Microarray analysis of global gene regulation in the Cry1Ab-resistant and Cry1Ab-susceptible strains of Diatraea saccharalis

BACKGROUND: Extensive adoption of transgenic Bt corn in recent years for stalk borer control has increased risk of resistance evolution in the target pest populations. A Bt‐resistant strain of the sugarcane borer, Diatraea saccharalis, was approximately 100‐fold more tolerant to Cry1Ab toxin than the susceptible counterpart. To gain a better understanding of the molecular mechanisms of Bt resistance, the Cry1Ab‐susceptible (Cry1Ab‐SS) and Cry1Ab‐resistant (Cry1Ab‐RR) strains of D. saccharalis were subjected to a microarray analysis. RESULTS: Results showed that the expression levels of many genes were significantly different between the Cry1Ab‐RR and Cry1Ab‐SS strains. Microarray analysis of 7145 cDNAs revealed 384 differentially expressed genes. A total of 273 genes were significantly upregulated 2–51.6‐fold, and 111 genes were significantly downregulated 2–22.6‐fold in the Cry1Ab‐RR strain. The upregulation of three potential resistance‐related genes, coding for a glutathione S‐transferase (GST), a chymotrypsin‐like protease (CHY) and a lipase (LP), was confirmed using real‐time PCR, indicating a reproducibility of the microarray data. Ontology analysis revealed that more than twice the number of metabolic‐related genes were upregulated compared with downregulated genes with the same biological function. Up to 35.2% of the upregulated genes in the resistant strain were associated with catalytic activity, while only 9.5% of the downregulated genes were related to the same catalytic molecular function. CONCLUSION: The large portion of metabolic‐ or catalytic‐related genes with significant upregulations indicated a potential large increase in metabolic or catalytic activities in the Cry1Ab‐RR strain. This cDNA microarray gene expression data could be used to characterize and identify new genes that may be associated with Bt resistance in D. saccharalis. Copyright © 2012 Society of Chemical Industry     

基于MaxEnt模型的检疫性害虫小蔗螟在中国的潜在分布预测

小蔗螟是重要的检疫性害虫,许多国家将其列入检疫性害虫名录,主要危害甘蔗、玉米等作物。本研究基于小蔗螟分布记录,结合环境数据,采用MaxEnt软件对其在中国潜在分布区进行预测。结果表明：小蔗螟在我国的高度和中度适生区主要分布在海南、云南、广东、广西及福建等。影响小蔗螟分布的最主要环境变量是最暖季平均降雨量和温度变化方差。阐明小蔗螟在我国潜在地理分布及限制其分布的环境变量,可为我国的检疫工作提供理论依据。     

Field‐evolved resistance to Bt maize in sugarcane borer (Diatraea saccharalis) in Argentina

BACKGROUND

Maize technologies expressing Bacillus thuringiensis (Bt) insecticidal proteins are widely used in Argentina to control sugarcane borer (Diatraea saccharalis Fabricius). Unexpected D. saccharalis damage was observed to Bt maize events TC1507 (expressing Cry1F) and MON 89034 × MON 88017 (expressing Cry1A.105 and Cry2Ab2) in an isolated area of San Luis Province. Diatraea saccharalis larvae were sampled from MON 89034 × MON 88017 fields in the area to generate a resistant strain (RR), which was subsequently characterized in plant and diet bioassays.

RESULTS

Survivorship of the RR strain was high on TC1507 leaf tissue, intermediate on MON 89034 × MON 88017, and low on MON 810 (expressing Cry1Ab). The RR strain had high resistance to Cry1A.105 (186.74‐fold) and no resistance to Cry2Ab2 in diet bioassays. These results indicate resistance to Cry1F and Cry1A.105 (and likely cross‐resistance between them) but not to Cry1Ab or Cry2Ab2. Resistance to MON 89034 × MON 88017 was functionally recessive. Reviews of grower records suggest that resistance initially evolved to Cry1F, conferring cross‐resistance to Cry1A.105, with low refuge compliance as the primary cause. A mitigation plan was implemented in San Luis that included technology rotation, field monitoring, and grower education on best management practices (BMPs) including refuges.

CONCLUSION

In the affected area, the resistance to Cry1F and Cry1A.105 is being managed effectively through use of MON 89034 × MON 88017 and MON 810 in combination with BMPs, and no spread of resistance to other regions has been observed. © 2017 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

F2 screening for resistance to pyramided Bacillus thuringiensis maize in Louisiana and Mississippi populations of Diatraea saccharalis (Lepidoptera: Crambidae)

BACKGROUND: In the mid‐southern region of the United States, sugarcane borer, Diatraea saccharalis (F.), is a major target pest of transgenic maize expressing Bacillus thuringiensis (Bt) proteins. Novel transgenic maize technologies containing two or more pyramided Bt genes for controlling lepidopteran pests have recently become commercially available. Insect resistance management (IRM) is an important issue in the sustainable use of Bt crop technologies. The objective of this study was to determine the frequency of resistance alleles in field populations of D. saccharalis to the new pyramided Bt maize technologies. RESULTS: A total of 382 F₂ family lines derived from 735 feral larvae/pupae of D. saccharalis collected from six locations in Louisiana and Mississippi during 2008 and 2009 were screened for resistance to three new Bt maize technologies: MON 89034, Genuity? VT Triple Pro? and SmartStax?. None of the 382 F₂ isoline families survived on the Bt maize leaf tissue for ≥ 12 days in the F₂ screen. The joint frequency for two‐ or three‐gene resistance models with 95% probability in these populations was estimated to be < 0.0063 to MON 89034 and < 0.003 to VT Triple Pro? and SmartStax?. CONCLUSION: These results suggest that the resistance allele frequency in D. saccharalis to the three pyramided Bt maize technologies is low in the mid‐southern region of the United States, which should meet the rare resistance assumption of the currently used IRM strategy for Bt maize. Copyright © 2011 Society of Chemical Industry     

Toxicity of extracts and isobutyl amides from Piper tuberculatum: potent compounds with potential for the control of the velvetbean caterpillar, Anticarsia gemmatalis

The isobutyl amides pellitorine (compound 1) and 4,5-dihydropiperlonguminine (compound 2) were extracted from the seeds of Piper tuberculatum Jacq. (Piperaceae) in yields of 6.10 and 4.45% respectively. The acute toxicities to the velvetbean caterpillar, Anticarsia gemmatalis (Hübner) (Lepidoptera: Noctuidae), of extracts of seeds, leaves and stems of P. tuberculatum, and of compounds 1 and 2, were evaluated by means of contact bioassays. The extracts caused 80% mortality when doses higher than 800.00 microg insect(-1) of extract of seeds, leaves and stems were administered to the velvetbean caterpillars. Compounds 1 and 2 showed 100% mortality at doses of 200 and 700 microg insect(-1) respectively. The LD(50) and LD(90) values were respectively 31.3 and 104.5 microg insect(-1) for compound 1, and 122.3 and 381.0 microg insect(-1) for compound 2. The potential value of extracts and amides derived from P. tuberculatum as efficient insecticides against velvetbean caterpillars is discussed.     

Susceptibility of field-collected populations of the southwestern corn borer,Diatraea grandiosella,to Bacillus thuringiensis

The susceptibilitity of newly hatched larvae of laboratory-adapted and field-collected populations of the Southwestern corn borer, Diatraea grandiosella (Dyar) (Lepidoptera: Crambidae), to a Bacillus thuringiensis protein (Cry1Ab) was examined using a larval feeding bioassay. D grandiosella populations were collected from five states: Missouri, Kansas, Texas, Tennessee and Kentucky. Using larval mortality as the end-point of the bioassay, the magnitude of differences in the susceptibility of the laboratory-adapted and the field-collected populations to Cry1Ab protein varied from one to 46 times depending on the time of observation and the standard of comparison (LC50 or LC95). However, significant differences in susceptibility to Cry1Ab protein among these populations were not detected when the comparisons were based on growth inhibition (EC50 or EC95); the magnitude of differences was less than fourfold. Either using larval mortality or larval growth inhibition, the results indicated that the field-collected populations of D grandiosella were susceptible to Cry1Ac, and differences in susceptibility may reflect natural variation among populations. The bioassay using larval growth inhibition offers advantages over that using larval mortality, including giving more accurate representation of the toxicological effects of the toxin.     

Aerial Spraying Trials in West Africa for Blackfly Control

Abstract

Organochlorine, organophosphate (o.p.) and carbamate insecticides have been extensively used in Australia to combat Lucilia cuprina Wied. the main initiator of fly-strike of sheep occurring wherever sheep are run on the continent. The improved mules operation, mid-season crutching and insecticides are important in the management of Merino sheep. Insecticides are particularly valuable against body strike, especially in young sheep, in years with intermittent rain during the warmer months. Resistance to larvicides developed in the blowfly over a period of 10y from 1957. The time between the introduction of various insecticides and development of resistance is considered and a comparison made between the emergence of resistance problems to the newer synthetic insecticides in the housefly in Denmark, and in the blowfly in Australia. To provide a complete history of insecticides against the blowfly reference is made to arsenicals, to which there was no suspicion of resistance until low order cross resistance was diagnosed in o.p.-carbamate resistant strains. Some seven years after their introduction DDT and γ BHC were replaced by cyclodiene insecticides in 1954/1955. Reasons are advanced to explain the non-emergence of resistance to DDT and BHC in that period. Resistance to dieldrin and aldrin developed in late 1957 after which diazinon was introduced. The resistance has a typical BHC/dieldrin resistance spectrum and is due to a semi-dominant gene which has persisted in the field in the absence of pressure from cyclodiene insecticides. Non-specific resistance to o.p. insecticides developed in two steps. Low order resistance, diagnosed in 1965, was supplemented by an additional resistance mechanism in 1966, three alleles on two chromosomes are involved. The carbamate, butacarb, was effective against o.p. resistant strains of the blowfly when introduced in the 1966/67 season. In 1967, resistance to butacarb was diagnosed and rapidly became widespread. For the past three fly seasons larvae have been used to monitor resistance levels to o.p. and carbamate insecticides; resistance factors to both these classes of insecticide are significantly higher in larvae than adult females. Resistance levels to o.p. insecticides have stabilised. By contrast resistance levels to butacarb have doubled. A combined o p.-carbamate resistance generally applies in field samples. Larvae from o.p.-carbamate resistant strains form artificial strikes earlier than susceptible larvae, particularly on sheep treated with butacarb. It is concluded that registered o.p. insecticides, but not butacarb, will still give considerable protection against fly-strike, providing the maximum levels of o.p. resistance, reached in laboratory selection programmes, are not exceeded in the field. Investigation into o.p. resistance in the species suggests that the resistance mechanisms place their carriers at a disadvantage in the absence of selection pressure. Measures to minimise the amount of insecticide used against the blowfly are therefore strongly advocated—these include: the improved mules operation, mid season crutching and good animal husbandry.