Tomato spotted wilt orthotospovirus(TSWV)causes substantial economic losses to vegetables and other crops.TSWV is mainly transmitted by thrips in a persistent and proliferative manner,and its most efficient vector is ...Tomato spotted wilt orthotospovirus(TSWV)causes substantial economic losses to vegetables and other crops.TSWV is mainly transmitted by thrips in a persistent and proliferative manner,and its most efficient vector is the western flower thrips,Frankliniella occidentalis(Pergande).In moving from the thrips midgut to the salivary glands in preparation for transmission,the virions must overcome multiple barriers.Although several proteins that interact with TSWV in thrips have been characterized,we hypothesized that additional thrips proteins interact with TSWV and facilitate its transmission.In the current study,67 F occidentalis proteins that interact with GN(a structural glycoprotein)were identified using a split-ubiquitin membrane-based yeast 2-hybrid(MbY2H)system.Three proteins,apolipoprotein-D(ApoD),orai-2-like(Orai),and obstructor-E-like isoform X2(Obst),were selected for further study based on their high abundance and interaction strength;their interactions with Gn were confirmed by MbY2H,yeastβ-galactosidase and luciferase complementation assays.The relative expressions of ApoD and Orai were significantly down-regulated but that of Obst was significantly up-regulated in viruliferous thrips.When interfering with Obst in larval stage,the TSWV acquisition rate in 3 independent experiments was significantly decreased by 26%,40%,and 35%,respectively.In addition,when Obst was silenced in adults,the virus titer was significantly decreased,and the TSWV transmission rate decreased from 66.7%to 31.9%using the leaf disk method and from 86.67%to 43.33%using the living plant method.However,the TSWV acquisition and transmission rates were not affected by interference with the ApoD or Orai gene.The results indicate that Obst may play an important role in TSWV acquisition and transmission in Frankliniella occidentalis.展开更多
The western flower thrips(WFT;Frankliniella occidentalis)is a mesophyll cell feeder that damages many crops.Management of WFT is complex due to factors such as high fecundity,short reproduction time,ability to feed on...The western flower thrips(WFT;Frankliniella occidentalis)is a mesophyll cell feeder that damages many crops.Management of WFT is complex due to factors such as high fecundity,short reproduction time,ability to feed on a broad range of host plants,and broad pesticide resistance.These challenges have driven research into developing alternative pest control approaches for WFT.This study analyzed the feasibility of a biological control-based strategy to manage WFT using RNA interference(RNAi)-mediated silencing of WFT endogenous genes.For the delivery of RNAi,we developed transgenic tomato lines expressing double-stranded RNA(dsRNA)of coatomer protein subunit epsilon(CopE)and Toll-like receptor 6(TLR6)from WFT.These genes are involved in critical biological processes of WFT,and their dsRNA can be lethal to these insects when ingested orally.Adult WFT that fed on the transgenic dsRNAexpressing tomato flower stalk showed increased mortality compared with insects that fed on wild-type samples.In addition,WFT that fed on TLR6 and CopE transgenic tomato RNAi lines showed reduced levels of endogenous CopE and TLR6 transcripts,suggesting that their mortality was likely due to RNAi-mediated silencing of these genes.Thus,our findings demonstrate that transgenic tomato plants expressing dsRNA of TLR6 and CopE can be lethal to F.occidentalis,suggesting that these genes may be deployed to control insecticide-resistant WFT.展开更多
Successful integrated pest management in protected crops implies an evalu-ation of the compatibility of pesticides and natural enemies(NE),as control strategies that only rely on one tactic can fail when pest populati...Successful integrated pest management in protected crops implies an evalu-ation of the compatibility of pesticides and natural enemies(NE),as control strategies that only rely on one tactic can fail when pest populations exceed NE activity or pests become resistant to pesticides.Nowadays in Almeria(Spain),growers release NE prior to transplanting or early in the crop cycle to favor their settlement before pest arrival because this improves biocontrol efficacy,although it extends pesticide exposure periods.The pur-pose of this research was to evaluate the compatibility of two applications of pesticides with key NE in 2-year trials inside tomato and sweet pepper commercial greenhouses:Nesidiocoris tenuis(Reuter)(Hemiptera:Miridae),Orius laevigatus(Say)(Hemiptera:Anthocoridae)and Amblyseius swirskii(Athias-Henriot)(Acari:Phytoseidac).In tomato,flubendiamide and chlorantraniliprole(IOBC category 1)were compatible with N.tenuis,but chlorpyrifos.:methyl and spinosad(IOBC categories 2-3),which efectively reduced Tiuta absoluta(Meyrick)(Lepidoptera:Gelechidae)density,compromised its predatory activity.In sweet pepper,chlorantraniliprole(IOBC category 1)was the only pesticide compatible with O.laevigatus while chlorantraniliprole,emamectin benzoate,spirote-tramat and pymetrozine were harmless(IOBC category 1)to Amblyseius swirskii,and sulfoxaflor slightly harmful(IOBC category 2)to this phytoseiid predator.展开更多
基金the China Agriculture Research System(CARS-24-C-02)the National Natural Science Foundation of China(32272548)+1 种基金the Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences(CAAS-ASTIP-IVFCAAS)the Hainan Major Science and Technology Project(ZDKJ2021007)。
文摘Tomato spotted wilt orthotospovirus(TSWV)causes substantial economic losses to vegetables and other crops.TSWV is mainly transmitted by thrips in a persistent and proliferative manner,and its most efficient vector is the western flower thrips,Frankliniella occidentalis(Pergande).In moving from the thrips midgut to the salivary glands in preparation for transmission,the virions must overcome multiple barriers.Although several proteins that interact with TSWV in thrips have been characterized,we hypothesized that additional thrips proteins interact with TSWV and facilitate its transmission.In the current study,67 F occidentalis proteins that interact with GN(a structural glycoprotein)were identified using a split-ubiquitin membrane-based yeast 2-hybrid(MbY2H)system.Three proteins,apolipoprotein-D(ApoD),orai-2-like(Orai),and obstructor-E-like isoform X2(Obst),were selected for further study based on their high abundance and interaction strength;their interactions with Gn were confirmed by MbY2H,yeastβ-galactosidase and luciferase complementation assays.The relative expressions of ApoD and Orai were significantly down-regulated but that of Obst was significantly up-regulated in viruliferous thrips.When interfering with Obst in larval stage,the TSWV acquisition rate in 3 independent experiments was significantly decreased by 26%,40%,and 35%,respectively.In addition,when Obst was silenced in adults,the virus titer was significantly decreased,and the TSWV transmission rate decreased from 66.7%to 31.9%using the leaf disk method and from 86.67%to 43.33%using the living plant method.However,the TSWV acquisition and transmission rates were not affected by interference with the ApoD or Orai gene.The results indicate that Obst may play an important role in TSWV acquisition and transmission in Frankliniella occidentalis.
基金supported by the Basic Science Research Program through the National Research Foundation(NRF),Ministry of Education,Korea(2021R1I1A1A01041938)a grant from the New Breeding Technologies Development Program,Rural Development Administration,Korea(PJ0165432022)supported in part by the BK21 Plus Program,Ministry of Education,Korea。
文摘The western flower thrips(WFT;Frankliniella occidentalis)is a mesophyll cell feeder that damages many crops.Management of WFT is complex due to factors such as high fecundity,short reproduction time,ability to feed on a broad range of host plants,and broad pesticide resistance.These challenges have driven research into developing alternative pest control approaches for WFT.This study analyzed the feasibility of a biological control-based strategy to manage WFT using RNA interference(RNAi)-mediated silencing of WFT endogenous genes.For the delivery of RNAi,we developed transgenic tomato lines expressing double-stranded RNA(dsRNA)of coatomer protein subunit epsilon(CopE)and Toll-like receptor 6(TLR6)from WFT.These genes are involved in critical biological processes of WFT,and their dsRNA can be lethal to these insects when ingested orally.Adult WFT that fed on the transgenic dsRNAexpressing tomato flower stalk showed increased mortality compared with insects that fed on wild-type samples.In addition,WFT that fed on TLR6 and CopE transgenic tomato RNAi lines showed reduced levels of endogenous CopE and TLR6 transcripts,suggesting that their mortality was likely due to RNAi-mediated silencing of these genes.Thus,our findings demonstrate that transgenic tomato plants expressing dsRNA of TLR6 and CopE can be lethal to F.occidentalis,suggesting that these genes may be deployed to control insecticide-resistant WFT.
基金This research was funded by Spanish Ministry of Sci-ence,Innovation and Universities projects AGL2013-47603-C2-1-R and AGL2017-83498-C2-2-R awarded to Elisa Viuela and Pilar Medina,along with post-doctoral fellowships awarded to Beatriz Dader(Span-ish Ministry of Science,Innovation and Universities FJCI-2016-28443 and Universidad Politecnica de Madrid PINV18XFWLGK24S2US6D).We deeply appreciate the cooperation of two farmers that allowed us to perform tri-als in their commercial greenhouses.We are indebted to Dr.Christina Elizabeth Pease for English revision and to Dr.Ismael Sanchez from INIA for help with statistics.
文摘Successful integrated pest management in protected crops implies an evalu-ation of the compatibility of pesticides and natural enemies(NE),as control strategies that only rely on one tactic can fail when pest populations exceed NE activity or pests become resistant to pesticides.Nowadays in Almeria(Spain),growers release NE prior to transplanting or early in the crop cycle to favor their settlement before pest arrival because this improves biocontrol efficacy,although it extends pesticide exposure periods.The pur-pose of this research was to evaluate the compatibility of two applications of pesticides with key NE in 2-year trials inside tomato and sweet pepper commercial greenhouses:Nesidiocoris tenuis(Reuter)(Hemiptera:Miridae),Orius laevigatus(Say)(Hemiptera:Anthocoridae)and Amblyseius swirskii(Athias-Henriot)(Acari:Phytoseidac).In tomato,flubendiamide and chlorantraniliprole(IOBC category 1)were compatible with N.tenuis,but chlorpyrifos.:methyl and spinosad(IOBC categories 2-3),which efectively reduced Tiuta absoluta(Meyrick)(Lepidoptera:Gelechidae)density,compromised its predatory activity.In sweet pepper,chlorantraniliprole(IOBC category 1)was the only pesticide compatible with O.laevigatus while chlorantraniliprole,emamectin benzoate,spirote-tramat and pymetrozine were harmless(IOBC category 1)to Amblyseius swirskii,and sulfoxaflor slightly harmful(IOBC category 2)to this phytoseiid predator.
