Frequency of Bacillus thuringiensis Cry1A.105 resistance alleles in field populations of the fall armyworm,Spodoptera frugiperda,in Louisiana and Florida

Fall armyworm, Spodoptera frugiperda (J.E. Smith), is a major pest of many crops and a cross-crop target of transgenic maize, cotton, and soybean containing Bacillus thuringiensis (Bt) genes. Some of the current Bt maize products for controlling lepidopteran species contain the Bt event MON 89034. The objective of this study was to determine the frequency of resistance alleles in field populations of S. frugiperda collected from Louisiana and Florida, U.S. to Cry1A.105, one of the two Bt genes in MON 89034. A total of 150 F₂ two-parent families of S. frugiperda were established using single-pair mating of field-collected individuals in 2011, which included 79 families from two locations in Louisiana and 71 families from one location in Florida. F₂ screen was conducted to detect resistance alleles in these families to Cry1A.105 protein in maize plants. Four out of the 79 Louisiana and 14 out of the 71 Florida families were identified to possess resistance alleles to the Cry1A.105 maize plants. Thus, the corresponding frequency of resistance alleles to Cry1A.105 maize was estimated to be 0.0158 with a 95% credibility interval (CI) of 0.0052–0.0323 for the Louisiana populations and 0.0559 with a 95% CI of 0.0319–0.0868 for the Florida populations. The resistant families survived on whole Cry1A.105 maize plants and demonstrated a significant level (>116-fold) of resistance to the Cry1A.105 protein in a diet-incorporated bioassay. These findings suggest that resistance allele frequency in S. frugiperda to single-gene Cry1A.105 maize in the U.S. southeast region apparently is not rare, most likely due to the selection of Cry1F resistance and its cross-resistance to Cry1A.105.     

Pest resistance to Cry1Ab Bt maize: Field resistance,contributing factors and lessons from South Africa

This paper documents the historical development of resistance of the African maize stem borer, Busseola fusca (Fuller) (Lepidoptera: Noctuidae) to Bt maize (Zea mays L.). This pest was one of the first to evolve resistance to Bt maize expressing Cry1Ab protein. A time-line of events and contributing factors are presented, from the commencement of efficacy testing through to the present situation, where the Cry1Ab toxin has lost its efficacy against B. fusca at many localities throughout the maize producing region, and single-gene Bt maize events often require insecticide treatments for which farmers are compensated. Significant levels of pest survival on Bt maize was observed in the first season after commercial release in 1998 and confirmed seven years later. Reduced selection pressure on the target pest is the objective of insect resistance management (IRM), and strategies to accomplish this should receive highest priority. Where resistance is prevalent, the only viable options to reduce selection pressure are withdrawal of the product and/or enforcement of high-dose/refuge requirements. The latter action may however be of no value under conditions where resistance is prevalent, since the value of refugia to an IRM strategy may be compromised. Remedial actions taken in South Africa included the propagation and enforcement of refuge compliance followed by the release of pyramided maize hybrids in 2011. These pyramids combine Cry1A.105 and Cry2Ab2 toxin-producing transgenes, replacing the ineffective single-transgene. However, it remains uncertain if cross-resistance occurs between Cry1A.105/Cry2Ab2 and the closely related Cry1Ab toxin, and for how long this pyramided event will endure. Cultivation of Cry1Ab-expressing hybrids continues in areas where resistance levels have been confirmed to be high. In retrospect, this case provides lessons regarding IRM, not only in South Africa, but wherever Bt crops are being introduced.     

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.     

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.     

Identification and Physicochemical Properties of the Novel Hemolysin(s) From Oral Secretions of Helicoverpa armigera (Lepidoptera: Noctuidae)

Hemolysins cause the lysis of invading organisms, representing major humoral immunity used by invertebrates. Hemolysins have been discovered in hemolymph of Helicoverpa armigera larvae as immune factors. As oral immunity is great important to clear general pathogens, we presumed that hemolysins may be present in oral secretions (OS). To confirm this hypothesis, we conducted four testing methods to identify hemolysin(s) in larval OS of H. armigera, and analyzed physicochemical properties of the hemolysin in comparison with hemolytic melittin of Apis mellifera (L.) (Hymenoptera: Apidae) venom. We found hemolysin(s) from OS of H. armigera for the first time, and further identified in other lepidopteran herbivores. It could be precipitated by ammonium sulfate, which demonstrates that the hemolytic factor is proteinaceous. Labial gland showed significantly higher hemolytic activity than gut tissues, suggesting that hemolysin of OS is mainly derived from saliva secreted by labial glands. Physicochemical properties of hemolysin in caterpillar’s OS were different from bee venom. It was noteworthy that hemolytic activity of OS was only partially inhibited even at 100°C. Hemolytic activity of OS was not inhibited by nine tested carbohydrates contrary to bee venom melittin. Moreover, effects of metal ions on hemolytic activity were different between OS and bee venom. We conclude that there is at least a novel hemolysin in OS of herbivorous insects with proposed antibacterial function, and its hemolytic mechanism may be different from melittin. Our study enriches understanding of the potential role of hemolysins in insect immunity and provides useful data to the field of herbivorous insect-pathogen research.     

Mechanism of Action and Therapeutic Potential of the β-Hairpin Antimicrobial Peptide Capitellacin from the Marine Polychaeta Capitella teleta

Among the most potent and proteolytically resistant antimicrobial peptides (AMPs) of animal origin are molecules forming a β-hairpin structure stabilized by disulfide bonds. In this study, we investigated the mechanism of action and therapeutic potential of the β-hairpin AMP from the marine polychaeta Capitella teleta, named capitellacin. The peptide exhibits a low cytotoxicity toward mammalian cells and a pronounced activity against a wide range of bacterial pathogens including multi-resistant bacteria, but the mechanism of its antibacterial action is still obscure. In view of this, we obtained analogs of capitellacin and tachyplesin-inspired chimeric variants to identify amino acid residues important for biological activities. A low hydrophobicity of the β-turn region in capitellacin determines its modest membranotropic activity and slow membrane permeabilization. Electrochemical measurements in planar lipid bilayers mimicking the E. coli membrane were consistent with the detergent-like mechanism of action rather than with binding to a specific molecular target in the cell. The peptide did not induce bacterial resistance after a 21-day selection experiment, which also pointed at a membranotropic mechanism of action. We also found that capitellacin can both prevent E. coli biofilm formation and destroy preformed mature biofilms. The marked antibacterial and antibiofilm activity of capitellacin along with its moderate adverse effects on mammalian cells make this peptide a promising scaffold for the development of drugs for the treatment of chronic E. coli infections, in particular those caused by the formation of biofilms.     

Effect of host plant resistance and insecticide on brown planthopper Nilaparvata lugens (Stål) and predator population development in the Mekong Delta, Vietnam

Crop yield and populations of brown planthopper Nilaparvata lugens (Stål), whitebacked planthopper Sogatella furcifera (Horvath) (Homoptera: Delphacidae), green leafhopper Nephotettix spp. (Homoptera: Cicadellidae), and hemipteran and spider predators were monitored on rice varieties susceptible, moderately resistant, and highly resistant to N. lugens, under insecticide-treated and insecticide-free conditions. Nilaparvata lugens outbreaks and hopperburned plants were observed only in plots of the insecticide-treated susceptible varieties. In insecticide-free plots, the yield of the susceptible variety was lower than those of resistant varieties in only one of four seasons. In plots of a moderately resistant and a highly resistant variety, populations of N. lugens, S. furcifera, Nephotettix spp., and predators were generally similar, and yields did not differ, under both insecticide-treated and insecticide-free conditions. In insecticide-free plots, predator-N. lugens ratios were generally higher on resistant varieties than susceptible varieties. Two conclusions can'be drawn from our results relevant to the question of what levels of N. lugens host plant resistance are appropriate for farmers who do not overuse insecticides. First, susceptible varieties will not necessarily be damaged by N. lugens, even when N. lugens outbreaks occur in adjacent plots. Second, moderate and high levels of N. lugens resistance do not appear to be incompatible with biological control of N. lugens or other homopteran pests.     

中国“槟榔黄化病”研究40年——病原、防控措施新进展

自1981年海南省屯昌县首次发现槟榔黄化病（yellow leaf disease of areca palm, YLD）以来,槟榔黄化问题日趋严重,现已成为制约中国槟榔产业可持续发展的最重要的因素。同时,鉴于生产中除槟榔黄化病外,炭疽病、细菌性叶斑病、椰心叶甲、干旱等一些其他因素也可引起槟榔黄化,以及“槟榔黄化病”病原或病因认识上存在混淆的问题,学界暂用“槟榔黄化现象”的表述。近年来,针对上述问题开展了一系列深入研究并取得突破性进展。本文首先简要回顾了中国槟榔黄化病的发生及病原方面的研究进展,介绍了另一种致黄关键新病害——由槟榔隐症病毒1（Areca palm velarivirus 1, APV1）引起的槟榔黄叶病毒病（areca palm leaf yellowing virus disease, ALYVD）,以及其他2种新发现的病毒病害——槟榔坏死环斑病毒病和槟榔坏死环斑病毒病的研究进展。对6个示范基地的病原检测结果表明,部分黄化植株被槟榔黄化植原体（areca palm yellow leaf phytoplasma, AYLP）或APV1单独感染,部分植株被AYLP和APV1复合感染。本文还探讨了槟榔黄化病研究中存在的病原分布不均、含量低引起的检测困难和田间诊断易混淆等问题,并对YLD、ALYVD、叶斑类病害、根腐病害、芽腐病、椰心叶甲、干旱、寒害、除草剂药害等9类因子引起的黄化症状特征进行了总结。进而分析了YLD和ALYVD 防控中存在的问题以及现阶段面临的紧迫形势,从防控策略和具体措施解读了《槟榔“黄化病“防控明白纸》,并指出了其有待进一步完善之处及防控中亟待解决的问题。最后,展望了YLD和ALYVD 2种致黄关键病害综合防控中亟待实施的措施。本文旨在让广大科研工作者和农技人员更好地了解槟榔“黄化病”方面的最新成果。