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The selective regulation of immune responses by matrix metalloproteinase MMP14 in Ostrinia furnacalis

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Insect Science 2023年第6期30卷 1622-1636页

作者： Kangkang Chen Shiqi Lu Jiahui Song Xiaoyi Dou Xiangyi Wei Xinyan Wang Xu Liu Congjing Feng Department of plant protection College of Plant ProtectionYangzhou UniversityYangzhouJiangsuChina Department of Entomology University of GeorgiaAthensGAUSA

Matrix metalloproteinases (MMPs) are crucial for tissue remodeling and immune responses in insects, yet it remains unclear how MMPs affect the various immune processes against pathogenic infections and whether the responses vary among insects. In this study, we used the lepidopteran pest Ostrinia furnacalis larvae to address these questions by examining the changes of immune-related gene expression and antimicrobial activity after the knockdown of MMP14 and bacterial infections. We identified MMP14 in O. furnacalis using the rapid amplification of complementary DNA ends (RACE), and found that it was conserved and belonged to the MMP1 subfamily. Our functional investigations revealed that MMP14 is an infection-responsive gene, and its knockdown reduces phenoloxidase (PO) activity and Cecropin expression, while the expressions of Lysozyme, Attacin, Gloverin, and Moricin are enhanced after MMP14 knockdown. Further PO and lysozyme activity determinations showed consistent results with gene expression of these immune-related genes. Finally, the knockdown of MMP14 decreased larvae survival to bacterial infections. Taken together, our data indicate that MMP14 selectively regulates the immune responses, and is required to defend against bacterial infections in O. furnacalis larvae. Conserved MMPs may serve as a potential target for pest control using a combination of double-stranded RNA and bacterial infection.

关键词： antimicrobial peptide cecropin lysozyme MMP14 phenoloxidase RNAi

The temporal-spatial expression and functional analysis of three gustatory receptor genes in Solenopsis invicta using sweet and bitter compounds

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Insect Science 2024年第2期31卷 448-468页

作者： Muhammad Irfan Waris Yanyuan Lei Guojun Qi Ziying Guan Abdul Rashied Jie Chen Lihua Lyu Institute of plant protection Guangdong Academy of Agricultural SciencesGuangdong Provincial Key Laboratory of High Technology for Plant ProtectionGuangzhouChina Department of Entomology College of Plant ProtectionSouth China Agricultural UniversityGuangzhouChina

The insect gustatory system participates in identifying potential food sources and avoiding toxic compounds.During this process,gustatory receptors(GRs)recognize feeding stimulant and deterrent compounds.However,the GRs involved in recognizing stimulant and deterrent compounds in the red imported fire ant,Solenopsis invicta,remain unknown.Therefore,we conducted a study on the genes SinvGR1,SinvGR32b,and SinvGR28a to investigate the roles of GRs in detecting feeding stimulant and deterrent compounds.In this current study,we found that sucrose and fructose are feeding stimulants and the bitter compound quinine is a feeding deterrent.The fire ant workers showed significant behavior changes to avoid the bitter taste in feeding stimulant compounds.Reverse transcription quantitative real-time polymerase chain reaction results from developmental stages showed that the SinvGR1,SinvGR32b,and SinvGR28a genes were highly expressed in fire ant workers.Tissue-specific expression profiles indicated that SinvGR1,SinvGR32b,and SinvGR28a were specifically expressed in the antennae and foreleg tarsi of workers,whereas SinvGR32b gene transcripts were also highly accumulated in the male antennae.Furthermore,the silencing of SinvGRI or SinvGR32b alone and the co-silencing of both genes disrupted worker stimulation and feeding on sucrose and fructose.The results also showed that SinvGR28a is required for avoiding quinine,as workers with knockdown of the SinvGR28a gene failed to avoid and fed on quinine.This study first identified stimulant and deterrent compounds of fire ant workers and then the GRs involved in the taste recognition of these compounds.This study could provide potential target gustatory genes for the control of the fire ant.

关键词： behavioral responses feeding assays gustatory receptors RNA interference Solenopsis invicta temporal-spatial expression

A Nicotiana benthamiana receptor-like kinase regulates Phytophthora resistance by coupling with BAK1 to enhance elicitin-triggered immunity

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Journal of Integrative plant Biology 2023年第6期65卷 1553-1565页

作者： Yifan Zhang Zhiyuan Yin Lei Pi Nan Wang Jinghao Wang Hao Peng Daolong Dou College of Plant Protection China Agricultural UniversityBeijing 100094China College of Plant Protection Nanjing Agricultural UniversityNanjing 210095China Institute of plant protection Jiangsu Academy of Agricultural SciencesNanjing 210014China Department of plant Pathology Washington State UniversityPullmanWashington 99164USA

Cell-surface-localized leucine-rich-repeat receptorlike kinases(LRR-RLKs)are crucial for plant immunity.Most LRR-RLKs that act as receptors directly recognize ligands via a large extracellular domain(ECD),whereas LRR-RLK that serve as regulators are relatively small and contain fewer LRRs.Here,we identified LRR-RLK regulators using high-throughput tobacco rattle virus(TRV)-based gene silencing in the model plant Nicotiana benthamiana.We used the cell-death phenotype caused by INF1,an oomycete elicitin that induces pattern-triggered immunity,as an indicator.By screening 33 small LRR-RLKs(≤6 LRRs)of unknown function,we identified ELICITIN INSENSITIVE RLK 1(NbEIR1)as a positive regulator of INF1-induced immunity and oomycete resistance.Nicotiana benthamiana mutants of eir1 generated by CRISPR/Cas9-editing showed significantly compromised immune responses to INF1 and were more vulnerable to the oomycete pathogen Phytophthora capsici.NbEIR1 associates with BRI1-ASSOCIATED RECEPTOR KINASE 1(NbBAK1)and a downstream component,BRASSINOSTEROIDSIGNALING KINASE 1(NbBSK1).NbBSK1 also contributes to INF1-induced defense and P.capsici resistance.Upon INF1 treatment,NbEIR1 was released from NbBAK1 and NbBSK1 in vivo.Moreover,the silencing of NbBSK1 compromised the association of NbEIR1 with NbBAK1.We also showed that NbEIR1 regulates flg22-induced immunity and associates with its receptor,FLAGELLIN SENSING 2(NbFLS2).Collectively,our results suggest that NbEIR1 is a novel regulatory element for BAK1-dependent immunity.NbBSK1-NbEIR1 association is required for maintaining the NbEIR1/NbBAK1 complex in the resting state.

关键词： BAK1 BSK1 INF1 leucine-rich repeat(LRR) LRR-RLK Oomycete

Multiple endocrine factors regulate nutrient mobilization and storage in Aedes aegypti during a gonadotrophic cycle

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Insect Science 2023年第2期30卷 425-442页

作者： Xiaoyi Dou Kangkang Chen Mark R.Brown Michael R.Strand Department of Entomology University of GeorgiaAthensGeorgia30602United States Department of plant protection College of Horticulture and Plant ProtectionYangzhou UniversityYangzhouJiangsu ProvinceChina

Anautogenous mosquitoes must blood feed on a vertebrate host to produce eggs.Each gonadotrophic cycle is subdivided into a sugar-feeding previtellogenic phase that produces primary follicles and a blood meal-activated vitellogenic phase in which large numbers of eggs synchronously mature and are laid.Multiple endocrine factors including juvenile hormone(JH),insulin-like peptides(ILPs),ovary ecdysteroidogenic hormone(OEH),and 20-hydroxyecdysone(20E)coordinate each gonadotrophic cycle.Egg formation also requires nutrients from feeding that are stored in the fat body.Regulation of egg formation is best understood in Aedes aegypti but the role different endocrine factors play in regulating nutrient mobilization and storage remains unclear.In this study,we report that adult female Ae.aegypti maintained triacylglycerol(TAG)stores during the previtellogenic phase of the first gonadotrophic cycle while glycogen stores declined.In contrast,TAG and glycogen stores were rapidly mobilized during the vitellogenic phase and then replenishment.Several genes encoding enzymes with functions in TAG and glycogen metabolism were differentially expressed in the fat body,which suggested regulation was mediated in part at the transcriptional level.Gain of function assays indicated that stored nutrients were primarily mobilized by adipokinetic hormone(AKH)while juvenoids and OEH regulated replenishment.ILP3 further showed evidence of negatively regulating certain lipolytic enzymes.Loss of function assays indicated AKH depends on the AKH receptor(AKHR)for function.Altogether,our results indicate that the opposing activities of different hormones regulate nutrient stores during a gonadotrophic cycle in Ae.aegypti.

关键词： adipokinetic hormone ecdysone endocrine glycogen insulin-like peptide juvenile hormone mosquito nutrient ovary ecdysteroidogenic hormone triacylglycerol

Species of the genus Steganopsis from China(Diptera:Lauxaniidae)

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Zoological Systematics 2023年第2期48卷 177-183页

作者： Li Shi Donghui Wang Ding Yang college of Horticulture and plant protection Inner Mongolia Agricultural UniversityHohhot 010018China College of Plant Protection China Agricultural UniversityBeijing 100094China

Species of the genus Steganopsis from China are revised,including a new species,namely S.mayanghensis Shi&Yang,sp.nov.,and a new recorded species,S.multilineata de Meijere,1924.A key to species of the genus Steganopsi... 详细信息

关键词： Lauxaniinae new species new record taxonomy

Detection of ryanodine receptor target-site mutations in diamide insecticide-resistant Spodoptera frugiperda in China

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Insect Science 2021年第3期28卷 639-648页

作者： Sheng-Lan Lv Yang Shi Jia-Cheng Zhang Pei Liang Lei Zhang Xi-Wu Gao College of Plant Protection China Agricultural UniversityBeijingChina

Spodoptera frugiperda(Lepidoptera:Noctuidae)is a widely distributed pest of corn.Since it invaded China in 2018,it has caused serious damage to local corn production.Chlorantraniliprole,an anthranilic diamide insecticide,has been widely used to control lepidopteran pests.Tetrachloropyramid is a new allosteric modulator insecticide devel-oped based on chlorantraniliprole,so it has a similar mechanism and insecticidal effect.In this study,we investigated resistance levels to chlorantraniliprole and tetrachloropyramid in S.frugiperda from 13 populations in China.Among the populations tested,the relative highest resistance to chlorantraniliprole occurred in the Guangzhou population,and the most susceptible to chlorantraniliprole was found in the Wuhan population.The lethal dosage LD50 value of the Guangzhou population against chlorantraniliprole was 27.8-fold higher than that of the Wuhan population.Minimal differences were observed among 5".frugiperda populations in terms of sensitivity to tetrachloropyramid.Heterozygous mutations at the 14734 site of the ryanodine receptor(RyR)were found,while no mutations were found in the G4891 site.The mutations were detected in only two of the 786 individuals analyzed one from the Qinzhou population and other from the Anshun population(frequency below 2%in both cases).There were no significant differences in the expression levels of RyR between Guangzhou and Wuhan populations.In summary,our results indicate that:(i)S.frugiperda has low resistance levels to diamide insecticides in China;and(ii)the differences in relative resistance among the 13 populations analyzed are not caused by the mutations in RyR or the expression of RyR.

关键词： diamide insecticides resistance resistance risk Spodoptera frugiperda

Genetic analysis and molecular detection of resistance to chlorpyrifos mediated by the A216S substitution in acetylcholinesterase-1 in the plant bug Apolygus lucorum

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Insect Science 2020年第6期27卷 1224-1232页

作者： Kai-Ran Zuo Yi-Hua Yang Yi-Dong Wu Shu-Wen Wu College of Plant Protection Nanjing Agricultural UniversityNanjingChina

The green plant bug Apolygus lucorum is a major pest of Bacillus thuringiensis cotton in China.Previously,we reported that chlorpyrifos resistance in a laboratory-selected strain of A.lucorum(BZ-R)is associated with the homozygosis of an allele in the ace-1 gene encoding an alanine to serine substitution at position 216 of acetylcholinesterase-1.Here we describe the results of crosses between the resistant BZ-R strain(41-fold to chlorpyrifos)and the unselected susceptible BZ-S strain homozygous for the wild type alanine allele at position 216.Resistance to chlorpyrifos was inherited as a semi-dominant trait mainly controlled by a single autosomal gene and co-segregates strongly but not completely with the serine substitution in ace-1.Synergism bioassays and enzyme assays showed that minor contributions to resistance are also made by enhanced cytochrome P450 and carboxylesterase activities.A survey of 25 field populations from five Chinese provinces showed strong positive correlations between 50%lethal concentration against chlorpyrifos and S216 allele and genotype frequencies,although the most tolerant populations still only show 40%-50%S216 allele frequencies.The results above provide important information for designing effective resistance monitoring and management strategies for A.lucorum in China.

关键词： acetylcholinesterase-1 Apolygus lucorum chlorpyrifos resistance inheritance

Knockout of three aminopeptidase N genes does not affect susceptibility of Helicoverpa armigera larvae to Bacillus thuringiensis Cry1A and Cry2A toxins

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Insect Science 2020年第3期27卷 440-448页

作者： Jing Wang Ya-Yun Zuo Ling-Li Li Hui Wang Shao-Yan Liu Yi-Hua Yang Yi-Dong Wu College of Plant Protection Nanjing Agricultural UniversityNanjingChina

Bacillus thuringiensis(Bt)insecticidal toxins have been globally utilized for control of agricultural insects through spraying or transgenic crops.Binding of Bt toxins to special receptors on midgut epithelial cells of target insects is a key step in the mode of action.Previous studies suggested aminopeptidase N1(APN1)as a receptor or putative receptor in several lepidopteran insects including Helicoverpa armigera through evidence from RNA interefence‐based gene silencing approaches.In the current study we tested the role of APNs in the mode of action of Bt toxins using clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR‐associated protein 9‐mediated gene knockout.Three APN genes(HaAPN1,HaAPN2 and HaAPN5)were individually knocked out in a susceptible strain(SCD)of H.armigera to establish three homozygous knockout strains.Qualitative in vitro binding studies indicated binding of Cry1Ac or Cry2Ab to midgut brush border membrane vesicles was not obviously affected by APN knockout.Bioassay results showed that none of the three knockouts had significant changes in susceptibility to Cry1A or Cry2A toxins when compared with the SCD strain.This suggests that the three HaAPN genes we tested may not be critical in the mode of action of Cry1A or Cry2A toxins in H.armigera.

关键词： aminopeptidase N Bacillus thuringiensis CRISPR/Cas9 Helicoverpa armigera receptor

Changes in fungal community and diversity in strawberry rhizosphere soil after 12 years in the greenhouse

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Journal of Integrative Agriculture 2019年第3期18卷 677-687页

作者： LI Wei-hua LIU Qi-zhi College of Plant Protection China Agricultural University

Soil fungi play a very important role in the soil ecological environment. In agricultural production, long-term monoculture and continuous cropping lead to changes in fungal community diversity. However, the effects of long-term monoculture and continuous cropping on strawberry plant health and fungal community diversity have not been elucidated. In this study, using high-throughput sequencing(HTS), we compared the fungal community and diversity of strawberry rhizosphere soil after various durations of continuous cropping(0, 2, 4, 6, 8, 10 and 12 years). The results showed that soil fungal diversity increased with consecutive cropping years. Specifically, the soil-borne disease pathogens Fusarium and Guehomyces were significantly increased after strawberry continuous cropping, and the abundance of nematicidal(Arthrobotrys) fungi decreased from the fourth year of continuous cropping. The results of correlation analysis suggest that these three genera might be key fungi that contribute to the changes in soil properties that occur during continuous cropping. In addition, physicochemical property analysis showed that the soil nutrient content began to decline after the fourth year of continuous cropping. Spearman's correlation analysis showed that soil pH, available potassium(AK) and ammonium nitrogen(NH_4^+-N) were the most important edaphic factors leading to contrasting beneficial and pathogenic associations across consecutive strawberry cropping systems.

关键词： fungal community soil-borne disease replanted strawberry rhizosphere soil agricultural soil ecology