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Insect-specific RNA virus affects the stylet penetration activity of brown citrus aphid(Aphis citricidus)to facilitate its transmission

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Insect Science 2024年第1期31卷 255-270页

作者： Xin An Qiaoying Gu Jing Wang Tengyu Chang Wei Zhang Jin-Jun Wang Jinzhi Niu key laboratory of entomology and pest control engineering College of Plant ProtectionAcademy of Agricultural ScienceSouthwest UniversityChongqingChina

Sap-sucking insects often transmit plant viruses but also carry insect viruses,which infect insects but not plants.The impact of such insect viruses on insect host biology and ecology is largely unknown.Here,we identified a novel insect-specific virus carried by brown citrus aphid(Aphis citricidus),which we tentatively named Aphis citricidus picornavirus(AcPV).Phylogenetic analysis discovered a monophyletic cluster with AcPV and other unassigned viruses,suggesting that these viruses represent a new family in order Picornavirales.Systemic infection with AcPV triggered aphid antiviral immunity mediated by RNA interference,resulting in asymptomatic tolerance.Importantly,we found that AcPV was transmitted horizontally by secretion of the salivary gland into the feeding sites of plants.AcPV influenced aphid stylet behavior during feeding and increased the time required for intercellular penetration,thus promoting its transmission among aphids with plants as an intermediate site.The gene expression results suggested that this mechanism was linked with transcription of salivary protein genes and plant defense hormone signaling.Together,our results show that the horizontal transmission of AcPV in brown citrus aphids evolved in a manner similar to that of the circulative transmission of plant viruses by insect vectors,thus providing a new ecological perspective on the activity of insect-specific viruses found in aphids and improving the understanding of insect virus ecology.

关键词： electrical penetration graph feeding behavior insect virus transmission Picornavirales salivary protein

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RNA interference in insects:the link between antiviral defense and pest control

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Insect Science 2024年第1期31卷 2-12页

作者： Jinzhi Niu Ruoyu Chen Jin-Jun Wang key laboratory of entomology and pest control engineering College of Plant ProtectionSouthwest UniversityChongqingChina key laboratory of Agricultural Biosafety and Green Production of Upper Yangtze River(Ministry of Education) Southwest UniversityChongqingChina

RNA interference(RNAi)is a form of gene silencing triggered by double-stranded RNA(dsRNA)that operates in all eukaryotic cells.RNAi has been widely investigated in insects to determine the underlying molecular mechanism,to investigate its role in systemic antiviral defense,and to develop strategies for pest control.When insect cells are infected by viruses,viral dsRNA signatures trigger a local RNAi response to block viral replication and generate virus-derived DNA that confers systemic immunity.RNAi-based insect pest control involves the application of exogenous dsRNA targeting genes essential for insect development or survival,but the efficacy of this approach has limited potency in many pests through a combination of rapid dsRNA degradation,inefficient dsRNA uptake/processing,and ineffective RNAi machinery.This could be addressed by dsRNA screening and evaluation,focusing on dsRNA design and off-target management,as well as dsRNA production and delivery.This review summarizes recent progress to determine the role of RNAi in antiviral defense and as a pest control strategy in insects,addressing gaps between our fundamental understanding of the RNAi mechanism and the exploitation of RNAi-based pest control strategies.

关键词： RNAi response systemic RNAi insecticidal dsRNA biological control off-targets

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Molting process revealed by the detailed expression profiles of RXR1/RXR2 and mining the associated genes in a spider mite, Panonychus citri

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Insect Science 2022年第2期29卷 430-442页

作者： Gang Li Xun-Yan Liu Guy Smagghe Jin-Zhi Niu Jin-Jun Wang key laboratory of entomology and pest control engineering College of Plant ProtectionInternational Joint Laboratory of China-Belgium on Sustainable Crop Pest ControlAcademy of Agricultural SciencesSouthwest UniversityChongqingChina Provincial key laboratory for Agricultural pest Management of Mountainous Regions Institute of EntomologyGuizhou UniversityGuiyangChina Department of Plants and Crops Faculty of Bioscience EngineeringGhent UniversityGhentBelgium

Spider mites have one ecdysone receptor(EcR)and multiple retinoid X receptors(RXRs).However,the function of these RXRs in spider mite development is unknown.Here,we screened the expression dynamics of two PcRXR isoforms at 4 h intervals in the deutonymphal stage of Panonychus citri.The results showed that PcEcR had an expression pattern similar to that of PcRXR2.For PcRXR1,its expression remained at a certain high level,when there was a decrease of both PcEcR and PcRXR2.In situ hybridization showed that PcRXR2 was detected in the central nervous mass,while the ecdysteroid biosynthesis gene PcSpo was mainly expressed at the edge of the central nervous mass.RNAi-based silencing of PcRXR1 or PcRXR2 showed the same phenotype as in mites with that of silencing PcEcR.Furthermore,RNA-seq was used to mine the genes associated with the expression dynamics of PcRXR1 or PcRXR2,which revealed that the heterodimer of EcR-RXR2 in spider mites might be linked with the cell autophagy and tissue remodeling during apolysis,and RXR1 might be linked with new epicuticle and exocuticle secretion during ecdysis.Taken together,these results increase our understanding of the regulation mechanism of ecdysteroid signal pathway in spider mite development.

关键词： ecdysone receptor(EcR) in situ hybridization retinoid X receptors RNA interference RNA-seq

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Discovery of a widespread presence bunyavirus that may have symbiont-like relationships with different species of aphids

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Insect Science 2022年第4期29卷 1120-1134页

作者： Xin An Wei Zhang Chao Ye Guy Smagghe Jin-Jun Wang Jinzhi Niu key laboratory of entomology and pest control engineering College of Plant ProtectionSouthwest UniversityChongqingChina International Joint laboratory of China–Belgium on Sustainable Crop pest control Academy of Agricultural SciencesSouthwest UniversityChongqingChina Department of Plants and Crops Faculty of Bioscience EngineeringGhent UniversityGhentBelgium

Aphids are important agricultural pests,vectors of many plant viruses and have sophisticated relationships with symbiotic microorganisms.Abundant asymptomatic RNA viruses have been reported in aphids due to the application of RNA-seq,but aphid-virus interactions remain unclear.Bunyavirales is the most abundant RNA virus order,which can infect mammals,arthropods,and plants.However,many bunyaviruses have specific hosts,such as insects.Here,we discovered 18 viruses from 10 aphid species by RNA-seq.Importantly,a widespread presence bunyavirus,Aphid bunyavirus 1(ABV-1),was determined to have a wide host range,infecting and replicating in all 10 tested aphid species.ABV-1 may be transmitted horizontally during feeding on plant leaves and vertically through reproduction.In a comparison of the physiological parameters of ABV-1high and ABV-1low strains of pea aphid,higher ABV-1 titers reduced the total nymphal duration and induced the reproduction.Moreover,viral titer significantly affected the lipid and protein contents in pea aphids.In summary,we proposed that ABV-1 may have stable symbiont-like relationships with aphids,and these observations may provide a new direction for studying bunyaviruses in aphids and establishing a model for virus-aphid interactions.

关键词： insec-specific virus pea aphid Phasmaviridae RNA virus RNAi

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Characterization of carotenoid biosynthetic pathway genes in the pea aphid(Acyrthosiphon pisum)revealed by heterologous complementation and RNA interference assays

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Insect Science 2022年第3期29卷 645-656页

作者： Bi-Yue Ding Xiu-Cheng Xie Feng Shang Guy Smagghe Jin-Zhi Niu Jin-Jun Wang key laboratory of entomology and pest control engineering College of Plant ProtectionSouthwest UniversityChongqingChina International Joint laboratory of China-Belgium on Sustainable Crop pest ControlAcademy of Agricultural SciencesSouthwest UniversityChongqingChina Department of Plants and Crops Faculty of Bioscience EngineeringGhent UniversityGhentBelgium

Carotenoids are involved in many essential physiological functions and are produced from geranylgeranyl pyrophosphate through synthase,desaturase,and cyclase activities.In the pea aphid(Acyrthosiphon pisum),the duplication of carotenoid biosynthetic genes,including carotenoid synthases/cyclases(ApCscA-C)and desaturases(ApCdeA-D),through horizontal gene transfer from fungi has been detected,and ApCdeB has known dehydrogenation functions.However,whether other genes contribute to aphid carotenoid biosynthesis,and its specific regulatory pathway,remains unclear.In the current study,functional analyses of seven genes were performed using heterologous complementation and RNA interference assays.The bifunctional enzymes ApCscA-C were responsible for the synthase of phytoene,and ApCscC may also have a cyclase activity.ApCdeA,ApCdeC,and ApCdeD had diverse dehydrogenation functions.ApCdeA catalyzed the enzymatic conversion of phytoene to neurosporene(three-step product),ApCdeC catalyzed the enzymatic conversion of phytoene to ζ-carotene(two-step product),and ApCdeD catalyzed the enzymatic conversion of phytoene to lycopene(four-step product).Silencing of ApCscs reduced the expression levels of ApCdes,and silencing these carotenoid biosynthetic genes reduced the α-,β-,and γ-carotene levels,as well as the total carotenoid level.The results suggest that these genes were activated and led to carotenoid biosynthesis in the pea aphid.

关键词： carotenoid biosynthesis heterologous complementary pea aphid RNA interference

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Genome-wide analysis of Panonychus citrimicro RNAs witha focus on potential insecticidal activity of 4 micro RNAs toeggs and nymphs

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Insect Science 2024年第2期31卷 354-370页

作者： Chuan-Zhen Li Yu-Hang Liu Deng Pan Meng-Hao Xia Qiang Zhang Yu-Chuang Li Guo-Rui Yuan Jin-Jun Wang Wei Dou key laboratory of entomology and pest control engineering College of Plant ProtectionAcademy of Agricultural SciencesSouthwestUniversityChongqingChina key laboratory of Agricultural Biosafety and Green Production of Upper Yangtze River(Ministry of Education) Academy of Agricultural SciencesSouthwest UniversityChongqingChina

Panonychus citriMcGregor(Acari:Tetranychidae),a destructive citrus pest,causes considerable annual economic losses due to its short lifespan and rapid resistancedevelopment.MicroRNA(miRNA)-induced RNA interference is a promising approachfor pest control because of endogenous regulation of pest growth and development.Tosearch for miRNAs with potential insecticidal activity inP.citri,genome-wide analysis ofmiRNAs at different developmental stages was conducted,resulting in the identificationof 136 miRNAs,including 73 known and 63 novel miRNAs.A total of 17 isomiRNAsand 12 duplicated miRNAs were characterized.MiR-1 and miR-252-5p were identified asreference miRNAs forP.citriandTetranychus urticae.Based on differential expressionanalysis,treatments with miR-let-7a and miR-315 mimics and the miR-let-7a antagomirsignificantly reduced the egg hatch rate and resulted in abnormal egg development.Over-expression or downregulation of miR-34-5p and miR-305-5p through feeding significantlydecreased the adult eclosion rate and caused molting defects.The 4 miRNAs,miR-let-7a,miR-315,miR-34-5p,and miR-305-5p,had important regulatory functions and insectici-dal properties in egg hatching and adult eclosion.In general,these data advance our un-derstanding of miRNAs in mite biology,which can assist future studies on insect-specificmiRNA-based green pest control technology.

关键词： growth and development microRNAs Panonychus citri referencemiRNAs regulation small RNA libraries

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Performance and Enzyme Activity of Beet Armyworm Spodoptera exigua (Hübner) (Lepidoptera:Noctuidae) Under Various Nutritional Conditions

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Agricultural Sciences in China 2011年第5期10卷 737-746页

作者： Helen Hull Sanders Chongqing Key Laboratory of Entomology and Pest Control Engineering Southwest University

We explored the influence of the artificial diets with different protein and glucose contents on larval development,fecundity and enzyme activities of the beet armyworm Spodoptera exigua （Hübner）.Our results suggested that development,fecundity and population growth index of beet armyworm increased with increased nutritional content in the artificial diets;however,when the yeast to cellulose content ratio reached 46.8：53.2,the fecundity and population growth had reached a maximum and additional protein did not yield additional growth.Additionally,3rd instar beet armyworm larvae fed on different artificial diets had increased enzymatic activities of acetylcholinesterase （AChE） with the increased nutritional content,but carboxylesterase （CarE） activities did not significantly change under variation in the nutritional content.

关键词： beet armyworm nutritional variation acetylcholinesterase carboxylesterase protein carbohydrate enzymeactivity

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Functional analysis of UG7207D3 associated with abamectin resistance in Tetranychus cinnabarinus(Boisduval)

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Insect Science 2020年第2期27卷 276-291页

作者： Meng-Yao Wang Xin-Yang Liu Li Shi Jia-Lu Liu Guang-Mao Shen Ping Zhang Wen-Cai Lu Lin He key laboratory of entomology and pest control engineering College of Plant ProtectionSouthwest UniversityChongqingChina Academy o f Agricultural Sciences Southwest UniversityChongqingChina

Uridine diphosphate(UDP)-glycosyltransferases(UGTS)are widely distributed within living organisms and share roles in biotransformation of various lipophilic endo-and xenobiotics with activated UDP sugars.In this study,it was found that the activity of UGTs in abamectin-resistant(AbR)strain was significantly higher(2.3-fod)than that in susceptible strain(SS)of Tetramychus cinnabarinus.Further analysis showed that 5-nitrouracil,the inhibitor ofUGTs,could enhance the lethal effect of abamectin on mites.From the previous microarray results,we found an UGT gene(UGT201D3)overexpressed in AbR strain.Quantitative PCR analysis showed that UGT20ID3 was highly expressed and more inducible with abamectin exposure in the AbR strain.After silencing the tran-scription of UGT201D3,the activity of UGTs was decreased and the susceptibility to abamectin was increased in AbR strain whereas it was not in sS.Furthermore,UGT201D3 gene was then succefully expressed in Escherichia coli.The recombinant UGT20ID3 exhibited a-naphthol activity(2.810.43 nmol mg protein/min),and the enzyme activity could be inhibited by abamectin(inhibitory concentration at 50%:57.50±3.54 μmol/L).High-performance liquid chromatography analysis demonstrated that the recombinant UGT201D3 could efctively deplete abamectin(15.77%±3.72%)incubating with 150 ug protein for 6 h.These results provided direct evidence that UGT201D3 was involved in abamectin resistance in T cinnabarinus.

关键词： abamectin resistance prokaryotic expression RNA interference Tetrumychus cimnabarins UDP-glycosy ltransferases

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Expression and functional characterization of odorant-binding protein 2 in the predatory mite Neoseiulus barkeri

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Insect Science 2023年第5期30卷 1493-1506页

作者： Yixia Wu Yaying Li Wenqiang Chu Tiandi Niu Xiaotian Feng Rongjiang Ma Huai Liu key laboratory of entomology and pest control engineering College of Plant ProtectionSouthwest UniversityBeibei DistrictChongqing400715 China

Olfaction plays a crucial role for arthropods in foraging,mating,and oviposition.The odorant-binding protein(OBP)gene is considered one of the most important olfactory genes.However,little is known about its functions in predatory mites.Here,we used Neoseiulus barkeri,an important commercialized natural pest control,to explore the chemosensory characteristics of OBP.In this study,N.barkeri was attracted by methyl salicylate(MeSA)and showed higher crawling speeds under MeSA treatment.Then,we identified and cloned an OBP gene named Nbarobp2 and analyzed its expression profiles in the predatory mite.Nbarobp2 was 663 bp,was highly expressed in larval and nymphal stages,and was significantly upregulated in N.barkeri under MeSA treatment.Nbarobp2 encoded 202 amino acid residues with a molecular weight of 23 kDa(after removing the signal peptide).Sequence comparisons revealed that the OBPs in Arachnida shared 6 conserved cysteine sites,but were distinguishable from the OBPs of Insecta on the phylogenetic tree.RNA interference,Western blotting,and binding affinity assays further proved that Nbarobp2 was involved in volatile perception in predatory mites.This study shed light on the functional characteristics of OBPs in predatory mites,providing a new insight for better biological control.

关键词： binding affinity crawling speed heterologous expression methyl salicylate olfactory behavior RNA interference

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GARP: A family of glycine and alanine-rich proteins that helps spider mites feed on plants

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Insect Science 2023年第5期30卷 1337-1351页

作者： Qin-Zhe Sun Xiao-Lin Li Yu-Fei Shi Yan-Chun Zhang Wen-Jie Chai Ruo-Yu Chen Jinzhi Niu Jin-Jun Wang key laboratory of entomology and pest control engineering College of Plant ProtectionAcademy of Agricultural SciencesSouthwest UniversityChongqingChina

Spider mites(Tetranychidae)are destructive agricultural pests which have evolved strategies to overcome plant defenses,such as the ability to puncture the leaves of their hosts to feed.The expression of many genes with unknown functions is altered during feeding,but little is known about the role of these genes in plant–mite interactions.Here,we identified 3 novel gene families through analysis of genomic and transcriptomic data from 3 spider mite species.These GARP family genes encode glycine and alanine-rich proteins;they are present in mites(Acariformes)but absent in ticks(Parasitiformes)in the subclass Acari,indicating that these genes have undergone a significant expansion in spider mites and thus play important adaptive roles.Transcriptomic analysis revealed that the expression of GARP genes is strongly correlated with feeding and the transfer to new hosts.We used RNA interference to silence GARP1d in the two-spotted spider mite Tetranychus urticae,which inhibited feeding and egg laying and significantly increased mortality when the mites were transferred to soybean shoots;a similar effect was observed after TuVATPase was silenced.However,no changes in mite mortality were observed after TuGARP1d-silenced mites were placed on an artificial diet,which was different from the effect of TuVATPase silencing.Our results indicate that GARP family members play important roles in mite–plant interactions.Additional studies are needed to clarify the mechanisms underlying these interactions.

关键词： feeding mite-plant interaction RNA interference Tetranychus urticae Tetranychus evansi

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