The body surface of crustaceans is covered with a sturdy shell.The growth and development of crustaceans are realized through molting.Neocaridina denticulata sinensis is a suitable candidate for crustacean scientific ...The body surface of crustaceans is covered with a sturdy shell.The growth and development of crustaceans are realized through molting.Neocaridina denticulata sinensis is a suitable candidate for crustacean scientific research.Two calcium-associated cuticular protein genes,named NdCAP-1 and NdCAP-2,were obtained from N.denticulata sinensis.Molecular docking simulated the binding effect of both proteins and calcium ions.Semi-quantitative reverse transcription PCR results show that NdCAP-1 is expressed in D_(2-4) stage,NdCAP-2 expressed in D_(2-4) and A-B stages,and both were significantly expressed in the cephalothorax cuticle and pereiopods.Then,it was revealed that NdCAP-1 and NdCAP-2 are regulated by NdEcR-mediated 20 E signaling pathways.Knockdown of NdCAP-1 and NdCAP-2 was observed to cause surface defects.The recombinant proteins(rNdCAP-1 and rNdCAP-2),obtained by prokaryotic expression,had calcium-binding and chitin-binding ability,inhibited formation of calcium carbonate precipitate.These results show that calcium-associated cuticular proteins play important roles in cuticle formation and calcification.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
The cuticle, an essential structure for insects, is produced from cuticular proteins and chitin via a series of biochemical reactions. Tweedle genes are important members of the cuticular protein family and have four ...The cuticle, an essential structure for insects, is produced from cuticular proteins and chitin via a series of biochemical reactions. Tweedle genes are important members of the cuticular protein family and have four conserved motifs binding to chitin. Tweedle family genes have been found to play a profound effect on cuticle development. Here, we report that the cuticular protein gene LmTwdll ofLocusta migratoria belongs to the Tweedle family. In situ hybridization showed that LmTwdll is localized to epidermal cells of the cuticle. The expression patterns of LmTwdll showed low expression in the cuticle during the early and middle stages of the fifth-instar nymphs; in contrast, its expression rapidly increased in the late stages of fifth-instar nymphs. We performed RNA interference to examine the function of LmTwdll in locusts. Silencing of LmTwdll resulted in high mortality during the molting process before the next stage. Also, the epicuticle of nymphs failed to molt, tended to be thinner and the arrangement of chitin in the procuticle appeared to be disordered compare to the control group. These results demonstrate that LmTwdl! plays a critical role in molting, which contributes to a better understanding of the distinct functions of the Tweedle family in locusts.展开更多
Thirteen cuticular protein （CP） families have been recognized in arthropods. In this study, 250 Anopheles sinensis CP genes were identified and named based on genome and transcriptome sequences. They were classified...Thirteen cuticular protein （CP） families have been recognized in arthropods. In this study, 250 Anopheles sinensis CP genes were identified and named based on genome and transcriptome sequences. They were classified into 10 families based on mo- tifs and phylogenetic analyses. In 11 other insect species, nine had CP numbers 〉 150 while Apis mellifera and Tribolium castaneum had CP numbers less than 52. The CPs of eight species occupied 〉 1.4% of the total genomic gene number, whereas in three species the CPs occupied 〈 1%. The phylogenies for each CP family in An. sinensis were constructed and discussed. The 250 CPs each had 1-8 exons with 144 CPs （57.6%） having two exons. The intron length ranged from 66--3888 bp with 174 introns （54.0%） being 66--100 bp long. Except for two CPs on two contigs, 248 CPs were mapped onto 28 scaffolds with 136 genes （54.4%） restricted to five scaffolds. A total of 107 CPs were clustered and located at 27 loci. The CPR family had the conserved motif GSYS- LVEPDGTVRTV. The RR- 1 subfamily had an additional 21 amino acid （aa） motifs with the YVADENGF sequence that is common in insects. The RR-2 subfamily had an additional 50 aa motifs with two additional regions RDGDWKG and G-x（3）-VV. A comparison with 115 orthologous counterparts of An. gambiae CPs suggested purifying selection for all of these genes. This study provides basic information useful for further studies on biological functions of An. sinensis CPs as well as for comparative genomics of insect CPs.展开更多
The acrostyle is a distinct anatomical region present on the cuticle at the inner face of the common food/salivary canal at the tip of aphid maxillary stylets. This conserved structure is of particular interest as it ...The acrostyle is a distinct anatomical region present on the cuticle at the inner face of the common food/salivary canal at the tip of aphid maxillary stylets. This conserved structure is of particular interest as it harbors the protein receptors of at least l plant virus, Cauliflower mosaic virus＇, and presumably has other roles in plant-insect interactions. Previously we reported immunolabeling of a highly conserved motif of cuticular proteins from the CPR family （named for the presence of a Rebers and Riddiford consensus） within the acrostyle. Here we report the development of novel tools to further study the proteomic composition of this region and to identify proteins involved in insect- virus interactions. Using a series of antibodies against cuticular proteins from the RR-2 subfamily, we identified additional peptides present within the acrostyle. Our results demonstrated that the acrostyle is a complex structure containing multiple domains of cuticular proteins accessible for interaction. In addition, an array of overlapping peptides,which covers the diversity of the majority of the RR-2 subfamily, was ~tevelopect as a generic tool to characterize cuticular protein/pathogen interactions. Upon probing this array with Cucumber mosaic virus particles, consensus peptide sequences from hybridizing peptides were identified. Use of these novel tools has extended our knowledge of the proteomic composition of insect maxillary stylets and identified sequences that could be involved in virus binding, thus contributing to further elucidation of the various properties and functions of the acrostyle.展开更多
Recently,parasitoid wasp species Microplitis mediator has evoked increasing research attention due to its possible use in the control of Lepidoptera insects.Because insect development involves changes in cuticle compo...Recently,parasitoid wasp species Microplitis mediator has evoked increasing research attention due to its possible use in the control of Lepidoptera insects.Because insect development involves changes in cuticle composition,identification and expression analysis of M.mediator cuticular proteins may clarify the mechanisms involved in parasite development processes.We found 70 cuticular proteins from the M.mediator transcrip-tome and divided them into seven distinct families.Expression profiling indicated that most of these cuticular protein genes have expression peaks specific for one particular developmental stage of M.mediator.Eggs and pupae have the highest number of tran-scriptionally active cuticular protein genes(47 and 52 respectively).Only 12 of these genes maintained high expression activity during late larval development.Functional analysis of two larval proteins,MmCPR3 and MmCPR 14,suggested their important role in the proper organization of the cuticle layers of larvac.During M.mediator larval development,normal cuticle formation can be supported by a limited number of cuticular proteins.展开更多
家蚕Bombyx mori不同品种对人工饲料的摄食性存在很大差异。为探讨食性差异的分子机理,本文基于对人工饲料摄食性不同的蚕品种(系)SAGE(serial analysis of gene expression)文库差异表达基因的分析,发掘了1条家蚕假定表皮蛋白(cuticula...家蚕Bombyx mori不同品种对人工饲料的摄食性存在很大差异。为探讨食性差异的分子机理,本文基于对人工饲料摄食性不同的蚕品种(系)SAGE(serial analysis of gene expression)文库差异表达基因的分析,发掘了1条家蚕假定表皮蛋白(cuticular protein hypothetical)基因BmCph2。采用半定量RT-PCR和实时荧光定量PCR方法,对BmCph2在不同摄食性蚕品种(系)不同发育时期的表达特征进行了研究。结果表明:BmCph2基因在家蚕幼虫眠期和起蚕期高表达,在胚胎期和幼虫将眠期几乎检测不到表达;在幼虫头部与全蚕的表达特征相似,而在中肠中表达活性很低,推测该基因表达可能与家蚕新表皮的形成有关。BmCph2在对人工饲料摄食性不同的蚕品种(系)中的表达存在较大差异,在摄食性好的高食性品种中表达量显著低于摄食性差的低食性品种;饲料和忌避剂的气味刺激及取食刺激对不同品种(系)该基因的表达有不同的影响,高食性蚕对诱导刺激比较敏感,而低食性蚕受影响较小,尤其是菁松A和菁松B的低食性品系几乎不受影响。本研究结果说明,BmCph2基因除可能参与表皮形成的同时,还与家蚕的食性有密切关系,但其具体机理有待于进一步研究。展开更多
基金Supported by the National Natural Science Foundation of China(Nos.32172954,32373121)the Key Research and Development Project of Hebei Province(No.22323201D)+2 种基金the Science and Technology Project of Hebei Education Department(No.ZD 2022093)the Natural Science Foundation of Hebei Province(Nos.D2022201003,D2023201002)the Hangzhou Qianjiang Special Expert fund for Prof.Jiquan ZHANG。
文摘The body surface of crustaceans is covered with a sturdy shell.The growth and development of crustaceans are realized through molting.Neocaridina denticulata sinensis is a suitable candidate for crustacean scientific research.Two calcium-associated cuticular protein genes,named NdCAP-1 and NdCAP-2,were obtained from N.denticulata sinensis.Molecular docking simulated the binding effect of both proteins and calcium ions.Semi-quantitative reverse transcription PCR results show that NdCAP-1 is expressed in D_(2-4) stage,NdCAP-2 expressed in D_(2-4) and A-B stages,and both were significantly expressed in the cephalothorax cuticle and pereiopods.Then,it was revealed that NdCAP-1 and NdCAP-2 are regulated by NdEcR-mediated 20 E signaling pathways.Knockdown of NdCAP-1 and NdCAP-2 was observed to cause surface defects.The recombinant proteins(rNdCAP-1 and rNdCAP-2),obtained by prokaryotic expression,had calcium-binding and chitin-binding ability,inhibited formation of calcium carbonate precipitate.These results show that calcium-associated cuticular proteins play important roles in cuticle formation and calcification.
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This research was supported by the National Natural Science Foundation of China Grants （Nos. 31472051, 31201710）, Natural Science Foundation of Shanxi Province of China （2014011028-3）, Fund Program for the Scientific Activities of Selected Returned Overseas Professionals in Shanxi Province, the Platform Project of Science and Technology in Shanxi Province （2015091010）, and National Basic Research Program of China （No. 2012CB114102）. We thank Dr. Feng Jiang for critical comments on the manuscript.
文摘The cuticle, an essential structure for insects, is produced from cuticular proteins and chitin via a series of biochemical reactions. Tweedle genes are important members of the cuticular protein family and have four conserved motifs binding to chitin. Tweedle family genes have been found to play a profound effect on cuticle development. Here, we report that the cuticular protein gene LmTwdll ofLocusta migratoria belongs to the Tweedle family. In situ hybridization showed that LmTwdll is localized to epidermal cells of the cuticle. The expression patterns of LmTwdll showed low expression in the cuticle during the early and middle stages of the fifth-instar nymphs; in contrast, its expression rapidly increased in the late stages of fifth-instar nymphs. We performed RNA interference to examine the function of LmTwdll in locusts. Silencing of LmTwdll resulted in high mortality during the molting process before the next stage. Also, the epicuticle of nymphs failed to molt, tended to be thinner and the arrangement of chitin in the procuticle appeared to be disordered compare to the control group. These results demonstrate that LmTwdl! plays a critical role in molting, which contributes to a better understanding of the distinct functions of the Tweedle family in locusts.
文摘Thirteen cuticular protein （CP） families have been recognized in arthropods. In this study, 250 Anopheles sinensis CP genes were identified and named based on genome and transcriptome sequences. They were classified into 10 families based on mo- tifs and phylogenetic analyses. In 11 other insect species, nine had CP numbers 〉 150 while Apis mellifera and Tribolium castaneum had CP numbers less than 52. The CPs of eight species occupied 〉 1.4% of the total genomic gene number, whereas in three species the CPs occupied 〈 1%. The phylogenies for each CP family in An. sinensis were constructed and discussed. The 250 CPs each had 1-8 exons with 144 CPs （57.6%） having two exons. The intron length ranged from 66--3888 bp with 174 introns （54.0%） being 66--100 bp long. Except for two CPs on two contigs, 248 CPs were mapped onto 28 scaffolds with 136 genes （54.4%） restricted to five scaffolds. A total of 107 CPs were clustered and located at 27 loci. The CPR family had the conserved motif GSYS- LVEPDGTVRTV. The RR- 1 subfamily had an additional 21 amino acid （aa） motifs with the YVADENGF sequence that is common in insects. The RR-2 subfamily had an additional 50 aa motifs with two additional regions RDGDWKG and G-x（3）-VV. A comparison with 115 orthologous counterparts of An. gambiae CPs suggested purifying selection for all of these genes. This study provides basic information useful for further studies on biological functions of An. sinensis CPs as well as for comparative genomics of insect CPs.
文摘The acrostyle is a distinct anatomical region present on the cuticle at the inner face of the common food/salivary canal at the tip of aphid maxillary stylets. This conserved structure is of particular interest as it harbors the protein receptors of at least l plant virus, Cauliflower mosaic virus＇, and presumably has other roles in plant-insect interactions. Previously we reported immunolabeling of a highly conserved motif of cuticular proteins from the CPR family （named for the presence of a Rebers and Riddiford consensus） within the acrostyle. Here we report the development of novel tools to further study the proteomic composition of this region and to identify proteins involved in insect- virus interactions. Using a series of antibodies against cuticular proteins from the RR-2 subfamily, we identified additional peptides present within the acrostyle. Our results demonstrated that the acrostyle is a complex structure containing multiple domains of cuticular proteins accessible for interaction. In addition, an array of overlapping peptides,which covers the diversity of the majority of the RR-2 subfamily, was ~tevelopect as a generic tool to characterize cuticular protein/pathogen interactions. Upon probing this array with Cucumber mosaic virus particles, consensus peptide sequences from hybridizing peptides were identified. Use of these novel tools has extended our knowledge of the proteomic composition of insect maxillary stylets and identified sequences that could be involved in virus binding, thus contributing to further elucidation of the various properties and functions of the acrostyle.
基金This work was supported by the National Key Plan for Scientific Research and Development of China(2017YFD0200400)and the National Key Plan for Scientific Research and Development of China(2016YFC1200603)+1 种基金the National Natural Science Foun-dation of China(31872298)Open Research Fund Pro-gram of State Key Laboratory ofIPM(Chinese IPM 1803).
文摘Recently,parasitoid wasp species Microplitis mediator has evoked increasing research attention due to its possible use in the control of Lepidoptera insects.Because insect development involves changes in cuticle composition,identification and expression analysis of M.mediator cuticular proteins may clarify the mechanisms involved in parasite development processes.We found 70 cuticular proteins from the M.mediator transcrip-tome and divided them into seven distinct families.Expression profiling indicated that most of these cuticular protein genes have expression peaks specific for one particular developmental stage of M.mediator.Eggs and pupae have the highest number of tran-scriptionally active cuticular protein genes(47 and 52 respectively).Only 12 of these genes maintained high expression activity during late larval development.Functional analysis of two larval proteins,MmCPR3 and MmCPR 14,suggested their important role in the proper organization of the cuticle layers of larvac.During M.mediator larval development,normal cuticle formation can be supported by a limited number of cuticular proteins.
文摘家蚕Bombyx mori不同品种对人工饲料的摄食性存在很大差异。为探讨食性差异的分子机理,本文基于对人工饲料摄食性不同的蚕品种(系)SAGE(serial analysis of gene expression)文库差异表达基因的分析,发掘了1条家蚕假定表皮蛋白(cuticular protein hypothetical)基因BmCph2。采用半定量RT-PCR和实时荧光定量PCR方法,对BmCph2在不同摄食性蚕品种(系)不同发育时期的表达特征进行了研究。结果表明:BmCph2基因在家蚕幼虫眠期和起蚕期高表达,在胚胎期和幼虫将眠期几乎检测不到表达;在幼虫头部与全蚕的表达特征相似,而在中肠中表达活性很低,推测该基因表达可能与家蚕新表皮的形成有关。BmCph2在对人工饲料摄食性不同的蚕品种(系)中的表达存在较大差异,在摄食性好的高食性品种中表达量显著低于摄食性差的低食性品种;饲料和忌避剂的气味刺激及取食刺激对不同品种(系)该基因的表达有不同的影响,高食性蚕对诱导刺激比较敏感,而低食性蚕受影响较小,尤其是菁松A和菁松B的低食性品系几乎不受影响。本研究结果说明,BmCph2基因除可能参与表皮形成的同时,还与家蚕的食性有密切关系,但其具体机理有待于进一步研究。
