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Gene cloning,protein expression,and enzymatic characterization of a double-stranded RNA degrading enzyme in Apolygus lucorum

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Insect Science 2024年第1期31卷 119-133页

作者： Jie-Yu Zhang Jing Zhao Keyan Zhu-Salzman Qin-Qin Ji Yi-Ping Jiang Liu-Bin Xiao De-Jin Xu Guang-Chun Xu Lin-Quan Ge Yong-An Tan College of plant protection Yangzhou UniversityYangzhouJiangsu ProvinceChina institute of plant protection Jiangsu Academy of Agricultural SciencesNanjingJiangsu ProvinceChina Department of Entomology Texas A&M UniversityCollege StationTXUSA Taizhou Customs of the People's Republic of China TaizhouJiangsu ProvinceChina

RNA interference(RNAi)is a powerful tool that post-transcriptionally silences target genes in eukaryotic ***,silencing efficacy varies greatly among different insect ***,we met with little success when attempting to knock down genes in the mirid bug Apolygus lucorum via dsRNA *** disappearance of double-stranded RNA(dsRNA)could be a potential factor that restricts RNAi ***,we found that dsRNA can be degraded in midgut fluids,and a dsRNase of ***(AldsRNase)was identified and *** alignment indicated that its 6 key amino acid residues and the Mg2+-binding site were similar to those of other insects’*** signal peptide and endonuclease non-specific domain shared high sequence identity with the brown-winged green stinkbug Plautia stali *** showed high salivary gland and midgut expression and was continuously expressed through the whole life cycle,with peaks at the 4th instar ecdysis in the whole *** purified AldsRNase protein obtained by heterologously expressed can rapidly degrade *** comparing the substrate specificity of AldsRNase,3 specific substrates(dsRNA,small interfering RNA,and dsDNA)were all degraded,and the most efficient degradation is ***,immunofluorescence revealed that AldsRNase was expressed in the cytoplasm of midgut *** cloning and functional study of AldsRNase,the enzyme activity and substrate specificity of the recombinant protein,as well as the subcellular localization of nuclease,the reason for the disappearance of dsRNA was explained,which was useful in improving RNAi efficiency in *** and related species.

关键词： Apolygus lucorum cloning dsRNase enzymatic activity protein expression

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A viral movement protein targets host catalases for 26S proteasome-mediated degradation to facilitate viral infection and aphid transmission in wheat

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Molecular plant 2024年第4期17卷 614-630页

作者： Shuyuan Tian Qingting Song Wenmei Zhou Jingke Wang Yanbin Wang Wei An Yunfeng Wu Lei Zhao State Key Laboratory for Crop Stress Resistance and High-Efficiency Production College of Plant ProtectionNorthwest A&F UniversityYanglingShaanxi 712100China Key Laboratory of Integrated Pest Management on Crops in Northwestern Loess Plateau Ministry of Agriculture and Rural AffairsCollege of Plant ProtectionNorthwest A&F UniversityYanglingShaanxi 712100China Key Laboratory of plant protection Resources and Pest Management Ministry of EducationCollege of Plant ProtectionNorthwest A&F UniversityYanglingShaanxi 712100China

The infection of host plants by many different viruses causes reactive oxygen species(Ros)accumulation and yellowing symptoms,but the mechanisms through which plant viruses counteract RoS-mediated immunity to facilitate infection and symptom development have not been fully *** plant viruses are transmitted by insect vectors in the field,but the molecular mechanisms underlying virus-host-insect interactions are *** this study,we investigated the interactions among wheat,barley yellow dwarf virus(BYDV),and its aphid vector and found that the BYDV movement protein(MP)interacts with both wheat catalases(CATs)and the 26S proteasomeubiquitin receptor non-ATPase regulatorysubunit2homolog(PSMD2)to facilitate the 26S proteasome-mediateddegradation of CATs,promotingviral infection,disease symptom development,and aphid *** of the BYDV MP gene in wheat enhanced the degradation of CATs,which leading to increased accumulation of ROS and thereby enhanced viral ***,transgenic wheat lines overexpressing BYDV MP showed significantly reduced proliferation of wingless aphids and an increased number of winged *** with this observation,silencing of CAT genes also enhanced viral accumulation and reduced the proliferation of wingless aphids but increased the occurrence of winged *** contrast,transgenic wheat plants overexpressing TaCAT1 exhibited the opposite changes and showed increases in grain size and weight upon infection with *** assays demonstrated that BYDV MP interacts with PSMD2 and promotes 26S proteasome-mediated degradation of TaCAT1 likely in a ubiquitination-independent ***,our study reveals a molecular mechanism by which a plant virus manipulates the Ros production system of host plants to facilitate viral infection and transmission,shedding new light on the sophisticated interactions among viruses,host plants,and insect vectors.

关键词： babarley yellow dwarf virus movement protein reactive oxygen species aphid catalase 26S proteasomeubiquitin receptor PSMD2

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Identification of an odorant receptor responding to sex pheromones in Spodoptera frugiperda extends the novel type-I PR lineage in moths

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Insect Science 2024年第2期31卷 489-502页

作者： Sai Zhang Emmanuelle Jacquin-Joly Nicolas Montagne Fang Liu Yang Liu Guirong Wang College of Horticulture and plant protection Yangzhou UniversityYangzhouChina State Key Laboratory for Biology of plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBejingChina institute of Ecology and En vironmental Sciences of Paris INRAESorbonne UniversityCNRSIRDUPECUniversity of ParisVersaillesFrance Shenzhen Branch Guangdong Laboratory for Lingnan Modern AgricultureGenome Analysis Laboratory of the Ministry of AgricultureAgricultural Genomics Institute at ShenzhenChinese Academy of Agricultural SciencesShenzhenChina

moths,pheromone receptors(PRs)are crucial for intraspecific sexual communication between males and *** PRs are considered as an ideal model for studying the evolution of insect PRs,and a large number of PRs have been identified and functionally characterized in different moth *** PRs were initially thought to fall into a single monophyletic clade in the odorant receptor(OR)family,but recent studies have shown that ORs in another lineage also bind type-I sex pheromones,which indicates that type-I PRs have multiple independent origins in the *** this study,we investigated whether ORs of the pest moth Spodoptera frugiperda belonging to clades closely related to this novel PR lineage may also have the capacity to bind type-I pheromones and serve as male *** the 7 ORs tested,only 1(SfruOR23)exhibited a male-biased expression ***,in vitro functional characterization showed that SfruOR23 could bind several type-I sex pheromone compounds with Z-9-tetradecenal(Z9-14:Ald),a minor component found in female sex pheromone glands,as the optimal *** addition,SfruOR23 also showed weak responses to plant volatile organic ***,we characterized an *** PR positioned in a lineage closely related to the novel PR clade,indicating that the type-I PR lineage can be extended in moths.

关键词： odorant receptor pheromone receptor lineage sex pheromone Spodoptera frugiperda

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Identification,characterization,and verification of miR399 target gene in grape

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Horticultural plant Journal 2024年第1期10卷 91-102页

作者： Maosong Pei Hainan Liu Tonglu Wei Huiying Jin Yihe Yu Mengting Ma Xiaomeng Song Rundong Dai Dalong Guo Henan Engineering Technology research Center of Quality Regulation and Controlling of Horticultural plants College of Horticulture and Plant ProtectionHenan University of Science and TechnologyLuoyangHenan 471023China

The microRNA miR399 plays an important role in phosphorus signal transduction pathways in ***,miR399 was shown to be closely associated with berry ripening in grape(Vitis vinifera).The objective of the present study was to elucidate the evolutionary characteristics of the miR399 gene family in grape and to verify the cleavage effect on the target *** miR399s were identified by miRNA sequencing and retrieval from the miRBase *** mature sequences and precursor sequences were subjected to phylogenetic analysis to reconstruct evolutionary trees,as well as secondary structure analysis of the precursor sequence,and prediction of target *** cisacting elements in the miR399 promoter were predicted and the cleavage effect of grape miR399b on its target genes was *** grape miR399 family comprised nine precursor sequences and nine mature *** precursor sequences formed a typical and stable stem—loop *** minimum folding free energy ranged from-55.70 kcal·mol^(-1)to-37.40 kcal·mol^(-1).Multiple sequence alignment revealed that the miR399 family was highly *** grape miR399 family was phylogenetically closely related to peach,apple,and citrus *** miR399s were predicted to target inorganic phosphate transporter 1—3,phospholipase D delta-like,and *** cleavage effect of grape miR399b on the target genes was verified by means of a dual-luciferase assay and 5’*** GUS staining showed that the promoter activity of miR399b was promoted by GA3treatment.

关键词： Grape miR399 Phosphorus metabolism Early ripening

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Hordedane diterpenoid phytoalexins restrict Fusarium graminearum infection but enhance Bipolaris sorokiniana colonization of barley roots

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Molecular plant 2024年第8期17卷 1307-1327页

作者： Yaming Liu Dario Esposto Lisa K.Mahdi Andrea Porzel Pauline Stark Hidayat Hussain Anja Scherr-Henning Simon Isfort Ulschan.Bathe Ivan F.Acosta Alga Zuccaro Gerd U.Balcke Alain Tissier Department of Cell and Metabolic Biology Leibniz Institute of Plant BiochemistryHalleGermany institute for plant Sciences Cluster of Excellence on Plant Sciences(CEPLAS)Cologne BiocenterUniversity of CologneCologneGermany Department of Bioorganic Chemistry Leibniz Institute of Plant BiochemistryHalleGermany Max Planck institute for plant Breeding research CologneGermany Horticultural Sciences University of FloridaGainesvilleFLUSA.

plant immunity is a multilayered process that includes recognition of patterns or effectors from pathogens to elicit defense *** include the induction of a cocktail of defense metabolites that typically restrict pathogen ***,we investigate the interaction between barley roots and the fungal pathogens Bipolaris sorokiniana(Bs)and Fusarium graminearum(Fg)at the metabolite *** identify hordedanes,a previously undescribed set of labdane-related diterpenoids with antimicrobial properties,as critical players in these *** of barley roots by Bs and Fg elicits hordedane synthesis from a 60o-kb gene *** reconstruction of the biosynthesis pathway in yeast and Nicotiana benthamiana produced several hordedanes,including one of the most functionally decorated products 19-β-hydroxy-hordetrienoic acid(19-OH-HTA).Barley mutants in the diterpene synthase genes of this cluster are unable to produce hordedanes but,unexpectedly,show reduced Bs *** contrast,colonization by Fusarium graminearum,another fungal pathogen of barley and wheat,is 4-fold higher in the mutants completely lacking ***,19-OH-HTA enhances both germination and growth of Bs,whereas it inhibits other pathogenic fungi,including *** of microscopy and transcriptomics data suggest that hordedanes delay the necrotrophic phase of *** together,these results show that adapted pathogens such as Bs can subvert plant metabolic defenses to facilitate root colonization.

关键词： barley Hordeum vulgare diterpenoid phytoalexins gene cluster pathogenic fungi Bipolaris sorokininiana Fusarium graminearum

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Sensitivity and resistance risk analysis of Didymella bryoniae populations to fluopyram

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Journal of Integrative Agriculture 2024年第7期23卷 2306-2317页

作者： Zhiwen Wu Xiaowei Cai Xuewei Mao Mingguo Zhou Yiping Hou College of plant protection Nanjing Agricultural UniversityNanjing 210095China

Fluopyram is an succinate dehydrogenase inhibitors(SDHI)fungicide that has been registered in China to control gummy stem blight(GSB)in watermelons for many ***,whether the field pathogens of GSB are still sensitive to fluopyram or not is ***,we collected 69 Didymella bryoniae isolates from the fields that usually use fluopyram to control GSB to determine the sensitivity *** EC_(50)(50%inhibition effect)values of fluopyram against *** ranged from 0.0691 to 0.3503μg mL^(–1) and the variation factor was *** mean EC_(50) value was(0.1579±0.0669)μg mL^(–1) and the curve of sensitivity was *** resistant strains were found in the isolates,which means that the pathogens were still sensitive to *** minimal inhibition concentration(MIC)of fluopyram against *** was 3μg mL^(–1).Four low-resistant mutants and two medium-resistant mutants were obtained using fungicide taming and the resistance of mutants could be inherited *** growth rate of mutants decreased significantly compared with that of wild-type strains while the biomass of most mutants was similar to that of wild-type *** sensitivity of most resistant mutants to various stresses was increased compared with that of wild-type *** virulence of mutants receded except for low-resistant mutant XN51FR-1,which had the same lesion area as XN51 on the watermelon *** results indicated that the fitness of resistant mutants was decreased compared with that of wild-type *** cross-resistance assay indicated that fluopyram-resistant mutants were positive cross-resistant to all six SDHI fungicides in this test but were still sensitive to fluazinam and *** the resistance risk of *** to fluopyram was *** addition,we found that the SdhB gene of low-resistant mutant XN30FR-1 had three new point mutations at positions K258N,A259P,and ***-resistant mutant XN52FR-1 showed a mutation at position H277N an

关键词： fluopyram D.bryoniae sensitivity risk assessment

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Development and identification of two novel wheat-rye 6R derivative lines with adult-plant resistance to powdery mildew and high-yielding potential

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The Crop Journal 2024年第1期12卷 308-313页

作者： Guohao Han Jing Wang Hanwen Yan Tiantian Gu Lijun Cao Shiyu Liu Xiuquan Li Yilin Zhou Jieru Fan Zhipeng Shi Hong Liu Lihui Li Diaoguo An Center for Agricultural Resources research Institute of Genetics and Developmental BiologyChinese Academy of SciencesShijiazhuang 050021HebeiChina The National Key Facility for Crop Gene Resources and Genetic Improvement Institute of Crop SciencesChinese Academy of Agricultural SciencesBeijing 100081China The State Key Laboratory for Biology of plant Disease and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing 100193China

Powdery mildew,caused by Blumeria graminis ***(Bgt),is a devastating disease that seriously threatens wheat yield and *** control this disease,host resistance is the most effective *** with the resistance genes from common wheat,alien resistance genes can better withstand infection of this highly variable *** of elite alien germplasm resources with powdery mildew resistance and other key breeding traits is an attractive strategy in wheat *** this study,three wheat-rye germplasm lines YT4-1,YT4-2,and YT4-3 were developed through hybridization between octoploid triticale and common wheat,out of which the lines YT4-1 and YT4-2 conferred adult-plant resistance(APR)to powdery mildew while the line YT4-3 was susceptible to powdery mildew during all of its growth *** genomic in situ hybridization,multi-color fluorescence in situ hybridization,multi-color GISH,and molecular marker analysis,YT4-1,YT4-2,and YT4-3 were shown to be cytogenetically stable wheat-rye 6R addition and T1RS.1BL translocation line,6RL ditelosomic addition and T1RS.1BL translocation line,and T1RS.1BL translocation line,*** with previously reported wheat-rye derivative lines carrying chromosome 6R,YT4-1 and YT4-2 showed stable APR without undesirable pleiotropic effects on agronomic ***,these novel wheat-rye 6R derivative lines are expected to be promising bridge resources in wheat disease breeding.

关键词： Powdery mildew Secale cereale Triticum aestivum Wheat-rye 6R derivatives Agronomic performance

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Invasion of fall armyworm led to the succession of maize pests in Southwest China

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Journal of Integrative Agriculture 2024年第4期23卷 1300-1314页

作者： Zezheng Fan Yifei Song Shengyuan Zhao Kongming Wu Agricultural Information institute Chinese Academy of Agricultural SciencesBeijing 100081China Guangdong Laboratory for Lingnan Modern Agriculture Guangzhou 510642China State Key Laboratory for Biology of plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing 100193China

The invasive fall armyworm Spodoptera frugiperda(***)invaded Asia in 2018,colonizing the tropical and southern subtropical regions as well as migrating with the monsoons into Northeast Asia during spring and *** has resulted in widespread infestations,with significant impacts on maize production in various Asian *** studies have shown that the invasion of this pest can alter the species relationships of maize pests,but the actual impact on maize pest management is still *** study investigated the changes in maize pest occurrence and pesticide use in the annual breeding areas of *** in Yunnan Province and the Guangxi Zhuang Autonomous Region of China during 2017-2021,based on surveys and interviews with small farmers in maize *** results showed that *** has emerged as the dominant species among maize pests after invasion and colonization,replacing traditional pests such as Ostrinia furnacalis,Spodoptera litura,Agrotis ypsilon,and Rhopalosiphum *** variety of pesticides used for maize pest control has changed from chlorpyrifos,lambda-cyhalothrin,and acetamiprid to emamectin benzoate-based pesticides with high effectiveness against ***,the frequency of maize pest chemical applications has increased from an average of 5.88 to 7.21 times per season,with the amounts of pesticides used in summer and autumn maize being significantly higher than in winter and spring maize,thereby increasing application costs by more than 35%.The results of this study clarified the impact of *** invasion on maize pest community succession and chemical pesticide use in tropical and south subtropical China,thereby providing a baseline for modifying the regional control strategies for maize pests after the invasion of this relatively new pest.

关键词： Spodoptera frugiperda annual breeding region sweet waxy corn economic evaluation

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A dicistrovirus increases pupal mortality in Spodoptera frugiperda by suppressing protease activity and inhibiting larval diet consumption

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Journal of Integrative Agriculture 2024年第8期23卷 2723-2734页

作者： Meixue Sun Tong Li Yingjie Liu Kenneth Wilson Xingyu Chen Robert I.Graham Xianming Yang Guangwei Ren Pengjun Xu Key Laboratory of Tobacco Pest Monitoring&Integrated Management Tobacco Research InstituteChinese Academy of Agricultural SciencesQingdao 266101China institute of plant protection Henan Academy of Agricultural SciencesZhengzhou 450000China Staff Development institute of China National Tobacco Corporation Zhengzhou 450000China Lancaster Environment Centre Lancaster UniversityLancaster LA14YWUK Department of Rural Land Use Scotland’s Rural CollegeCraibstone CampusAberdeen AB01ABUK State Key Laboratory for Biology of plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing 100193China

Understanding interactions between viruses and their hosts is conducive to enabling better application of viruses as biocontrol *** viruses carried by parasitic wasps enhance the parasitic efficiency of wasp-larvae by protecting them against the immune system of their Lepidopteran ***,the relationship between prey pests and viruses found in predatory natural enemies remains ***,we report the interaction between Arma chinensis virus-1(AcV-1),originally isolated from a predatory natural enemy,Arma chinensis(Hemiptera:Pentatomidae),and one of its prey species,Spodoptera frugiperda(Lepidoptera:Noctuidae).The results showed that the AcV-1 virus appeared harmful to the novel host *** by inhibiting larval diet consumption and increasing pupal ***,sequencing data indicated that the virus altered the gene expression profiles of *** analysis showed that the proteasome and phagosome pathways related to protein degradation and immune response were significantly *** the expression levels of digestive enzyme genes did not change significantly,the total protease activity of AcV-1 virus-positive individuals was significantly decreased,suggesting that the virus inhibited diet consumption of *** via the down-regulation of digestive enzyme *** results indicate that a virus initially isolated in a predatory natural enemy can decrease the fitness of its prey *** virus was found to impact the host proteasome and phagosome pathways related to protein degradation and immunity,providing a potential mechanism to enhance controlling efficiency.

关键词： Arma chinensis virus-1 diet consumption fitness transcriptome protease activity

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Cytospora pyri promotes Erwinia amylovora virulence by providing metabolites and hyphae

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Journal of Integrative Agriculture 2024年第9期23卷 3045-3054页

作者： Tong Shen Mengdi Ye Yeping Xu Bohan Ding Hongtao Li Li Zhang Jun Wang Yanli Tian Baishi Hu Youfu Zhao College of plant protection/Key Laboratory of Integrated Management of Crop Diseases and Pests Nanjing Agricultural UniversityNanjing 210095China Key Laboratory of plant Quarantine Pests Monitoring and Control Ministry of Agriculture and Rural AffairsNanjing 210095China plant protection and Quarantine Station of Xinjiang Uygur Autonomous Region Urumqi 830001China Xinjiang Key Laboratory of Agricultural Biosafety Institute of Plant ProtectionXinjiang Academy of Agricultural SciencesUrumqi 830013China College of Agriculture Xinjiang Agricultural UniversityUrumqi 830052China plant Pathology Department Washington State UniversityProsserWA 24106USA

Bacterial-fungal interactions are widespread in *** observed that pear orchards affected by Cytospora pyri(formerly Valsa pyri)were often accompanied with Erwinia ***,the relationship of the two pathogens was *** objective of this study was to determine whether the synergistic effect exists between *** and *** first analyzed the coexistence frequencies of *** and *** in pear *** of the two pathogens,growth,physical interactions,amylovoran production,and expression of genes for amylovoran biosynthesis were *** results showed that *** and *** could coexist on the same lesion and caused much more severe *** also found that *** could physically attach to *** and the expression of amylovoran biosynthesis genes were up-regulated with fungal metabolite *** results indicate that *** and *** can cooperatively interact,which provides *** with an opportunity to promote bacterial dispersal and production of virulence factor in ***.

关键词： pear fire blight Erwinia amylovora Valsa canker amylovoran synergy

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