Fitness costs of Cry1F resistance in fall armyworm,Spodoptera frugiperda

Transgenic corn, Zea mays L., expressing the Bacillus thuringiensis Berliner (Bt) protein Cry1F has been registered for Spodoptera frugiperda (J. E. Smith) control since 2003 in the USA. Unexpected damage to Cry1F corn was reported in 2006 in Puerto Rico, and Cry1F resistance in S. frugiperda from Puerto Rico was documented. The study of fitness costs associated with insect resistance to Bt insecticidal proteins is important for understanding resistance evolution and for evaluating resistance management practices used to mitigate resistance to transgenic corn. Currently, no studies have addressed the fitness costs associated with Cry1F resistance in S. frugiperda. In this study, susceptible and resistant strains with similar genetic background and their reciprocal crosses were used to estimate Cry1F resistance fitness costs. Comparisons between life‐history traits and population growth rates of homozygous susceptible, heterozygous and homozygous resistant S. frugiperda were used to determine whether the resistance is associated with fitness costs. Major fitness costs were not apparent in either heterozygotes or homozygous resistant insects. However, there was a slight indication of hybrid vigour in the heterozygotes. Additionally, two lines in which the frequency of the resistant alleles was fixed at 0.5 were followed for seven generations, after which the frequency of resistant alleles slightly decreased in both lines. The lack of strong fitness costs associated with Cry1F resistance in S. frugiperda indicates that initial allele frequencies may be higher than expected in field populations and will tend to remain stable in field populations in the absence of selection pressure (e.g. Puerto Rico).     

Rapid selection and characterization of Cry1F resistance in a Brazilian strain of fall armyworm

Transgenic maize (Zea mays L., Poaceae) event TC1507, producing the Cry1F protein of Bacillus thuringiensis Berliner, has been used for management of the fall armyworm, Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae), in Brazil since 2009. A strain of S. frugiperda, obtained from field collections of larvae in TC1507 maize in Minas Gerais state in 2010, was selected in the laboratory for resistance to Cry1F using leaves of TC1507 maize in two selection regimes. Continuous exposure of larvae to Cry1F was more effective than exposure for 6, 8, and 10 days in the selection of resistant S. frugiperda individuals. With only four generations of laboratory selection, a strain with high levels of resistance to Cry1F was obtained, as indicated by the survival of insects reared on leaves of TC1507 maize plants and by the more than 300‐fold resistance level measured in bioassays with the purified Cry1F protein. Importantly, reciprocal crosses between control and the Cry1F‐selected strains revealed that the resistance is autosomal and incompletely recessive, and the response obtained in the backcross of the F₁ generation with the resistant strain was consistent with simple monogenic inheritance. Additionally, there were no apparent fitness costs associated with resistance either for survival or larval growth on non‐Bt maize leaves. Our findings provide experimental evidence for rapid evolution of Cry1F resistance in S. frugiperda in the laboratory and further reinforce the potential of this species to evolve field resistance to the TC1507 maize as previously reported. The resistant strain isolated in this study provides an opportunity to estimate the resistance allele frequency in the field and to determine the biochemical and molecular basis of the resistance, which should provide further information to assist in the resistance management of S. frugiperda on transgenic maize producing B. thuringiensis proteins.     

Plant defense against fall armyworm in micro‐sympatric maize (Zea mays ssp. mays) and Balsas teosinte (Zea mays ssp. parviglumis)

Maize [Zea mays L. ssp. mays (Poaceae)] was domesticated from Balsas teosinte (Zea mays ssp. parviglumis Iltis & Doebley) in present‐day Mexico. Fall armyworm, Spodoptera frugiperda JE Smith (Lepidoptera: Noctuidae), is among the most important pests of maize in Mexico and Central America. We compared the strength of plant defenses against fall armyworm between micro‐sympatric landrace maize and Balsas teosinte in the field and laboratory. The field comparison, conducted in Mexico, consisted of comparing the frequency of fall armyworm infestation between young maize and Balsas teosinte plants in dryland agricultural fields in which Balsas teosinte grew as a weed. The laboratory comparison contrasted the performance of fall armyworm larvae provided a diet of leaf tissue excised from maize or Balsas teosinte plants that were intact or had been primed by larval feeding. In the field, maize plants were more frequently infested with fall armyworm than Balsas teosinte plants: over 3 years and three fields, maize was infested at a ca. 1.8‐fold greater rate than Balsas teosinte. In the laboratory, larval growth, but not survivorship, was differently affected by feeding on maize vs. Balsas teosinte, and on primed vs. intact plants. Specifically, survivorship was ca. 98%, and did not differ between maize and Balsas teosinte, nor between primed and intact plants. Larvae grew less on intact vs. primed maize, and similarly on intact vs. primed Balsas teosinte; overall, growth was 1.2‐fold greater on maize compared to Balsas teosinte, and on primed compared to intact plants. Parallel observations showed that the differences in growth could not be attributed to the amount of leaf tissue consumed by larvae. We discuss our results in relation to differences in the strength of plant defenses between crops and their ancestors, the relevance of unmanaged Balsas teosinte introgression in the context of fall armyworm defenses in maize, and whether greater growth of larvae on primed vs. intact plants signifies herbivore offense.     

First report of the fall armyworm,Spodoptera frugiperda (Lepidoptera: Noctuidae), natural enemies from Africa

The fall armyworm (FAW), Spodoptera frugiperda, is a major pest of maize in North and South America. It was first reported from Africa in 2016 and currently established as a major invasive pest of maize. A survey was conducted to explore for natural enemies of the fall armyworm in Ethiopia, Kenya and Tanzania in 2017. Smallholder maize farms were randomly selected and surveyed in the three countries. Five different species of parasitoids were recovered from fall armyworm eggs and larvae, including four within the Hymenoptera and one Dipteran. These species are new associations with FAW and were never reported before from Africa, North and South America. In Ethiopia, Cotesia icipe was the dominant larval parasitoid with parasitism ranging from 33.8% to 45.3%, while in Kenya, the tachinid fly, Palexorista zonata, was the primary parasitoid with 12.5% parasitism. Charops ater and Coccygidium luteum were the most common parasitoids in Kenya and Tanzania with parasitism ranging from 6 to 12%, and 4 to 8.3%, respectively. Although fall armyworm has rapidly spread throughout these three countries, we were encouraged to see a reasonable level of biological control in place. This study is of paramount importance in designing a biological control program for fall armyworm, either through conservation of native natural enemies or augmentative release.     

Feeding of fall armyworm,Spodoptera frugiperda,on Bt transgenic cotton and its isoline

Studies on insect food intake and utilization are important for determining the degree of insect/plant association and host species’ resistance, and also for helping design pest management programs by providing estimates of potential economic losses, techniques for mass breeding of insects, and identifying physiological differences between species. We studied the feeding and development of fall armyworm, Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae), on transgenic (Bt) and non‐transgenic (non‐Bt) cotton. The larvae of S. frugiperda fed on Bt cotton had a longer development period (23.0 days) than those fed on non‐Bt cotton (20.2 days). Survivorship of S. frugiperda larvae fed on Bt cotton (74.1%) was lower than that of larvae fed on non‐Bt cotton (96.7%). Pupal weight of larvae fed on Bt cotton (0.042 g) was lower than that of larvae fed on non‐Bt cotton (0.061 g). The cotton cultivar significantly affected food intake, feces production, metabolization, and food assimilation by S. frugiperda larvae. However, it did not affect their weight gain. Intake of Bt‐cotton leaf (0.53 g dry weight) per S. frugiperda larva was lower than the intake of non‐Bt‐cotton leaf (0.61 g dry weight). Larvae fed on Bt‐cotton leaves produced less feces (0.25 g dry weight) than those fed on non‐Bt‐cotton leaves (0.37 g dry weight). Weight gain per S. frugiperda larva fed on Bt‐cotton leaves (0.058 g dry weight) was similar to the weight gain for larvae fed on non‐Bt‐cotton leaves (0.056 g dry weight). The cotton cultivar significantly affected the relative growth, consumption, and metabolic rates, as well as other nutritional indices: the figures were lower for larvae fed on Bt‐cotton leaves than for larvae fed on non‐transgenic cotton leaves.     

Effect of Cry1F maize on the behavior of susceptible and resistant Spodoptera frugiperda and Ostrinia nubilalis

Understanding the behavior of pests targeted with Bacillus thuringiensis Berliner (Bt) crops is important to define resistance management strategies. Particularly the study of larval movement between plants is important to determine the feasibility of refuge configurations. Exposure to Bt maize, Zea mays L. (Poaceae), has been suggested to increase larval movement in lepidopteran species but few studies have examined the potential for resistance to interact with behavioral responses to Bt toxins. Choice and no‐choice experiments were conducted with Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae) and Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) to determine whether Cry1F resistance influences neonate movement. Leaf discs of Cry1F maize and the corresponding isoline were used to characterize behavioral responses. In both experiments, the location (on or off of plant tissues) and mortality of susceptible and Cry1F resistant neonates was recorded for 5 days, but the analysis of larvae location was performed until 7 h. Our results indicated no strong difference between resistant and susceptible phenotypes in S. frugiperda and O. nubilalis, although a small percentage of susceptible neonates in both species abandoned maize tissue expressing Cry1F. However, significant behavioral differences were observed between species. Ostrinia nubilalis exhibited increased movement between leaf discs, whereas S. frugiperda selected plant tissue within the first 30 min and remained on the chosen plant regardless of the presence of Cry1F. Spodoptera frugiperda reduced larval movement may have implications to refuge configuration. This study represents the first step toward understanding the effects of Cry1F resistance on Lepidoptera larval behavior. Information regarding behavioral differences between species could aid in developing better and more flexible resistance management strategies.     

Larval dispersal of the invasive fall armyworm,Spodoptera frugiperda,the exotic stemborer Chilo partellus,and indigenous maize stemborers in Africa

Larval dispersal either through ballooning or crawling results in a redistribution of the insect population and infestations within and between plants. In addition, invasive species, such as the fall armyworm (FAW), Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae), and the exotic stemborer Chilo partellus (Swinhoe) (Lepidoptera: Crambidae), may displace indigenous stemborers on maize in Africa. To test whether larval dispersal activity may play a role in the displacement of indigenous stemborers, larval dispersal was compared between FAW, C. partellus, and the indigenous species Busseola fusca (Fuller) and Sesamia calamistis (Hampson) (both Lepidoptera: Noctuidae). Twenty potted maize plants were infested with one batch of eggs either from stemborers (B. fusca, S. calamistis, or C. partellus) or from FAW and monitored in the greenhouse for ballooning activities. After egg hatching, both ballooning and non-ballooning larvae were identified according to species and counted. FAW neonate larvae had greater potential for ballooning off than stemborers, irrespective of species. For each species, more females dispersed than males, and their survival rate was higher than that of non-ballooning larvae. In addition, plant-to-plant larval movements were studied using 6.25-m² plots of caged maize in a completely randomized design with five replicates. FAW was found to have wider dispersal and plant damage potential than any of the stemborer species. In conclusion, in contrast to C. partellus, the invasive characteristic of FAW can be explained, in part, by its higher larval dispersal activity compared to stemborers. This difference in larval dispersal might also be considered in sampling plans for monitoring pest density in the field.     

Physical leaf defenses – altered by Zea life‐history evolution,domestication, and breeding – mediate oviposition preference of a specialist leafhopper

Plant anti‐herbivore defenses are known to be affected by life‐history evolution, as well as by domestication and breeding in the case of crop species. A suite of plants from the maize genus Zea (Poaceae) and the specialist herbivore Dalbulus maidis (DeLong & Wolcott) (Hemiptera: Cicadellidae) were used to test the hypothesis that anti‐herbivore defenses are affected by plant life‐history evolution and human intervention through domestication and breeding for high yield. The suite of plants included a maize (Zea mays ssp. mays L.) commercial hybrid, a maize landrace, two populations of the annual Balsas teosinte (Z. mays ssp. parviglumis Iltis & Doebley), and perennial teosinte (Z. diploperennis Iltis, Doebley & Guzman). Leaf toughness, pubescence, and oviposition preference were compared among the suite of host plants looking for effects of transitions in life history (i.e., from perennial to annual life cycle), domestication (i.e., from wild annual to domesticated annual), and breeding (i.e., from landrace to hybrid maize) on defense against D. maidis. Results on leaf toughness suggested that the life‐history and domestication transitions weakened the plant's resistance to penetration by the mouthparts and ovipositor of D. maidis, whereas results on pubescence suggested that this putative defense was strengthened with the breeding transition, contrary to expectations. Results on oviposition preference of D. maidis coincided with the expectation that life‐history and domestication transitions would lead to preference for Balsas teosinte over perennial teosinte, and of landrace maize over Balsas teosinte. Also, a negative correlation suggested that oviposition preference is significantly influenced by leaf toughness. Overall, the results suggested that Zea defenses against the specialist herbivore D. maidis were variably affected by plant life‐history evolution, domestication, and breeding, and that chemical defense may play a role in Zea defense against D. maidis because leaf toughness and pubescence only partially explained its host preferences.     

Identification and expression profiling of Spodoptera litura OR18, a gene with a potential role in olfactory recognition

The full‐length cDNA of the gene SlituOR18, encoding a candidate olfactory receptor in the antennae of the tobacco cutworm, Spodoptera litura (Fabricius) (Lepidoptera Noctuidae), was identified through homology cloning strategies using RACE (rapid amplification of cDNA ends) and PCR (polymerase chain reaction). The protein of SlituOR18 shared >80% sequence identity and the same seven transmembrane domains with other olfactory receptor 18 (OR18) proteins sequenced from noctuid moths. SlituOR18 also had a similar structure to SlituORco, encoding the olfactory co‐receptor in S. litura. The sequence between transmembrane segments IV and V was longer than other sequences between transmembrane segments, and the N‐terminus was intracytoplasmic. Analysis by qRT‐PCR showed SlituOR18 was predominantly expressed in adult moths and there was higher expression in female antennae than in male antennae. Weak expression of SlituOR18 was observed in eggs and no expression was observed in the antennae of fourth instars and 5‐day‐old pupae. In situ hybridization experiments revealed that expression of these receptor types was clearly restricted to the bases of trichodea‐type olfactory sensilla (sensillum trichodeum), that are known to contain neurons sensitive to food odor or pheromones. Our data demonstrated that both SlituORco and SlituOR18 showed diurnal changes in their relative expression level. Expression of SlituOR18 varied among geographic populations of S. litura that had been trapped in the field using synthetic pheromone lures. The mRNA expression level of SlituOR18 was similar among S. litura populations from Sichuan, Guangxi, and Hunan, and higher than in populations from Shanghai and Ningbo. We suggest that OR18 could play a critical role in olfaction in noctuid moths and is a potential target for novel pest management strategies in the future.     

The limited role of differential fractionation in genome content variation and function in maize (Zea mays L.) inbred lines

Maize is a diverse paleotetraploid species with considerable presence/absence variation and copy number variation. One mechanism through which presence/absence variation can arise is differential fractionation. Fractionation refers to the loss of duplicate gene pairs from one of the maize subgenomes during diploidization. Differential fractionation refers to non‐shared gene loss events between individuals following a whole‐genome duplication event. We investigated the prevalence of presence/absence variation resulting from differential fractionation in the syntenic portion of the genome using two whole‐genome de novo assemblies of the inbred lines B73 and PH207. Between these two genomes, syntenic genes were highly conserved with less than 1% of syntenic genes being subject to differential fractionation. The few variably fractionated syntenic genes that were identified are unlikely to contribute to functional phenotypic variation, as there is a significant depletion of these genes in annotated gene sets. In further comparisons of 60 diverse inbred lines, non‐syntenic genes were six times more likely to be variable than syntenic genes, suggesting that comparisons among additional genome assemblies are not likely to result in the discovery of large‐scale presence/absence variation among syntenic genes.     

Preference of Bt‐resistant and susceptible Busseola fusca moths and larvae for Bt and non‐Bt maize

The sustainability of genetically engineered insecticidal Bacillus thuringiensis Berliner (Bt) maize, Zea mays L. (Poaceae), is threatened by the evolution of resistance by target pest species. Several Lepidoptera species have evolved resistance to Cry proteins expressed by Bt maize over the last decade, including the African maize stem borer, Busseola fusca (Fuller) (Lepidoptera: Noctuidae). The insect resistance management (IRM) strategy (i.e., the high‐dose/refuge strategy) deployed to delay resistance evolution is grounded on certain assumptions about the biology and ecology of a pest species, for example, the interactions between the insect pest and crop plants. Should these assumptions be violated, the evolution of resistance within pest populations will be rapid. This study evaluated the assumption that B. fusca adults and larvae select and colonize maize plants at random, and do not show any preference for either Bt or non‐Bt maize. Gravid female B. fusca moths of a resistant and susceptible population were subjected to two‐choice oviposition preference tests using stems of Bt and non‐Bt maize plants. Both the number of egg batches as well as the total number of eggs laid on each stem were recorded. The feeding preference of Bt‐resistant and susceptible neonate B. fusca larvae were evaluated in choice test bioassays with whorl leaf samples of specific maize cultivars. Although no differential oviposition preference was observed for either resistant or susceptible female moths, leaf damage ratings indicated that neonate larvae were able to detect Bt toxins and that they displayed feeding avoidance behaviour on Bt maize leaf samples.     

Comparison of pheromone trap design and lures for Spodoptera frugiperda in Togo and genetic characterization of moths caught

Fall armyworm, Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae), is a pest of grain and vegetable crops endemic to the Western Hemisphere that has recently become widespread in sub‐Saharan Africa and has appeared in India. An important tool for monitoring S. frugiperda in the USA is pheromone trapping, which would be of value for use with African populations. Field experiments were conducted in Togo (West Africa) to compare capture of male fall armyworm using three commercially available pheromone lures and three trap designs. The objectives were to identify optimum trap × lure combinations with respect to sensitivity, specificity, and cost. Almost 400 moths were captured during the experiment. Differences were found in the number of S. frugiperda moths captured in the various trap designs and with the three pheromone lures, and in the number of non‐target moths captured with each lure. The merits of each trap × lure combination are discussed with respect to use in Africa. A nearly equal number of COI‐CS (161) and COI‐RS (158) moths was captured with no differences found in COI marker proportions among traps or lures. However, the diagnostic rice strain marker Tpi was rarely found. Overall, the genetic characterization of the pheromone trap collections indicated a consistent distribution of genetic markers from 2016 to 2017, suggesting a population at or near equilibrium.     

Differential response of antioxidative systems of maize (Zea mays L.) roots cell walls to osmotic and heavy metal stress

An analysis of peroxidase and ascorbate oxidase activity, phenolic content and antioxidant capacity of isolated maize root cell walls was performed in controls and plants stressed with polyethylene glycol (PEG) or heavy metals, zinc or copper. Peroxidase activity (oxidative and peroxidative) was more pronounced in the ionic than in the covalent cell wall fraction. PEG induced an increase and Zn²⁺ a decrease of both ionically bound peroxidase activities. In the covalent fraction, Cu²⁺ decreased oxidative and increased peroxidative activity of peroxidase. Isoelectric focusing of ionically bound proteins and activity staining for peroxidase demonstrated increased intensities and appearance of new acidic isoforms, especially in Zn²⁺ and PEG treatments. Most pronounced basic isoforms (pI ~ 7.5) in controls, decreased in intensity or completely disappeared in stressed plants. Ascorbate oxidase activity was significantly increased by PEG and decreased by Zn²⁺ treatments, and highly correlated with peroxidase activity. Antioxidant capacity and total phenolics content increased in heavy metal‐treated and decreased in PEG‐treated plants. Analysis of individual phenolic components revealed p‐coumaric and ferulic acids, as the most abundant, as well as ferulic acid dimers, trimers and tetramers in the cell walls; their quantity increased under stress conditions. Results presented demonstrate the existence of diverse mechanisms of plant response to different stresses.     

Quantifying rates of random mating in western corn rootworm emerging from Cry3Bb1‐expressing and refuge maize in field cages

The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), is the most significant pest of field maize, Zea mays L. (Poaceae), in the USA. Maize plants expressing Bt toxins targeting the corn rootworm complex have been widely adopted and are the primary insecticidal control measure for this pest in North America. Insect resistance management tactics using various refuge structures have been adopted to ensure Bt products will retain durability. An assumption of the refuge strategy is that males and females emerging from Bt and refuge plantings mate randomly; this has not been tested in the field. We conducted cage studies using field populations of WCR in Indiana, USA, to generate empirical field data on mating rates between beetles emerging from Cry3Bb1‐expressing Bt and refuge maize plants. Two refuge configurations were tested; all refuge plants were labeled using the stable isotope ¹⁵N. This mark persists in adult beetles after eclosion, allowing for collection and analysis of isotopic ratios of all beetles. Additional data collected included adult emergence rates, timing and sex ratios for each of the treatments, and head capsule size and dry weights of beetles collected. Treatment had a significant effect on dry weight; mean dry weight decreased in Bt‐only treatments. Fisher's exact test of proportions of mating pairs of refuge and Bt insects indicated that mating was not random in 20% strip refuges and 5% seed blend treatments. We found high percentages of beetles that fed on Bt‐expressing plants as larvae, suggesting that mating between resistant beetles may not be rare even if random mating did occur.     

Effects of defoliation on the resistance and tolerance of rice,Oryza sativa,to root injury by the rice water weevil,Lissorhoptrus oryzophilus

Understanding plant‐mediated interactions in agricultural systems may facilitate the development of novel and improved management practices, which is important, as management of these insects is currently heavily reliant on insecticides. The fall armyworm, Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae, Prodeniini), is a sporadic pest of rice fields in the southern USA. In southwestern Louisiana, this defoliating insect typically attacks rice early in the growth season, before fields are flooded. Defoliation by fall armyworm larvae may trigger increased expression of plant defenses, which may result in increased resistance to subsequent herbivores. The rice water weevil (RWW), Lissorhoptrus oryzophilus Kuschel (Coleoptera: Curculionidae, Stenopelmini), enters rice fields as an adult both before and after flooding, but oviposition and larval infestation occur only after fields are flooded. RWW may be affected by changes in plant resistance caused by fall armyworm defoliation before flooding. The objectives of this study were to investigate the plant‐mediated effects of natural and artificial defoliation on population densities of RWW larvae after flooding and on the ability of rice plants to compensate for root injury by RWW larvae. In the 2015 season, fall armyworm defoliation before flooding resulted in reduced RWW densities after flooding. However, in 2016 no significant effects of fall armyworm defoliation on densities of RWW larvae were detected. Similarly, mechanical defoliation of rice before flooding did not affect RWW densities after flooding. Although lowest yields were observed in plots subjected to both root injury and defoliation, there was little evidence of a greater than additive reduction in yields from simultaneous injury. These results suggest a lack of plant‐mediated interactions among these two pests in rice in the southern USA.     

Resistance of the fall armyworm,Spodoptera frugiperda,to transgenic Bacillus thuringiensis Cry1F corn in the Americas: lessons and implications for Bt corn IRM in China

The fall armyworm, Spodoptera frugiperda (J.E. Smith), is a major target pest of Bt crops (e.g., corn, cotton, and soybean) in North and South America. This pest has recently invaded Africa and Asia including China and the invasion has placed a great threat to the food security in many countries of these two continents. Due to the extensive use of Bt crops, practical resistance of S. frugiperda to Cry1F corn (TC 1507) with field control problems has widely occurred in Puerto Rico, Brazil, Argentina, and the mainland United States. Analyzing data generated from decade-long studies showed that several factors might have contributed to the wide development of the resistance. These factors include (1) limited modes of action of Bt proteins used in Bt crops; (2) cross-resistance among Cry1 proteins; (3) use of nonhigh dose Bt crop traits; (4) that the resistance is complete on Bt corn plants; (5) abundant in initial Cry1F resistance alleles; and (6) lack of fitness costs/recessive fitness costs of the resistance. The long-term use of Bt crop technology in the Americas suggests that Bt corn can be an effective tool for controlling S. frugiperda in China. IRM programs for Bt corn in China should be as simple as possible to be easily adopted by small-scale growers. The following aspects may be considered in its Bt corn IRM programs: (1) use of only “high dose” traits for both S. frugiperda and stalk borers; (2) developing and implementing a combined resistance monitoring program; (3) use “gene pyramiding” as a primary IRM strategy; and (4) if possible, Bt corn may not be planted in the areas where S. frugiperda overwinters. Lessons and experience gained from the global long-term use of Bt crops should have values in improving IRM programs in the Americas, as well as for a sustainable use of Bt corn technology in China.     

不同类型春玉米灌浆期间籽粒中内源激素IAA、GA、ZR、ABA含量的变化

田间种植的高淀粉（‘郑单19’）、高油（‘通油1号’）和普通型（‘吉单209’）春玉米在籽粒灌浆过程中,‘郑单19’的IAA在授粉后28d达到峰值,‘吉单209’和‘通油1号’的峰值出现在35d;在3个杂交种中,‘郑单19’的IAA峰值最大,‘吉单209’次之,‘通油1号’最低;整个籽粒灌浆期间,‘通油1号’和‘吉单209’的GA含量高于‘郑单19’,在灌浆后期,‘通油1号’仍保持较高的GA含量;‘通油1号’ZR含量的峰值最高,直到后期仍保持较高的水平;‘郑单19’在籽粒灌浆前期的ABA含量较低,但在后期的含量较高。     

Processing of the maize Bt toxin in the gut of Mythimna unipuncta caterpillars

Mythimna unipuncta Haworth (Lepidoptera: Noctuidae) is a well‐known moth species whose larvae can cause devastating damage to some Poaceae crops, including maize (Zea mays L.). The low susceptibility to the Bacillus thuringiensis Berliner (Bt) toxin observed in L6 larvae of this species has been the object of several studies. This study aimed to clarify whether the toxin eliminated from the content of the peritrophic membrane is degraded or excreted and whether the effects of the Bt toxin depend on the doses ingested. To this end, L6 larvae were fed on diets with different amounts of lyophilized Bt or non‐Bt maize leaves. The effect of the Bt concentrations on larval development was measured and the fate of the toxin in the larval tissues was tracked. Results indicated that the larvae of M. unipuncta fed on the various diets showed few differences in weight gain, duration of development, or pupal weight between sublethal Bt concentrations. The larvae rapidly excreted a large part of the toxin ingested, whereas inside the peritrophic membrane the toxin was eliminated, degraded, or sequestered at a rate that increased with the dose and the duration of feeding. As a consequence, little toxin reached the midgut epithelium and therefore the binding sites of the toxin. Moreover, larvae fed on the Bt toxin recovered quickly when they were transferred to a non‐Bt diet.     

Leaf biomechanical properties as mechanisms of resistance to black cutworm (Agrotis ipsilon) among Poa species

Biomechanical properties can be important parameters in resistance of plants to herbivorous insects. As plants age, however, there can be dramatic changes in physical defenses that can then influence their susceptibility to insect herbivores. We measured changes in leaf biomechanical properties during ontogeny of Poa species and the relationship of these changes to the development of a generalist herbivore, the black cutworm, Agrotis ipsilon Hufnagel (Lepidoptera: Noctuidae), was investigated. Larvae were reared on two representative age classes, i.e., young (<60 days after planting) and old (>1 year after planting), of foliage in laboratory assays. Foliage generally reaches a peak fracture force between 80 and 109 days after planting depending on grass type. Foliage from old plants was significantly tougher than that of young plants, and black cutworm larvae reared on foliage from young plants gained significantly (ca. four times) more weight than those fed on foliage from old Poa plants. In addition, fracture force has a negative relationship with black cutworm development. Plant fiber, particularly neutral detergent fiber accounted for 65 and 46% of the variation in fracture force and larval development, respectively. These results provide additional insight into how plant ontogeny influences physical defenses to an insect herbivore in a grass system. Likewise, this is seemingly the first study to suggest a mechanism for host plant resistance to black cutworm. Plant fiber may be a useful trait to explore in plant improvement programs in which black cutworm is a primary pest (e.g., managed turfgrass).     

Yield,development, and quality response of dual‐toxin Bt cotton to Helicoverpa spp. infestations in Australia

To verify current thresholds for Bollgard II^® cotton in Australia, the impact of Helicoverpa spp. (Lepidoptera: Noctuidae) larvae on yield, development, and quality under various infestation intensities and durations, and stages of growth, was tested using small plot field experiments over two seasons. Infestation with up to 80 eggs m^?1 of Helicoverpa armigera (Hübner) and Helicoverpa punctigera Wallengren showed that species, infestation level, and stage of growth had no significant effect on yields of seed‐cotton or lint and on maturity and fibre quality. The duration of infestation of white flowers with H. punctigera neonates (maximum of every day for up to 4 weeks) had no impact on the yield of seed‐cotton or lint, maturity, and fibre quality, but when 100% of flowers were infested (compared with 0 or 50%), seed‐cotton and lint yields were significantly reduced and maturity was delayed. Infestation with up to 18 medium H. armigera larvae m^?1 at several plant stages did not significantly affect yields of seed‐cotton and lint, maturity, and fibre quality. A heliocide spray applied on a commercial farm at the current threshold resulted in a significantly higher lint yield, compared with a farm where no spray was applied. In conclusion, Bollgard II^® cotton is highly resistant to Helicoverpa spp. infestation.