Henosepilachna vigintioctopunctata is an emerging horticultural pest of Solanaceae plants in Asia.Here,we employed transcriptome sequencing of three female and three male H.vigintioctopunctata libraries to identify se...Henosepilachna vigintioctopunctata is an emerging horticultural pest of Solanaceae plants in Asia.Here,we employed transcriptome sequencing of three female and three male H.vigintioctopunctata libraries to identify sex-related genes.We recorded 281599294 bp of clean reads,and de novo assembly generated 85206 unigenes,with N_(50) of 1311 bp and average length of 763 bp.We found 5002 genes highly expressed in males and 2179 genes highly expressed in females.Expression profiles of six unigenes specific to females,six unigenes specific to males,and ten unigenes common to both sexes were verified using semi-quantitive RT-PCR;results showed expression levels of these 22 unigenes were relatively consistent with the transcriptome analysis.Using all unigenes as references,we identified 6657 putative simple sequence repeats.The transcriptome sequences and gene expression profiles of female and male H.vigintioctopunctata help explore genetic mechanisms of sexual dimorphism.展开更多
Six different indigenous plants were screened for antifeedant and insecticidal activity against fourth instar larvae of Epilachna beetle, Henosepilachna vigintioctopunctata, which is a severe pest on brinjal. Among th...Six different indigenous plants were screened for antifeedant and insecticidal activity against fourth instar larvae of Epilachna beetle, Henosepilachna vigintioctopunctata, which is a severe pest on brinjal. Among the plants screened, Achyranthes aspera showed higher activity against the selected pest. Ethyl acetate extracts of A. aspera showed higher antifeedant index and insecticidal activity against fourth instar larvae of H. vigintioctopunctata. Preliminary phytochemical analysis revealed that the presence of alkaloid and quinines in the ethyl acetate extracts indicate higher percentage of activities. Hence, it may suggest its use for controlling the vegetables insect pest, H. vigintioctopunctata.展开更多
RNA interference(RNAi)has emerged as a powerful tool for developing novel management strategies for controlling insect pests.The 28-spotted ladybeetle,Henosepilachna vigintioctopunctata is one of the most important pe...RNA interference(RNAi)has emerged as a powerful tool for developing novel management strategies for controlling insect pests.The 28-spotted ladybeetle,Henosepilachna vigintioctopunctata is one of the most important pests attacking solanaceous plants in Asia.In this study,the potential of dietary RNAi to manage H.vigintioctopunctata was investigated using both in vitro synthesized and bacterially expressed double-stranded RNAs(dsRNAs)of HvvATPase A and HvvATPase E.The expression levels of HvvATPase A and HvvATPase E were higher in Malpighian tubules than in other tissue types.The silencing of HvvATPase A and HvvATPase E led to significant mortality in H.vigintioctopunctata larvae.In addition,the ingestion of HvvATPase A and HvvATPase E significantly deterred feeding behavior and subsequently arrested the development of H.vigintioctopunctata.Notably,the bacterially expressed dsRNAs consistently caused higher mortality in larvae and adults.Finally,the nontarget effects of the dsRNAs of H.vigintioctopunctata on the predatory ladybeetle Propylaea japonica were evaluated.P.japonica 1st instar larvae were administered vATPase A and vATPase E dsRNAs from H.vigintioctopunctata and P.japonica under the worst-case scenario,in which dsGFP served as negative control.There were significant effects of dsHvvATPase A on P.japonica at the transcriptional level but not at the organismal level,whereas dsHvvATPase E did not effect P.japonica at either the transcriptional or the organismal level.Collectively,the results of the study suggest that HvvATPase A and HvvATPase E can act as novel molecular targets for the control of H.vigintioctopunctata.展开更多
Henosepilachna vigintioctopunctata is a serious insect pest which attacks a large number of nightshades and cucurbits in Asian countries,Brazil and Australia.Prolonged application of traditional pesticides has caused ...Henosepilachna vigintioctopunctata is a serious insect pest which attacks a large number of nightshades and cucurbits in Asian countries,Brazil and Australia.Prolonged application of traditional pesticides has caused environmental pollution and exerted deleterious effects on human health.Finding new approaches with high target specificity and low environmental contamination has become an urgent task.RNA interference(RNAi)induced by double-stranded RNA(dsRNA)is expected to be applicable to managing this pest.Here we evaluated the effects of Escherichia co/Z-expressed dsRNAs targeting ecdvsone receptor(EcR)gene via dietary delivery in laboratory and foliar spraying in a greenhouse.The target transcript was successfully knocked down when the 4th-instar larvae had fed on potatofoliage dipped with dsEcR in a laboratory bioassay.Around 85%of the HvEcR RNAi larvae remained as prepupae or became abnormal pupae,and failed to emerge into adults.Ingestion of ds£c7?-immersed foliage by the 3rd-instar larvae effectuated a comparable RNAi response and brought about more severe defects:all the resultant larvae arrested development,remained as prepupae and finally died.For assay in the greenhouse,a ds£c7?-contained E.coli suspension was directly sprayed to the foliage of greenhouse-growing potato plants and the 3rd-and 4th-instar larvae were transferred to the leaves.High RNAi efficacy was obtained and identical RNAi phenotypes were observed in treated larvae.In addition,spraying dsEcR reduced leaf damage.Our results indicate a possibility of practical application of dsEcR as an environmentally friendly RNA pesticide to control H.vigintioctopunctata larvae.展开更多
RNA interference(RNAi)techniques have emerged as powerful tools that facilitate development of novel management strategies for insect pests,such as Henosepilachna vigintioctopunctata(Coleoptera:Coccinellidae),which is...RNA interference(RNAi)techniques have emerged as powerful tools that facilitate development of novel management strategies for insect pests,such as Henosepilachna vigintioctopunctata(Coleoptera:Coccinellidae),which is a major pest of solanaceous plants in Asia.In this study,the potential of oral delivery of in vvYro-synthesized and bacterially expressed double-stranded H.vigintioctopunctata lesswright(Iwr)gene(dsHvlwr)to manage of H.vigintioctopunctata was investigated.Our results showed that the gene Hvlwr had a 480-bp open reading frame and encoded a 160-amino acid protein.Hvlwr expression levels were greater in the fat body than other tissue types.Hvlwr silencing led to greater H.vigintioctopunctata mortality rates and appeared to be time-and partially dose-dependent,likely as a result of the number of hemocytes increasing with dsRNA concentration,but decreasing with time.Bacterially expressed dsHvlwr that was applied to leaf discs caused 88%,66%,and 36%mortality in 1st instars,3rd instars,and adults after 10,10,and 14 d,respectively;when applied to living plants,there was greater mortality in 1 st and 3rd instars,but there was no effect on adults.Furthermore,dsHvlwr led to improved plant protection against H.vigintioctopunctata.Our study shows an effective dietary RNAi response in H.vigintioctopunctata and that Hvlwr is a promising RNAi target gene for control of this pest species.展开更多
RNA interference(RNAi)has emerged as a powerful technology for pest management.Previously,we have shown that plastid-mediated RNAi(PM-RNAi)can be utilized to control the Colorado potato beetle,an insect pest in the Ch...RNA interference(RNAi)has emerged as a powerful technology for pest management.Previously,we have shown that plastid-mediated RNAi(PM-RNAi)can be utilized to control the Colorado potato beetle,an insect pest in the Chrysomelidae family;however,whether this technology is suitable for controlling pests in the Coccinellidae remained unknown.The coccinellid 28-spotted potato ladybird(Henosepilachna vigintioctopunctata;HV)is a serious pest of solanaceous crops.In this study,we identified three efficient target genes(β-Actin,SRP54,and SNAP)for RNAi using in vitro double-stranded RNAs(dsRNAs)fed to HV,and found that dsRNAs targetingβ-Actin messenger RNA(dsACT)induced more potent RNAi than those targeting the other two genes.We next generated transplastomic and nuclear transgenic potato(Solanum tuberosum)plants expressing HV dsACT.Long dsACT stably accumulated to up to 0.7%of the total cellular RNA in the transplastomic plants,at least three orders of magnitude higher than in the nuclear transgenic plants.Notably,the transplastomic plants also exhibited a significantly stronger resistance to HV,killing all larvae within 6 d.Our data demonstrate the potential of PM-RNAi as an efficient pest control measure for HV,extending the application range of this technology to Coccinellidae pests.展开更多
基金the National Key R and DProgram of China(Grant No.2017YFD0200900)the Guangdong Province Universities and Colleges Pearl River Scholar Funded Scheme(2017).
文摘Henosepilachna vigintioctopunctata is an emerging horticultural pest of Solanaceae plants in Asia.Here,we employed transcriptome sequencing of three female and three male H.vigintioctopunctata libraries to identify sex-related genes.We recorded 281599294 bp of clean reads,and de novo assembly generated 85206 unigenes,with N_(50) of 1311 bp and average length of 763 bp.We found 5002 genes highly expressed in males and 2179 genes highly expressed in females.Expression profiles of six unigenes specific to females,six unigenes specific to males,and ten unigenes common to both sexes were verified using semi-quantitive RT-PCR;results showed expression levels of these 22 unigenes were relatively consistent with the transcriptome analysis.Using all unigenes as references,we identified 6657 putative simple sequence repeats.The transcriptome sequences and gene expression profiles of female and male H.vigintioctopunctata help explore genetic mechanisms of sexual dimorphism.
文摘Six different indigenous plants were screened for antifeedant and insecticidal activity against fourth instar larvae of Epilachna beetle, Henosepilachna vigintioctopunctata, which is a severe pest on brinjal. Among the plants screened, Achyranthes aspera showed higher activity against the selected pest. Ethyl acetate extracts of A. aspera showed higher antifeedant index and insecticidal activity against fourth instar larvae of H. vigintioctopunctata. Preliminary phytochemical analysis revealed that the presence of alkaloid and quinines in the ethyl acetate extracts indicate higher percentage of activities. Hence, it may suggest its use for controlling the vegetables insect pest, H. vigintioctopunctata.
基金supported by the National Key R&D Program of China(2017YFD0200900)National Natural Science Foundation of China(31972269)GDUPS(2017).
文摘RNA interference(RNAi)has emerged as a powerful tool for developing novel management strategies for controlling insect pests.The 28-spotted ladybeetle,Henosepilachna vigintioctopunctata is one of the most important pests attacking solanaceous plants in Asia.In this study,the potential of dietary RNAi to manage H.vigintioctopunctata was investigated using both in vitro synthesized and bacterially expressed double-stranded RNAs(dsRNAs)of HvvATPase A and HvvATPase E.The expression levels of HvvATPase A and HvvATPase E were higher in Malpighian tubules than in other tissue types.The silencing of HvvATPase A and HvvATPase E led to significant mortality in H.vigintioctopunctata larvae.In addition,the ingestion of HvvATPase A and HvvATPase E significantly deterred feeding behavior and subsequently arrested the development of H.vigintioctopunctata.Notably,the bacterially expressed dsRNAs consistently caused higher mortality in larvae and adults.Finally,the nontarget effects of the dsRNAs of H.vigintioctopunctata on the predatory ladybeetle Propylaea japonica were evaluated.P.japonica 1st instar larvae were administered vATPase A and vATPase E dsRNAs from H.vigintioctopunctata and P.japonica under the worst-case scenario,in which dsGFP served as negative control.There were significant effects of dsHvvATPase A on P.japonica at the transcriptional level but not at the organismal level,whereas dsHvvATPase E did not effect P.japonica at either the transcriptional or the organismal level.Collectively,the results of the study suggest that HvvATPase A and HvvATPase E can act as novel molecular targets for the control of H.vigintioctopunctata.
基金This research was supported by the N ational Key R&D Program of China(2017YFD0200900)China A griculture Research System(CARS-09-P22).
文摘Henosepilachna vigintioctopunctata is a serious insect pest which attacks a large number of nightshades and cucurbits in Asian countries,Brazil and Australia.Prolonged application of traditional pesticides has caused environmental pollution and exerted deleterious effects on human health.Finding new approaches with high target specificity and low environmental contamination has become an urgent task.RNA interference(RNAi)induced by double-stranded RNA(dsRNA)is expected to be applicable to managing this pest.Here we evaluated the effects of Escherichia co/Z-expressed dsRNAs targeting ecdvsone receptor(EcR)gene via dietary delivery in laboratory and foliar spraying in a greenhouse.The target transcript was successfully knocked down when the 4th-instar larvae had fed on potatofoliage dipped with dsEcR in a laboratory bioassay.Around 85%of the HvEcR RNAi larvae remained as prepupae or became abnormal pupae,and failed to emerge into adults.Ingestion of ds£c7?-immersed foliage by the 3rd-instar larvae effectuated a comparable RNAi response and brought about more severe defects:all the resultant larvae arrested development,remained as prepupae and finally died.For assay in the greenhouse,a ds£c7?-contained E.coli suspension was directly sprayed to the foliage of greenhouse-growing potato plants and the 3rd-and 4th-instar larvae were transferred to the leaves.High RNAi efficacy was obtained and identical RNAi phenotypes were observed in treated larvae.In addition,spraying dsEcR reduced leaf damage.Our results indicate a possibility of practical application of dsEcR as an environmentally friendly RNA pesticide to control H.vigintioctopunctata larvae.
基金Special thanks to Dr.Xuguo Zhou(University of Kentucky)for his valuable suggestion on preparing this manuscript.This research was supported by the National Key R&D Program of China(2017YFD0200900)National Natural Science Foundation of China(31972269),and GDUPS(2017).The granting agencies had no role in the study design,data collection and analysis,decision to publish,or preparation of the manuscript.
文摘RNA interference(RNAi)techniques have emerged as powerful tools that facilitate development of novel management strategies for insect pests,such as Henosepilachna vigintioctopunctata(Coleoptera:Coccinellidae),which is a major pest of solanaceous plants in Asia.In this study,the potential of oral delivery of in vvYro-synthesized and bacterially expressed double-stranded H.vigintioctopunctata lesswright(Iwr)gene(dsHvlwr)to manage of H.vigintioctopunctata was investigated.Our results showed that the gene Hvlwr had a 480-bp open reading frame and encoded a 160-amino acid protein.Hvlwr expression levels were greater in the fat body than other tissue types.Hvlwr silencing led to greater H.vigintioctopunctata mortality rates and appeared to be time-and partially dose-dependent,likely as a result of the number of hemocytes increasing with dsRNA concentration,but decreasing with time.Bacterially expressed dsHvlwr that was applied to leaf discs caused 88%,66%,and 36%mortality in 1st instars,3rd instars,and adults after 10,10,and 14 d,respectively;when applied to living plants,there was greater mortality in 1 st and 3rd instars,but there was no effect on adults.Furthermore,dsHvlwr led to improved plant protection against H.vigintioctopunctata.Our study shows an effective dietary RNAi response in H.vigintioctopunctata and that Hvlwr is a promising RNAi target gene for control of this pest species.
基金supported by grants from the Natural Science Foundation of Hubei Province(2020CFA012)the National Natural Science Foundation of China(32271912)to J.Z。
文摘RNA interference(RNAi)has emerged as a powerful technology for pest management.Previously,we have shown that plastid-mediated RNAi(PM-RNAi)can be utilized to control the Colorado potato beetle,an insect pest in the Chrysomelidae family;however,whether this technology is suitable for controlling pests in the Coccinellidae remained unknown.The coccinellid 28-spotted potato ladybird(Henosepilachna vigintioctopunctata;HV)is a serious pest of solanaceous crops.In this study,we identified three efficient target genes(β-Actin,SRP54,and SNAP)for RNAi using in vitro double-stranded RNAs(dsRNAs)fed to HV,and found that dsRNAs targetingβ-Actin messenger RNA(dsACT)induced more potent RNAi than those targeting the other two genes.We next generated transplastomic and nuclear transgenic potato(Solanum tuberosum)plants expressing HV dsACT.Long dsACT stably accumulated to up to 0.7%of the total cellular RNA in the transplastomic plants,at least three orders of magnitude higher than in the nuclear transgenic plants.Notably,the transplastomic plants also exhibited a significantly stronger resistance to HV,killing all larvae within 6 d.Our data demonstrate the potential of PM-RNAi as an efficient pest control measure for HV,extending the application range of this technology to Coccinellidae pests.
