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Alanine-substituted mutant on Gly^(373) and Asn^(375) of Cry1Ai-h-loop 2 causes reduction in both toxicity and binding against Helicoverpa armigera

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Journal of Integrative Agriculture 2019年第5期18卷 1064-1071页

作者： LIU Yu-xiao ZHOU Zi-shan LIANG Ge-mei SONG Fu-ping ZHANG Jie State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant Protection Chinese Academy of Agricultural Sciences

Cry1Ai-h-loop 2 is a mutant of Cry1Ai constructed by exchanging loop 2 from Cry1Ah protein and shows insecticidal activity against Helicoverpa armigera. The toxicity of Cry1 Ai-h-loop 2, in contrast to the very low toxicity of Cry1Ai, is closely associated with the eleven residues in the loop 2 region. To characterize the key sites of loop 2 in Cry1Ai-h-loop 2, alaninesubstituted mutants were generated. The toxicity of these mutants against H. armigera indicated that dual-mutant on Gly373 and Asn375 caused a significant decrease in toxic activity. ELISA binding and competition binding assays demonstrated that the reduction of toxicity in the mutant of interest was correlated with decreased binding affinity.

关键词： Bacillus thuringiensis Cry1Ai Domain Ⅱ-loop2 Helicoverpa armigera binding affinity

Comparative transcriptomic analysis of larval and adult Malpighian tubules from the cotton bollworm Helicoverpa armigera

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Insect Science 2018年第6期25卷 991-1005页

作者： Yi-Yang Yuan Mei Li Fan Fan Xing-Hui Qiu State Key Laboratory of Integrated Management of Pest Insects and Rodents Institute of ZoologyChinese Academy of SciencesBeijingChina College of Plant Protection Hebei Agricultural UniversityBaodingHebei ProvinceChina

Malpighian tubules (MTs)are moregulatory organs of insects.However, usually considered the key excretory and os- increasing evidence has suggested that MTs perform many more functions than just osmoregulation.Until now,the molecular and physiological functions of MTs in the cotton bollworm (Helicoverpa armigera),a very important agricultural pest,are largely unknown.In this study,the transcriptomes of H. armigera MTs from larvae,male adults and female adults were sequenced using RNA-Seq technology,and comparative analyses of transcriptomes between two life stages (larval and adult)and between adult sexes were conducted.We generated a total of 84 643 high-quality unigenes,and identified a large number of abundant transcripts putatively encoding proteins involved in diuresis,detoxification,immunity,carbohydrate transport and metabolism,development and reproduction.We found that the expression pattern of unigenes was relatively similar between female and male adult MTs,but different between larval and adult MTs.Our data suggest that insect MTs may take multiple physiological fimctions as versatile organs.The extensive alterations in gene expression in MTs occurred from larvae to adults reflect an ecological adaptation to different feeding habits.Sexual dimorphism in the cotton bollworm is somewhat indicated by the transcriptional difference of genes related to carbohydrate metabolism,detoxification,immunity and reproduction in the MTs of male and female adults.

关键词： ecological adaptation Helicoverpa armigera Malpighian tubules sexual dimorphism transcriptome

Identification of the 2-tridecanone cis-acting element in the promoter of cytochrome P450 CYP6B7 in Helicoverpa armigera

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Insect Science 2018年第6期25卷 959-968页

作者： Li Xu Dong-Zhi Li Yuan-Yuan Luo Jian-Ying Qin Li-Hong Qiu College of Science China Agricultural UniversityBeijingChina

The expression level of cytochrome P450 genes in insects can be induced by plant allelochemicals,which is important for insects to adapt to host plants.Cytochrome P450CYP6B 7has been reported to be involved in pyethroid insecticide resistance in Heli- coverpa armigera,and its transcription level was induced by some inducers.Currently,the regulatory mechanism of the induced expression of CYP6B7remains unknown,although it is very important for understanding the detoxification mechanism to allelochemicals in host plants.The objective of the present study was to investigate the eis-acting ele- ment in the promoter of CYP6B7 mediating the inducible up-regulation of CYP6B7in H.armigera by 2-tridecanone.The promoter region of CYP6B7was cloned by genome walking technique and analyzed by transient transfeetion assay.Progressive 5'deletion of the promoter region of CYP6B7revealed that the relative luciferase activity of construct -320/+232could be significantly induced by 2-trideeanone.Further stepwise deletion between -320 and -238 bp found that construct -292/+232 could also be significantly induced by 2-tridecanone,but the adjacent construct -256/+232could not,suggesting the essential role of the sequence between -292 and --257 bp for 2-tridecanone induction. Nucleotide mutations between -292 and -281 bp had no influence on the induction ef- fect by 2-tridecanone,but nucleotide mutations between -280 and -257 bp significantly decreased the induction effect.These results demonstrated that the cis-acting element for 2-trideeanone induction was between -280 and -257 bp in the promoter of CYP6B7.

关键词： cis-acting element cytochrome P450 CYP6B7 Helicoverpa armigera induction 2-tridecanone

Progress in understanding hormonal regulation during the postembryonic development of Helicoverpa armigera

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Journal of Integrative Agriculture 2020年第6期19卷 1417-1428页

作者： ZHAO Xiao-fan Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology School of Life SciencesShandong UniversityQingdao 266237P.R.China

Lepidoptera,with 0.16 million species,is the second largest order of insecta.This order includes silk worms,butterflies,and many agricultural pests.The cotton bollworm,Helicoverpa armigera(Hubner)is one of the pests in Lepidoptera that seriously harms cotton plant and other crops in China and other countries.This pest develops resistance to chemical insecticides rapidly.Bacillus thuringiensis(Bt)transgenic cotton plants are developed to control H.armigera population in the field.However,duri ng the past years,the rapid evoluti on of Bt toxi n resista nee is observed in H.armigera in tran sge nic cott on fields.New approaches for the development of new envir on mentally friendly insecticides to control H.armigera have become necessary,and the molecular mechanisms underlying the development and physiological processes of this species need to be further understood.Considerable progress in the study of H.armigera development and physiology has been achieved in the last decade.This mini-review summarizes the main fin dings on the molecular mechanisms of horm onal regulation of the development of H.armigera to present new target genes for developing new approaches to control the pest.

关键词： Helicoverpa armigera Lepidoptera hormonal regulation development pest control

Polycalin is involved in the action mechanism of Cry2Aa toxin in Helicoverpa armigera(Hübner)

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Journal of Integrative Agriculture 2019年第3期18卷 627-635页

作者： WANG Bing-jie WANG Ya-nan WEI Ji-zhen LIU Chen CHEN Lin Myint Myint Khaing LIANG Ge-mei State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant Protection Chinese Academy of Agricultural Sciences Environment and Plant Protection Institute Chinese Academy of Tropical Agricultural Sciences Key Laboratory of Integrated Pest Management of Tropical Crops Ministry of Agriculture

Receptor proteins on the brush border membrane of the insect midgut epithelium are involved in the mode of action of insecticidal Cry proteins from Bacillus thuringiensis(Bt). Polycalin has been identified as a binding protein of the Bt Cry1 Ac toxin in several Lepidoptera including Helicoverpa armigera, but its role in the action mechanism of Cry2 Aa is still unclear. In this study, we investigated the binding characteristics of polycalin from the midgut of H. armigera with Cry2 Aa and its role in the toxicity of Cry2 Aa. The results demonstrated that heterologously expressed H. armigera polycalin peptide could bind with Cry2 Aa with high affinity(K_d=32 nmol L^(–1)). The toxicity of Cry2 Aa decreased by 27% after H. armigera larvae ingested polycalin antisera. These results suggested that polycalin could be a potential functional receptor for Cry2 Aa, and it plays an important role in the susceptibility of H. armigera to Cry2 Aa.

关键词： Helicoverpa armigera polycalin insecticidal toxicity antisera binding affinity

Screening of High Virulent Isolate of Beauveria spp. against Helicoverpa assulta and Changes in Protective Enzymes Activities of Infected Larvae

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Plant Diseases and Pests 2019年第2期10卷 15-18,21页

作者： Zhang Guochao Liu Mingke Zhou Jianyun Chen Dexin Yan Fangfang Xu Chuangtao Jiang Lianqiang Li Maoye Shandong Tobacco Research Institute Corporation School of Plant Protection Anhui Agricultural University Guiyang Branch of Guizhou Tobacco Company Qingzhou Tobacco Research Institute China National Tobacco Corporation Sichuan Panzhihua Tobacco Company Sichuan Luzhou Tobacco Company Sichuan Liangshan Tobacco Company

Eleven isolates of Beauveria spp., including B. bassiana(Bb062) newly isolated from Helicoverpa assulta, and other seven B. bassiana isolates and three B. brongniartii isolates that were originally isolated from different geographic origins and various hosts, were tested against the 3^(rd) instar larvae of H. assulta. The protective enzyme activity in the 3^(rd) larvae of H. assulta infected by highly virulent isolate was also assayed. The results showed that the isolate Bb062 had the highest virulence to the 3^(rd) instar larvae among eleven isolates of Beauveria spp., with a corrected mortality reaching 91.07% within 10 d post treatment, and the LT_(50) was 4.67 d. After inoculated with three concentrations(1.0 ×10~6, 1.0 ×10~7 and 1.0×10~8 conidia/mL) of Bb062 conidial suspension, the accumulative mortality of H. assulta larvae increased with the increase of concentration and observation time, and the LC_(50) was 1.82×10~7 conidia/mL. Activities of superoxide dismutase(SOD), peroxidase(POD) and catalase(CAT) in H. assulta larvae first increased rapidly then dropped sharply within 72 h. Bb062 showed high virulence to H. assulta and could inhibit activities of protective enzymes. Therefore, it will be a promising biocontrol agent against H. assulta.

关键词： Helicoverpa assulta Beauveria spp. Virulence Protective enzymes

Forecasting Helicoverpa populations in Australia： A comparison of regression based models and a bioclimatic based modelling approach

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Insect Science 2005年第1期12卷 45-56页

作者： MYRONP.ZALUCKI MICHAELJ.FURLONG SchoolofIntegrativeBiology TheUniversityofQueenslandStLuciaBrisbaneQueensland4072Australia

Long-term forecasts of pest pressure are central to the effective management of many agricultural insect pests. In the eastern cropping regions of Australia, serious infestations of Helicoverpa punctigera (Wallenglen) and H. armigera (Hübner)(Lepidoptera:Noctuidae) are experienced annually. Regression analyses of a long series of light-trap catches of adult moths were used to describe the seasonal dynamics of both species. The size of the spring generation in eastern cropping zones could be related to rainfall in putative source areas in inland Australia. Subsequent generations could be related to the abundance of various crops in agricultural areas, rainfall and the magnitude of the spring population peak. As rainfall figured prominently as a predictor variable, and can itself be predicted using the Southern Oscillation Index (SOI), trap catches were also related to this variable. The geographic distribution of each species was modelled in relation to climate and CLIMEX was used to predict temporal variation in abundance at given putative source sites in inland Australia using historical meteorological data. These predictions were then correlated with subsequent pest abundance data in a major cropping region. The regression-based and bioclimatic-based approaches to predicting pest abundance are compared and their utility in predicting and interpreting pest dynamics are discussed.

关键词：澳大利亚种群数量预测方法生物气候学迁移 Helicoverpa 害虫防治种群地理分布

Influences of insecticides on toxicity and cuticular penetration of abamectin in Helicoverpa armigera

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Insect Science 2005年第2期12卷 109-119页

作者： QIANGWANG JIA-ANCHENG ZHONG-MINLIU SHENG-GANWU XUE-PINGZHAO CHANG-XINGWU InstituteofAppliedEntomology ZhejiangUniversityChina//InstituteforQualityandStandardofAgriculturalProductsZhejiangAcademyofAgriculturalSciencesChina//ZhejiangKeyLabofFoodSafetyHangzhouChina InstituteofAppliedEntomology ZhejiangUniversityChina CentreforHorticultureandPlantSciences UniversityofWesternsydneyFlawkesburyRichmondNewSouthWalesAustralia InstituteforQualityandStandardofAgriculturalProducts ZhejiangAcademyofAgriculturalSciencesChina InstituteforQualityandStandardofAgriculturalProducts ZhejiangAcademyofAgriculturalSciencesChina//ZhejiangKeyLabofFoodSafetyHangzhouChina

Synergistic actions for mixtures of abamectin with other insecticides in some insect pests were evaluated, and the possible synergistic mechanism was studied by the comparison in toxicity and cuticular penetration of abamectin between with and without other insecticides or synergists in Helicoverpa armigera larvae. The results of bioassay showed that horticultural mineral oil (HMO), hexaflumuron, chlorpyrifos, and some other insecticides were synergistic to abamectin with 152.0-420.0 of co-toxicity coefficient(CTC) in some agricultural insect pests. In topical application tests, HMO or piperonyl butoxide (PBO) increased the toxicity of abamectin in larvae of H. armigera, but the mortality was not affected by s,s,s-tributylphorotrithioate (DEF) and triphenylphosphate (TPP). The synergistic action of HMO was obviously higher than PBO, and when treated simultaneously with abamectin, HMO gave a more significant synergism than if treated 2 hours ahead. The highest synergistic effect (SE) was found in the mixture of ‘abamectin+HMO (1:206)'. The mortality did not increase or the toxicity drop, when a synergist or HMO was added into the mixture of ‘abamectin+HMO' or ‘abamectin+synergist', respectively. Results from the isotope tracing experiments showed that HMO significantly enhanced the penetration of ^3H-abamectin through the cuticle of H.armigera larvae, which resulted in the synergism of the mixture. The cuticular penetration of ^3H-abamectin was not accumulatively affected by chlorpyrifos, nor by hexaflumuron,though there was an inhibition within 30 seconds or 1 hour after treated by these two chemicals respectively. Results suggested that the synergism of abamectin mixed with hexaflumuron or chlorpyrifos might be related to inhibition of metabolic enzymes or target sites in the larvae.

关键词：杀虫剂混用剂增效剂毒性表皮渗透能力 Helicoverpa armigera abamectin

Partial purification of Cytochrome P450 from Helicoverpa armigera （Hübner）

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Insect Science 2005年第4期12卷 313-317页

作者： CAI-HONG YU XI-WU GAO Department of Entomology China Agricultural University Beijing China College of Chemistry and Environment Engineering China University of Mining and Technology BeijingChina.

从胖身体和棉花一种蛾的幼虫的中间的勇气的细胞色素 P450 (Cyt-P450 ) 蛋白质， Helicoverpa 有佩带徽章权利的人一，被一套纯化过程部分分别地包括微分离心分离，小伙子的增溶，由木钉降水的蛋白质降水和 DE-32 柱层析净化。Cyt-P450 ... 详细信息

从胖身体和棉花一种蛾的幼虫的中间的勇气的细胞色素 P450 (Cyt-P450 ) 蛋白质， Helicoverpa 有佩带徽章权利的人一，被一套纯化过程部分分别地包括微分离心分离，小伙子的增溶，由木钉降水的蛋白质降水和 DE-32 柱层析净化。Cyt-P450 被 COdifference 光谱和 SDS 页的方法检测。从胖身体 ofH 的净化的 solubilized 微体的部分。有佩带徽章权利的人一多于 17 褶层被净化。三个蛋白质乐队被 SDS 页与分子的群众检测 70 600， 63 300 和 571 200Da。有 63 300 和 571 200Da 的分子的质量的蛋白质是 Cyt-P450 的 isozymes，是可能的。

关键词：细胞色素P450 净化方法棉铃虫 Helicoverpa armigera 昆虫学光谱分析分子机制