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Assessment of the potential toxicity of insecticidal compounds to Peristenus spretus,a parasitoid of mirid bugs

Journal of Integrative Agriculture 2022年第5期21卷 1424-1435页

作者： ZHAO Man LI Yun-he NIU Lin-lin CHEN Lin liang ge-mei State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing 100193P.R.China College of Plant Protection Henan Agricultural UniversityZhengzhou 450002P.R.China

With the increased cultivation of Bt crops in China,Apolygus lucorum and other mirid bugs have emerged as important agricultural pests because they are insensitive to the Bt *** addition,the reduction of pesticide applications after planting Bt crops also increases the severity of mirid bug *** spretus is a parasitoid of mirid nymphs,but its sensitivity to Bt proteins is not *** the current study,we developed a dietary exposure assay to assess the effects of Bt proteins(Cry1Ab,Cry1Ac,Cry1F,Cry2Aa,and Cry2Ab)on *** adults using a diet consisting of a 10% honey solution with or without Bt proteins at 400μg g^(-1) *** results showed that the survival and reproduction of *** adults were reduced by the cysteine protease inhibitor E-64(a positive control)but were not affected by any of the five Bt *** activities of digestive,detoxifying,and antioxidant enzymes in *** were also unaffected by diets containing the Cry proteins,but they were significantly affected by the diet containing *** then developed a tritrophic bioassay to determine the effects of the five Bt proteins on *** larvae and *** this assay,*** nymphs fed an artificial diet containing Cry proteins were used as the hosts for *** results of the tri-trophic assay indicated that neither the pupation rate nor the eclosion rate of the *** parasitoids were significantly affected by the presence of high concentrations of Bt proteins in the parasitized *** *** overall results indicate that these two assays can be used to evaluate the toxicity of insecticidal compounds to *** and that the tested Cry proteins are not toxic to ***.

关键词： transgenic crops plant bugs parasitic natural enemy Cry proteins non-target risk assessment

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Performance of Three Adelphocoris spp.(Hemiptera: Miridae) on Flowering and Non-flowering Cotton and Alfalfa

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Journal of Integrative Agriculture 2014年第8期13卷 1727-1735页

作者： GAO Zhen PAN Hong-sheng LIU Bing LU Yan-hui liang ge-mei State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant Protection Chinese Academy of Agricultural Sciences

The genus Adelphocoris（Hemiptera： Miridae） is a group of key insect pests in cotton fields in China that includes three dominant species： A. suturalis（Jakovlev）, A. lineolatus（Goeze） and A. fasciaticollis（Reuter）. Previous field surveys have found that adults of these Adelphocoris species usually move onto specific host plants when the plants enter the flowering stage. To determine the potential trade-offs for this host-plant preference behavior, the performance of these three Adelphocoris spp. on flowering and non-flowering cotton and alfalfa were compared in the laboratory. The results showed that Adelphocoris spp. had significantly higher nymphal developmental and survival rates, along with increased adult longevity and fecundity on flowering cotton and alfalfa than on non-flowering plants of either species. In addition, compared with cotton plants, alfalfa generally promoted better performance of these three Adelphocoris species, especially for A. lineolatus. Simple correlation analysis indicated that female adult longevity was positively correlated to male adult longevity and female fecundity, and female fecundity was positively correlated to nymphal development and survival rates. This study demonstrated a positive correlation between adult preference and offspring/adult performance for all three Adelphocoris species, with no evidence of any trade-offs for this preference for flowering host plants. These findings support the hypothesis that hemimetabolous insects such as mirid bugs have positive adult preference-adult/nymphal performance relationships, which is likely due to the similar feeding habits and nutritional requirements of adults and nymphs.

关键词： Adelphocoris lineolatus Adelphocoris suturalis Adelphocoris fasciaticollis preference-performance relationship cotton alfalfa flowering stage

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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

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Characterization and functional analysis of β-1,3-galactosyltransferase involved in Cry1Ac resistance from Helicoverpa armigera(Hübner)

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Journal of Integrative Agriculture 2015年第2期14卷 337-346页

作者： ZHANG Li-li liang ge-mei GAO Xi-wu CAO Guang-chun GUO Yu-yuan College of Agriculture and Biotechnology China Agricultural University State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant Protection Chinese Academy of Agricultural Sciences

Carbohydrate chains are the principal antigens by which Bacillus thuringiensis（Bt） identify receptor proteins. The interaction between the antigen and Bt causes a pore in the membrane of midgut epithelial cells of insects. Receptor proteins, such as aminopeptidase N and alkaline phosphatase, are glycoproteins. Cadherin is another cell surface receptor protein which has potential glycosylation sites. Glycosyltransferase is very important for the synthesis and modification of receptor proteins. It can indirectly influence the function of Bt. The 1 950 bp full-length c DNA encoding β-1,3-galactosyltransferase was cloned from the the midgut of Helicoverpa armigera by degenerative PCR combined with RACE techniques（GAL-Harm, gen Bank accession no.： GQ904195.1） with two potential N-glycosylation sites（^157NNTI^160 and ^272NKTL^275）. Protein sequence alignments revealed that H. armigera β-1,3-galactosyltransferase shared high identity with β-1,3-galactosyltransferase in other insect species. The expression level of the β-1,3-galactosyltransferase gene in Cry1Ac-resistant H. armigera larvae was 9.2-fold higher than that in susceptible strain. The function of β-1,3-galactosyltransferase was investigated using RNAi technique. The result showed Cry1 Ac enhanced the toxicity against the si RNA-treated larvae compared with non-si RNA-treated ones, which indicated β-1,3-galactosyltransferase played an important role for the insecticidal toxicity of Cry1 Ac in H. armigera.

关键词： β-1 3-galactosyltransferase Cry1Ac resistance Helicoverpa armigera RNAi

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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

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Comparative Analysis of Cry1Ac Toxin Oligomerization and Pore Formation Between Bt-Susceptible and Bt-Resistant Helicoverpa armigera Larvae

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Journal of Integrative Agriculture 2012年第11期11卷 1845-1851页

作者： LI Yi-ping WU Jun-xiang LIU Chen-xi YUAN Xiang-qun WU Kong-ming liang ge-mei GUO Yu-yuan Key Laboratory of Applied Entomology " Key Laboratory of Plant Protection Resources and Pest Management Ministry of Education~ College of Plant Protection Northwest Agriculture and Forestry University Yangling 712100 P.R.China State Key Laboratory for Biology of Plant Diseases and Insect Pests/Institute of Plant Protection Chinese Academy of Agricultural Sciences Beijing 100193 P.R.China

With the long-term use of Bacillus thuringiensis （Bt） insecticide and expansion of CrylA-expressing transgenic plants, some insect pests have developed resistance to Bt in open fields, greenhouses, and in the laboratory. Bt resistance is complex and there appear to be different ways for resistance development. Understanding the Bt resistance mechanisms is critical to prolong its usefulness. In this article, Bt receptors, the cadherin and aminopeptidase N （APN）, in brush border membrane vesicles （/3BMV） of Helicoverpa armigera were examined in both CrylAc-susceptible （96S） and CrylAc- resistant （LF120） strains, to compare CrylAc toxin oligomerization and pore formation in these two strains. CrylAc toxin oligomerization and pore formation in these two strains were compared. Results showed that cadherin and aminopeptidase N proteins could express normally in both susceptible and resistant 11. armigera strains. The ability to form CrylAc oligomers and ion channels on BBMVs was also not significantly different between these two strains.

关键词： Helicoverpa armigera Bt resistance oligomerization pore formation

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The Binding Characterization of Cry Insecticidal Proteins to the Brush Border Membrane Vesicles of Helicoverpa armigera, Spodoptera exigua, Spodoptera litura and Agrotis ipsilon

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Journal of Integrative Agriculture 2013年第9期12卷 1598-1605页

作者： LU Qiong CAO Guang-chun ZHANG Li-li liang ge-mei GAO Xi-wu ZHANG Yong-jun GUO Yu-yuan State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural Sciences College of Agriculture and Biotechnology China Agricultural University

Cry toxins produced by Bacillus thuringiensis （Bt） are effective biological insecticides against certain insect species. However, there are potential risks of the evolved resistance of insects to Cry toxin owing to decreased binding of toxins to target sites in the brush border membranes of the larva midgut. The Cry toxins with different binding sites in the larval midgut have been considered to be a good combination to deploy in delaying resistance evolution. Bioassay results demonstrated that the toxicity of different Cry toxins ranked differently for each species. The toxicity ranking was CrylAc〉CrylAb〉Cry2Ab for Helicoverpa armigera, CrylB〉CrylC〉Cry2Ab for Spodoptera exigua, and Cry2Ab〉CrylB〉 CrylC for S. litura. Only Cry2Ab was toxic to Agrotis ipsilon. Binding experiments were performed with ^125I-CrylAb, ^125I-CrylAc, ^125I-CrylB, ^125I-CrylC, ^125I-Cry2Ab and the brush border membranes vesicles （BBMV） from H. armigera, S. exigua, S. litura and A. ipsilon. The binding of CrylAb and CrylAc was shown to be saturable by incubating with increasing concentrations of H. armigera BBMV （Kd=（45.00＋2.01） nmol L-1 and （12.80＋0.18） nmol L^-1, respectively; Bmax=（54.95±l.79） ng and （55.44±0.91） ng, separately）. The binding of CrylB was shown to be saturable by incubating with increasing concentrations ofS. exigua BBMV （Kd=（23.26±1.66） nmol L^-1; Bmax=（65.37±1.87） ng）. The binding of ^125I-Cry toxins was shown to be non-saturable by incubating with increasing concentrations of S. litura and A. ipsilon BBMV. In contrast, CrylB and CrylC showed some combination with the BBMV orS. litura, and a certain amount of Cry2Ab could bind to the BBMV ofA. ipsilon. These observations suggest that a future strategy could be devised for the focused combination of specific cry genes in transgenic crops to control target pests, widen the spectrum of insecticide effectiveness and postpone insect resistance evolution.

关键词： Cry toxin toxicity binding assays 1251 labeling BBMV

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Toxicity and binding analyses of Bacillus thuringiensis toxin Vip3A in Cry1Ac-resistant and-susceptible strains of Helicoverpa armigera(Hübner)

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Journal of Integrative Agriculture 2015年第2期14卷 347-354页

作者： ZHANG Qian CHEN Li-zhen LU Qiong ZHANG Yan liang ge-mei State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant Protection Chinese Academy of Agricultural Sciences Institute of Cotton Research Hebei Academy of Agriculture and Forestry Sciences College of Plant Science and Technology Huazhong Agricultural University

The Bacillus thuringiensis vegetative insecticidal protein, Vip3 A, represents a new family of Bt toxin and is currently applied to commercial transgenic cotton. To determine whether the Cry1Ac-resistant Helicoverpa armigera is cross-resistant to Vip3 Aa protein, insecticidal activities, proteolytic activations and binding properties of Vip3 Aa toxin were investigated using Cry1Ac-susceptible（96S） and Cry1Ac-resistant H. armigera strain（Cry1Ac-R）. The toxicity of Vip3 Aa in Cry1Ac-R slightly reduced compared with 96 S, the resistance ratio was only 1.7-fold. The digestion rate of full-length Vip3 Aa by gut juice extracts from 96 S was little faster than that from Cry1Ac-R. Surface plasmon resonance（SPR） showed there was no significant difference between the binding affinity of Vip3 Aa and BBMVs between 96 S and Cry1Ac-R strains, and there was no significant competitive binding between Vip3 Aa and Cry1 Ac in susceptible or resistant strains. So there had little cross-resistance between Vip3 Aa and Cry1 Ac,Vip3A＋Cry proteins maybe the suitable pyramid strategy to control H. armigera in China in the future.

关键词： vegetative insecticidal proteins(Vip3Aa) Bacillus thuringiensis Helicoverpa armigera cross-resistance binding affinity

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Expression of Aminopeptidase N1(APN1),the Main Receptor Protein for Bacillus thuringiensis CrylA Toxin from Helicoverpa armigera Larval Midgut in Trichoplusia ni cells

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Agricultural Sciences in China 2008年第3期7卷 329-335页

作者： CHANG Hong-lei liang ge-mei WANG Gui-rong YU Hong-kun GUO Yu-yuan WU Kong-ming State Key Laboratory for Biology of Plant Disease and Insect Pests Institute of Plant Protection Chinese Academy of Agricultural Sciences Beijing 100094 P.R.China

The aim of this article is to successfully express the Bt （Bacillus thuringiensis） toxin receptor protein located on the internal membrane of larval midgut of cotton bollworm （Helicoverpa armigera Hübner） within eukaryotic expression system, which is one of the key links for clarifying the relationship between receptor and Bt resistance. The fragments of aminopeptidase N1 （APN1） gene without signal peptide in the susceptible and the resistant H. armigera were cloned separately using PCR method, and were separately cloned into pUC 19 vector. After sequencing the gene, the fragments encoding for APN1 without signal peptide were cloned into the Bac-to-Bac baculovirus expression system with transfer vector pFastBacHTB under the polyhedron gene promoter. The recombinant transposing plasmid pFastBacHTB/APN1 was screened and then transformed into Escherichia coli DH10Bac. It was cultured in LB medium, which contained Te, Kan, ge, X-gal, and IPTG. The resulting recombinant bacmid was transfected into cells of the insect Trichoplusia ni and recombinant baculoviruse was obtained. The lysate of cells infected with recombinant baculoviruse was analyzed by SDS-PAge and blot analysis. The results showed that the recombinant baculoviruse was fully capable of expressing APN1. The APN1 gene successfully expressed in T. ni cell established the base for continuing the research on its function and relationshio of resistance with Bt.

关键词： APN1 baculovirus expression vector Trichoplusia ni expression

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A Fragment of Cadherin-Like Protein Enhances Bacillus thuringiensis Cry1B and Cry1C Toxicity to Spodoptera exigua(Lepidoptera:Noctuidae)

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Journal of Integrative Agriculture 2012年第4期11卷 628-638页

作者： LU Qiong ZHANG Yongojun CAO Guang-chun ZHANG Li-li liang ge-mei LU Yan-hui WU Kong-ming GAO Xi-wu GUO Yu-yuan College of Agriculture and Biotechnology China Agricultural University Beijing 100193 P.R.China State Key Laboratory for Biology of Plant Diseases and Insect Pests/Institute of Plant Protection Chinese Academy of Agricultural Sciences Beijing 100193 P.R. China

Cry toxins produced by Bacillus thuringiensis（Bt） are effective biological insecticides against certain insect *** this study,bioassay results indicated that Cry1B and Cry1C were toxic to Spodoptera *** also identified a cadherin-like gene in *** that could enhance the toxicity of Cry1B and *** cadherin-like gene identified from the larvae midgut tissue was cloned by reverse transcription polymerase chain reaction（RT-PCR） and rapid amplification of cDNA ends（RACE）.The full-length cDNA of the gene consisted of 5 220 bp encoding 1 740 amino acid with a predicted molecular mass of 196 *** search analysis showed that the predicted amino acid sequence had a high sequence identity to the published sequences of cadherin-like proteins from other Lepidoptera *** expression of the cadherin-like gene detected by qRT-PCR analysis revealed that the cadherin-like gene was mainly present in the gut of 4th instar larvae and during different life *** results suggested that the commercial development of this synergist has the potential to enhance Cry1B and Cry1C toxicity against Lepidoptera insects.

关键词： Bacillus thuringiensis cadherin-like Spodoptera exigua enhancement toxicity

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