Helicoverpa zea and Bt cotton in the United States

Helicoverpa zea (Boddie), the bollworm or corn earworm, is the most important lepidopteran pest of Bt cotton in the United States. Corn is the preferred host, but the insect feeds on most flowering crops and wild host plants. As a cotton pest, bollworm has been closely linked to the insecticide-resistance prone Heliothis virescens (F.), tobacco budworm. Immature stages of the two species are difficult to separate in field environments. Tobacco budworm is very susceptible to most Bt toxins, and Bt cotton is considered to be "high dose." Bollworm is less susceptible to Bt toxins, and Bt cotton is not "high dose" for this pest. Bt cotton is routinely sprayed with traditional insecticides for bollworm control. Assays of bollworm field populations for susceptibility to Bt toxins expressed in Bt cotton have produced variable results since pre-deployment of Bt cottons in 1988 and 1992. Analyses of assay response trends have been used by others to suggest that field resistance has evolved to Bt toxins in bollworm, but disagreement exists on definitions of field resistance and confidence of variable assay results to project changes in susceptibility of field populations. Given historical variability in bollworm response to Bt toxins, erratic field control requiring supplemental insecticides since early field testing of Bt cottons, and dramatic increases in corn acreage in cotton growing areas of the Southern US, continued vigilance and concern for resistance evolution are warranted.     

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.     

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.     

华北棉区棉铃虫种群动态与其为害特征分析

本文述及棉铃虫为害特性及其与为害后棉株的补偿作用关系。根据棉株生命表资料,蕾期棉株蕾花的脱落主要系由棉铃虫为害所致。而铃期蕾铃的脱落,则主要系由棉株的生理条件。在蕾期,前三周内蕾花被害脱落后,棉株的补偿作用是很高的,从第四周起补偿作用逐渐下降,而到铃期阶段时补偿作用降至很低水平。因此蕾期棉株遭受一定数量以内的棉铃虫为害,而使蕾花脱落后,反而可促使棉花增产。根据棉铃虫生命表资料,从卵期至三龄末幼虫期的累计死亡率占总死亡率的90%以上,结合幼虫死亡率与不同龄期为害水平分析,棉铃虫对棉株的主要为害阶段系在2—3龄期,其对果实的为害量占总为害的65%。而棉株被害蕾铃的田间空间分布型系遵从负二项分布,期公共K值为3.4626。     

The halo effect: suppression of pink bollworm on non-bt cotton by bt cotton in china

In some previously reported cases, transgenic crops producing insecticidal proteins from Bacillus thuringiensis (Bt) have suppressed insect pests not only in fields planted with such crops, but also regionally on host plants that do not produce Bt toxins. Here we used 16 years of field data to determine if Bt cotton caused this "halo effect" against pink bollworm (Pectinophora gossypiella) in six provinces of the Yangtze River Valley of China. In this region, the percentage of cotton hectares planted with Bt cotton increased from 9% in 2000 to 94% in 2009 and 2010. We found that Bt cotton significantly decreased the population density of pink bollworm on non-Bt cotton, with net decreases of 91% for eggs and 95% for larvae on non-Bt cotton after 11 years of Bt cotton use. Insecticide sprays targeting pink bollworm and cotton bollworm (Helicoverpa armigera) decreased by 69%. Previously reported evidence of the early stages of evolution of pink bollworm resistance to Bt cotton in China has raised concerns that if unchecked, such resistance could eventually diminish or eliminate the benefits of Bt cotton. The results reported here suggest that it might be possible to find a percentage of Bt cotton lower than the current level that causes sufficient regional pest suppression and reduces the risk of resistance.     

Improvement of pest resistance in transgenic tobacco plants expressing dsRNA of an insect-associated gene EcR

The adoption of pest-resistant transgenic plants to reduce yield loss and pesticide utilization has been successful in the past three decades. Recently, transgenic plant expressing double-stranded RNA (dsRNA) targeting pest genes emerges as a promising strategy for improving pest resistance in crops. The steroid hormone, 20-hydroxyecdysone (20E), predominately controls insect molting via its nuclear receptor complex, EcR-USP. Here we report that pest resistance is improved in transgenic tobacco plants expressing dsRNA of EcR from the cotton bollworm, Helicoverpa armigera, a serious lepidopteran pest for a variety of crops. When H. armigera larvae were fed with the whole transgenic tobacco plants expressing EcR dsRNA, resistance to H. armigera was significantly improved in transgenic plants. Meanwhile, when H. armigera larvae were fed with leaves of transgenic tobacco plants expressing EcR dsRNA, its EcR mRNA level was dramatically decreased causing molting defects and larval lethality. In addition, the transgenic tobacco plants expressing H. armigera EcR dsRNA were also resistant to another lepidopteran pest, the beet armyworm, Spodoptera exigua, due to the high similarity in the nucleotide sequences of their EcR genes. This study provides additional evidence that transgenic plant expressing dsRNA targeting insect-associated genes is able to improve pest resistance.     

棉铃虫田间种群抗药性的生态学干扰和生化机制研究

从生态遗传学角度和抗性机制两个方面研究了作物布局和解毒酶系对棉铃虫高效氯氰菊酯抗药性的影响,通过田间采样,测定了姜堰,如皋,兴化3个地区的棉铃虫对高效氯氰菊酯的抗药性。结果表明,作物布局对棉铃虫菊酯抗性的发展具有明显的影响,寄主相对单一的棉花连片种植区（兴化）棉田棉铃虫对高效氯氰菊酯的抗性水平最高,是棉花春玉米混栽区（如皋）棉田棉铃虫抗药性的3．5倍,在邻近春玉米种植区的稻棉区（姜堰）,棉田棉铃虫的抗药性水平,是4－5km外玉米田棉铃虫的4．8倍,研究同时表明,四代棉铃虫的抗药性比3代棉铃虫的抗药性下降了2．3倍,棉铃虫离体解毒酶和乙酰胆碱酯酶活性测定表明,棉铃虫的羧酸酯酶,谷胱甘肽S－转移酶和乙酰胆碱酯酶活性与棉铃虫对高效氯氰菊酯的抗药性水平有显著的相关性,对姜堰南部棉田和如振戴庄棉田棉铃虫的活体增效试验表明,多功能氧化酶是抗性棉铃虫对高效氯氰菊酯重要的解毒酶系,增效醚可分别增效22．79倍和12．33倍,羧酸酯酶对抗性棉铃虫的解毒代谢也有部分贡献,磷酸三苯酯可分别增效4．41倍和3．70倍。     

Diminishing Returns from Increased Percent Bt Cotton: The Case of Pink Bollworm

Regional suppression of pests by transgenic crops producing insecticidal proteins from Bacillus thuringiensis (Bt) has been reported in several cropping systems, but little is known about the functional relationship between the ultimate pest population density and the pervasiveness of Bt crops. Here we address this issue by analyzing 16 years of field data on pink bollworm (Pectinophora gossypiella) population density and percentage of Bt cotton in the Yangtze River Valley of China. In this region, the percentage of cotton hectares planted with Bt cotton increased from 9% in 2000 to 94% in 2009 and 2010. We find that as the percent Bt cotton increased over the years, the cross-year growth rate of pink bollworm from the last generation of one year to the first generation of the next year decreased. However, as the percent Bt cotton increased, the within-year growth rate of pink bollworm from the first to last generation of the same year increased, with a slope approximately opposite to that of the cross-year rates. As a result, we did not find a statistically significant decline in the annual growth rate of pink bollworm as the percent Bt cotton increased over time. Consistent with the data, our modeling analyses predict that the regional average density of pink bollworm declines as the percent Bt cotton increases, but the higher the percent Bt cotton, the slower the decline in pest density. Specifically, we find that 95% Bt cotton is predicted to cause only 3% more reduction in larval density than 80% Bt cotton. The results here suggest that density dependence can act against the decline in pest density and diminish the net effects of Bt cotton on suppression of pink bollworm in the study region. The findings call for more studies of the interactions between pest density-dependence and Bt crops.     

湖北棉区转Bt基因棉对棉铃虫的控制作用

2000-2001年通过田间系统调查,表明转Bt基因棉（品种为GK19）在湖北江汉平原棉区对棉铃虫抗性稳定。试验设3个处理：转Bt基因棉化防田（使用化学农药控制害虫）、转Bt基因棉自控田（依靠天敌控制害虫）及常规棉对照田（利用综合防治措施控制害虫）。从棉铃虫的第2代到第5代整个发生期内,即使在不进行化学防治的情况下,棉铃虫在Bt棉田的发生量也保持在极低的水平(最高百株虫量为12头)。室内饲养结果表明,转Bt基因棉对棉铃虫的生长发育（幼虫体重、蛹重）有较为明显的影响,使6龄幼虫体重减少25.6%,蛹重减少18.2%。棉铃虫幼虫取食转Bt基因棉组织后,发育迟缓,相对于常规棉喂养的整个发育历期延长17 d,使棉铃虫在田间的危害减少至少一个世代。另外,接虫试验表明,棉铃虫幼虫在常规棉上的取食时间是转Bt基因棉株上的6.1倍,极大地减轻了棉铃虫的危害程度。     

Spring phenology of cotton bollworm affects wheat yield

Climate change has changed numerous species phenologies. Understanding the asynchronous responses between pest insects and host plants to climate change is helpful in improving integrated pest management. It is necessary to use long‐term data to analyze the effects of climate change on cotton bollworm and wheat anthesis. Data for cotton bollworm, wheat yield, and wheat anthesis collected since 1990 were analyzed using linear regression and partial least‐squares regression, as well as the Mann–Kendall test. The results showed that warmer temperatures in the spring advanced the phenologies of cotton bollworm and wheat anthesis, but the phenology changes in overwintering cotton bollworm were faster than those in wheat anthesis, and the eclosion period of overwintering was prolonged, resulting in an increase in overwintering adult abundance. This might lead to more first‐generation larvae and subsequent wheat damage. An early or late first‐appearance date significantly affected the eclosion days. The abrupt changes of phenologies in cotton bollworm, wheat anthesis, and climate were asynchronous, but the abrupt phenology changes occurred after or around the climate abrupt change, especially after or around the abrupt changes of temperature in March and April. The expansion of asynchronous responses in the change rate of wheat anthesis and overwintering cotton bollworm would likely decrease wheat yield due to climate warming in the future. Accumulated temperature was the major affecting factor on the first eclosion date (t₁), adult abundance, and eclosion days. Temperatures in March and April and precipitation in the winter mainly affected the prepeak date (t₂), peak date (t₃), and postpeak date (t₄), respectively, and these factors indirectly affected wheat yield. Thus, the change in the spring phenology of the cotton bollworm and wheat anthesis, and hence wheat yield, was affected by climate warming.     

Engineered Repressible Lethality for Controlling the Pink Bollworm, a Lepidopteran Pest of Cotton

The sterile insect technique (SIT) is an environmentally friendly method of pest control in which insects are mass-produced, irradiated and released to mate with wild counterparts. SIT has been used to control major pest insects including the pink bollworm (Pectinophora gossypiella Saunders), a global pest of cotton. Transgenic technology has the potential to overcome disadvantages associated with the SIT, such as the damaging effects of radiation on released insects. A method called RIDL (Release of Insects carrying a Dominant Lethal) is designed to circumvent the need to irradiate insects before release. Premature death of insects’ progeny can be engineered to provide an equivalent to sterilisation. Moreover, this trait can be suppressed by the provision of a dietary antidote. In the pink bollworm, we generated transformed strains using different DNA constructs, which showed moderate-to-100% engineered mortality. In permissive conditions, this effect was largely suppressed. Survival data on cotton in field cages indicated that field conditions increase the lethal effect. One strain, called OX3402C, showed highly penetrant and highly repressible lethality, and was tested on host plants where its larvae caused minimal damage before death. These results highlight a potentially valuable insecticide-free tool against pink bollworm, and indicate its potential for development in other lepidopteran pests.     

Sustained susceptibility of pink bollworm to Bt cotton in the United States

Evolution of resistance by pests can reduce the benefits of transgenic crops that produce toxins from Bacillus thuringiensis (Bt) for insect control. One of the world's most important cotton pests, pink bollworm (Pectinophora gossypiella), has been targeted for control by transgenic cotton producing Bt toxin Cry1Ac in several countries for more than a decade. In China, the frequency of resistance to Cry1Ac has increased, but control failures have not been reported. In western India, pink bollworm resistance to Cry1Ac has caused widespread control failures of Bt cotton. By contrast, in the state of Arizona in the southwestern United States, monitoring data from bioassays and DNA screening demonstrate sustained susceptibility to Cry1Ac for 16 y. From 1996-2005, the main factors that delayed resistance in Arizona appear to be abundant refuges of non-Bt cotton, recessive inheritance of resistance, fitness costs associated with resistance and incomplete resistance. From 2006-2011, refuge abundance was greatly reduced in Arizona, while mass releases of sterile pink bollworm moths were made to delay resistance as part of a multi-tactic eradication program. Sustained susceptibility of pink bollworm to Bt cotton in Arizona has provided a cornerstone for the pink bollworm eradication program and for integrated pest management in cotton. Reduced insecticide use against pink bollworm and other cotton pests has yielded economic benefits for growers, as well as broad environmental and health benefits. We encourage increased efforts to combine Bt crops with other tactics in integrated pest management programs.     

Cotton bollworm resistance to Bt transgenic cotton: A case analysis

Cotton bollworm (Helicoverpa armigera) is one of the most serious insect pests of cotton. Transgenic cotton expressing Cry toxins derived from a soil bacterium, Bacillus thuringiensis (Bt), has been produced to target this pest. Bt cotton has been widely planted around the world, and this has resulted in efficient control of bollworm populations with reduced use of synthetic insecticides. However, evolution of resistance by this pest threatens the continued success of Bt cotton. To date, no field populations of bollworm have evolved significant levels of resistance; however, several laboratory-selected Cry-resistant strains of H. armigera have been obtained, which suggests that bollworm has the capacity to evolve resistance to Bt. The development of resistance to Bt is of great concern, and there is a vast body of research in this area aimed at ensuring the continued success of Bt cotton. Here, we review studies on the evolution of Bt resistance in H. armigera, focusing on the biochemical and molecular basis of Bt resistance. We also discuss resistance management strategies, and monitoring programs implemented in China, Australia, and India.     

环渤海湾地区棉铃虫成虫的卵巢发育特点及与迁飞行为的关系

1996－2000年期间先后在河北省廊坊市、北京市西郊和辽宁省辽阳市系统研究了不同代别棉铃虫成虫的卵巢发育特点及与季节性迁飞行为的关系,研究结果表明环渤海湾南部的河北省廊坊市和北部的辽宁省辽阳市第1代棉铃虫成虫种群主要由外地迁人,第2－3代种群多数年份主要来源于当地,但部分年份亦有外地种群大量迁入,北京市西郊空中诱捕的棉铃虫1－3代成虫多为产卵前期个体,具迁飞昆虫的生殖生理特征,表明棉铃虫成虫迁飞活动存在于各个代别。     

Effects of cotton cultivar on fitness costs associated with resistance of pink bollworm (Lepidoptera: Gelechiidae) to Bt cotton

Fitness costs associated with insect resistance to transgenic crops producing toxins from Bacillus thuringiensis (Bt) reduce the fitness on non-Bt refuge plants of resistant individuals relative to susceptible individuals. Because costs may vary among host plants, choosing refuge cultivars that increase the dominance or magnitude of costs could help to delay resistance. Specifically, cultivars with high concentrations of toxic phytochemicals could magnify costs. To test this hypothesis, we compared life history traits of three independent sets of pink bollworm, Pectinophora gossypiella (Saunders), populations on two cotton cultivars that differed in antibiosis against this cotton pest. Each set had an unselected susceptible population, a resistant population derived by selection from the susceptible population, and the F1 progeny of the susceptible and resistant populations. Confirming previous findings with pink bollworm feeding on cotton, costs primarily affected survival and were recessive on both cultivars. The magnitude of the survival cost did not differ between cultivars. Although the experimental results did not reveal differences between cultivars in the magnitude or dominance of costs, modeling results suggest that differences between cultivars in pink bollworm survival could affect resistance evolution. Thus, knowledge of the interaction between host plants and fitness costs associated with resistance to Bt crops could be helpful in guiding the choice of refuge cultivars.     

转Bt基因抗虫棉田棉铃虫消长规律及危害特点

通过对转 Bt基因抗虫棉田棉铃虫的系统调查研究表明 :(1)转 Bt基因抗虫棉对棉铃虫蛾、卵无抗性表现 ,田间落卵量与常规棉田无明显差异 ,相反 ,二代棉铃虫百株累计卵量倒比常规棉处理区增加了 2 16～ 2 74粒 ,增长幅度为 30 %～ 38% ;(2 )转 Bt基因抗虫棉对二代棉铃虫幼虫的抗性及控制作用较强 ,基本不用进行药剂防治 ,但对三代棉虫幼虫的抗性控制效果明显下降 ,田间 3龄以上幼虫平均达2 0～ 4 0头 /百株 ,多者达 6 0头以上 ,仍需进行必要的药剂防治 ;(3)棉铃虫在转 Bt基因抗虫棉株上的危害特点与常规棉株有明显的差异 ,表现在幼虫和危害症状均较隐蔽 ,不易被发现。     

Effects of refuge contamination by transgenes on Bt resistance in pink bollworm (Lepidoptera: Gelechiidae)

Refuges of non-Bacillus thuringiensis (Bt) cotton, Gossypium hirsutum L., are used to delay Bt resistance in pink bollworm, Pectinophora gossypiella (Saunders) (Lepidoptera: Gelechiidae), a pest that eats cotton seeds. Contamination of refuges by Bt transgenes could reduce the efficacy of this strategy. Previously, three types of contamination were identified in refuges: 1) homozygous Bt cotton plants, with 100% of their seeds producing the Bt toxin Cry1Ac; 2) hemizygous Bt plants with 70-80% of their seeds producing Cry1Ac; and 3) non-Bt plants that outcrossed with Bt plants, resulting in bolls with Cry1Ac in 12-17% of their seeds. Here, we used laboratory bioassays to examine the effects of Bt contamination on feeding behavior and survival of pink bollworm that were resistant (rr), susceptible (ss), or heterozygous for resistance (rs) to Cry1Ac. In choice tests, rr and rs larvae did not differ from ss in preference for non-Bt versus Bt seeds. Survival of rr and rs also did not differ from ss on artificial outcrossed bolls (a mixture of 20% Bt and 80% non-Bt cotton seeds). On artificial hemizygous Bt bolls (70% Bt seeds) and homozygous Bt bolls (100% Bt seeds), rr had higher survival than ss, although rs and ss did not differ. In a simulation model, levels of refuge contamination observed in the field had negligible effects on resistance evolution in pink bollworm. However, in hypothetical simulations where contamination conferred a selective advantage to rs over ss individuals in refuges, resistance evolution was accelerated.     

Suppressing resistance to Bt cotton with sterile insect releases

Genetically engineered crops that produce insecticidal toxins from Bacillus thuringiensis (Bt) are grown widely for pest control. However, insect adaptation can reduce the toxins' efficacy. The predominant strategy for delaying pest resistance to Bt crops requires refuges of non-Bt host plants to provide susceptible insects to mate with resistant insects. Variable farmer compliance is one of the limitations of this approach. Here we report the benefits of an alternative strategy where sterile insects are released to mate with resistant insects and refuges are scarce or absent. Computer simulations show that this approach works in principle against pests with recessive or dominant inheritance of resistance. During a large-scale, four-year field deployment of this strategy in Arizona, resistance of pink bollworm (Pectinophora gossypiella) to Bt cotton did not increase. A multitactic eradication program that included the release of sterile moths reduced pink bollworm abundance by >99%, while eliminating insecticide sprays against this key invasive pest.     

常规棉花粉和转Cry1Ac+CpTI棉花粉对拟澳洲赤眼蜂繁殖和存活的影响

在室内评价了常规棉花粉和转Cry1Ac CpTI棉(Bt棉)花粉通过不同处理方式作为食物源对拟澳洲赤眼蜂Trichogrammaconfusum寿命、寄生卵数、子代羽化数和性比等繁殖和存活特征的影响。共设计7个饲喂处理,即不饲喂(UNFED)、水(W)、水 常规棉花粉(W P)、水 Bt棉花粉(W BtP)、10%蜂蜜水(H)、10%蜂蜜水 常规棉花粉(H P)和10%蜂蜜水 转Bt棉花粉(H BtP)。在提供或不提供米蛾Corcyracephalonica卵条件下,经W P或W BtP饲喂的雌蜂寿命与W或UNFED间无显著差异,但均显著短于H和H P或H BtP饲喂的雌蜂,而经H P或H BtP饲喂的雌蜂寿命又均显著长于经H饲喂的雌蜂(p<0.05)。比较每雌寄生卵数和子代羽化数来看,各处理明显分为3类,即UNFED、W、W P和W BtP为最低,H为居中,H P和H BtP为最高。比较每雌子代性比来看,各处理也明显分为3类,即H、H P和H BtP为最低,W P和W BtP为居中,UNFED和W为最高。饲喂W P和W BtP的雌蜂寿命、寄生卵粒数、子代羽化数和性比分别与W BtP和H BtP饲喂间无显著差异(p<0.05)。因而,棉花花粉需与蜂蜜组合才能成为拟澳洲赤眼蜂实现其最大存活和繁殖力的食物;Bt棉花粉对拟澳洲赤眼蜂无影响。