Effect of Bt genetic engineering on indirect defense in cotton via a tritrophic interaction

We present a tritrophic analysis of the potential non-intended pleiotropic effects of cry1Ac gene derived from Bacillus thurigiensis (Bt) insertion in cotton (DeltaPine 404 Bt Bollgard® variety) on the emission of herbivore induced volatile compounds and on the attraction of the egg parasitoid Trichogramma pretisoum (Hymenoptera: Trichogrammatidae). Both the herbivore damaged Bt variety and its non-Bt isoline (DeltaPine DP4049 variety) produced volatiles in higher quantity when compared to undamaged plants and significantly attracted the egg parasitoids (T. pretiosum) when compared to undamaged plants. However, Trichogramma pretiosum did not differentiate between the transgenic and nontransgenic varieties, suggesting that the ratios between the compounds released by herbivory damaged -Bt cotton and herbivory damaged-nonBt cotton did not change significantly. Finally, no detrimental effect of the Bt genetic engineering was detected related to the volatile compounds released by Bollgard cotton on the behavior of the natural enemy studied.     

不同类型品种棉花上棉蚜适生性及种群动态

通过田间调查、室内网罩盆栽苗测定选择性等方法,考察了常规棉（泗棉3号,石远321）、杂交抗虫棉（辽棉19号,鲁棉研18号）、转单价基因抗虫棉（国抗12号,中棉所32）和转双价抗虫棉（SGK321,中棉所41）4种类型8个品种棉花上棉蚜的适生性及种群动态。结果表明： 棉蚜在各棉花品种上的种群动态有明显差异（P<0.05）,单株蚜量以转单价基因抗虫棉中棉32上最高,常规棉泗棉3号上最低,分别为297.81头/株和76.88头/株。棉蚜对4种类型棉花品种的选择性有明显差异（P<0.05）,其中对转单价基因抗虫棉有很强的选择性。根据棉蚜实验种群的参数判断,其在不同品种棉花上的生长发育、存活及繁殖存在显著差异： 若虫发育历期常规棉石远321上最长（6.46天）,双价棉中棉所41上最短（5.75天）; 存活率转单价基因抗虫棉中棉32上最高（88.21%）,双价棉SGK321上最低（76.46%）; 单雌产蚜量杂交抗虫棉辽棉19上最大（44.48头）,双价棉SGK321上最小（33.51头）; 内禀增长率转单价基因抗虫棉中棉32上最高（0.3695）,双价棉中棉所41上最低（0.3389）。综合评价,棉蚜的生存和繁殖适合性在转单价基因抗虫棉上最高,在双价棉上最低。     

Feeding and Dispersal Behavior of the Cotton Leafworm,Alabama argillacea (Hübner) (Lepidoptera: Noctuidae), on Bt and Non-Bt Cotton: Implications for Evolution and Resistance Management

The host acceptance of neonate Alabama argillacea (Hübner) (Lepidoptera: Noctuidae) larvae to Bt cotton plants exerts a strong influence on the potential risk that this pest will develop resistance to Bt cotton. This will also determine the efficiency of management strategies to prevent its resistance such as the “refuge-in-the-bag” strategy. In this study, we assessed the acceptance of neonate A. argillacea larvae to Bt and non-Bt cotton plants at different temperatures during the first 24 h after hatching. Two cotton cultivars were used in the study, one a Bt DP 404 BG (Bollgard) cultivar, and the other, an untransformed isoline, DP 4049 cultivar. There was a greater acceptance by live neonate A. argillacea larvae for the non-Bt cotton plants compared with the Bt cotton plants, especially in the time interval between 18 and 24 h. The percentages of neonate A. argillacea larvae found on Bt or non-Bt plants were lower when exposed to temperatures of 31 and 34°C. The low acceptance of A. argillacea larvae for Bt cotton plants at high temperatures stimulated the dispersion of A. argillacea larvae. Our results support the hypothesis that the dispersion and/or feeding behavior of neonate A. argillacea larvae is different between Bt and non-Bt cotton. The presence of the Cry1Ac toxin in Bt cotton plants, and its probable detection by the A. argillacea larvae tasting or eating it, increases the probability of dispersion from the plant where the larvae began. These findings may help to understand how the A. argillacea larvae detect the Cry1Ac toxin in Bt cotton and how the toxin affects the dispersion behavior of the larvae over time. Therefore, our results are extremely important for the management of resistance in populations of A. argillacea on Bt cotton.     

Stylet penetration behaviors of the cotton aphid Aphis gossypii on transgenic Bt cotton

Stylet penetration behaviors of cotton aphids Aphis gossypii Glover on a transgenic cotton line "GK-12" expressing Bt toxic protein of Cry1A (Bt cotton) and a non-Bt conventional cotton line "Simian-3" (CK cotton) were recorded with the direct current electrical penetration graph (DC-EPG) technique. Cotton aphids reared on Bt cotton (abbreviated as Bt-aphids) and its parental non-Bt control line (CK-aphids) for more than 20 generations each, were used for recordings on two cotton lines. Among 47 selected parameters reflecting the activities of aphid stylets within plant tissues, there were eight parameters of CK-aphids showing significant differences between the performances of CK-aphids on Bt cotton and CK cotton, while for Bt-aphids, all the parameters were statistically equal between the performances on the two cotton lines. All parameters with significant differences indicated that CK-aphids could penetrate into Bt cotton more easily, but the phloem saps of Bt cotton were not as good as those of regular cotton for CK-aphids. Based on the present results, we concluded that there were some factors in Bt cotton affecting penetration behaviors of CK-aphids, but it just took several generations for CK-aphids to completely adapt Bt cotton, and Bt-aphids could feed on two cotton lines without difficulty.     

Impact of transgenic cotton plants on a non-target pest, Aphis gossypii Glover

Abstract.  1. Transgenic crops have shown great promise for the control of target pest insects, but in some cases they can also influence non-target species. This study investigated the impact of Bt and Bt+CpTI transgenic cottons on the non-target cotton aphid, Aphis gossypii Glover, by comparing life-table parameters, feeding behaviour, and the fluctuating asymmetry of morphological traits of aphids reared on transgenic cotton and those on untransformed control plants.
2. Aphids on the Bt+CpTI cotton showed a shorter reproductive duration and maximum lifespan, lower survival rates and potential maximum fecundity, and an earlier occurrence of peak daily mortality in the first or second generation. However, the aphid population soon developed fitness and overcame the negative effect in the second or third generation. The aphids on the Bt cotton had significantly longer reproductive durations in the first generation, higher survival rates in the third generation, and apparently larger potential maximum fecundity in all three generations.
3. The percentages of accumulated duration of feeding waveforms E1 and E2 were significantly lower in aphids on the Bt+CpTI cotton than in those on the Bt or control cotton, whereas the frequencies of moving and finding feeding sites, and probe behaviour were significantly higher.
4. Fluctuating asymmetry in three morphological characters of aphids reared on transgenic and control cotton was detected. The fluctuating asymmetry value of the third segment of antenna in aphids on Bt+CpTI cotton was significantly higher than that of aphids on Bt or control cotton. Based on the fluctuating asymmetry value, the stress of cotton on the aphids could be ranked as Bt+CpTI cotton > Bt cotton > control cotton.     

Vertical and Temporal Distribution of Aphis gossypii Glover and Coccinellid Populations on Different Chilli (Capsicum annuum) Varieties

The vertical and temporal distribution of an aphid, Aphis gossypii Glover, and the coccinellid populations on six chilli varieties were studied. The total number of apterous aphid per plant stratum was significantly different among plant strata of a particular variety (treatment) as well as among the treatments. Generally, the total number of aphids was significantly greater in the lower stratum than in the middle and upper strata. However, the varieties with erect and open plant architecture (Kulai and MC 11) had significantly less number of apterous aphids at all strata as compared to varieties with compact or prostrate plant architectures. There was a significant difference in the total number of coccinellids per plant strata among the treatments but not within a treatment. The distribution of apterous aphid populations varied significantly among sampling periods and treatments. The temporal distribution of coccinellids showed a similar trend as that of apterous aphids. The total number of alate aphids caught per week was significantly different among the sampling periods. However, its population was significantly greater during the early season and gradually declined as the season progressed except during June 18 to 24. The importance of recording the most observed coccinellids species, which limit the aphid populations at each particular plant stratum per variety, and the conditions that favor natural enemies are also discussed.     

Leaf surface factors of transgenic Bt cotton associated with the feeding behaviors of cotton aphids: A case study on non-target effects

The present paper reports case study results of the risk assessment of transgenic Bt cotton on a non-target pest, cotton aphid, Aphis gossypii. Several types of techniques, i.e., electrical penetration graph (EPG), light and electron microscopy, bioassays and chemical analysis, were applied to investigate physical and chemical leaf factors of 2 transgenic Bt cotton lines (GK12 and GK19) and their pa-rental non-Bt cotton line (Simian3) associated with searching and feeding behaviors of cotton aphids on leaves or leaf extracts of cotton plants. EPG results showed that there were some differences among behaviors of cotton aphids on 2 Bt cotton and 1 non-Bt cotton lines. Cotton aphids performed similarly to leaf surface extracts from 3 cotton lines; and leaf surface chemicals, mainly volatiles and waxes, were almost identical in the components and concentrations among the cotton lines. However, three cotton lines were quite different from each other in the densities of certain kinds of covering trichomes. Therefore, the relationships between the physical characteristics and the searching behaviors of cotton aphids on the three cotton lines were constructed as the regression equations. Glandular trichomes and covering trichomes with 5 branches influenced the cotton aphids' searching behaviors effectively; and other trichomes with other branches affected aphids in varying ways. These results demonstrated that leaf surface physical factors of transgenic Bt cotton lines different from their parental non-Bt line could affect the penetration behaviors of non-target cotton aphids. Cotton aphids penetrate and feed more easily on two Bt cotton lines than on the non-Bt cotton line.     

Tri-trophic interactions between Bt cotton, the herbivore Aphis gossypii Glover (Homoptera: Aphididae), and the predator Chrysopa pallens (Rambur) (Neuroptera: Chrysopidae)

Tri-trophic impacts of transgenic Bacillus thuringiensis (Bt) cotton GK12 and NuCOTN 99B were studied using a predator, the great lacewing Chrysopa pallens (Rambur), and its prey, the cotton aphid Aphis gossypii Glover, in laboratory feeding experiments. The parental nontransgenic cotton cultivar of GK12 was used as control. The predator was fed with uniform (aphids from a single cultivar) or mixed prey (aphids from the three cotton cultivars provided on alternate days). Mortality and development of the immature stages, pupal body mass, adult sex ratio, fecundity, and egg viability of C. pallens were measured. When fed GK12-originated aphid prey, pupal body mass of C. pallens was significantly higher than that of the control, more females emerged, and these females laid significantly more eggs. Other parameters were not impacted. Females emerging from larvae maintained on NuCOTN 99B-originated prey laid fewer eggs than those maintained on GK12. Other measurements did not differ significantly between the two Bt cotton cultivars. Compared with the control, mixed feeding significantly prolonged pupal development time and increased pupal body mass and percentage of females but did not affect other parameters. These results indicate that C. pallens is sensitive to aphid prey from different cotton cultivars. Transgenic Bt cotton GK12-originated aphid prey has no adverse impact on survival, development, and fecundity of C. pallens. Between the two Bt cotton cultivars, NuCOTN 99B-originated aphid prey provided to C. pallens in the larval stage may lower female fecundity. Mixed feeding of C. pallens with the two Bt cotton-originated prey and non-Bt prey may have some adverse impacts on pupal development.     

用微卫星引物PCR分析棉蚜不同蚜型的DNA多态性

用微卫星引物PCR方法, 对棉蚜Aphis gossypii干母、干雌、孤雌蚜(有翅迁飞孤雌蚜和无翅孤雌蚜)、性母、性雌蚜和雄蚜的DNA多态性进行分析。 结果表明：① 棉蚜的有性蚜型和无性蚜型之间在遗传上有较大的差异; ② 性母与性雌蚜和雄蚜的遗传关系都十分接近,说明同一性母既产性雌蚜,也产雄蚜;③ 干母和干雌的遗传关系很近, 但孤雌蚜已与二者有分化。     

转RRM2基因棉和其亲本苗蚜种群数量及其优势天敌的动态规律

【目的】转基因作物的大规模种植,可能会对人类健康和生态环境造成影响。因此,商业化种植之前,评价环境安全性十分必要。【方法】以转基因(RRM2)高产棉为实验品种,受体材料中棉所12为对照品种,分别于2013年和2014年连续2年对2种棉田的苗期蚜虫及其几种主要捕食性天敌的田间种群数量进行系统的田间调查,并比较它们在这2种不同棉田间的差异。【结果】与中12相比,转RRM2基因棉苗期无翅蚜的发生数量显著增加,有翅蚜迁入棉田的数量也有所增加,但二者差异不显著;2个棉花品系间棉蚜的几种主要捕食性天敌发生数量也无明显差异。【结论】与亲本材料相比,转高产棉花苗蚜数量显著增加,但其捕食性天敌数量增加不明显。本研究为转RRM2高产基因棉花环境安全评价技术的完善提供了理论依据。     

棉蚜寄主专化型及其形成的行为机理

通过生活在甜瓜和棉花上的棉蚜Aphisgossypii Glover的行为,研究棉蚜的寄主专化型及其形成的行为机理。生物学观察显示： 两类棉蚜在寄主植物相互交换以后,定居数显著减少,棉花蚜型棉蚜的繁殖系数及若虫存活率显著下降,说明棉蚜存在甜瓜蚜型和棉花蚜型两种寄主专化型。通过刺探电位技术研究棉蚜的取食行为,以探索其寄主专化型形成的行为机理。结果表明： 甜瓜蚜型棉蚜在棉花上的取食行为容易被中断,但其口针定位韧皮部的能力并没有显著削弱;而棉花蚜型棉蚜在甜瓜上的取食行为受到更大的影响,口针无法顺利定位至韧皮部,并在2 h内根本无法在筛管内取食。生物学观察和EPG取食行为分析都显示： 与甜瓜蚜型棉蚜相比,棉花蚜型棉蚜对寄主的要求更严格-寄主专化程度更高,对寄主的利用率更高。     

Transgenic Bt maize and Rhopalosiphum padi (Hom., Aphididae) performance

Abstract.  1. The population abundance and age structure of Rhopalosiphum padi , one of the most common maize aphid species, on transgenic Bt (expressing the Cry1Ab protein) and non-Bt isogenic maize was studied in commercial plots during three crop seasons.
2. A higher density of aphids, particularly alates and young nymphs, occurred in Bt plots at very young maize development stages, corresponding to the settlement period, in the 3 years studied. Possible causes are discussed. After this period, there were no differences between Bt and non-Bt maize.
3. Mortality, development, and reproduction of the offspring of alate forms of R. padi and the offspring of different generations of apterous forms fed with Bt maize were evaluated in the laboratory under controlled conditions.
4. The developmental and pre-reproductive times of the offspring of the first generation of alatae were shorter and the intrinsic rate of natural increase ( r _m ) higher when aphids fed on Bt maize. The opposite occurs with the offspring of the first generation of apterous mothers, which have lower nymphal and adult mortality, shorter developmental and pre-reproductive times, a higher effective fecundity rate, and greater r _m , when fed on non-Bt maize. The differences in aphid development on the two cultivars may be linked to changes in host-plant quality due to pleiotropic effects of the genetic modification.
5. No differences on aphid mortality, developmental and pre-reproductive times, fecundity, and r _m were found between the offspring of apterous aphids maintained on Bt or non-Bt maize for several generations.     

Predation of cotton bollworm by green lacewings in the presence of cotton aphid as alternative prey on transgenic Bt and conventional cotton

Experiments were conducted in small arenas and on whole plants to explore the effect of cotton aphids, Aphis gossypii Glover (Hemiptera: Aphididae), as alternative prey on the predation of Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) larvae by green lacewing larvae, Mallada signatus Schneider (Neuroptera: Chrysopidae). Transgenic Bt (Bollgard II^®) and conventional cotton plants were included to explore potential differences in the predator's performance on these cotton types. In small arenas, the presence of 20 aphids reduced predation on H. armigera larvae by 22% (from 5.5 to 3.3 of 10) by a single lacewing larva over a 24‐h period. The presence of H. armigera reduced predation on aphids by ca. 29% (from 16.8 to 11.0 of 20) over 24 h. On whole plants, the presence of alternative prey had no effect on the number of H. armigera larvae or aphids remaining after 3 days. The presence of H. armigera larvae alone, without the predator, caused a 24% reduction in the numbers of aphids on conventional, but not on Bt cotton plants. The combination of Bt cotton and lacewing larvae caused a 96.6% removal of early‐stage H. armigera larvae, a statistically significant increase over the addition of the proportions (91.6%) removed by each factor measured separately, providing evidence of synergism. These studies suggest that the presence of aphids as alternative prey would not necessarily disrupt the predation by green lacewing on larvae of H. armigera, especially on Bt cotton.     

棉花苗期棉蚜的二项式抽样设计及其精度分析

基于棉花苗期棉蚜（Ａｐｈｉｓ ｇｏｓｓｙｐｉｉ）的２４组调查数据,利用每样方虫口不超过数阈值Ｔ（为０,１,２,…,９,１０,１５,２０,３０）头蚜虫的植株比例（ＰＴ）与种群密度（ｍ,头／株）的关系,拟合经验关系式１ｎ（ｍ）＝α＋ｂ１ｎ〔－１ｎ（ＰＴ）〕设计二项式抽样。通过对不同数阈值Ｔ的决定系数（ｒ＾２）、估计方差（Ｖａｒ（ｍ））和抽样精度（ｄ估计）等进行综合分析,结果表明该蚜虫在数阈值Ｔ为１５时     

Reproductive biology of two nontarget insect species, Aphis gossypii (Homoptera: Aphididae) and Orius sauteri (Hemiptera: Anthocoridae), on Bt and non-Bt cotton cultivars

Transgenic Bt cotton, engineered to continuously produce activated delta-endotoxins of the soil bacteria Bacillus thuringiensis, holds great promise in controlling Helicoverpa armigera and other lepidopteran pests. However, it also may impact the invertebrate community, which needs to be clarified. The effects of Bt cotton on two nontarget insects, Aphis gossypii and Orius sauteri, were assessed under semifield and laboratory conditions. Mean total duration of nymphal stages of A. gossypii was shorter (5.9 versus 6.3 d), and rm was higher (0.418 versus 0.394) on conventional Simian 3 (the most frequently planted non-Bt cotton in northern China) than on Bt transgenic NuCOTN 33B (the first Bt cotton commercially planted in China). Mean duration of fourth-instar O. sauteri was significantly longer on transgenic GK-12 (3.7 d) than on NuCOTN 33B (3.2 d), but no different from Simian 3. Mean total mortality was significantly lower on Simian 3 (3.7%) than on GK-12 (14.8%). During the fourth instar, the predator consumed a significantly higher number of prey on Simian 3 (202.3 prey) than on NuCOTN 33B (159.0), whereas the mean total number of A. gossypii prey consumed during the nymphal stage was significantly higher on Simian 3 (336.8 prey) and GK-12 (330.3 prey) than on NuCOTN 33B (275.7). No detrimental effects were detected on development (nymphs, adults, and progeny eggs), fecundity, longevity, and egg viability of O. sauteri on Bt cotton aphids compared with non-Bt cotton aphids. These results suggest that Bt cotton cultivars GK-12 and NuCOTN 33B have no direct effect on nontargets A. gossypii and O. sauteri. Germplasm divergence may account for the negative effects observed on A. gossypii and O. sauteri when reared on NuCOTN 33B or NuCOTN 33B-fed aphids. The biological meanings of the small difference observed between GK-12 and Simian 3 on survival of O. sauteri will require close monitoring over longer time periods.     

Field and laboratory evaluations of transgenic cottons expressing one or two Bacillus thuringiensis var. kurstaki Berliner proteins for management of noctuid (Lepidoptera) pests

Field studies were conducted from 1999 to 2001 to evaluate the efficacy of the transgenic cotton, Gossypium hirsutum (L.), genotype, Bollgard II (Monsanto 15985), which expresses two Bacillus thuringiensis Berliner (Bt) proteins (Cry1Ac + Cry2Ab) that are active against lepidopterous pests. Bollgard II was compared with Bollgard (DP50B), which expresses only one Bt protein (Cry1Ac), and, in all tests, the conventional variety, DP50, was used as a non-Bt control. Larval populations of the bollworm, Helicoverpa zea (Boddie), and the soybean looper, Pseudoplusia includens (Walker), were significantly lower in Bollgard II than in Bollgard and conventional cotton, and the proportion of fruit damaged by H. zea was also lower. Fall armyworm, Spodoptera frugiperda (J. E. Smith), populations were lower in Bollgard II than in Bollgard, although not significantly. Field tests were supplemented with laboratory bioassays in 2001 to compare mortality of S. frugiperda, and beet armyworms, Spodoptera exigua (Hübner), feeding on these genotypes. Mortality of both species was significantly greater on Bollgard II plant material than on either Bollgard or conventional cotton. This study demonstrated that the dual-toxin Bollgard II genotype is highly effective against lepidopterous pests that are not adequately controlled by the current single-toxin Bollgard varieties. If toxin expression in future Bollgard II varieties remains consistent with that of Monsanto 15985, supplemental insecticides will be reduced, and may be eliminated for lepidopterous pests in South Carolina.     

Cytopathology, Mode of Aphid Transmission and Search for the Casual Agent of Cotton Bunchy Top Disease

The cytopathological effects of cotton bunchy top (CBT) disease and its mode of transmission by Aphis gossypii Glover (cotton aphid), were studied. CBT infection affected the leaf epidermal layer producing a loose, ruptured and rough surface morphology with many stomata closed and misshapen. Roots of CBT‐infected plants showed reduced growth, small knots and a dark brown appearance. A single aphid per plant was capable of transmitting CBT at 5%, whereas three aphids per plant transmitted CBT to 50% of the cotton seedlings and 20 aphids per plant transmitted the disease agent to 80% of the cotton seedlings. Aphis gossypii acquired CBT after a minimum acquisition access period of 5 min and transmitted the agent after a minimum inoculation access period of 1 h. Both alate and apterous aphids and nymph instars 2, 3 and 4 of A. gossypii transmitted CBT. This preliminary data suggest that A. gossypii transmits CBT in a semi‐persistent manner. Myzus persicae Sulz (green peach aphid) was unable to transmit CBT. A comprehensive attempt to isolate the CBT agent, using a range of virological techniques including double‐stranded RNA extraction, two‐dimensional gel electrophoresis for viroid, circular DNA test, nanovirus polymerase chain reaction (PCR), luteovirus PCR and enzyme‐linked immunosorbent assay, phytoplasma test, nucleoprotein purification and electron microscopy, was unsuccessful, raising the possibility that CBT may be caused by a unique new pathogen.     

麦长管蚜和禾谷缢管蚜对小麦挥发物的触角电位反应

采用活体蚜虫测定法,利用EAG技术比较分析了麦长管蚜Sitobion avenae和禾谷缢管蚜Rhopalosiphum padi有翅及无翅成蚜对小麦挥发物及麦蚜取食诱导挥发物组分的嗅觉反应,揭示了两种麦蚜的嗅觉变异特点。结果表明：麦长管蚜对水杨酸甲酯、反-2-己烯醛、反-2-己烯醇、-6-甲基-5-庚烯-2-酮和-6-甲基-5-庚烯-2醇的反应较强,禾谷缢管蚜对水杨酸甲酯、反-3-己烯乙酸酯、-6-甲基-5-庚烯-2-酮和-6-甲基-5-庚烯-2-醇的反应较强,并得到了剂量反应曲线。麦长管蚜的有翅和无翅成蚜对6-甲基-5-庚烯-2-酮、反-2-己烯醇和水杨酸甲酯的反应差异显著;禾谷缢管蚜的有翅和无翅成蚜对反-2-己烯醇、辛醛、里那醇、水杨酸甲酯和反-3-己烯乙酸酯的EAG反应差异显著,其原因与禾谷缢管蚜迁移及转主为害的生物学特性有关。     

Field efficacy and seasonal expression profiles for terminal leaves of single and double Bacillus thuringiensis toxin cotton genotypes.

Examination of commercial Cry1Ac transgenic Bacillus thuringiensis Berliner (Bt) cotton varieties (Bollgard, Monsanto, St. Louis, MO) and an experimental Cry1Ac + Cry2Ab transgenic Bt cotton variety (Bollgard II, Monsanto) for lepidopteran field efficacy was conducted during the 2000 growing season. In addition, a commercially available (Envirologix, Portland, ME) quantification assay (ELISA) was used to measure and profile the expression levels of Cry proteins in two of these varieties ['DP50B, Bollgard'; 'DP50BII, Bollgard II' (Delta & Pine Land, Scott, MS)]. Populations of beet army worms, Spodoptera exigua (Hübner), and soybean loopers, Pseudoplusia includens (Walker), were significantly lower (P < 0.05) in Bollgard II plots compared with Bollgard. Population numbers for fall army worms, Spodoptera frugiperda (J. E. Smith), and salt marsh caterpillars, Estigmene acrea (Drury), were lower in Bollgard II plots compared with Bollgard but means did not differ significantly (P > 0.05). Single and dual-toxin genotypes remained superior (P < 0.05) compared with conventional cotton against the tobacco budworm, Heliothis virescens (F.). The addition of Cry2Ab had no significant (P > 0.05) impact on Cry1Ac expression in Bollgard II compared with Cry1Ac expression in Bollgard. Furthermore, throughout the season Cry2Ab was present at much higher levels in the plant compared with Cry1Ac for Bollgard II plants. Possible species-specific reasons for increased efficacy of Bollgard II over Bollgard are discussed.