因抗虫作物在对靶标害虫有效控制的同时 ,一些对杀虫蛋白不敏感的非靶标害虫有加重危害的趋势 ,由于种植转基因抗虫作物 ,减少了化学农药的使用 ,客观上也使非靶标害虫种群数量上升 ,这对转基因抗虫作物害虫综合治理提出了新的要求。靶标害虫数量的减少直接影响了害虫天敌种群数量 ,靶标害虫取食转基因抗虫作物后发育迟缓 ,也间接影响了天敌昆虫的生长发育 ,转基因抗虫作物的花粉或花蜜是一些重要经济昆虫如蜜蜂、熊蜂和一些寄生蜂 ,甚至捕食性天敌的食物来源 ,或花粉飘落到一些鳞翅目昆虫如家蚕或重要蝶类昆虫的寄主植物上 ,直接或间接对这些昆虫造成一定影响。目前大多数研究表明转基因抗虫作物对非靶标昆虫 ,特别是对有益昆虫没有明显的不利影响 ,也有研究报道认为对某些有益昆虫有一定的不良影响。这为深入开展转基因抗虫作物的生态安全     

Field performance and seasonal changes in the efficacy against Helicoverpa armigera (Hübner) of transgenic cotton expressing the insecticidal protein vip3A

1 Three years of field experiments in Eastern Australia were carried out on transgenic cotton (Gossypium hirsutum L.) event Cot102 expressing the insecticidal protein gene vip3A from Bacillus thuringiensis to evaluate performance against Helicoverpa armigera Hübner. Efficacy, defined as the capacity of plant tissues to induce larval mortality, was determined with a well‐validated leaf bioassay fortnightly through the growth cycle of the cotton in each season. 2 Cot102 plants proved highly efficacious against H. armigera, particularly early in the season, although their efficacy declined as the season progressed, in a manner similar to, but not as dramatic as, that observed with commercial Cry1Ac expressing cotton (Bollgard or Ingard cotton). 3 Field surveys indicated that very few larvae survived beyond first instar on intact growing plants. 4 In one season efficacy declined for a period of approximately 20 days after a cool wet period, suggesting that this may have had a detrimental effect on the expression or efficacy of the gene, but this will need to be verified in further replicated trials. 5 Quantitative enzyme‐linked immunosorbent assays indicated that there was no dramatic reduction in production of the vip3A protein during growth and maturation of the crop, suggesting that other host plant factors were affecting the efficacy of the insecticidal protein in the insect gut. 6 These data indicate that Cot102 cotton would provide a useful alternative to Bollgard cotton but, given the similar lytic mode of action of vip3A proteins in the insect midgut, there may be similar inherent vulnerabilities to resistance evolution for these proteins if used alone. Pyramiding of the vip3A trait with a second insecticidal gene would appear to be a high priority for achieving sustainable deployment against H. armigera or similar susceptible species.     

转基因作物中2种除草剂抗性基因aad1和dmo的PCR检测方法

【背景】抗除草剂转基因作物是全球种植面积最大的一类转基因植物,以除草剂抗性基因作为检测靶标的分子鉴定方法的研究与应用,对转基因生物安全的检测与监测有重要意义。【方法】根据除草剂抗性基因aad1和dmo的核苷酸序列设计PCR检测引物,并进行PCR反应体系优化、方法特异性、灵敏度、再现性等方面的测试,分别建立aad1基因和dmo基因的特异性PCR检测方法。【结果】建立的PCR检测方法在56~64℃的退火温度范围内均能获得一致性结果,具有良好的稳健性。该方法可将含有aad1基因和dmo基因的转基因作物与其他转基因作物区分开,其灵敏度可分别达到20个拷贝和40个拷贝。通过将aad1基因和dmo基因的检测引物放入同一管PCR反应体系中,还能在一次PCR中同时检测这2个靶标基因,双重PCR的检测灵敏度与单一PCR一致。【结论与意义】建立的分子方法可精准检测出含有aad1基因和dmo基因的转基因作物,具有特异性强、灵敏度高的特点,为抗除草剂转基因作物的筛选检测提供了可靠的技术支撑。     

Stable expression of Gal/GalNAc lectin of Entamoeba histolytica in transgenic chloroplasts and immunogenicity in mice towards vaccine development for amoebiasis

Chloroplast genetic engineering offers several advantages, including high levels of transgene expression, transgene containment via maternal inheritance and multigene engineering in a single transformation event. Entamoeba histolytica infects 50 million people, causing about 100 000 deaths annually, but there is no approved vaccine against this pathogen. LecA , a potential target for blocking amoebiasis, was expressed for the first time in transgenic plants. Stable transgene integration into chloroplast genomes and homoplasmy were confirmed by polymerase chain reaction and Southern blot analyses. LecA expression was evaluated by Western blots and quantified by enzyme-linked immunosorbent assay (up to 6.3% of total soluble protein or 2.3 mg LecA/g leaf tissue). Subcutaneous immunization of mice with crude extract of transgenic leaves resulted in higher immunoglobulin G titres (up to 1 : 10 000) than in previous reports. An average yield of 24 mg of LecA per plant should produce 29 million doses of vaccine antigen per acre of transgenic plants. Such high levels of expression and immunogenicity should facilitate the development of a less expensive amoebiasis vaccine.     

ToMV-Resistant transgenic tomato as a material for the first field experiment of genetically engineered plants in Japan

Summary An F₁ hybrid betweenLycopersicon esculentum andL. peruvianum was transformed using a Ti-plasmid binary vector with a coat protein gene cDNA of an attenuated tomato mosaic virus (ToMV) strain L₁₁A which was expressible by the 35S promoter of cauliflower mosaic virus (CaMV). A transgenic plant which expressed the most resistance to ToMV was chosen as a material to be tested in a nonisolated greenhouse and in the field. This transgenic tomato plant was propagated by cutting. In the first test using an isolated greenhouse and in the second test conducted in an nonisolated greenhouse, no major morphologic and physiologic differences were found between the transgenic plants and the nontransgenic control plants. Also, there was no evidence that the transgenic plants produced any new hazardous substances. Both the transgenic and the nontransgenic plants were self-sterile, and crossing of the cultivated species with pollen of these plants produced few seeds. These features of the transgenic plants satisfied the requirements for a small scale field test. The field test of the transgenic plants are in progress. Presented in the Session-in-Depth “Field Test Requirements and Performance of Transgenic Plants” at the 1991 World Congress on Cell and Tissue Culture, Anaheim, California, June 16–20, 1991.     

Stable transformation of the cotton plastid genome and maternal inheritance of transgenes

Chloroplast genetic engineering overcomes concerns of gene containment, low levels of transgene expression, gene silencing, positional and pleiotropic effects or presence of vector sequences in transformed genomes. Several therapeutic proteins and agronomic traits have been highly expressed via the tobacco chloroplast genome but extending this concept to important crops has been a major challenge; lack of 100 homologous species-specific chloroplast transformation vectors containing suitable selectable markers, ability to regulate transgene expression in developing plastids and inadequate tissue culture systems via somatic embryogenesis are major challenges. We employed a Double Gene/Single Selection (DGSS) plastid transformation vector that harbors two selectable marker genes (aphA-6 and nptII) to detoxify the same antibiotic by two enzymes, irrespective of the type of tissues or plastids; by combining this with an efficient regeneration system via somatic embryogenesis, cotton plastid transformation was achieved for the first time. The DGSS transformation vector is at least 8-fold (1 event/2.4 bombarded plates) more efficient than Single Gene/Single Selection (SGSS) vector (aphA-6; 1 event per 20 bombarded plates). Chloroplast transgenic lines were fertile, flowered and set seeds similar to untransformed plants. Transgenes stably integrated into the cotton chloroplast genome were maternally inherited and were not transmitted via pollen when out-crossed with untransformed female plants. Cotton is one of the most important genetically modified crops ($ 120 billion US annual economy). Successful transformation of the chloroplast genome should address concerns about transgene escape, insects developing resistance, inadequate insect control and promote public acceptance of genetically modified cotton.     

PCR detection of genetically modified soya and maize in foodstuffs

The detection of genetically modified foodstuffs is becoming both a food sales and legal necessity. This study reports a rapid DNA extraction/PCR-based method for the detection of genetically modified soya (GMS) and maize (GMM) in mixed samples of transgenic and unmodified soybeans and maize kernels, and a variety of processed samples including soya flour, soya protein isolates, extruded defatted soya, acid- and alcohol-precipitated soya concentrates, soya lecithin, maize grits, seasoned corn puffs and salted corn chips. The presence of GMS DNA was determined with two pairs of primers directed towards different GMS target sequences and GMM by one primer pair. In addition, a multiplex PCR reaction which utilises an internal positive control was developed for both genetically modified organisms (GMOs). Results indicated that the methods are sensitive and specific enough to detect GMS down to a level of 0.01% dry weight in single-product PCRs and 0.1% in multiplex PCRs and GMM down to 0.001% dry weight in single-product PCRs and 0.01% in multiplex PCR. The methods are considered to represent a viable route for the commercial detection of GMS and GMM in foodstuffs.     

转Bt基因抗虫作物对鳞翅目非靶标昆虫生态影响的研究进展

任何转基因作物在进入商业化应用之前都必须经过严格的环境风险评价。评价转基因作物特别是抗虫作物对农田重要非靶标节肢动物的生态影响是其中一项重要内容。当前,全球种植的转基因抗虫作物大多表达对鳞翅目害虫具有活性的Cry1或Cry2类杀虫蛋白。由于非靶标鳞翅目昆虫如斑蝶、家蚕等与靶标害虫具有较近的亲缘关系,其幼虫可能同样对这类杀虫蛋白敏感。因此,这类转基因抗虫作物对非靶标鳞翅目害虫的潜在影响引起了研究者的广泛关注。在总结国内外相关研究数据的基础上,系统分析了转基因抗虫作物对非靶标蝶类和蚕类昆虫的潜在影响,获得以下结论：虽然蚕类和蝶类昆虫对Cry1或Cry2类杀虫蛋白敏感,但在自然条件下这类非靶标昆虫暴露于Cry杀虫蛋白的水平很低,抗鳞翅目害虫转基因作物的种植不可能显著影响田间蝶类昆虫的种群密度,也不会给我国的蚕丝产业带来负面影响。     

Long-term resistance to tomato spotted wilt virus in transgenic tobacco cultivars expressing the viral nucleoprotein gene: greenhouse and field tests

We examined the resistance phenotype of a large number of transgenic tobacco plants originating from 12 commercial (Nicotiana tabacum) cultivars expressing the sense form of the nucleoprotein (N) gene of L3, a Bulgarian isolate of tomato spotted wilt virus (TSWV). The analysis revealed that transgenic plants are completely protected against the homologous L3 isolate of TSWV irrespective of whether or not they contain detectable levels of translational product. The effectiveness of protection against the virus was investigated upon mechanical inoculation under greenhouse conditions and in field trials. Non-segregating resistant lines were selected and the inheritance of the resistance to TSWV was analysed in successive generations (R₃–R₆). Extensive tests under controlled conditions and two-year field trials proved that the resistance to TSWV is stable in different environments and is a stably inherited trait.     

我国转基因植物研发形势及发展战略

当前,发达国家及跨国种业集团在功能基因组学、转基因技术和转基因产品研发方面进展显著,转基因产业发展势头强劲,已成为近年来持续保持高速增长的新兴产业。我国高度重视转基因植物研发及产业化,整体水平领先于发展中国家,但与国际转基因生物产业快速发展和我国衣业发展对转基因产品的需求相比,在转基因技术和产品创新、产业化机制以及支撑条件等方面尚存在较多制约因素。基于系统比较分析,建议我国进一步加强转基因植物研发能力建设,夯实转基因育种基础,突破转基因核心技术,培育转基因植物新品种,加强产学研紧密结合,培育具有自主创新能力和市场竞争力的大型企业,与此同时加强科普宣传,营造良好的社会氛围,推进我国生物育种战略性新兴产业的快速发展。     

Relevance of genetically modified crops in light of future environmental and legislative challenges to the agri-environment

A key challenge for countries like Ireland up to 2030 is to produce sufficient supplies of food, feed and fuel, without compromising on public health or negatively impacting the environment. As we progress through the technology era, certain agricultural technologies [e.g. genetically modified (GM) crops] have been championed to maximise production while minimising environmental impact. Yet, multiple arguments have been made to counter such a claim, which has led to a polarisation of opinions and a plethora of generic commentaries being made in regard to the impact of this technology. Yet, few studies within the European Union (EU) have conducted a critical needs analysis to assess the potential of specific GM traits in light of issues, such as climate change, increased environmental legislation (e.g. EU Water Framework, Nitrates Directive, proposed reform to the Pesticide Directive and Common Agricultural Policy reform), mitigating biodiversity loss and sustainable biofuel production. The goal of this study is to collate a register of GM traits such that a list of potential GM crops could be prioritised against the backdrop of the challenges facing the tillage sector. Clearly, the crops with the most significant potential for genetic modification are those that are grown widely and/or receive high applications of pesticides and fertilisers (e.g. potato, wheat, barley and maize). GM traits with significant agronomic potential include late blight resistant potato, Fusarium head blight resistant wheat and Septoria resistant wheat and herbicide‐tolerant winter oilseed rape and maize. Following on from these, crops with enhanced nitrogen‐use efficiency could provide significant input to the tillage sector in light of EU‐based restrictions on nitrogen usage, crops with elevated protein content could offset the costs of imported animal feed and crops with modified oil content/lignocellulose composition could assist in biodiesel/bioenergy production at a regional level. This study is relevant to other European countries that cultivate similar crops and like Ireland, are facing multiple challenges to their tillage sector in the near future.     

Assessment of Real-time PCR Based Methods for Quantification of Pollen-mediated Gene Flow from GM to Conventional Maize in a Field Study

Maize is one of the main crops worldwide and an increasing number of genetically modified (GM) maize varieties are cultivated and commercialized in many countries in parallel to conventional crops. Given the labeling rules established e.g. in the European Union and the necessary coexistence between GM and non-GM crops, it is important to determine the extent of pollen dissemination from transgenic maize to other cultivars under field conditions. The most widely used methods for quantitative detection of GMO are based on real-time PCR, which implies the results are expressed in genome percentages (in contrast to seed or grain percentages). Our objective was to assess the accuracy of real-time PCR based assays to accurately quantify the contents of transgenic grains in non-GM fields in comparison with the real cross-fertilization rate as determined by phenotypical analysis. We performed this study in a region where both GM and conventional maize are normally cultivated and used the predominant transgenic maize Mon810 in combination with a conventional maize variety which displays the characteristic of white grains (therefore allowing cross-pollination quantification as percentage of yellow grains). Our results indicated an excellent correlation between real-time PCR results and number of cross-fertilized grains at Mon810 levels of 0.1–10%. In contrast, Mon810 percentage estimated by weight of grains produced less accurate results. Finally, we present and discuss the pattern of pollen-mediated gene flow from GM to conventional maize in an example case under field conditions.     

农业活动及转基因作物对农田生物多样性的影响

农田生物多样性是生态系统生物多样性的重要组成部分,但较少受到关注.近50年来,由于农业活动引起的环境污染、生境破碎和单一化种植等严重威胁着农田生物多样性.为了了解各因素对农田生物多样性的影响程度,优化农田管理措施,以提高农作物产量并降低环境影响,本文综述了种植方式、地膜覆盖、农药和化肥使用等农业活动及转基因作物对我国农田生物多样性的影响.农药和化肥的过度使用对农田生物多样性的影响最大;而转基因作物对农田生物多样性的影响受诸多因素影响,如携带的转基因性状等.需要加强转基因作物生态环境影响评价研究,特别是对农田生物多样性的潜在影响.农业生产活动应当与农田生物多样性保护密切结合,不仅有利于提高农作物产量,同时也可减少对环境的负面影响.     

Outcrossing potential between 11 important genetically modified crops and the Chilean vascular flora

The potential impact of genetically modified (GM) crops on biodiversity is one of the main concerns in an environmental risk assessment (ERA). The likelihood of outcrossing and pollen‐mediated gene flow from GM crops and non‐GM crops are explained by the same principles and depend primarily on the biology of the species. We conducted a national‐scale study of the likelihood of outcrossing between 11 GM crops and vascular plants in Chile by use of a systematized database that included cultivated, introduced and native plant species in Chile. The database included geographical distributions and key biological and agronomical characteristics for 3505 introduced, 4993 native and 257 cultivated (of which 11 were native and 246 were introduced) plant species. Out of the considered GM crops (cotton, soya bean, maize, grape, wheat, rice, sugar beet, alfalfa, canola, tomato and potato), only potato and tomato presented native relatives (66 species total). Introduced relative species showed that three GM groups were formed having: a) up to one introduced relative (cotton and soya bean), b) up to two (rice, grape, maize and wheat) and c) from two to seven (sugar beet, alfalfa, canola, tomato and potato). In particular, GM crops presenting introduced noncultivated relative species were canola (1 relative species), alfalfa (up to 4), rice (1), tomato (up to 2) and potato (up to 2). The outcrossing potential between species [OP; scaled from ‘very low’ (1) to ‘very high’ (5)] was developed, showing medium OPs (3) for GM–native relative interactions when they occurred, low (2) for GMs and introduced noncultivated and high (4) for the grape‐Vitis vinifera GM–introduced cultivated interaction. This analytical tool might be useful for future ERA for unconfined GM crop release in Chile.