Wing dimorphism is regarded as an important phenotypic plasticity involved in the migration and reproduction of aphids.However,the signal transduction and regulatory mechanism of wing dimorphism in aphids are still un...Wing dimorphism is regarded as an important phenotypic plasticity involved in the migration and reproduction of aphids.However,the signal transduction and regulatory mechanism of wing dimorphism in aphids are still unclear.Herein,the optimal environmental conditions were first explored for inducing winged offspring of green peach aphid,and the short photoperiod was the most important environmental cue to regulate wing dimorphism.Compared to 16 L:8 D photoperiod,the proportion of winged offspring increased to 90%under 8 L:16 D photoperiod.Subsequently,5 differentially expressed microRNAs(miRNAs)in aphids treated with long and short photoperiods were identified using small RNA sequencing,and a novel miR-3040 was identified as a vital miRNA involved in photoperiod-mediated wing dimorphism.More specifically,the inhibition of miR-3040 expression could reduce the proportion of winged offspring induced by short photoperiod,whereas its activation increased the proportion of winged offspring under long photoperiod.Meanwhile,the expression level of miR-3040 in winged aphids was about 2.5 times that of wingless aphids,and the activation or inhibition of miR-3040 expression could cause wing deformity,revealing the dual-role regulator of miR-3040 in wing dimorphism and wing development.In summary,the current study identified the key environmental cue for wing dimorphism in green peach aphid,and the first to demonstrate the dual-role regulator of miR-3040 in photoperiod-mediated wing dimorphism and wing development.展开更多
The ecdysone-induced transcription factor E93 in model insects plays multiple roles in the insect metamorphosis processes,such as remodeling larval tissues and determining adult tissue formation.The knockdown of E93in...The ecdysone-induced transcription factor E93 in model insects plays multiple roles in the insect metamorphosis processes,such as remodeling larval tissues and determining adult tissue formation.The knockdown of E93in insects leads to incomplete metamorphosis,suggesting that E93 is a potential target for pest control.In this study,the HaE93 gene in the cotton bollworm Helicoverpa armigera,a polyphagous pest of various commercial crops worldwide,was identified and found to have high expression in the egg,prepupal,and pupal stages.The injection of ds HaE93 induced about 60%mortality in H.armigera at the larval-pupal stage.About 30%survived but showed delayed pupation and abnormal wings,and the females developed reduced ovaries.Therefore,about 90%of the HaE93 knockdown individuals failed to reproduce before they died.The results of qRT-PCR showed that the expression levels of ecdysone primary-response genes,chitin synthesis-related genes,and wing and ovary development-related genes were reduced in HaE93 knockdown H.armigera.These results indicated that HaE93plays a critical role in larva-pupa-adult metamorphosis and the development of the cuticle,wing,and ovary in female H.armigera by regulating the expression of the associated genes.Bioassays of ds HaE93 administered by either oral delivery or injection showed similar knockdown results,which suggested that HaE93 can be used as a target gene for the RNAi control of the pest H.armigera.展开更多
The tanning hormone,Bursicon,is a neuropeptide secreted by the insect nervous system that functions as a heterodimer composed of Burs-αand Burs-βsubunits.It plays a critical role in the processes of cuticle tanning ...The tanning hormone,Bursicon,is a neuropeptide secreted by the insect nervous system that functions as a heterodimer composed of Burs-αand Burs-βsubunits.It plays a critical role in the processes of cuticle tanning and wing expansion in insects.In this study,we successfully identified the AcBurs-αand AcBurs-βgenes in Aphis citricidus.The open reading frames of AcBurs-αand AcBurs-βwere 480 and 417 bp in length,respectively.Both AcBurs-αand AcBurs-βexhibited 11 conserved cysteine residues.AcBurs-αand AcBurs-βwere expressed during all developmental stages of A.citricidus and showed high expression levels in the winged aphids.To investigate the potential role of AcBurs-αand AcBurs-βin wing development,we employed RNA interference(RNAi)techniques.With the efficient silencing of AcBurs-α(44.90%)and AcBurs-β(52.31%),malformed wings were induced in aphids.The proportions of malformed wings were 22.50%,25.84%,and 38.34%in dsAcBurs-α-,dsAcBur-β-,and dsAcBurs-α+dsAcBur-β-treated groups,respectively.Moreover,feeding protein kinase A inhibitors(H-89)also increased the proportion of malformed wings to 30.00%.Feeding both double-stranded RNA and inhibitors(H-89)significantly downregulated the wing development-related genes nubbin,vestigial,notch and spalt major.Silence of vestigial through RNAi also led to malformed wings.Meanwhile,the exogenous application of 3 hormones that influence wing development did not affect the expression level of AcBursicon genes.These findings indicate that AcBursicon genes plays a crucial role in wing development in A.citricidus;therefore,it represents a potential molecular target for the control of this pest through RNAi-based approaches.展开更多
Wings are an important flight organ of insects.Wing development is a complex process controlled by a series of genes.The flightless wing pad transforms into a mature wing with the function of migratory flight during t...Wings are an important flight organ of insects.Wing development is a complex process controlled by a series of genes.The flightless wing pad transforms into a mature wing with the function of migratory flight during the nymphto-adult metamorphosis.However,the mechanism of wing morphogenesis in locusts is still unclear.This study analyzed the microstructures of the locust wing pads at pre-eclosion and the wings after eclosion and performed the comparative transcriptome analysis.RNA-seq identified 25,334 unigenesand 3,430 differentially expressed genes(DEGs)(1,907 up-regulated and 1,523 down-regulated).The DEGs mainly included cuticle development(LmACPs),chitin metabolism(Lm Idgf4),lipid metabolism-related genes,cell adhesion(Integrin),zinc finger transcription factors(LmSalm,LmZF593 andLmZF521),and others.Functional analysis based on RNA interference and hematoxylin and eosin(H&E)staining showed that the three genes encoded zinc finger transcription factors are essential for forming wing cuticle and maintaining morphology in Locusta migratoria.Finally,the study found that the LmSalm regulates the expression of LmACPs in the wing pads at pre-eclosion,and LmZF593 and LmZF521 regulate the expression of LmIntegrin/LmIdgf4/LmHMT420 in the wings after eclosion.This study revealed that the molecular regulatory axis controls wing morphology in nymphal and adult stages of locusts,offering a theoretical basis for the study of wing development mechanisms in hemimetabolous insects.展开更多
RNA interference(RNAi)has developed rapidly as a potential“green”pest management strategy.At present,most studies have focused on the screening of aphid lethal genes,whereas only a few studies have been conducted on...RNA interference(RNAi)has developed rapidly as a potential“green”pest management strategy.At present,most studies have focused on the screening of aphid lethal genes,whereas only a few studies have been conducted on wing development,which is crucial for aphid migration and plant-virus dissemination.Here,the Myzus persicae genes vestigial(vg)and Ultrabithorax(Ubx)related to wing development,were cloned.These two genes were expressed in various tissues of 3rd-instar winged aphids.The mRNA level of vg was high in 3rd-instar nymphs,whereas the expression level of Ubx was high in adults.The nanocarrier-mediated delivery system delivered double-stranded RNAs for aphid RNAi using topical and root applications.The expression levels of vg and Ubx were downregulated by 44.0%and 36.5%,respectively,using the topical application.The simultaneous RNAi of the two target genes caused 63.3%and 32.2%wing aberration rates using topical and root applications,respectively.The current study provided a promising method for controlling aphid migration to alleviate the spread of insect transmitted plant diseases.展开更多
The Notch signaling pathway plays a central role in the development of various organisms.However,dysregulation of microRNAs(miRNAs),which are crucial regulators of gene expression,can disrupt signaling pathways at all...The Notch signaling pathway plays a central role in the development of various organisms.However,dysregulation of microRNAs(miRNAs),which are crucial regulators of gene expression,can disrupt signaling pathways at all stages of development.Although Notch signaling is involved in wing development in Drosophila,the mechanism underlying miRNA-based regulation of the Notch signaling pathway is unclear.Here,we report that loss of Drosophila miR-252 increases the size of adult wings,whereas the overexpression of miR-252 in specific compartments of larval wing discs leads to patterning defects in the adult wings.The miR-252 overexpression-induced wing phenotypes were caused by aberrant Notch signaling with intracellular accumulation of the full-length Notch receptor during development,which could be due to defects in intracellular Notch trafficking associated with its recycling to the plasma membrane and autophagy-mediated degradation.Moreover,we identified Rab6 as a direct target of miR-252-5p;Rab6 encodes a small Ras-like GTPase that regulates endosomal trafficking pathways.Consistent with this finding,RNAi-mediated downregulation of Rab6 led to similar defects in both wing patterning and Notch signaling.Notably,co-overexpression of Rab6 completely rescued the wing phenotype associated with miR-252 overexpression,further supporting that Rab6 is a biologically relevant target of miR-252-5p in the context of wing development.Thus,our data indicate that the miR-252-5p-Rab6 regulatory axis is involved in Drosophila wing development by controlling the Notch signaling pathway.展开更多
Since it takes a long time to design an aircraft, the succession to or the acquirement of knowledges and technologies is a key to provide to aircraft design high quality and productivity. This paper describes a comput...Since it takes a long time to design an aircraft, the succession to or the acquirement of knowledges and technologies is a key to provide to aircraft design high quality and productivity. This paper describes a computer support system for wing design using a new tool based on an object-oriented approach. The system is programmed in a module of objects which represent design tasks or design knowledges. The modulated program gives advantages to maintain and extend the system easily. The system provides to users a flexible support with excellent user interface which consists of mouse, menu-driven system, and window system.展开更多
Cuticular proteins(CPs)are major components of the insect cuticle-associated organs such as integument and wings,although the importance of CPs for wing development and function in hemimetabolous insects remains under...Cuticular proteins(CPs)are major components of the insect cuticle-associated organs such as integument and wings,although the importance of CPs for wing development and function in hemimetabolous insects remains understudied.In the present study,a wing cuticular protein LmACP8 was identified from Locusta migratoria,which belongs to the RR-2 subfamily of cuticular protein R&R consensus(CPR)chitin-binding proteins.LmACP8 was mainly expressed in the wing pads and showed high expression levels before ecdysis of third-,fourth-,and fifth-instar nymphs,with its encoded protein located in the procuticle of wing pads and adult wings.Depletion of LmACP8 by RNA interference markedly reduced the amount of its protein,which consequently caused abnormal wing morphogenesis in the transition from nymph to adult of L.migratoria.We further demonstrated that the abnormal morphogenesis was caused by severe damage of the endocuticle in the wings.LmACP8 was suppressed by 20-hydroxyecdysone(20 E)in vivo,however,its expression was significantly up-regulated after knocking down the hormone receptor gene LmHR39.Thus,the LmACP8 that is negatively regulated by the LmHR39-mediated 20 E signaling pathway is involved in wing development during the nymph to adult transition.展开更多
Insect wings are subject to strong selective pressure,resulting in the evolution of remarkably diverse wing morphologies that largely determine flight capacity.However,the genetic basis and regulatory mechanisms under...Insect wings are subject to strong selective pressure,resulting in the evolution of remarkably diverse wing morphologies that largely determine flight capacity.However,the genetic basis and regulatory mechanisms underlying wing size and shape development are not well understood.The silkworm Bombyx mori micropterous(mp)mutant exhibits shortened wing length and enlarged vein spacings,albeit without changes in total wing area.Thus,the mp mutant comprises a valuable genetic resource for studying wing de-velopment.In this study,we used molecular mapping to identify the gene responsible for the mp phenotype and designated it Bmmp.Phenotype-causing mutations were identified as indels and single nucleotide polymorphisms in noncoding regions.These mutations resulted in decreased Bmmp messenger RNA levels and changes in transcript isoform composition.Bmmp null mutants were generated by clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9 and exhibited changed wing shape,similar to mp mutants,and significantly smaller total wing area.By examining the expression of genes critical to wingdevelopment inwildtype and Bmmp null mutants,we found that Bmmp exerts its function by coordinately modulating anterior-posterior and proximal-distal axes development.We also studied a Drosophila mp mutant and found that Bmmp is functionally conserved in Drosophila.The Drosophila mp mutant strain exhibits curly wings of reduced size and a complete loss of flight capacity.Our results increase our understanding of the mechanisms underpinning insect wing development and reveal potential targets for pest control.展开更多
Wings are an important flight organ of insects and their morphogenesis depends on a series of cell-to-cell and cell-to-extracellular matrix interactions.Integrin as a transmembrane protein receptor mediates cell-to-ce...Wings are an important flight organ of insects and their morphogenesis depends on a series of cell-to-cell and cell-to-extracellular matrix interactions.Integrin as a transmembrane protein receptor mediates cell-to-cell adhesion,cell-to-extracellular matrix interactions and signal transduction.In the present study,we characterized an integrin gene that encodes integrinβ-PS protein in Locusta migratoria.Lmlntegrinβ-PS is highly expressed in the wing pads and the middle stages of 5th instar nymphs.Immunohisto-chemical analysis revealed that the Lmlntegrinβ-PS protein was localized at the cell base of the two layers of wings.After suppression of Lmlntegrinβ-PS by RNA interference,the wing pads or wings were unable to form normally,with a blister wing appearance during nymph to nymph transition and nymph to adult transition.We further found that the dorsal and ventral epidermis of the wings after dsLmlntegrinβ-PS injection were improperly connected and formed huge cavities revealed by hematoxylin and eosin staining.Furthermore,the morphology and structure of the wing cuticle was significantly disturbed which affected the stable arrangement and attachments of the wing epidermis.Moreover,the expression of related cell adhesion genes was significantly decreased in LmIntegrinβ-PS-suppressed L.migratoria,suggesting that Lmlntegrinβ-PS is required for the morphogenesis and development of wings during molting by stabilizing cell adhesion and maintaining the cytoskeleton of these cells.展开更多
基金the National Natural Science Foundation of China(32030012)National Key Research and Development Program of the Ministry of Science and Technology(2022YFD1401800).
文摘Wing dimorphism is regarded as an important phenotypic plasticity involved in the migration and reproduction of aphids.However,the signal transduction and regulatory mechanism of wing dimorphism in aphids are still unclear.Herein,the optimal environmental conditions were first explored for inducing winged offspring of green peach aphid,and the short photoperiod was the most important environmental cue to regulate wing dimorphism.Compared to 16 L:8 D photoperiod,the proportion of winged offspring increased to 90%under 8 L:16 D photoperiod.Subsequently,5 differentially expressed microRNAs(miRNAs)in aphids treated with long and short photoperiods were identified using small RNA sequencing,and a novel miR-3040 was identified as a vital miRNA involved in photoperiod-mediated wing dimorphism.More specifically,the inhibition of miR-3040 expression could reduce the proportion of winged offspring induced by short photoperiod,whereas its activation increased the proportion of winged offspring under long photoperiod.Meanwhile,the expression level of miR-3040 in winged aphids was about 2.5 times that of wingless aphids,and the activation or inhibition of miR-3040 expression could cause wing deformity,revealing the dual-role regulator of miR-3040 in wing dimorphism and wing development.In summary,the current study identified the key environmental cue for wing dimorphism in green peach aphid,and the first to demonstrate the dual-role regulator of miR-3040 in photoperiod-mediated wing dimorphism and wing development.
基金supported by the National Natural Science Foundation of China(32001912 and 32370525)the Henan Provincial Natural Science Foundation,China(232300420012)。
文摘The ecdysone-induced transcription factor E93 in model insects plays multiple roles in the insect metamorphosis processes,such as remodeling larval tissues and determining adult tissue formation.The knockdown of E93in insects leads to incomplete metamorphosis,suggesting that E93 is a potential target for pest control.In this study,the HaE93 gene in the cotton bollworm Helicoverpa armigera,a polyphagous pest of various commercial crops worldwide,was identified and found to have high expression in the egg,prepupal,and pupal stages.The injection of ds HaE93 induced about 60%mortality in H.armigera at the larval-pupal stage.About 30%survived but showed delayed pupation and abnormal wings,and the females developed reduced ovaries.Therefore,about 90%of the HaE93 knockdown individuals failed to reproduce before they died.The results of qRT-PCR showed that the expression levels of ecdysone primary-response genes,chitin synthesis-related genes,and wing and ovary development-related genes were reduced in HaE93 knockdown H.armigera.These results indicated that HaE93plays a critical role in larva-pupa-adult metamorphosis and the development of the cuticle,wing,and ovary in female H.armigera by regulating the expression of the associated genes.Bioassays of ds HaE93 administered by either oral delivery or injection showed similar knockdown results,which suggested that HaE93 can be used as a target gene for the RNAi control of the pest H.armigera.
基金supported by the National Natural Science Foundation of China(32272526,32302338,and 32020103010)Natural Science Foundation of Chongqing,China(CSTB2022NSCQ-MSX0750)China Agriculture Research System of the Ministry of Finance and the Ministry of Agriculture and Rural Affairs.
文摘The tanning hormone,Bursicon,is a neuropeptide secreted by the insect nervous system that functions as a heterodimer composed of Burs-αand Burs-βsubunits.It plays a critical role in the processes of cuticle tanning and wing expansion in insects.In this study,we successfully identified the AcBurs-αand AcBurs-βgenes in Aphis citricidus.The open reading frames of AcBurs-αand AcBurs-βwere 480 and 417 bp in length,respectively.Both AcBurs-αand AcBurs-βexhibited 11 conserved cysteine residues.AcBurs-αand AcBurs-βwere expressed during all developmental stages of A.citricidus and showed high expression levels in the winged aphids.To investigate the potential role of AcBurs-αand AcBurs-βin wing development,we employed RNA interference(RNAi)techniques.With the efficient silencing of AcBurs-α(44.90%)and AcBurs-β(52.31%),malformed wings were induced in aphids.The proportions of malformed wings were 22.50%,25.84%,and 38.34%in dsAcBurs-α-,dsAcBur-β-,and dsAcBurs-α+dsAcBur-β-treated groups,respectively.Moreover,feeding protein kinase A inhibitors(H-89)also increased the proportion of malformed wings to 30.00%.Feeding both double-stranded RNA and inhibitors(H-89)significantly downregulated the wing development-related genes nubbin,vestigial,notch and spalt major.Silence of vestigial through RNAi also led to malformed wings.Meanwhile,the exogenous application of 3 hormones that influence wing development did not affect the expression level of AcBursicon genes.These findings indicate that AcBursicon genes plays a crucial role in wing development in A.citricidus;therefore,it represents a potential molecular target for the control of this pest through RNAi-based approaches.
基金This work was supported by the National Key R&D Program of China(2022YFD1700200)the National Natural Science Foundation of China(31970469)+2 种基金earmarked fund for Modern Agro-industry Technology Research System,China(2023CYJSTX01-20)the Fund for Shanxi“1331 Project”,Chinathe Scientific and Technological Innovation Programs of Higher Education Institutions in Shanxi,China(2022Y032)。
文摘Wings are an important flight organ of insects.Wing development is a complex process controlled by a series of genes.The flightless wing pad transforms into a mature wing with the function of migratory flight during the nymphto-adult metamorphosis.However,the mechanism of wing morphogenesis in locusts is still unclear.This study analyzed the microstructures of the locust wing pads at pre-eclosion and the wings after eclosion and performed the comparative transcriptome analysis.RNA-seq identified 25,334 unigenesand 3,430 differentially expressed genes(DEGs)(1,907 up-regulated and 1,523 down-regulated).The DEGs mainly included cuticle development(LmACPs),chitin metabolism(Lm Idgf4),lipid metabolism-related genes,cell adhesion(Integrin),zinc finger transcription factors(LmSalm,LmZF593 andLmZF521),and others.Functional analysis based on RNA interference and hematoxylin and eosin(H&E)staining showed that the three genes encoded zinc finger transcription factors are essential for forming wing cuticle and maintaining morphology in Locusta migratoria.Finally,the study found that the LmSalm regulates the expression of LmACPs in the wing pads at pre-eclosion,and LmZF593 and LmZF521 regulate the expression of LmIntegrin/LmIdgf4/LmHMT420 in the wings after eclosion.This study revealed that the molecular regulatory axis controls wing morphology in nymphal and adult stages of locusts,offering a theoretical basis for the study of wing development mechanisms in hemimetabolous insects.
基金We would like to acknowledge the National Natural Science Foundation of China(32030012 and 31900363)Beijing Natural Science Foundation(6204043)National Key Research and Development Program of China(2018YFD0200804).
文摘RNA interference(RNAi)has developed rapidly as a potential“green”pest management strategy.At present,most studies have focused on the screening of aphid lethal genes,whereas only a few studies have been conducted on wing development,which is crucial for aphid migration and plant-virus dissemination.Here,the Myzus persicae genes vestigial(vg)and Ultrabithorax(Ubx)related to wing development,were cloned.These two genes were expressed in various tissues of 3rd-instar winged aphids.The mRNA level of vg was high in 3rd-instar nymphs,whereas the expression level of Ubx was high in adults.The nanocarrier-mediated delivery system delivered double-stranded RNAs for aphid RNAi using topical and root applications.The expression levels of vg and Ubx were downregulated by 44.0%and 36.5%,respectively,using the topical application.The simultaneous RNAi of the two target genes caused 63.3%and 32.2%wing aberration rates using topical and root applications,respectively.The current study provided a promising method for controlling aphid migration to alleviate the spread of insect transmitted plant diseases.
基金supported by the National Research Foundation of Korea(NRF)grant funded by the Korea Government(MSIT)(2021R1F1A1059864 to D-H.L.and 2021R1A2C1006353 to Y.S.L.)a Korea University Grant(to Y.S.L.).
文摘The Notch signaling pathway plays a central role in the development of various organisms.However,dysregulation of microRNAs(miRNAs),which are crucial regulators of gene expression,can disrupt signaling pathways at all stages of development.Although Notch signaling is involved in wing development in Drosophila,the mechanism underlying miRNA-based regulation of the Notch signaling pathway is unclear.Here,we report that loss of Drosophila miR-252 increases the size of adult wings,whereas the overexpression of miR-252 in specific compartments of larval wing discs leads to patterning defects in the adult wings.The miR-252 overexpression-induced wing phenotypes were caused by aberrant Notch signaling with intracellular accumulation of the full-length Notch receptor during development,which could be due to defects in intracellular Notch trafficking associated with its recycling to the plasma membrane and autophagy-mediated degradation.Moreover,we identified Rab6 as a direct target of miR-252-5p;Rab6 encodes a small Ras-like GTPase that regulates endosomal trafficking pathways.Consistent with this finding,RNAi-mediated downregulation of Rab6 led to similar defects in both wing patterning and Notch signaling.Notably,co-overexpression of Rab6 completely rescued the wing phenotype associated with miR-252 overexpression,further supporting that Rab6 is a biologically relevant target of miR-252-5p in the context of wing development.Thus,our data indicate that the miR-252-5p-Rab6 regulatory axis is involved in Drosophila wing development by controlling the Notch signaling pathway.
文摘Since it takes a long time to design an aircraft, the succession to or the acquirement of knowledges and technologies is a key to provide to aircraft design high quality and productivity. This paper describes a computer support system for wing design using a new tool based on an object-oriented approach. The system is programmed in a module of objects which represent design tasks or design knowledges. The modulated program gives advantages to maintain and extend the system easily. The system provides to users a flexible support with excellent user interface which consists of mouse, menu-driven system, and window system.
基金the National Key R&D Program of China(2017YFD0200900)the National Natural Science Foundation of China(31702067 and 31970469)+1 种基金the National Natural Science Foundation of China–Deutsche Forschungsgemeinschaft of Germany(31761133021)the Scientific and Technological Innovation Programs of Higher Education Institutions in Shanxi,China(2019L0033)。
文摘Cuticular proteins(CPs)are major components of the insect cuticle-associated organs such as integument and wings,although the importance of CPs for wing development and function in hemimetabolous insects remains understudied.In the present study,a wing cuticular protein LmACP8 was identified from Locusta migratoria,which belongs to the RR-2 subfamily of cuticular protein R&R consensus(CPR)chitin-binding proteins.LmACP8 was mainly expressed in the wing pads and showed high expression levels before ecdysis of third-,fourth-,and fifth-instar nymphs,with its encoded protein located in the procuticle of wing pads and adult wings.Depletion of LmACP8 by RNA interference markedly reduced the amount of its protein,which consequently caused abnormal wing morphogenesis in the transition from nymph to adult of L.migratoria.We further demonstrated that the abnormal morphogenesis was caused by severe damage of the endocuticle in the wings.LmACP8 was suppressed by 20-hydroxyecdysone(20 E)in vivo,however,its expression was significantly up-regulated after knocking down the hormone receptor gene LmHR39.Thus,the LmACP8 that is negatively regulated by the LmHR39-mediated 20 E signaling pathway is involved in wing development during the nymph to adult transition.
基金supported by the National Natural Science Foundation of China(awards U20A2058 and 31830094).
文摘Insect wings are subject to strong selective pressure,resulting in the evolution of remarkably diverse wing morphologies that largely determine flight capacity.However,the genetic basis and regulatory mechanisms underlying wing size and shape development are not well understood.The silkworm Bombyx mori micropterous(mp)mutant exhibits shortened wing length and enlarged vein spacings,albeit without changes in total wing area.Thus,the mp mutant comprises a valuable genetic resource for studying wing de-velopment.In this study,we used molecular mapping to identify the gene responsible for the mp phenotype and designated it Bmmp.Phenotype-causing mutations were identified as indels and single nucleotide polymorphisms in noncoding regions.These mutations resulted in decreased Bmmp messenger RNA levels and changes in transcript isoform composition.Bmmp null mutants were generated by clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9 and exhibited changed wing shape,similar to mp mutants,and significantly smaller total wing area.By examining the expression of genes critical to wingdevelopment inwildtype and Bmmp null mutants,we found that Bmmp exerts its function by coordinately modulating anterior-posterior and proximal-distal axes development.We also studied a Drosophila mp mutant and found that Bmmp is functionally conserved in Drosophila.The Drosophila mp mutant strain exhibits curly wings of reduced size and a complete loss of flight capacity.Our results increase our understanding of the mechanisms underpinning insect wing development and reveal potential targets for pest control.
基金the National Key R&D Program of China(2017YFD0200900)National Natural Science Foundation of China(31702067,31970469)+3 种基金NSFC-DFG(31761133021)Scientific and Technological Innovation Programs of Higher Education Institutions in Shanxi(2019L0033)Key Research and Development Program of Shanxi Province,China(201803D221004-5)2018 Special Talents Projects in Shanxi Province,China(201805D211019).
文摘Wings are an important flight organ of insects and their morphogenesis depends on a series of cell-to-cell and cell-to-extracellular matrix interactions.Integrin as a transmembrane protein receptor mediates cell-to-cell adhesion,cell-to-extracellular matrix interactions and signal transduction.In the present study,we characterized an integrin gene that encodes integrinβ-PS protein in Locusta migratoria.Lmlntegrinβ-PS is highly expressed in the wing pads and the middle stages of 5th instar nymphs.Immunohisto-chemical analysis revealed that the Lmlntegrinβ-PS protein was localized at the cell base of the two layers of wings.After suppression of Lmlntegrinβ-PS by RNA interference,the wing pads or wings were unable to form normally,with a blister wing appearance during nymph to nymph transition and nymph to adult transition.We further found that the dorsal and ventral epidermis of the wings after dsLmlntegrinβ-PS injection were improperly connected and formed huge cavities revealed by hematoxylin and eosin staining.Furthermore,the morphology and structure of the wing cuticle was significantly disturbed which affected the stable arrangement and attachments of the wing epidermis.Moreover,the expression of related cell adhesion genes was significantly decreased in LmIntegrinβ-PS-suppressed L.migratoria,suggesting that Lmlntegrinβ-PS is required for the morphogenesis and development of wings during molting by stabilizing cell adhesion and maintaining the cytoskeleton of these cells.