Nine species in the subgenus Zeugodacus Hendel were known to occur in Yunnan. We add seven more species to the fauna of Yunnan. Among them, one species, Bactrocera (Zeugodacus) vultus Hardy, 1973, is reported from Chi...Nine species in the subgenus Zeugodacus Hendel were known to occur in Yunnan. We add seven more species to the fauna of Yunnan. Among them, one species, Bactrocera (Zeugodacus) vultus Hardy, 1973, is reported from China for the first time. This species is redescribed in detail and its male genitalia are illustrated and described for the first time. A key to the species of the subgenus Zeugodacus from Yunnan is provided.展开更多
The initiation of sex differentiation in insects is regulated by primary sex determination signals.In the Medfly Ceratitis capitata and other tephritids,Maleness-on-the-Y(MoY) is the master gene for male sex determina...The initiation of sex differentiation in insects is regulated by primary sex determination signals.In the Medfly Ceratitis capitata and other tephritids,Maleness-on-the-Y(MoY) is the master gene for male sex determination.However,the primary signal in Zeugodacus cucurbitae(Coquillett),a very destructive tephritid pest across the world,remains ambiguous.In this study,we have isolated and characterized the Medfly MoY homolog in Z.cucurbitae,ZcMoY.ZcMOY protein shows high sequence conservation to its homologs in Bactrocera species.ZcMoY transcription begins and peaks at very early embryonic stages and then becomes undetectable except the testes and heads of day 1 male adults.Silencing ZcMoY in early embryos by RNAi causes abnormal external genitalia and interior reproductive organs,giving rise to intersexes and feminization of XY individuals.The expression pattern and knockdown phenotypes of ZcMoY indicate that ZcMoY plays a key role in regulating sex determination of Z.cucurbitae males.Our findings will help the understanding of sex determination in Z.cucurbitae and facilitate the development of genetic sexing strains in its biological control.展开更多
The stripped fruit fly, Bactrocera （Zeugodacus） scutellata （Hendel）, is one of the most dangerous quarantine insect pests in China. Due to its wide distribution and heavy damage, it has caused serious economic los...The stripped fruit fly, Bactrocera （Zeugodacus） scutellata （Hendel）, is one of the most dangerous quarantine insect pests in China. Due to its wide distribution and heavy damage, it has caused serious economic losses in fruit and vegetable production in China. This paper assembles research progresses in mor- phological characteristics, classification status, host ranges, geographical distribution, population dynamics, molecular ecology and biology of B. scutellata, and put forward the following research prospects as well.展开更多
transformer is a switch gene for sex determination in many insects, which cooperates with transformer-2 that is expressed in both sexes to regulate female differentiation, particularly in dipterans. Zeugodacus cucurbi...transformer is a switch gene for sex determination in many insects, which cooperates with transformer-2 that is expressed in both sexes to regulate female differentiation, particularly in dipterans. Zeugodacus cucurbitae(Coquillett) is a very destructive pest worldwide, however, its sex determination pathway remains largely uncharacterized. Here, we show that the female sex ratio is sharply reduced with knockdown of either transformer or transformer-2 by RNA interference in early embryos of Z. cucurbitae. Most of the males grown from the embryos with transient transformer and transformer-2 suppression mated with wild-type females and produced mixed sex progeny, with one exception that produced only female progeny, and all of the few remaining males failed to mate with wild-type females and produced no progeny. The exceptional male and those males with mating failure were XX pseudomales as determined by the detection of Y chromosome-linked Maleness-on-the-Y, indicating that most XX pseudomales are not viable. The phenotypes of transformer and transformer-2 suggest that they play a key role in regulating sex determination and are required for female sexual development of Z. cucurbitae. Our results will be beneficial to the understanding of sex determination in Z. cucurbitae and can facilitate the development of genetic sexing strains for its biological control.展开更多
In the organisms with XX/XY sex chromosomes,Y chromosome is unique to males and plays an important role in male reproductive development.The study of Y chromosome genes will contribute to the development of pest genet...In the organisms with XX/XY sex chromosomes,Y chromosome is unique to males and plays an important role in male reproductive development.The study of Y chromosome genes will contribute to the development of pest genetic prevention and control technology.In this study,we identified 9 Y chromosome genes in Zeugodacus tau(Diptera:Tephritidae),including gene 16222.Protein structure analysis showed that 16222 was highly similar to odorant binding protein,and thus gene 16222 was named obp-Y.Obp-Y knockout(KO)significantly reduced hatching rate of offspring.Sperm detection results showed that obp-Y KO did not affect sperm number in the testes or sperm transfer during mating.We further examined the storage of sperms in females,and found that sperms in females mating with wild-type males began to transfer from spermathecal ducts to the spermathecae at hour 0 after the end of mating(AEM),and at 0-24 h AEM,the sperm count in the spermathecae gradually increased.However,no sperms were observed in spermathecae of females mating with mutant males at hours 0,4,8,24 and 48 AEM.In summary,this study revealed that Y chromosome gene obp-Y was necessary for the storage of sperms in females.Our findings not only provide theoretical basis for elucidating the function of the Y chromosome,but also offer a molecular target for the genetic control over Z.tau.展开更多
Previous studies have shown oviposition deterring properties of 8 coconut free fatty acid(CFFA)compounds on fruit flies with different key deterrent components for different species.Here we evaluated oviposition deter...Previous studies have shown oviposition deterring properties of 8 coconut free fatty acid(CFFA)compounds on fruit flies with different key deterrent components for different species.Here we evaluated oviposition deterrence of CFFA using laboratory 2-choice bioassays against Zeugodacus cucurbitae,determined key-bioactive deterrent compounds,and evaluated their behavioral mode.Unlike other reported fruit fly species,CFFA mixture increased Z.cucurbitae oviposition when directly applied on an oviposition substrate.When tested individually in subsequent tests,4 compounds(caprylic,capric,oleic,and linoleic acids)significantly reduced the oviposition(“negative-compounds”),1 compound(stearic acid)had no effect(“neutral-compound”),and 3 compounds(lauric,myristic,and palmitic acids)stimulated the oviposition(“positive-compounds”).The 4-component negative-compound blend was effective at reducing oviposition.However,adding stearic acid to the 4-component blend(5-component blend,5c)further reduced oviposition.Adding any of the positive-compounds to the 5c resulted in loss of oviposition deterrence,suggesting the 5c as the key deterrent component blend.The blend was also effective in no-choice assays and when applied on cucumbers,a preferred host of Z.cucurbitae.When given a choice,Z.cucurbitae made 48.5%fewer visits,spent 39%less time,and oviposited 88.2%fewer eggs per min on 5c treated pumpkin agar than on control agar,suggesting that the 5c blend has both spatial repellency and contact deterrence.Given that all compounds are registered food additives and generally regarded as safe,this blend has potential application in behavioral control strategies,such as push-pull,to protect host fruit against Z.cucurbitae.展开更多
Fruit flies usually harbor diverse communities of bacteria in their digestive systems,which are known to play a significant role in their fitness.However,little information is available on Zeugodacus tau,a polyphagous...Fruit flies usually harbor diverse communities of bacteria in their digestive systems,which are known to play a significant role in their fitness.However,little information is available on Zeugodacus tau,a polyphagous pest worldwide.This study reports the first extensive analysis of bacterial communities in different life stages and their effect on the development and reproduction of laboratory-reared Z tan.Cultured bacteria were identified using the conventional method and all bacteria were identified by highthroughput technologies(16S ribosomal RNA gene sequencing of V3-V4 region).A total of six bacterial phyla were identified in larvae,pupae,and male and female adult flies,which were distributed into 14 classes,32 orders,58 families and 96 genera.Proteobacteria was the most represented phylum in all the stages except larvae.Enterobacter,Klebsiella,Providencia,and Pseudomonas were identified by conventional and next-generation sequencing analysis in both male and female adult flies,and Enterobacter was found to be the main genus.After being fed with antibiotics from the first instar larvae,bacterial diversity changed markedly in the adult stage.Untreated flies laid eggs and needed 20 days before oviposition while the treated flies showed ovary development inhibited and were not able to lay eggs,probably due to the alteration of the microbiota.These findings provide the cornerstone for unexplored research on bacterial function in Z tau,which will help to develop an environmentally friendly management technique for this kind of harmful insect.展开更多
Zeugodacus tau(Walker)is an invasive pest.The sterile insect technique is an environment-friendly method for pest control.Understanding the mechanism of sex determination will contribute to improving efficiency of thi...Zeugodacus tau(Walker)is an invasive pest.The sterile insect technique is an environment-friendly method for pest control.Understanding the mechanism of sex determination will contribute to improving efficiency of this technique.In this study,we identified the transformer(tra)gene in Z.tau.One female-specific and two male-specific isoforms of tra were found in Z.tau,and the male-specific splicing pattern of tra was found to occur 5 h after egg laying.We performed transcriptome sequencing at 1 h(E1),5 h(E5),and 9 h(E9)after egg laying and obtained high-quality transcriptome libraries of early embryo development.We identified 13297 and 11713 differentially expressed genes(DEGs)from E5 versus E1 and E9 versus E1 comparisons,respectively.To explore the potential functions of the DEGs during embryonic development,Gene Ontology,Clusters of Orthologous Groups of proteins,and Kyoto Encyclopedia of Genes and Genomes analyses were performed.Twenty-six genes potentially involved in sex determination or differentiation,including Maleness-on-the-Y(MoY),were identified in Z.tau.To verify the transcriptome results,15 genes were selected for quantitative real-time PCR validation.The results were consistent with the transcriptome sequencing results.Moreover,U2 small nuclear riboprotein auxiliary factor(U2AF-50),female lethal d(fl(2)d),and virilizer(vir)were highly expressed at E5,indicating that they may be related to the sex-specific splicing of tra.Further functional analysis is needed to confirm this speculation.Our data provide an insight into the mechanism underlying sex determination and differentiation in tephritid species.展开更多
Accurate species-level identifications underpin many aspects of basic and applied biology;however,identifications can be hampered by a lack of discriminating morphological characters,taxonomic expertise or time.Molecu...Accurate species-level identifications underpin many aspects of basic and applied biology;however,identifications can be hampered by a lack of discriminating morphological characters,taxonomic expertise or time.Molecular approaches,such as DNA"barcoding"of the cytochrome c oxidase(COI)gene,are argued to overcome these issues.However,nuclear encoding of mitochondrial genes(numts)and poor amplification success of suboptimally preserved specimens can lead to erroneous identifications.One insect group for which these molecular and morphological problems are significant are the dacine fruit flies(Diptera:Tephritidae:Dacini).We addressed these issues associated with COI barcoding in the dacines by first assessing several"universal"COI primers against public mitochondrial genome and numt sequences for dacine taxa.We then modified a set of four primers that more closely matched true dacine COI sequence and amplified two overlapping portions of the COI barcode region.Our new primers were tested alongside universal primers on a selection of dacine species,including both fresh preserved and decades-old dry specimens.Additionally,Bactrocera tiyoni mitochondrial and nuclear genomes were compared to identify putative numts.Four numt clades were identified,three of which were amplified using existing universal primers.In contrast,our new primers preferentially amplified the"true"mitochondrial COI barcode in all dacine species tested.The new primers also successfully amplified partial barcodes from dry specimens for which full length barcodes were unobtainable.Thus we recommend these new primers be incorporated into the suites of primers used by diagnosticians and quarantine labs for the accurate identification of dacine species.展开更多
文摘Nine species in the subgenus Zeugodacus Hendel were known to occur in Yunnan. We add seven more species to the fauna of Yunnan. Among them, one species, Bactrocera (Zeugodacus) vultus Hardy, 1973, is reported from China for the first time. This species is redescribed in detail and its male genitalia are illustrated and described for the first time. A key to the species of the subgenus Zeugodacus from Yunnan is provided.
基金supported by the the National Natural Science Foundation of China (31702059, 31860523, and 31660339)the Hainan Provincial Natural Science Foundation of China (321CXTD435)。
文摘The initiation of sex differentiation in insects is regulated by primary sex determination signals.In the Medfly Ceratitis capitata and other tephritids,Maleness-on-the-Y(MoY) is the master gene for male sex determination.However,the primary signal in Zeugodacus cucurbitae(Coquillett),a very destructive tephritid pest across the world,remains ambiguous.In this study,we have isolated and characterized the Medfly MoY homolog in Z.cucurbitae,ZcMoY.ZcMOY protein shows high sequence conservation to its homologs in Bactrocera species.ZcMoY transcription begins and peaks at very early embryonic stages and then becomes undetectable except the testes and heads of day 1 male adults.Silencing ZcMoY in early embryos by RNAi causes abnormal external genitalia and interior reproductive organs,giving rise to intersexes and feminization of XY individuals.The expression pattern and knockdown phenotypes of ZcMoY indicate that ZcMoY plays a key role in regulating sex determination of Z.cucurbitae males.Our findings will help the understanding of sex determination in Z.cucurbitae and facilitate the development of genetic sexing strains in its biological control.
基金Supported by National Natural Science Foundation of China(41401065)Applied Fundamental Research Project of Yunnan Province(2015FB109)
文摘The stripped fruit fly, Bactrocera （Zeugodacus） scutellata （Hendel）, is one of the most dangerous quarantine insect pests in China. Due to its wide distribution and heavy damage, it has caused serious economic losses in fruit and vegetable production in China. This paper assembles research progresses in mor- phological characteristics, classification status, host ranges, geographical distribution, population dynamics, molecular ecology and biology of B. scutellata, and put forward the following research prospects as well.
基金supported by the Hainan Provincial Natural Science Foundation of China(321CXTD435)the National Natural Science Foundation of China(31860523,31660339,31702059,and 32260665)the National Key R&D Program of China(2022YFC2601400)。
文摘transformer is a switch gene for sex determination in many insects, which cooperates with transformer-2 that is expressed in both sexes to regulate female differentiation, particularly in dipterans. Zeugodacus cucurbitae(Coquillett) is a very destructive pest worldwide, however, its sex determination pathway remains largely uncharacterized. Here, we show that the female sex ratio is sharply reduced with knockdown of either transformer or transformer-2 by RNA interference in early embryos of Z. cucurbitae. Most of the males grown from the embryos with transient transformer and transformer-2 suppression mated with wild-type females and produced mixed sex progeny, with one exception that produced only female progeny, and all of the few remaining males failed to mate with wild-type females and produced no progeny. The exceptional male and those males with mating failure were XX pseudomales as determined by the detection of Y chromosome-linked Maleness-on-the-Y, indicating that most XX pseudomales are not viable. The phenotypes of transformer and transformer-2 suggest that they play a key role in regulating sex determination and are required for female sexual development of Z. cucurbitae. Our results will be beneficial to the understanding of sex determination in Z. cucurbitae and can facilitate the development of genetic sexing strains for its biological control.
基金supported by National Key R&D Program of China(No.2021YFC2600400)China Agriculture Research System of MOF and MARA(CARS-26)Hubei Hongshan Laboratory.
文摘In the organisms with XX/XY sex chromosomes,Y chromosome is unique to males and plays an important role in male reproductive development.The study of Y chromosome genes will contribute to the development of pest genetic prevention and control technology.In this study,we identified 9 Y chromosome genes in Zeugodacus tau(Diptera:Tephritidae),including gene 16222.Protein structure analysis showed that 16222 was highly similar to odorant binding protein,and thus gene 16222 was named obp-Y.Obp-Y knockout(KO)significantly reduced hatching rate of offspring.Sperm detection results showed that obp-Y KO did not affect sperm number in the testes or sperm transfer during mating.We further examined the storage of sperms in females,and found that sperms in females mating with wild-type males began to transfer from spermathecal ducts to the spermathecae at hour 0 after the end of mating(AEM),and at 0-24 h AEM,the sperm count in the spermathecae gradually increased.However,no sperms were observed in spermathecae of females mating with mutant males at hours 0,4,8,24 and 48 AEM.In summary,this study revealed that Y chromosome gene obp-Y was necessary for the storage of sperms in females.Our findings not only provide theoretical basis for elucidating the function of the Y chromosome,but also offer a molecular target for the genetic control over Z.tau.
基金supported in part by funding from Plant Protection Act 7721 funding to Dong H.Cha,Amy Roda,Junwei Zhu,and Paul Kendra(#2021-6.1171)Jia-Wei Tay and Dong H.Cha(AP22PPQS&T00C067)a CRADA with Marron Bio Innovations,Inc to Dong H.Cha and Junwei Zhu(#58-2040-0-004)。
文摘Previous studies have shown oviposition deterring properties of 8 coconut free fatty acid(CFFA)compounds on fruit flies with different key deterrent components for different species.Here we evaluated oviposition deterrence of CFFA using laboratory 2-choice bioassays against Zeugodacus cucurbitae,determined key-bioactive deterrent compounds,and evaluated their behavioral mode.Unlike other reported fruit fly species,CFFA mixture increased Z.cucurbitae oviposition when directly applied on an oviposition substrate.When tested individually in subsequent tests,4 compounds(caprylic,capric,oleic,and linoleic acids)significantly reduced the oviposition(“negative-compounds”),1 compound(stearic acid)had no effect(“neutral-compound”),and 3 compounds(lauric,myristic,and palmitic acids)stimulated the oviposition(“positive-compounds”).The 4-component negative-compound blend was effective at reducing oviposition.However,adding stearic acid to the 4-component blend(5-component blend,5c)further reduced oviposition.Adding any of the positive-compounds to the 5c resulted in loss of oviposition deterrence,suggesting the 5c as the key deterrent component blend.The blend was also effective in no-choice assays and when applied on cucumbers,a preferred host of Z.cucurbitae.When given a choice,Z.cucurbitae made 48.5%fewer visits,spent 39%less time,and oviposited 88.2%fewer eggs per min on 5c treated pumpkin agar than on control agar,suggesting that the 5c blend has both spatial repellency and contact deterrence.Given that all compounds are registered food additives and generally regarded as safe,this blend has potential application in behavioral control strategies,such as push-pull,to protect host fruit against Z.cucurbitae.
基金We thank the other members of Plant Quarantine and Invasion Biology Laboratory,China Agricultural University(CAUPQL)who helped us during this study.The research was supported by the National Natural Science Foundation Project of China(31801802)Fundamental Research Funds for the Central Universities,China(2019 TC060).
文摘Fruit flies usually harbor diverse communities of bacteria in their digestive systems,which are known to play a significant role in their fitness.However,little information is available on Zeugodacus tau,a polyphagous pest worldwide.This study reports the first extensive analysis of bacterial communities in different life stages and their effect on the development and reproduction of laboratory-reared Z tan.Cultured bacteria were identified using the conventional method and all bacteria were identified by highthroughput technologies(16S ribosomal RNA gene sequencing of V3-V4 region).A total of six bacterial phyla were identified in larvae,pupae,and male and female adult flies,which were distributed into 14 classes,32 orders,58 families and 96 genera.Proteobacteria was the most represented phylum in all the stages except larvae.Enterobacter,Klebsiella,Providencia,and Pseudomonas were identified by conventional and next-generation sequencing analysis in both male and female adult flies,and Enterobacter was found to be the main genus.After being fed with antibiotics from the first instar larvae,bacterial diversity changed markedly in the adult stage.Untreated flies laid eggs and needed 20 days before oviposition while the treated flies showed ovary development inhibited and were not able to lay eggs,probably due to the alteration of the microbiota.These findings provide the cornerstone for unexplored research on bacterial function in Z tau,which will help to develop an environmentally friendly management technique for this kind of harmful insect.
基金This study was supported by the National Key R&D Program of China(no.2019YFD1002100)the China Agriculture Research System of MOF and MARA(no.CARS-26)+2 种基金the National Natural Science Foundation of China(no.31872931)Scientific Research Plan Project of Education Department of Hubei Province,China(no.B2019329)Fundamental Research Funds for the Central Universities(nos.2662019PY055 and 2662015PY129).
文摘Zeugodacus tau(Walker)is an invasive pest.The sterile insect technique is an environment-friendly method for pest control.Understanding the mechanism of sex determination will contribute to improving efficiency of this technique.In this study,we identified the transformer(tra)gene in Z.tau.One female-specific and two male-specific isoforms of tra were found in Z.tau,and the male-specific splicing pattern of tra was found to occur 5 h after egg laying.We performed transcriptome sequencing at 1 h(E1),5 h(E5),and 9 h(E9)after egg laying and obtained high-quality transcriptome libraries of early embryo development.We identified 13297 and 11713 differentially expressed genes(DEGs)from E5 versus E1 and E9 versus E1 comparisons,respectively.To explore the potential functions of the DEGs during embryonic development,Gene Ontology,Clusters of Orthologous Groups of proteins,and Kyoto Encyclopedia of Genes and Genomes analyses were performed.Twenty-six genes potentially involved in sex determination or differentiation,including Maleness-on-the-Y(MoY),were identified in Z.tau.To verify the transcriptome results,15 genes were selected for quantitative real-time PCR validation.The results were consistent with the transcriptome sequencing results.Moreover,U2 small nuclear riboprotein auxiliary factor(U2AF-50),female lethal d(fl(2)d),and virilizer(vir)were highly expressed at E5,indicating that they may be related to the sex-specific splicing of tra.Further functional analysis is needed to confirm this speculation.Our data provide an insight into the mechanism underlying sex determination and differentiation in tephritid species.
文摘Accurate species-level identifications underpin many aspects of basic and applied biology;however,identifications can be hampered by a lack of discriminating morphological characters,taxonomic expertise or time.Molecular approaches,such as DNA"barcoding"of the cytochrome c oxidase(COI)gene,are argued to overcome these issues.However,nuclear encoding of mitochondrial genes(numts)and poor amplification success of suboptimally preserved specimens can lead to erroneous identifications.One insect group for which these molecular and morphological problems are significant are the dacine fruit flies(Diptera:Tephritidae:Dacini).We addressed these issues associated with COI barcoding in the dacines by first assessing several"universal"COI primers against public mitochondrial genome and numt sequences for dacine taxa.We then modified a set of four primers that more closely matched true dacine COI sequence and amplified two overlapping portions of the COI barcode region.Our new primers were tested alongside universal primers on a selection of dacine species,including both fresh preserved and decades-old dry specimens.Additionally,Bactrocera tiyoni mitochondrial and nuclear genomes were compared to identify putative numts.Four numt clades were identified,three of which were amplified using existing universal primers.In contrast,our new primers preferentially amplified the"true"mitochondrial COI barcode in all dacine species tested.The new primers also successfully amplified partial barcodes from dry specimens for which full length barcodes were unobtainable.Thus we recommend these new primers be incorporated into the suites of primers used by diagnosticians and quarantine labs for the accurate identification of dacine species.
