转Bt基因抗虫作物对非靶标害虫的影响

转基因作物的长期大面积种植, 在为农业生产带来惠益的同时, 对农业生态系统的健康和稳定可能会产生潜在的影响。转基因作物表达的Bt蛋白对靶标害虫起到较好的控制效果, 而对Bt蛋白不敏感的非靶标害虫种群可能会迅速发展起来, 对作物造成为害。随着抗虫转基因作物的连续多年种植, 科学家们对于田间杀虫剂施用量的增减看法不尽一致。通过总结已有的研究报道, 本文以Bt玉米和Bt棉花为例, 分析了大田中非靶标害虫暴发的现状, 以及暴发的主要原因(如杀虫剂的使用、害虫天敌减少和物种替代)。在生产实践中, 抗虫作物的长期大面积释放导致广谱杀虫剂施用量减少, 田间非靶标害虫数量上升。因此今后需要继续开展更多的研究来综合评估种植转Bt基因作物产生的长期潜在影响, 优化害虫防治措施, 避免非靶标害虫暴发。     

抗虫转基因植物对非靶标节肢动物生态影响的研究进展

重组DNA技术的发展为培育高效的抗虫作物提供了前所未有的便利条件。通过转基因技术,全世界已培育出众多转基因抗虫植物品系。其中,表达苏云金芽孢杆菌(Bt)基因的作物品系如Bt棉花和Bt玉米已在很多国家大规模种植,在害虫控制方面发挥了重要的作用。转基因抗虫作物可能带来的生态风险问题,如对农田非靶标节肢动物的潜在影响,一直受到相关研究者及民众的广泛关注。至今,已有大量研究论文发表。本文在总结、归纳前人研究的基础上,阐述了从实验室到田间多层次评价转基因抗虫作物对非靶标生物影响的一般研究程序和方法,并简要综述了Bt玉米和Bt棉花2种已商业化种植的转基因抗虫作物对农田非靶标节肢动物生态影响的研究进展。现有研究表明:当前种植的Bt作物所表达的Cry蛋白杀虫专一性非常强,对农田非靶标节肢动物没有毒性;且Bt作物的利用降低了广谱化学杀虫剂的施用量,从而提高了非专一性害虫天敌的种群密度,加强了对害虫的控制,并有效地保护了生态环境和农民健康。因此,Bt作物可以作为害虫综合防治(IPM)的一个策略,结合其他防治措施可加强对害虫的有效控制。     

棉铃虫对转Bt基因抗虫棉花的抗性机制及治理

棉铃虫是危害棉花最严重的害虫之一. 作为生物技术产品, 转Bt杀虫基因棉花产生的Cry毒素对棉铃虫有高效毒杀作用. Bt棉花已在世界范围内商业化种植, 通过有效控制棉铃虫种群数量, 而显著减少了化学农药的用量. 尽管没有发现棉铃虫田间种群对Bt棉花产生高水平抗性, 但室内持续筛选已培育出多个高水平抗性品系, 表明存在棉铃虫对Bt棉花产生抗性的风险. 鉴于棉铃虫对Bt棉花产生抗性可能对Bt棉花利用价值的影响, 国内外近10年来对此进行了系统深入地研究. 本文综述了棉铃虫对Bt棉花抗性的生物化学和分子机制、抗性治理与监测技术的最新研究进展, 并分析了中国、澳大利亚和印度等国家棉铃虫对Bt棉花的抗性治理策略.     

转基因棉花主要靶标害虫的抗性发展及抗性治理策略研究

转基因棉花的大面积种植有效的控制了棉铃虫Helicoverpa armigera、红铃虫Pectinophora gossypiella 等靶标害虫的危害,然而抗性监测结果显示,转Bt基因棉田的棉铃虫耐受性正逐年提高,抗性问题已成为影响持续利用转Bt基因棉花的主要因素,发展新的转基因棉花势在必行.新的基因或蛋白的选择应以对靶标害虫高效、不易产生抗性,且与现在广泛应用的基因或蛋白无交互抗性为原则.本文综述了转Bt基因棉花的主要靶标害虫对Bt抗性的研究进展,及其与新抗虫转基因棉花的关系,并讨论了抗性治理策略的发展历程.     

靶标害虫对Bt玉米的抗性发展和治理策略

全生育期有效表达Bt杀虫蛋白的转基因抗虫玉米为靶标害虫的防治提供了新途径。但是,靶标害虫抗性种群的发展严重威胁了转基因抗虫玉米的可持续应用。截止到2018年,已经有13例报道表明靶标害虫对转基因抗虫玉米产生了田间抗性;5例监测结果表明靶标种群没有降低对Bt玉米的敏感性,其中包括转vip3Aa玉米。抗性治理策略成功的关键主要包括:Bt杀虫蛋白的高剂量表达、靶标害虫的隐性遗传、初始抗性等位基因频率较低、不完全抗性、适合度代价等。当抗性为非隐性遗传时,可以通过增加庇护所的种植面积达到延缓抗性发展的目的。     

Bt棉花害虫综合治理研究前沿

自1996年以来,全球Bt(Bacillus thuringiensis)棉花应用规模迅速增长,目前已占棉花种植总面积的60%左右,主要种植国家包括美国、澳大利亚、中国、印度和巴基斯坦等。大量研究表明,Bt棉花的大面积种植有效控制了多种靶标害虫的发生危害,从而大幅度减少了化学杀虫剂的使用量;化学杀虫剂的减少使用导致一些非靶标害虫的发生危害明显加重。针对Bt棉花生产中呈现出的害虫新问题,各国分别发展了由农业防治、生物防治、化学防治等不同措施构成的防控技术体系。     

Advance in insect pest management in Bt cotton worldwide

自1996年以来,全球Bt(Bacillus thuringiensis)棉花应用规模迅速增长,目前已占棉花种植总面积的60％左右,主要种植国家包括美国、澳大利亚、中国、印度和巴基斯坦等.大量研究表明,Bt棉花的大面积种植有效控制了多种靶标害虫的发生危害,从而大幅度减少了化学杀虫剂的使用量;化学杀虫剂的减少使用导致一些非靶标害虫的发生危害明显加重.针对Bt棉花生产中呈现出的害虫新问题,各国分别发展了由农业防治、生物防治、化学防治等不同措施构成的防控技术体系.     

转抗虫基因植物生态安全性研究进展

转抗虫基因植物如Bt棉花等已在美国、中国和澳大利亚等国家大规模商业化种植 ,有关转抗虫基因植物潜在的生态风险已引起广泛的关注。该文综述了转抗虫基因植物研究应用现状与安全性研究进展。主要内容包括 :转抗虫基因植物的种类及其对靶标害虫的抗性 ,对非靶标害虫和天敌发生的影响 ,对农田生态系统生物多样性的影响 ,靶标昆虫的抗性治理及转抗虫基因植物的基因漂移等     

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     

Cry1Ie毒素胁迫下亚洲玉米螟的抗性发展及汰选种群对其他Bt毒素的交互抗性

亚洲玉米螟Ostrinia furnacalis (Guenée) 是危害玉米的重要害虫之一, 转Bt基因抗虫玉米为其防治提供了新的途径。然而, 靶标害虫产生抗性将严重阻碍Bt制剂及转Bt基因抗虫玉米的持续应用。明确害虫对转Bt基因玉米表达的毒素蛋白的抗性演化, 对于制定科学有效的抗性治理策略具有重要的理论和实际意义。本实验通过人工饲料汰选法研究了Bt Cry1Ie毒素胁迫下亚洲玉米螟的抗性发展及汰选14代的种群对其他Bt毒素（Cry1Ab, Cry1Ac和Cry1Fa）的交互抗性, 并观察了Cry1Ie蛋白胁迫对亚洲玉米螟生物学的影响。结果表明： 随着汰选压不断提高, 亚洲玉米螟种群对Cry1Ie毒素的敏感性逐渐下降。汰选14代后, 种群对Cry1Ie毒素的抗性水平提高了23倍。然而, Cry1Ab, Cry1Ac和Cry1Fa对所获Cry1Ie汰选种群的毒力与对敏感种群的毒力相比没有显著差异, 说明Cry1Ie汰选没有引起亚洲玉米螟对Cry1Ab, Cry1Ac和Cry1Fa毒素产生交互抗性。同时, 与敏感种群相比, Cry1Ie汰选14代的种群幼虫平均发育历期延长5.7 d, 蛹重减轻13.7%, 单雌产卵量下降40.0%。本研究结果说明, 大面积单一种植转cry1Ie基因抗虫玉米, 可能引起亚洲玉米螟产生抗性; 亚洲玉米螟Cry1Ie抗性种群对Cry1Ab, Cry1Ac和Cry1Fa没有交互抗性, 含有cry1Ie和cry1Ab, cry1Ac或cry1F双/多基因抗虫玉米, 可作为靶标害虫抗性治理的重要策略。     

Bt crop effects on functional guilds of non-target arthropods: a meta-analysis

Background

Uncertainty persists over the environmental effects of genetically-engineered crops that produce the insecticidal Cry proteins of Bacillus thuringiensis (Bt). We performed meta-analyses on a modified public database to synthesize current knowledge about the effects of Bt cotton, maize and potato on the abundance and interactions of arthropod non-target functional guilds.

Methodology/Principal Findings

We compared the abundance of predators, parasitoids, omnivores, detritivores and herbivores under scenarios in which neither, only the non-Bt crops, or both Bt and non-Bt crops received insecticide treatments. Predators were less abundant in Bt cotton compared to unsprayed non-Bt controls. As expected, fewer specialist parasitoids of the target pest occurred in Bt maize fields compared to unsprayed non-Bt controls, but no significant reduction was detected for other parasitoids. Numbers of predators and herbivores were higher in Bt crops compared to sprayed non-Bt controls, and type of insecticide influenced the magnitude of the difference. Omnivores and detritivores were more abundant in insecticide-treated controls and for the latter guild this was associated with reductions of their predators in sprayed non-Bt maize. No differences in abundance were found when both Bt and non-Bt crops were sprayed. Predator-to-prey ratios were unchanged by either Bt crops or the use of insecticides; ratios were higher in Bt maize relative to the sprayed non-Bt control.

Conclusions/Significance

Overall, we find no uniform effects of Bt cotton, maize and potato on the functional guilds of non-target arthropods. Use of and type of insecticides influenced the magnitude and direction of effects; insecticde effects were much larger than those of Bt crops. These meta-analyses underscore the importance of using controls not only to isolate the effects of a Bt crop per se but also to reflect the replacement of existing agricultural practices. Results will provide researchers with information to design more robust experiments and will inform the decisions of diverse stakeholders regarding the safety of transgenic insecticidal crops.     

Six Years after the Commercial Introduction of Bt Maize in Spain: Field Evaluation, Impact and Future Prospects

We carried out a 6-year-field evaluation to assess potential hazards of growing Compa, a transgenic Bt maize variety based on the transformation event CG 00256-176. Two categories of hazards were investigated: the potential of the target corn borer Sesamia nonagrioides to evolve resistance to Bt maize and effects on non-target organisms. In order to address the first hazard, dispersal capacity of the corn borer was measured and our results indicated that larvae move to plants other than those onto which the female oviposited - even to plants in adjacent rows - in remarkable numbers and they do so mostly at a mature age, suggesting that mixing Bt and non-Bt seeds in the same field would not be a very useful deployment strategy to delay/prevent resistance. In addition, adults move among fields to mate and males may do so for up to 400 m. Three different aspects of potential non-target effects were investigated: sub-lethal effects on the target S. nonagrioides, effects on non-target maize pests, and effects on maize-dwelling predators. Larvae collected in Bt fields at later growth stages, in which event 176 Bt maize expresses Bt toxin at sub-lethal concentrations, had longer diapause and post-diapause development than larvae collected in non-Bt fields, a feature that might lead to a certain isolation between populations in both type of fields and accelerate Bt resistance evolution. Transgenic maize did not have a negative impact on non-target pests in the field; more aphids and leafhoppers but similar numbers of cutworms and wireworms were counted in Bt versus non-Bt fields; in any case differences in damage or yield were recorded. We observed no difference in the numbers of the most relevant predators in fields containing transgenic or no transgenic maize.     

Increased frequency of pink bollworm resistance to Bt toxin Cry1Ac in China

Transgenic crops producing insecticidal proteins from Bacillus thuringiensis (Bt) kill some key insect pests, but evolution of resistance by pests can reduce their efficacy. The main approach for delaying pest adaptation to Bt crops uses non-Bt host plants as "refuges" to increase survival of susceptible pests. To delay evolution of pest resistance to transgenic cotton producing Bt toxin Cry1Ac, the United States and some other countries have required refuges of non-Bt cotton, while farmers in China have relied on "natural" refuges of non-Bt host plants other than cotton. The "natural" refuge strategy focuses on cotton bollworm (Helicoverpa armigera), the primary target of Bt cotton in China that attacks many crops, but it does not apply to another major pest, pink bollworm (Pectinophora gossypiella), which feeds almost entirely on cotton in China. Here we report data showing field-evolved resistance to Cry1Ac by pink bollworm in the Yangtze River Valley of China. Laboratory bioassay data from 51 field-derived strains show that the susceptibility to Cry1Ac was significantly lower during 2008 to 2010 than 2005 to 2007. The percentage of field populations yielding one or more survivors at a diagnostic concentration of Cry1Ac increased from 0% in 2005-2007 to 56% in 2008-2010. However, the median survival at the diagnostic concentration was only 1.6% from 2008 to 2010 and failure of Bt cotton to control pink bollworm has not been reported in China. The early detection of resistance reported here may promote proactive countermeasures, such as a switch to transgenic cotton producing toxins distinct from Cry1A toxins, increased planting of non-Bt cotton, and integration of other management tactics together with Bt cotton.     

Early detection of field-evolved resistance to Bt cotton in China: cotton bollworm and pink bollworm

Transgenic crops producing Bacillus thuringiensis (Bt) toxins kill some major insect pests, but pests can evolve resistance and thereby reduce the effectiveness of such Bt crops. The main approach for slowing pest adaptation to Bt crops uses non-Bt host plants as "refuges" to increase survival of susceptible pests. To delay evolution of pest resistance to cotton producing Bt toxin Cry1Ac, several countries have required refuges of non-Bt cotton, while farmers in China have relied on "natural" refuges of non-Bt host plants other than cotton. This strategy is designed for cotton bollworm (Helicoverpa armigera), which attacks many crops and is the primary target of Bt cotton in China, but it does not apply to pink bollworm (Pectinophora gossypiella), which feeds almost entirely on cotton in China. Here we review evidence of field-evolved resistance to Cry1Ac by cotton bollworm in northern China and by pink bollworm in the Yangtze River Valley of China. For both pests, results of laboratory diet bioassays reveal significantly decreased susceptibility of field populations to Cry1Ac, yet field control failures of Bt cotton have not been reported. The early detection of resistance summarized here may spur countermeasures such as planting Bt cotton that produces two or more distinct toxins, increased planting of non-Bt cotton, and integration of other management tactics together with Bt cotton.     

Evolution of resistance to transgenic crops: interactions between insect movement and field distribution

The refuge strategy is designed to delay evolution of pest resistance to transgenic crops producing Bacillus thuringiensis Berliner (Bt) toxins. Movement of insects between Bt crops and refuges of non-Bt crops is essential for the refuge strategy because it increases chances that resistant adults mate with susceptible adults from refuges. Conclusions about optimal levels of movement for delaying resistance are not consistent among previous modeling studies. To clarify the effects of movement on resistance evolution, we analyzed simulations of a spatially explicit model based partly on the interaction of pink bollworm, Pectinophora gossypiella (Saunders), with Bt cotton. We examined resistance evolution as a function of insect movement under 12 sets of assumptions about the relative abundance of Bt cotton (50 and 75%), temporal distribution of Bt cotton and refuge fields (fixed, partial rotation, and full rotation), and spatial distribution of fields (random and uniform). The results show that interactions among the relative abundance and distribution of refuges and Bt cotton fields can alter the effects of movement on resistance evolution. The results also suggest that differences in conclusions among previous studies can be explained by differences in assumptions about the relative abundance and distribution of refuges and Bt crop fields. With fixed field locations and all Bt cotton fields adjacent to at least one refuge, resistance evolved slowest with low movement. However, low movement and fixed field locations favored rapid resistance evolution when some Bt crop fields were isolated from refuges. When refuges and Bt cotton fields were rotated to the opposite crop type each year, resistance evolved fastest with low movement. Nonrecessive inheritance of resistance caused rapid resistanceevolution regardless of movement rate. Confirming previous reports, results described here show that resistance can be delayed effectively by fixing field locations and distributing refuges uniformly to ensure that Bt crop fields are not isolated from refuges. However, rotating fields provided better insect control and reduced the need for insecticide sprays.     

Contamination and management of resistance evolution to high-dose transgenic insecticidal crops

For maize and cotton, transgenic varieties that express toxins derived from Bacillus thuriengensis (Bt) are now planted in several countries. To slow resistance evolution, the “high-dose/refuge” strategy is broadly implemented in which resistance is recessive and some fields (or areas within fields) are planted exclusively with Bt crops and other fields planted exclusively with non-transgenic refuge crops for susceptible insects. This strategy, however, could potentially be undermined by contamination. Here, we investigate general models of resistance evolution for high-dose events in which fields are contaminated due to the inadvertent mixing of seeds, volunteer plants, or pollen flow between Bt and non-Bt varieties coupled with seed-saving by farmers. Contamination of the refuge by Bt plants increases selection for resistance, thereby speeding resistance evolution. Nonetheless, in most situations this effect is small. Contamination of Bt fields by non-transgenic plants might be expected to have the opposite effect and always reduce the rate of resistance evolution. While this is often the case, it is not always so. If larvae move among plants within a field, then high movement rates may reverse the effect of contamination of Bt fields to slow resistance evolution. Furthermore, if the dispersal rates of adult females between Bt and refuge fields are low, then contamination of Bt fields may speed resistance. These results suggest that contamination has the potential to undermine the efficacy of the high-dose/refuge strategy, yet depending upon the particular pest and situation, contamination may not be a concern.     

Aphid Honeydew Quality as a Food Source for Parasitoids Is Maintained in Bt Cotton

Bt-transgenic cotton has proven to be highly efficient in controlling key lepidopteran pests. One concern with the deployment of Bt cotton varieties is the potential proliferation of non-target pests. We previously showed that Bt cotton contained lower concentrations of insecticidal terpenoids as a result of reduced caterpillar damage, which benefited the aphid Aphis gossypii. It is thus important that non-target herbivores are under biological control in Bt cotton fields. The induction or lack of induction of terpenoids could also influence the quality of aphid honeydew, an important food source for beneficial insects. We therefore screened A. gossypii honeydew for cotton terpenoids, that are induced by caterpillars but not the aphids. We then tested the influence of induced insect-resistance of cotton on honeydew nutritional quality for the aphid parasitoid Lysiphlebus testaceipes and the whitefly parasitoid Eretmocerus eremicus. We detected the cotton terpenoids gossypol and hemigossypolone in A. gossypii honeydew. Although a feeding assay demonstrated that gossypol reduced the longevity of both parasitoid species in a non-linear, dose-dependent manner, the honeydew was capable of sustaining parasitoid longevity and reproduction. The level of caterpillar damage to Bt and non-Bt cotton had no impact on the quality of honeydew for the parasitoids.These results indicate that the nutritional quality of honeydew is maintained in Bt cotton and is not influenced by induced insect resistance.     

Spatial processes in the evolution of resistance in Helicoverpa zea (Lepidoptera: Noctuidae) to Bt transgenic corn and cotton in a mixed agroecosystem: a biology-rich stochastic simulation model

A simulation model is developed to examine the role of spatial processes in the evolution of resistance in Helicoverpa zea populations to Bt corn and Bt cotton. The model is developed from the stochastic spatially explicit Heliothis virescens model described by Peck et al. (1999), to accommodate a spatial mix of two host crops (corn and cotton), and to reflect the agronomic practices, as well as the spatial and temporal population dynamics of H. zea, in eastern North Carolina. The model suggests that selection for resistance is more intense in Bt cotton fields than in Bt corn fields. It further suggests that local gene frequencies are highly dependent on local deployment levels of Bt crops despite the high mobility of the adult insects. Region-wide average gene frequencies depend on the region-wide level of Bt deployment, so incomplete technology adoption slows the rate of resistance evolution. However, on a local scale, H. zea populations in clusters of fields in which Bt use is high undergo far more rapid evolution than populations in neighboring clusters of fields in which Bt use is low. The model suggests that farm-level refuge requirements are important for managing the risk of resistance. The model can be used as an aid in designing plans for monitoring for resistance by suggesting the appropriate distribution of monitoring locations, which should focus on areas of highest Bt crop deployment. The findings need to be placed in the context of the input parameters, many of which are uncertain or highly variable in nature, and therefore, a thorough sensitivity analysis is warranted.     

The Cultivation of Bt Corn Producing Cry1Ac Toxins Does Not Adversely Affect Non-Target Arthropods

Transgenic corn producing Cry1Ac toxins from Bacillus thuringiensis (Bt) provides effective control of Asian corn borer, Ostrinia furnacalis (Guenée), and thus reduces insecticide applications. However, whether Bt corn exerts undesirable effects on non-target arthropods (NTAs) is still controversial. We conducted a 2-yr study in Shangzhuang Agricultural Experiment Station to assess the potential impact of Bt corn on field population density, biodiversity, community composition and structure of NTAs. On each sampling date, the total abundance, Shannon''s diversity index, Pielou''s evenness index and Simpson''s diversity index were not significantly affected by Bt corn as compared to non-Bt corn. The “sampling dates” had a significant effect on these indices, but no clear tendencies related to “Bt corn” or “sampling dates X corn variety” interaction were recorded. Principal response curve analysis of variance indicated that Bt corn did not alter the distribution of NTAs communities. Bray-Curtis dissimilarity and distance analysis showed that Cry1Ac toxin exposure did not increase community dissimilarities between Bt and non-Bt corn plots and that the evolution of non-target arthropod community was similar on the two corn varieties. The cultivation of Bt corn failed to show any detrimental evidence on the density of non-target herbivores, predators and parasitoids. The composition of herbivores, predators and parasitoids was identical in Bt and non-Bt corn plots. Taken together, results from the present work support that Bt corn producing Cry1Ac toxins does not adversely affect NTAs.