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cuticular protein LmTwdll is involved in molt development of the migratory locust

Insect Science 2016年第4期23卷 520-530页

作者： Tian-Qi Song Mei-Ling Yang Yan-Li Wang Qing Liu Hui-Min Wang Jie Zhang Tao Li Research Institute of Applied Biology Shanxi University Taiyuan and State Key Laboratory of Integrated Management of Pest Insects and Rodents Institute of Zoology Chinese Academy of Sciences Beijing China Beijing Institutes of Life Science Chinese Academy of Sciences Beijing China

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.

关键词： cuticular protein Lm Twdll Loeusta migratoria molting RNA interference

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cuticular protein defective Bamboo mutant of Bombyx mori is sensitive to environmental stresses

Cuticular protein defective Bamboo mutant of Bombyx mori is ...

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中国蚕学会第十届青年学术研讨会

作者： Gao Xiong Xiaoling Tong Zhengwen Yan Hai Hu Xiaohui Duan Chunlin Li Minjin Han Cheng Lu Fangyin Dai State Key Laboratory of Silkworm Genome Biology Southwest University Key Laboratory of Sericultural Biology and Genetic Breeding Ministry of AgricultureCollege of BiotechnologySouthwest University

Insect cuticle acts as a primary protective barrier against environment stresses that may directly impact the insect ***,we report the mechanical defense function of a structural cuticular protein,BmorCPH24,to environmental stresses using a silkworm Bamboo（Bo） mutant with this gene ***（UV） irradiation and topical application of an acetone insecticide were used as environmental stresses to determine the differences in susceptibility between Bo and wild-type *** irradiation resulted in a sunburn phenotype in the Bo strains earlier than the wild-type indicating the sensitivity of *** malondialdehyde（MDA） content and a lower survival ratio were also observed in the Bo *** with deltamethrin revealed that Bo larvae were more sensitive to insecticides than the ***,cuticle analysis by microsection revealed thinner cuticle and a significant decrease in the endocuticle layer（～64.0%） in *** results suggest that BmorCPH24 mutation can lead to deficiency in resources required to construct the cuticle in Bo resulting in thin cuticle and reduced resistance to UV and *** results provide us new insight into the role of structural cuticular proteins in insect cuticle against environment stresses.

关键词： Bombyx mori Cuticle thickness cuticular protein Environmental stress Resistance

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cuticular protein defective Bamboo mutant of Bombyx mori is sensitive to environmental stresses

Cuticular protein defective Bamboo mutant of Bombyx mori is ...

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中国蚕学会第十四届暨国家蚕桑产业技术体系家（柞）蚕遗传育种学术研讨会

Insect cuticle acts as a primary protective barrier against environment stresses that may directly impact the insect body. Here, we report the mechanical defense function of a structural cuticular protein, BmorCPH24, to environmental stresses using a silkworm Bamboo mutant(Bo) with this gene mutation. Ultraviolet(UV) irradiation and topical application of an acetone insecticide were used as environmental stresses to determine the differences in susceptibility between Bo and wild-type larvae. UV irradiation resulted in a sunburn phenotype in the Bo strains earlier than the wild-type indicating the sensitivity of Bo. Higher malondialdehyde(MDA) content and a lower survival ratio were also observed in the Bo strains. Treatment with deltamethrin revealed that Bo larvae were more sensitive to insecticides than the wild-type. Furthermore, cuticle analysis by microsection revealed thinner cuticle and a significant decrease in the endocuticle layer(～64.0%) in Bo. These results suggest that BmorCPH24 mutation can lead to deficiency in resources required to construct the cuticle in Bo resulting in thin cuticle and reduced resistance to UV and insecticides. These results provide us new insight into the role of structural cuticular proteins in insect cuticle against environment stresses.

关键词： Bombyx mori UV Insecticide resistance cuticular protein Cuticle thickness

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Body Shape and Coloration of Silkworm Larvae Are Influenced by a Novel cuticular protein

Body Shape and Coloration of Silkworm Larvae Are Influenced ...

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中国蚕学会第十届青年学术研讨会

作者： Gao Xiong Xiaoling Tong Tingting Gai Chunlin Li Liang Qiao Antónia Monteiro Hai Hu Minjin Han Xin Ding Songyuan Wu Zhonghuai Xiang Cheng Lu Fangyin Dai State Key Laboratory of Silkworm Genome Biology Southwest University Key Laboratory of Sericultural Biology and Genetic Breeding Ministry of AgricultureCollege of BiotechnologySouthwest University Institute of Entomology and Molecular Biology College of Life SciencesChongqing Normal University Department of Biological Sciences National University of Singapore Yale-NUS College

The genetic basis of body shape and coloration patterns on caterpillars is often assumed to be regulated separately,but it is possible that common molecules affect both types of trait *** we examine the genetic basis of a spontaneous cuticle defect in silkworm,where larvae exhibit a bamboo-like body shape and decreased *** performed linkage mapping and mutation screening to determine the gene product that affects body shape and coloration *** these mutant larvae we identified a null mutation in BmorCPH24,a gene encoding a cuticular protein with low complexity *** expression analyses showed that BmorCPH24 is expressed in the larval epidermis ***-mediated knockdown and CRISPR/Cas9-mediated knockout of BmorCPH24 produced the abnormal body shape and the inhibited pigment typical of the mutant *** addition,our results showed that BmorCPH24 may be involved in the synthesis of endocuticle and its disruption-induced apoptosis of epidermal cells that accompanied the reduced expression of R＆R-type larval cuticle protein genes and pigmentation gene Wnt1 Strikingly,BmorCPH24,a fast-evolving gene,has evolved a new function responsible for the assembly of silkworm larval cuticle and has evolved to be an indispensable factor maintaining the larval body shape and its coloration *** is the first study to identify a molecule whose pleiotropic function affects the development of body shape and color patterns in insect larvae.

关键词： apoptosis body shape color patterns cuticular protein novel gene

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Body shape and coloration of silkworm larvae are influenced by a novel cuticular protein

Body shape and coloration of silkworm larvae are influenced ...

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中国蚕学会第十四届暨国家蚕桑产业技术体系家（柞）蚕遗传育种学术研讨会

作者： Gao Xiong Xiaoling Tong Tingting Gai Chunlin Li Liang Qiao Antónia Monteiro Hai Hu Minjin Han Xin Ding Songyuan Wu Zhonghuai Xiang Cheng Lu Fangyin Dai State Key Laboratory of Silkworm Genome Biology College of Biotechnology Southwest University Key Laboratory of Sericultural Biology and Genetic Breeding Ministry of Agriculture College of Biotechnology Southwest University Institute of Entomology and Molecular Biology College of Life Sciences Chongqing Normal University Department of Biological Sciences National University of Singapore Yale-NUS College

The genetic basis of body shape and coloration patterns on caterpillars is often assumed to be regulated separately, but it is possible that common molecules affect both types of trait simultaneously. Here we examine the genetic basis of a spontaneous cuticle defect in silkworm, where larvae exhibit a bamboo-like body shape and also decreased pigmentation. We performed linkage mapping and mutation screening to determine the gene product that affects body shape and coloration simultaneously. In these mutant larvae we identified a null mutation in BmorCPH24, a gene encoding a cuticular protein with low complexity sequence. Spatiotemporal expression analyses showed that BmorCPH24 is expressed in the larval epidermis post-ecdysis. RNAi-mediated knockdown and CRISPR/Cas9-mediated knockout of BmorCPH24 produced the abnormal body shape and the inhibited pigment typical of the mutant phenotype. In addition, our results showed that BmorCPH24 may be involved in the synthesis of endocuticle and its disruption induced apoptosis of epidermal cells which accompanied the reduced expression of R&R-type larval cuticle protein genes and pigmentation gene Wnt1. Strikingly, BmorCPH24, a fast-evolving gene, has evolved a new function responsible for the assembly of silkworm larval cuticle and has evolved to be an indispensable factor maintaining the larval body shape and its coloration pattern. This is the first study to identify a molecule whose pleiotropic function affects the development of body shape and color patterns in insect larvae.

关键词： body shape color patterns cuticular protein apoptosis novel gene

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Cloning and Expression pattern of a cuticular protein Gene（CP2-130） in Antheraea pernyi during embryonic development

Cloning and Expression pattern of a Cuticular Protein Gene（...

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第十二届家（柞）蚕遗传育种暨良种繁育学术研讨会

作者：马月月 Lei Piao Wang Yong 秦利 College of Bioscience and Biotechnology Shenyang Agricultural University Liaoning Engineering and Technology Resource Center for Insect Resource

Bivoltine tussah silkworm variety "Xuan Da No.1" was used to investigate the changes in embryo development in Antheraea pernyi. In this study, we collected the eggs at different embryonic development stages. Using 2D-PAGE, a changing protein was selected and identified by LC-MS/MS technology. This protein is similar to CP2-130 motif protein through BLASTp searching. The full sequence of this gene was cloned based on transcriptome data using RT-PCR technology, and this gene including an open reading frame（ORF） of 516 bp coding for a theoretical 171 amino acids protein. The deduced amino acid sequence has a molecular weight of 18.85 k Da（MW） and an isoelectric point（p I） of 6.11. The changes in the progress of embryonic development, semi-RT-PCR was used to detect the gene expression, control to the β-actin gene, Ap CP2-130 gene was not detected in the early stages（in 2 d and 4 d）, and gradually increased in the middle and later period. In this research, we have analysed one kind of cuticular protein and express pattern during different embryonic stages. This study would help us to learn more about the embryonic mechanism in A. pernyi.

关键词： Antheraea pernyi Embryo development cuticular protein 2D-PAGE Gene clone

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Body shape and coloration of silkworm larvae are influenced by a novel cuticular protein

Body shape and coloration of silkworm larvae are influenced ...

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中国蚕学会第九届青年学术研讨会

The genetic basis of body shape and coloration patterns on caterpillars is often assumed to be regulated separately,but it is possible that common molecules affect both types of trait *** we examine the genetic basis of a spontaneous cuticle defect in silkworm,where larvae exhibit a bamboo-like body shape and also decreased *** performed linkage mapping and mutation screening to determine the gene product that affects body shape and coloration *** these mutant larvae we identified a null mutation in BmorCPH24,a gene encoding a cuticular protein with low complexity *** expression analyses showed that BmorCPH24 is expressed in the larval epidermis ***-mediated knockdown and CRISPR/Cas9-mediated knockout of BmorCPH24 produced the abnormal body shape and the inhibited pigment typical of the mutant *** addition,our results showed that BmorCPH24 may be involved in the synthesis of endocuticle and its disruption induced apoptosis of epidermal cells which accompanied the reduced expression of R＆R-type larval cuticle protein genes and pigmentation gene ***,BmorCPH24,a fast-evolving gene,has evolved a new function responsible for the assembly of silkworm larval cuticle and has evolved to be an indispensable factor maintaining the larval body shape and its coloration *** is the first study to identify a molecule whose pleiotropic function affects the development of body shape and color patterns in insect larvae.

关键词： body shape color patterns cuticular protein apoptosis novel gene

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Mutation of a cuticular protein,BmorCPR2,alters larval body shape and adaptability in silkworm,Bombyx mori

Mutation of a cuticular protein,BmorCPR2,alters larval body ...

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中国遗传学会第九次全国会员代表大会暨学术研讨会

作者： Qiao Liang Xiong Gao Wang Ri-xin He Song-zhen Chen Jie Tong Xiao-ling Hu Hai Li Chun-lin Gai Ting-ting Xin Ya-qun Liu Xiao-fan Chen Bin Xiang Zhong-huai Lu Cheng Dai Fang-yin State Key Laboratory of Silkworm Genome Biology:Key Laboratory for Sericulture Functional Genomics and Biotechnology of Agricultural Ministry Southwest University Institute of Entomology and Molecular Biology College of Life SciencesChongqing Normal University State Key Laboratory of Silkworm Genome Biology

cuticular proteins(CPs) are crucial components of the insect *** numerous genes encoding cuticular proteins have been identified in known insect genomes to date, their functions in maintaining insect body shape and adaptability remain largely *** the current study,positional cloning led to the identification of a gene encoding a RRl-type cuticular protein,BmorCPR2,highly expressed in larval chitin-rich tissues and at the mulberry leaf-eating stages,which is responsible for the silkworm stony *** the Dazao-stony strain,the BmorCPR2 allele is a null mutation with significantly lower expression,compared to the wild-type Dazao *** BmorCPR2 in the stony mutant lost chitin binding ability,leading to reduced chitin content in larval cuticle,limitation of cuticle extension,abatement of cuticle tensile properties,and aberrant ratio between internodes and intersegmental *** variations induce a significant decrease in cuticle capacity to hold the growing internal organs in the larval development process,resulting in whole body stiffness,tightness and hardness,bulging intersegmental folds,and serious defects in larval *** our knowledge,this is the first study to report the corresponding phenotype of stony in insects caused by mutation of RR1-type cuticular *** findings collectively shed light on the specific role of cuticular proteins in maintaining normal larval body shape,and will aid in the development of pest control strategies for the management of Lepidoptera.

关键词： cuticular protein Silkworm stony mutant Body shape Adaptability Chitin

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Proteomic composition of the acrostyle＂ Novel approaches to identify cuticular proteins involved in virus-insect interactions

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Insect Science 2017年第6期24卷 990-1002页

作者： Craig Graham Webster Maelle Thillier Elodie Pirolles Bastien Cayrol Stephane Blanc Marilyne Uzest INRA UMR 0385 BGPI CIRAD-INRA-Montpellier SupAgro Campus International de Baillarguet Montpellier Cedex France

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.

关键词： acrostyle aphid cuticular protein immunolabeling peptide array stylet

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Identification and temporal expression profiles of cuticular proteins in the endoparasitoid wasp,Microplitis mediator

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Insect Science 2020年第5期27卷 998-1018页

作者： Olga Volovych Zhe Lin Jie Du Hong Jiang Zhen Zou State Key Laboratory of Integrated Management of Pest Insects and Rodents Institute of ZoologyChinese Academy of SciencesBeijingChina University of Chinese Academy of Sciences BeijingChina

Recently,parasitoid wasp species Microplitis mediator has evoked increasing research attention due to its possible use in the control of Lepidoptera *** insect development involves changes in cuticle composition,identification and expression analysis of *** cuticular proteins may clarify the mechanisms involved in parasite development *** found 70 cuticular proteins from the *** transcrip-tome and divided them into seven distinct *** profiling indicated that most of these cuticular protein genes have expression peaks specific for one particular developmental stage of *** 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 *** analysis of two larval proteins,MmCPR3 and MmCPR 14,suggested their important role in the proper organization of the cuticle layers of *** *** larval development,normal cuticle formation can be supported by a limited number of cuticular proteins.

关键词： cuticular protein expression pattern Microplis mediator transcriptome

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