Interaction between host plant resistance and biological activity of Bacillus thuringiensis in managing the pod borer Helicoverpa armigera in chickpea

The legume pod borer Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae) has developed high levels of resistance to conventional insecticides, and therefore, efforts are being made to develop transgenic chickpea expressing toxin genes from the bacterium Bacillus thuringiensis (Bt) for controlling this pest. However, there is an apprehension that acid exudates in chickpea might interfere with the biological activity of Bt. Therefore, we studied the biological activity of Bt (Biolep^R) on four chickpea genotypes with different levels of resistance to H. armigera under field conditions, and by incorporating lyophilized leaf and pod tissue into the artificial diet with and without Bt. The pH of the acid exudates varied from 2.1 to 2.9, and malic and oxalic acids were the major components of the acid exudates in different chickpea genotypes. There was no survival of H. armigera larvae in chickpea plants sprayed with 0.1, 0.2 and 0.5% Bt. There was a significant reduction in larval survival, larval and pupal weights and fecundity, and prolongation of larval and pupal periods in chickpea plots sprayed with Bt (0.05%) as compared to the unsprayed plots. Biological activity of Bt was lower on artificial diets with leaf or pod powder of chickpea genotypes, which might be because of a low intake of Bt toxins due to the antifeedant effects of acid exudates in the chickpea or reduction in biological activity of Bt due to the interaction of biochemical constituents in chickpea with the Bt toxins. Larval survival, larval and pupal weights, pupation and adult emergence were significantly lower on diets with leaf or pod powder of the H. armigera-resistant genotypes than on the susceptible check. Chickpea genotypes with resistance to H. armigera acted in concert with Bt to cause adverse effects on the survival and development of this insect. The results suggested that development of transgenic chickpeas expressing toxin genes form Bt will be quite effective for controlling of the pod borer, H. armigera.     

Susceptibility of Helicoverpa armigera from different host plants in northern China to Bacillus thuringiensis toxin Cry1Ac

Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) is a serious pest of cotton and many other crops in northern China. To evaluate the contribution of alternative hosts as an effective refuge for transgenic cotton expressing the Bacillus thuringiensis (Bt) Cry1Ac toxin, the susceptibility to this toxin was measured in progeny derived from field-collected H. armigera larvae and pupae from different hosts in the Xiajin’s region of the Shandong Province in northern China. During 2008-2010, progeny from a total of 258,56,184 and 160 single-pair crosses derived from wheat (first-generation), Bt cotton (second-generation), Bt cotton (third-generation), and corn (third-generation) were screened on Cry1Ac diets, respectively. Based on relative average development rates (RADR) of H. armigera larvae in these F₁ tests, the second and third-generation moths emerging from Bt cotton fields were more tolerant to the Bt toxin than the first and third-generation moths emerging from wheat and corn each year. These results suggest that there is significant variation in susceptibility to Bt toxins among H. armigera populations derived from different host crops. Alternate crops, such as corn, that maintain Bt susceptible populations of H. armigera could be used as refugia to minimize the evolution of resistance to Bt cotton.     

Relationships between Cry1Ac and Cry2Ab protein expression in field-grown Bollgard II® cotton and efficacy against Helicoverpa armigera and Helicoverpa punctigera (Lepidoptera: Noctuidae)

Seven-day laboratory bioassays with first-instar Helicoverpa armigera and Helicoverpa punctigera larvae were conducted using leaves from field-grown Bollgard II^® cotton during the 2009/10 season and larval mortality and development recorded. Leaves were from three farms in the St George region and two fields on one farm at Emerald in eastern Australia. The Cry1Ac and Cry2Ab content of leaves from the same samples was determined (see Knight et al., 2013) and the relationships between Cry protein content and larval mortality and development were examined. The Cry1Ac protein content was between 3.98 and 12.08 μg/g during the growing season, while Cry2Ab content ranged between 300.6 and 953.3 μg/g. Cry1Ac and Cry2Ab content of leaves were highly correlated (r = 0.8276, P < 0.001). Seven-day mortality of H. punctigera larvae was close to 100% throughout the season. H. armigera mortality was close to 100% early in the season, but fell to ∼65% by mid-to late February in the laboratory bioassays. Fitting three-dimensional non-linear models associating Cry1Ac and Cry2Ab content with H. armigera and H. punctigera mortality elucidated the relative importance of the two proteins in determining larval mortality; for this analysis, data were pooled with data from an isoline study to provide better sampling of the three-dimensional surface being modelled. For both Helicoverpa species, the fitted mortality response to the Cry1Ac protein was close to its maximum at protein concentrations above ∼3 μg/g Cry1Ac. For H. punctigera, response to the Cry2Ab protein was close to maximal once Cry2Ab was greater than ∼200 μg/g. In contrast, the fitted H. armigera mortality response to Cry2Ab increased steadily with concentration up to ∼1200 μg/g Cry2Ab. These responses led to markedly different response surfaces for the two species; H. punctigera mortality was close to 100% at most places on the response surface, while for H. armigera the response surface showed stronger increases in mortality with concentration for Cry2Ab than for Cry1Ac. These results can be interpreted as meaning that at the plant-expressed range of concentrations in Bollgard II cotton the two proteins are approximately equally important for H. punctigera but that changes in Cry2Ab content more strongly influences changes in larval mortality in H. armigera than does Cry1Ac, with Cry1Ac contributing a consistent 40–45% mortality for concentrations above 3 μg/g. For H. armigera, there was no evidence of either synergism or antagonism between Cry1Ac and Cry2Ab proteins (P > 0.05) but this aspect was not testable for H. punctigera because mortality was mostly close to 100%.     

Frequency of Bt resistance alleles in Helicoverpa armigera in 2007-2009 in the Henan cotton growing region of China

Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) is a key insect pest of cotton in the Henan cotton growing region of China. In this region, cotton is grown on small acreages in rich agricultural landscapes, contrary to cropping systems in the United States or Australia. Under such cropping regimes, naturally occurring refuges (with non-Bt plants) may be sufficient to prevent H. armigera resistance development to Bt toxins. In order to gain a timely understanding of the evolution of resistance of H. armigera to Bt toxin after continuous cultivation of Bt cotton for c. 10 years, we assessed the frequency of alleles conferring resistance to Cry1Ac toxin in field populations of H. armigera sampled from Xinxiang County in Henan province during 2007-2009. Screening F₁ and F₂ generations from isofemale lines, derived from female moths trapped in the field, were used with a discriminating dose of Cry1Ac diet to estimate the frequency of resistance alleles. Totals of 625, 516 and 488 isofemale lines were screened for the F₁ generation in 2007, 2008 and 2009, respectively. Resistance gene frequency in Xinxiang fluctuated between 0.0000 and 0.0005, and it did not increase significantly from 2007 to 2009. Based on the relative average development rates (RADR) of H. armigera larvae in F₁ tests, no substantial increase in Cry1Ac tolerance was found in the Xinxiang region over the 3-yr period.     

Relative abundance and damage by target and non-target insects on Bollgard and BollgardII cotton cultivars

Bollgard^® and BollgardII^® cotton cultivars were evaluated for their efficacy for control of bollworms and their effects on sucking insects and the abundance of natural enemies under bollworm insecticide protected and non-insecticide protected field conditions during the 2004 and 2005 cropping seasons. Bollgard cultivars are genetic transforms that produce Cry1Ac toxins from Bacillus thuringiensis (Bt) to control American bollworm, Helicoverpa armigera. Second generation BollgardII cultivars contain 2 toxins, Cry1Ac and Cry2Ab, and are more effective in controlling a broader range of caterpillar species. Bollgard cultivars were free of H. armigera damage until harvest under insecticide protected as well as non-protected conditions. Both BollgardII and Bollgard cultivars were infested with larvae of spotted bollworm, Earias vitella, and spiny bollworm, Earias insulana, at later crop growth stages. Neither BollgardII nor Bollgard cultivars were free of square (fruiting body), open boll and loculi damage. Bollworm damage did not reach economic threshold levels up to harvest. Densities of sucking insects (Amrasca biguttula biguttula, Bemisia tabaci, Aphis gossypi, and Thrips tabaci), of the foliage feeder Myllocerus undecimpustulatus and of predators (Chrysoperla spp., Orius spp., Coccinella spp., Brumus spp., Vespa spp., Lycosa spp., and Aranews spp.) were similar on Bollgard, BollgardII and conventional cultivars. The time of the first appearance of bollworms, sucking insects and predators on Bt cotton did not vary from conventional cotton varieties. Insecticidal protection based on economic threshold levels resulted in significant reductions in bollworm damage on conventional cultivars.     

Impacts of transgenic Bt cotton on a non-target pest, Apolygus lucorum (Meyer-Dür) (Hemiptera: Miridae), in northern China

Transgenic Bt (Bacillus thuringiensis) cotton has been effectively used to control the cotton bollworm, Helicoverpa armigera (Hübner) in China. However, in recent years, following the wide commercialization of Bt cotton in northern China there have been frequent outbreaks of the non-target pest Apolygus lucorum (Meyer-Dür). To clarify how transgenic cotton contributes to these outbreaks, a four-year field investigation of population dynamics and laboratory life table studies were carried out from 2007 to 2010 to evaluate the impact of two transgenic cotton cultivars (SGK321 expressing Cry1Ac + CpTI and GK12 expressing Cry1Ac) and their corresponding parental non-transgenic lines (Shiyuan321 and Simian3) on A. lucorum. There were no significant differences in the population densities of A. lucorum found in Bt cotton and non-Bt cotton plots, whether one compared those that had received insecticide treatments or those that had not. However, population densities of A. lucorum were significantly lower in pesticide treated plots than in controls. Furthermore, there were no significant differences in the net reproductive rates, generation times or intrinsic rates of increase of A. lucorum when reared on either Bt or non-Bt cotton cultivars. These results suggest that Bt cotton has no direct positive or negative effects on the biology of A. lucorum, so the most logical explanation for the observed outbreaks is the decrease in pesticide applications following the commercial release of Bt cotton.     

Significance of host-plant phenology in the dynamics and pest incidence of the cotton bollworm,Heliothis armigera Hübner (Lepidoptera: Noctuidae), in Western Tanzania

Heliothis armigera is the most important cotton pest in Tanzania, and also attacks several food crops grown in a relay-intercropping system practised by small-scale farmers in Western Tanzania. The level and timing of H. armigera infestations were studied on its main alternative host plants: maize, sorghum, cotton, chickpea, tomato and the wild host Cleome sp. The attractive stages of these hosts overlap throughout the year. The expansion of chickpea and the introduction of tomato production has enabled the pest to survive better during the dry season, thus bridging an otherwise unfavourable period. The recent increased popularity of maize as a food crop, coupled with the introduction of compact and semi-compact panicle sorghum varieties instead of the traditional open-panicle sorghum cultivars, together with the expansion of cotton acreage, has increased the severity of attacks on sorghum and cotton. Effective control of the pest on early-sown sorghum offers the possibility of reducing numbers that subsequently attack early-season cotton. Although early-sown maize and sorghum can generate heavy populations which attack cotton in February–March, later crops flowering in March–April can usefully divert the pest away from cotton. Some of the alternative host plants, notably the weed, Cleome sp., are important sources of pesticide-susceptible strains of the pest population, which may decrease the likelihood of pesticide-resistant strains developing.     

Transgenic Bt corn varietal resistance against the Mexican rice borer,Eoreuma loftini (Dyar) (Lepidoptera: Crambidae) and implications to sugarcane

The Mexican rice borer, Eoreuma loftini (Dyar), attacks crops including corn, Zea mays L., rice, Oryza sativa L., sorghum, Sorghum bicolor (L.) Moench, and sugarcane, Saccharum spp. Strongly resistant varieties of any kind, native or otherwise, have not been identified. A field plot corn variety test using two transgenic Bt varieties, Pioneer 31G71, expressing the Cry1F insecticidal protein, and Golden Acres 28V81, expressing the Cry1A.105, Cry2Ab2, and Cry3Bb1 insecticidal proteins, and two non-Bt controls, Dekalb DKC 69-72 and BH Genetics 9050, all four commonly grown in the Lower Rio Grande Valley of Texas, showed that, although oviposition preference was not affected, 28V81 resisted larval stalk boring to the extent that Mexican rice borer injury was almost non-existent. Pioneer 31G71 was infested nearly as much as the controls, but larval development to adulthood was reduced by ≈70%. Rearing larvae on 5, 50, 500, and 5000 μg of corn leaf tissue per ml of artificial diet showed that, while the three lowest concentrations did not affect larval growth and development, the high concentration of 28V81 reduced survivorship to the pupal stage, decreased weight of 4-wk-old larvae, and prolonged development to pupation. Lower numbers of pheromone trap-captured adults at the edges of commercial Bt and non-Bt corn fields showed that populations were lower at the Bt cornfields, suggesting a lesser rate of adult production. Because corn is a preferred host plant over sugarcane, sorghum or rice, use of resistant transgenic Bt corn varieties will likely protect the crop from the substantial injury that can be caused by the pest. This study also suggests that Bt genes might result in similarly strong resistance when inserted in other vulnerable crops such as sugarcane.     

Toxicity of several δ-endotoxins of Bacillus thuringiensis against the cotton pest Earias insulana (Lepidoptera: Noctuidae)

The toxicity of nine Bacillus thuringiensis Cry proteins against neonate Earias insulana larvae was tested using a mixture of crystals and spores. The mean lethal concentration (LC₅₀) of Cry1Ac was 1.99 μg/ml. Cry1Fa, Cry1Ca, Cry1Ja and Cry2Aa were more active than Cry1Ac, with LC₅₀ values of 0.22, 0.24, 0.29, 0.43 μg/ml, respectively. Cry1Da and Cry1Aa were considerably less active than Cry1Ac. The remaining proteins, Cry1Ba and Cry1Ab, displayed no activity. Relative potencies were also calculated. Cry1Ja and Cry1Fa were significantly more active (7.72 and 5.71 times, respectively) than Cry1Ac, while Cry1Ca was significantly (1.95 times) more active than Cry2Aa.     

Feeding disruption tests for monitoring the frequency of larval lepidopteran resistance to Cry1Ac, Cry1F and Cry1Ab

An alternative to traditional larval lepidopteran resistance-monitoring bioassays was developed. Feeding disruption tests were developed for detecting insects resistant to three Bacillus thuringiensis (Bt) proteins: Cry1Ac, Cry1F and Cry1Ab. The assays rely on a diagnostic dose of Bt toxin in 100-μl hydratable meal pads of artificial diet containing blue indicator dye. The assay was formatted as a portable (palm-sized) plastic plate containing an array of 16 test wells, each containing a single hydratable meal pad with one insect added per well. The diagnostic dose was the concentration of Bt in meal pad rehydration solution that reduced 24 h dyed fecal production of Bt-susceptible neonates to ≤2 fecal pellets per larva. Bt-resistant neonates were able to consume the diagnostic dose of the insecticidal protein and produce >2 blue fecal pellets. The feces were distinctly visible on the white background of the feeding disruption test plate. Diagnostic doses were determined with lab-strain Bt-susceptible Heliothis virescens and Helicoverpa zea. For H. virescens, the diagnostic doses were 10, 20 and 15 μg/ml for Cry1Ac, Cry1F and Cry1Ab, respectively. For H. zea, the diagnostic doses were 40, 200 and 500 μg/ml, respectively. The assays were validated against a lab-strain of Bt-resistant H. virescens and with susceptible larval H. virescens collected as eggs from field-grown tobacco in North Carolina.     

Comparative susceptibility of Ostrinia furnacalis, Ostrinia nubilalis and Diatraea saccharalis (Lepidoptera: Crambidae) to Bacillus thuringiensis Cry1 toxins

Transgenic corn hybrids that express toxins from Bacillus thuringiensis (Bt) are highly effective against the European corn borer, Ostrinia nubilalis (Hübner), and the closely related Asian corn borer, Ostrinia furnacalis (Guenée). Since the registration of Bt corn hybrids in the U.S. in 1996, there has been a great deal of information generated on O. nubilalis. However, relatively little information exists for O. furnacalis. To help determine whether the information generated for O. nubilalis can be leveraged for decisions regarding the use of transgenic Bt corn against O. furnacalis, experiments were designed to determine whether the pattern of sensitivity to various Bt Cry1 toxins is similar between the two species. Test insects included laboratory-reared O. furnacalis originating from Malaysia, a Bt-susceptible laboratory colony of O. nubilalis maintained at the University of Nebraska-Lincoln (UNL) and an out-group consisting of the sugarcane borer, Diatraea saccharalis (F.), from Louisiana which represents a different genus from the same family. O. furnacalis and O. nubilalis exhibited a similar pattern of susceptibility to all the Cry1 toxins and were highly susceptible to the range of Bt toxins tested including Cry1Aa, Cry1Ab, Cry1Ac and Cry1F. Both of the Ostrinia species were more tolerant to Cry1Ba compared with D. saccharalis, although sensitivity of O. furnacalis was intermediate and did not differ significantly from that of O. nubilalis and D. saccharalis. D. saccharalis was also susceptible to the range of toxins tested but unlike the two Ostrinia species, was more tolerant to Cry1F and more susceptible to Cry1Ba. These results indicate that both of the Ostrinia corn borer species are similar in sensitivity to the Cry1Aa, Cry1Ab, Cry1Ac, Cry1Ba and Cry1F toxins, thus suggesting shared toxin receptors and mechanisms of toxicity for the two species.     

Low susceptibility of Spodoptera cosmioides,Spodoptera eridania and Spodoptera frugiperda (Lepidoptera: Noctuidae) to genetically-modified soybean expressing Cry1Ac protein

Spodoptera cosmioides (Walker), Spodoptera eridania (Stoll) and Spodoptera frugiperda (J. E. Smith) have caused significant damage on soybean Glycine max (L.) Merrill in Brazil. Genetically-modified MON 87701 × MON 89788 soybean that expresses the Cry1Ac protein is potentially an alternative tool for the management of these species. Purified protein bioassays were done to evaluate the susceptibility of S. cosmioides, S. eridania and S. frugiperda to Cry1Ac protein. The level of efficacy of the Bt soybean plants in controlling these species was measured through laboratory and greenhouse trials under high artificial insect infestations. The biology of these insects was evaluated over their development cycles to understand their life history when fed on Bt soybean. Purified Cry1Ac protein at the maximum concentration tested (100 μg Cry1Ac mL⁻¹ diet) resulted in low mortality of S. cosmioides and S. eridania (<13%) and intermediate mortality of S. frugiperda (50%). No significant effects of the Bt soybean plants were observed in the life table parameters of S. cosmioides and S. eridania. However, S. frugiperda fed on Bt soybean plants had a prolonged larval stage (by 5 days), reduced larvae viability, increased mean generation time (by 8 days) and reduced intrinsic rate of increase. In general, the Bt soybean plants showed poor control of Spodoptera species when evaluated by leaf-disc bioassay and greenhouse trials. Consequently, other control tactics must be used in combination with MON 87701 × MON 89788 soybean in the field for the efficient management of S. cosmioides, S. eridania and S. frugiperda.     

Relative Fitness of Helicoverpa armigera (Lepidoptera: Noctuidae) on Seven Host Plants: A Perspective for IPM in Brazil

The cotton bollworm Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) is a widespread pest of many cultivated and wild plants in Europe, Africa, Asia, and Australia. In 2013, this species was reported in Brazil, attacking various host crops in the midwestern and northeastern regions of the country and is now found countrywide. Aiming to understand the effects of different host plants on the life cycle of H. armigera, we selected seven species of host plants that mature in different seasons and are commonly grown in these regions: cotton (Gossypium hirsutum, “FM993”), corn (Zea mays, “2B587”), soybean (Glycine max, “99R01”), rattlepods (Crotalaria spectabilis), millet (Pennisetum glaucum, “ADR300”), sorghum (Sorghum bicolor, “AGROMEN70G35”), and cowpea (Vigna unguiculata, “SEMPRE VERDE”). The development time of immatures, body weight, survivorship, and fecundity of H. armigera were evaluated on each host plant under laboratory conditions. The bollworms did not survive on corn, millet, or sorghum and showed very low survival rates on rattlepods. Survival rates were highest on soybean, followed by cotton and cowpea. The values for relative fitness found on soybean, cotton, cowpea, and rattlepods were 1, 0.5, 0.43, and 0.03, respectively. Survivorship, faster development time, and fecundity on soybean, cotton, and cowpea were positively correlated. Larger pupae and greater fecundity were found on soybean and cotton. The results indicated that soybean, cotton, and cowpea are the most suitable plants to support the reproduction of H. armigera in the field.     

Adaptation of Habrobracon hebetor (Hymenoptera: Braconidae) to Rearing on Ephestia kuehniella (Lepidoptera: Pyralidae) and Helicoverpa armigera (Lepidoptera: Noctuidae)

Food characteristics strongly regulate digestive enzymatic activity of insects through direct influences on their midgut mechanisms. Insect performance is better on diets that contain nutrients in proportions that fit its digestive enzymes. Little is known about the influences of rearing history on parasitism success of Habrobracon hebetor Say. This research focused on the effect of nutrient regulation on survival, development, and parasitism of H. hebetor. Life history and digestive enzyme activity of fourth-stage larvae of H. hebetor were studied when reared on Ephestia kuehniella Zeller. This parasitoid was then introduced to Helicoverpa armigera (Hübner), and above-mentioned parameters were also studied in the first and fourth generations after transfer. In term of parasitism success, H. hebetor preferred E. kuehniella over He. armigera. When the first and fourth generations of He. armigera-reared H. hebetor were compared, the rearing history affected the life history and enzymatic activity of the parasitoid. A better performance of H. hebetor was achieved after it was reared on He. armigera for the four generations. Because, digestive α-amylase and general protease of the parasitoid were matched with the new host, it used reserve energy for a better performance. Thus, a better performance of H. hebetor could be obtained when the parasitoid was reared on its original host for at least four generations.     

Acute toxicity and sublethal effects of methoxyfenozide and thiodicarb on survival,development and reproduction of Helicoverpa armigera (Lepidoptera: Noctuidae)

The cotton bollworm Helicoverpa armigera (Hubner) is one of the most destructive pest insects in Iran and many other countries. In this study, lethal and sublethal effects of methoxyfenozide, and thiodicarb were evaluated against H. armigera larvae that fed on insecticide-treated artificial diet. The effects of methoxyfenozide and thiodicarb were assessed in 3rd instars. Methoxyfenozide and thiodicarb showed LC₅₀ values of 4 and 639 mg a.i./ml, respectively, in H. armigera larvae. Sublethal effects on development, adult longevity, and reproduction were observed in H. armigera larvae that survived exposure to an LC₃₀ of the insecticides. Larvae that were exposed to an LC₃₀ concentration of methoxyfenozide exhibited lower pupal weight and increased larval and pupal developmental times compared with thiodicarb treated larvae or control larvae. Adults that were exposed as larvae to an LC₃₀ concentration of methoxyfenozide or thiodicarb showed reduced fecundity (35% and 30%, respectively) compared to control adults. The tested insecticides significantly reduced adult longevity. The longevity of adult females that as larvae were treated with an LC₃₀ concentration of methoxyfenozide or thiodicarb was reduced by 28% and 23%, respectively, in comparison to control females. We predict that the combination of lethal and sublethal effects of the insecticides, especially methoxyfenozide, will induce significant effects on field population dynamics of H. armigera.     

Manual topping decreases bollworm infestations in cotton cultivation in Mali

In sub-Saharan areas of Africa, cotton growers no longer cut the shoot tips from plants (topping), although manual topping was promoted at the start of the 20th century to improve yield and, surprisingly, to reduce pest incidence. In these areas, the bollworms Helicoverpa armigera Hübner, Earias spp., and Diparopsis watersi Rothschild are responsible for the majority of cotton yield losses, and the use of pyrethroids has resulted in resistance in field populations of H. armigera. In the face of these problems and given the scarcity of literature on the effects of topping on pest control, we assessed bollworm infestation levels in 12 trials comparing manual topping and non-topping cotton plots in Mali over a six-year period (2002, 2003, and 2005 to 2008). Topping was performed at the emergence of the 15th sympodial branch or at 10 days after the first flower opening. Our results showed no significant difference in seed cotton yields between topped and non-topped cotton. Bollworm infestations (all species) were always lower on topped cotton and 7 out of 12 trials showed significantly lower infestations on topped cotton. In plots of topped cotton, we recorded an average of 56% fewer H. armigera larvae, 68% fewer Earias spp. larvae, and 71% fewer D. watersi larvae with respectively 5, 4, and 3 out of 12 trials with significant differences in favor of topping and no significant difference in favor of non-topping. To our knowledge, our study is the first to report decreases in D. watersi larval infestation with cotton topping. Further studies are required to understand the mechanisms involved in these effects and to ensure that topping is economically attractive for farmers.     

Pest-damage relationships for Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) on vegetative soybean

The response of vegetative soybean (Glycine max) to Helicoverpa armigera feeding was studied in irrigated field cages over three years in eastern Australia to determine the relationship between larval density and yield loss, and to develop economic injury levels. Rather than using artificial defoliation techniques, plants were infested with either eggs or larvae of H. armigera, and larvae allowed to feed until death or pupation. Larvae were counted and sized regularly and infestation intensity was calculated in Helicoverpa injury equivalent (HIE) units, where 1 HIE was the consumption of one larva from the start of the infestation period to pupation. In the two experiments where yield loss occurred, the upper threshold for zero yield loss was 7.51 ± 0.21 HIEs and 6.43 ± 1.08 HIEs respectively. In the third experiment, infestation intensity was lower and no loss of seed yield was detected up to 7.0 HIEs. The rate of yield loss/HIE beyond the zero yield loss threshold varied between Experiments 1 and 2 (−9.44 ± 0.80 g and −23.17 ± 3.18 g, respectively). H. armigera infestation also affected plant height and various yield components (including pod and seed numbers and seeds/pod) but did not affect seed size in any experiment. Leaf area loss of plants averaged 841 and 1025 cm²/larva in the two experiments compared to 214 and 302 cm²/larva for cohort larvae feeding on detached leaves at the same time, making clear that artificial defoliation techniques are unsuitable for determining H. armigera economic injury levels on vegetative soybean. Analysis of canopy leaf area and pod profiles indicated that leaf and pod loss occurred from the top of the plant downwards. However, there was an increase in pod numbers closer to the ground at higher pest densities as the plant attempted to compensate for damage. Defoliation at the damage threshold was 18.6 and 28.0% in Experiments 1 and 2, indicating that yield loss from H. armigera feeding occurred at much lower levels of defoliation than previously indicated by artificial defoliation studies. Based on these results, the economic injury level for H. armigera on vegetative soybean is approximately 7.3 HIEs/row-metre in 91 cm rows or 8.0 HIEs/m².     

Damage and survivorship of fall armyworm (Lepidoptera: Noctuidae) on transgenic field corn expressing Bacillus thuringiensis Cry proteins

Field corn, Zea mays L., plants expressing Cry1Ab and Cry1F insecticidal crystal (Cry) proteins of Bacillus thuringiensis (Bt) Berliner are planted on considerable acreage across the Southern region of the United States. The fall armyworm, Spodoptera frugiperda (J.E. Smith), is an economically important pest during the mid-to-late season on non-Bt and some commercial Bt corn hybrids. The objective of this study was to quantify foliar injury and survivorship of fall armyworm on transgenic corn lines expressing Cry1Ab or Cry1F Bt proteins. Corn lines/hybrids expressing Cry1Ab, Cry1F, and a conventional non-Bt cultivar were evaluated against artificial infestations of fall armyworm in field trials. Larvae (second instars) of fall armyworm were placed on corn plants (V8-V10 stages). Leaf injury ratings were recorded 14 d after infestation. Hybrids expressing Cry1F had significantly lower feeding injury ratings than non-Bt corn plants. Development and survivorship of fall armyworm on Bt corn lines/hybrids were also evaluated in no-choice laboratory assays by offering freshly harvested corn leaf tissue to third instars. Transgenic corn hybrids expressing Cry1Ab or Cry1F significantly reduced growth, development, and survivorship of fall armyworm compared to those offered non-Bt corn tissue. However, 25-76% of third instars offered Bt corn leaf tissues successfully pupated and emerged as adults. These results suggest Cry1Ab has limited effects on fall armyworm; whereas Cry1F demonstrated significant reductions in foliar injury and lower survivorship compared to that on non-Bt corn tissues. Although fall armyworm is not considered a primary target for insect resistance management by the U.S. Environmental Protection Agency, these levels of survivorship could impact selection pressures across the farmscape, especially when considering that transgenic Bt cotton cultivars express similar Cry (Cry1Ac or Cry1F) proteins.     

Resistance of Bt-maize (MON810) against the stem borers Busseola fusca (Fuller) and Chilo partellus (Swinhoe) and its yield performance in Kenya

A study was conducted to assess the performance of maize hybrids with Bt event MON810 (Bt-hybrids) against the maize stem borer Busseola fusca (Fuller) in a biosafety greenhouse (BGH) and against the spotted stem borer Chilo partellus (Swinhoe) under confined field trials (CFT) in Kenya for three seasons during 2013–2014. The study comprised 14 non-commercialized hybrids (seven pairs of near-isogenic Bt and non-Bt hybrids) and four non-Bt commercial hybrids. Each plant was artificially infested twice with 10 first instar larvae. In CFT, plants were infested with C. partellus 14 and 24 days after planting; in BGH, plants were infested with B. fusca 21 and 31 days after planting. In CFT, the seven Bt hybrids significantly differed from their non-Bt counterparts for leaf damage, number of exit holes, percent tunnel length, and grain yield. When averaged over three seasons, Bt-hybrids gave the highest grain yield (9.7 t ha⁻¹), followed by non-Bt hybrids (6.9 t ha⁻¹) and commercial checks (6 t ha⁻¹). Bt-hybrids had the least number of exit holes and percent tunnel length in all the seasons as compared to the non-Bt hybrids and commercial checks. In BGH trials, Bt-hybrids consistently suffered less leaf damage than their non-Bt near isolines. The study demonstrated that MON810 was effective in controlling B. fusca and C. partellus. Bt-maize, therefore, has great potential to reduce the risk of maize grain losses in Africa due to stem borers, and will enable the smallholder farmers to produce high-quality grain with increased yield, reduced insecticide inputs, and improved food security.     

The competitive ability of different chickpea (Cicer arietinum) genotypes against Polygonum aviculare under field conditions

Polygonum aviculare L. is a troublesome weed in chickpea cultivated in the Mediterranean environment of Central Italy. A 2-year field study was carried out to evaluate the competitive ability and the yield response of different chickpea genotypes against P. aviculare. Experimental treatments consisted in six chickpea genotypes (Alto Lazio, C1017, C133, C134, C6150 stable lines and cultivar Sultano) cultivated in weed-free conditions and with P. aviculare at four densities (4, 8, 16, 32 plants m⁻²). The competitive ability of chickpea against P. aviculare was assessed on the basis of (i) the relative biomass total (RBT); (ii) the competitive balance index (Cb), and (iii) the competitive index (CI). The chickpea seed yield in weed-free conditions ranged from 2.6 to 2.1 t ha⁻¹ of DM and was higher in C6150 and Sultano. P. aviculare caused an average chickpea seed yield loss of 14, 46, 74 and 88% at the density of 4, 8, 16, 32 plants m⁻² compared to the weed-free crop. The relationship between the P. aviculare density and the percentage of chickpea yield loss was described by the rectangular hyperbola model with the asymptote constrained to 100% maximum yield loss. The estimated coefficient I (yield loss per unit density as density approaches zero) was lower in C133, Sultano, and C1017. RBT was higher than 1 in all chickpea genotypes at 4 plants m⁻² of P. aviculare, while at higher P. aviculare densities it was similar to 1 suggesting that there is no resource use complementarity between chickpea and the weed. Generally, at the density of 50 plants m⁻² the chickpea crop was more competitive than P. aviculare at 4 plants m⁻² (Cb > 0), equally competitive at 8 plants m⁻² (Cb = 0), and less competitive at 16 and 32 plants m⁻² (Cb < 0). No chickpea genotype achieved the objective of combining a high seed yield potential and a great competitive ability against P. aviculare. C6150 and Sultano had a high seed yield production in weed-free conditions, but they were poorly competitive against P. aviculare at intermediate and high weed infestation, while C1017 showed a satisfactory level of Cb and CI at all P. aviculare densities although its seed yield was the lowest in weed-free conditions. However, the results suggest that, from an agronomical point of view, P. aviculare plant density should be less than 4 plant m⁻² in order to prevent severe chickpea seed yield loss in field conditions.