We analyzed 20 chemosensory protein （CSP） genes of the silkworm Bombyx mori. We found a high number of retrotransposons inserted in introns. We then analyzed expression of the 20 BrnorCSP genes across tissues using ...We analyzed 20 chemosensory protein （CSP） genes of the silkworm Bombyx mori. We found a high number of retrotransposons inserted in introns. We then analyzed expression of the 20 BrnorCSP genes across tissues using quantitative real-time polymerase chain reaction （PCR）. Relatively low expression levels of BmorCSPs were found in the gut and fat body tissues. We thus tested the effects of endectocyte insecticide abamectin （B 1 a and Blb avermectins） on BmorCSP gene expression. Quantitative real-time PCR experi- ments showed that a single brief exposure to insecticide abamectin increased dramatically CSP expression not only in the antennae but in most tissues, including gut and fat body. Furthermore, our study showed coordinate expression of CSPs and metabolic cytochrome P450 enzymes in a tissue-dependent manner in response to the insecticide. The function of CSPs remains unknown. Based on our results, we suggest a role in detecting xenobiotics that are then detoxified by cytochrome P450 anti-xenobiotic enzymes.展开更多
Chemosensory proteins (CSPs) are ubiquitous soluble small proteins isolated from sensory organs thought to be involved in chemical communication. Here we report the first cDNA of CSPs, called Ac-ASP3, cloned and chara...Chemosensory proteins (CSPs) are ubiquitous soluble small proteins isolated from sensory organs thought to be involved in chemical communication. Here we report the first cDNA of CSPs, called Ac-ASP3, cloned and characterized from antennas of adult worker bees in Chinese honeybee, Apis cerana cerana. The Ac-ASP3 cDNA comprises 2 exons, with an ORF of 393-bp encoding 130 aa. Protein signature analyses show that the protein consists of four conserved cysteines and a signal peptide with 19 aa in the N-terminal sequence. The deduced protein sequence shares high homology with Am-ASP3 of Apis mellifera and low similarity with other species of insects. Immunocytochemical localization shows that Ac-ASP3 is only specifically expressed on the antenna contact chemosensilla such as sensilla trichodea B and sensilla basiconica, whereas Ac-ASP3 is scarcely expressed on olfactory chemosensilla such as sensilla placodea. Real-time PCR of Ac-ASP3 transcripts shows that Ac-ASP3 is highly expressed on wings and legs, but expression is lower on antenna. Temporal expression patterns suggest that Ac-ASP3 is expressed during the period of pupa and adults from 1-d to 6-d stages when bees act as house bees, cleaning the comb and taking care of the queen and larvae in comb. The above evidence suggests that Ac-ASP3 is unique in species and is generally not involved in olfaction during searching for honey and pollen. Rather, the protein seems to function in recognition of chemosensory substances on bees' cuticle and mechanical movement of antenna.展开更多
A chemosensory protein named HarmCSP5 in cotton bollworm Helicoverpa armigera （Hvbner） was obtained from antennal eDNA libraries and expressed in Escherichia coll. The real time quantitative PCR （RT-qPCR） results ...A chemosensory protein named HarmCSP5 in cotton bollworm Helicoverpa armigera （Hvbner） was obtained from antennal eDNA libraries and expressed in Escherichia coll. The real time quantitative PCR （RT-qPCR） results indicated that HarmCSP5 gene was mainly expressed in male and female antennae but also expressed in female legs and wings. Competitive binding assays were performed to test the binding affinity of recombinant HarmCSP5 to 60 odor molecules including some cotton volatiles. The resules showed that HarmCSP5 showed strong binding abilities to 4-ehtylbenzaldehyde and 3,4-dimethlbenz aldehyde, whereas methyl phenylacetate, 2-decanone, 1-pentanol, carvenol, isobomeol, nerolidol, 2- nonanone and ethyl heptanoate have relatively weak binding affinity. Moreover, the predicted 3D model of HarmCSP5 consists of six α-helices located among residues 33-38 （αl）, 40-48 （α2）, 62-72 （α3）, 80-96 （α4）, 98-108 （α5）, and 116-119 （α6）, two pairs of disulfide bridges Cys49-Cys55, Cys75-Cys78. The two amino acid residues, Ile94 and Trpl01, may play crucial roles in HarmCSP5 binding with ligands and need further study for confirmation.展开更多
Chemosensory proteins(CSPs) perform several functions in insects.This study performed the gene expression,ligand-binding,and molecular docking assays on the EforCSP3 identified in the parasitoid wasp Encarsia formosa,...Chemosensory proteins(CSPs) perform several functions in insects.This study performed the gene expression,ligand-binding,and molecular docking assays on the EforCSP3 identified in the parasitoid wasp Encarsia formosa,to determine whether EforCSP3 functions in olfaction,especially in host location and host preference.The results showed that EforCSP3 was highly expressed in the female head,and its relative expression was much higher in adults than in other developmental stages.The fluorescence binding assays suggested that the EforCSP3 exhibited high binding affinities to a wide range of host-related volatiles,among which dibutyl phthalate,1-octene,β-elemene,and tridecane had the strongest binding affinity with EforCSP3,besides α-humulene and β-myrcene,and should be assessed as potential attractants.Protein structure modeling and molecular docking predicted the amino acid residues of EforCSP3possibly involved in volatile binding.α-Humulene and β-myrcene attracted E.formosa in a previous study and exhibited strong binding affinities with EforCSP3 in the current study.In conclusion,EforCSP3 may be involved in semiochemical reception by E.formosa.展开更多
文摘We analyzed 20 chemosensory protein （CSP） genes of the silkworm Bombyx mori. We found a high number of retrotransposons inserted in introns. We then analyzed expression of the 20 BrnorCSP genes across tissues using quantitative real-time polymerase chain reaction （PCR）. Relatively low expression levels of BmorCSPs were found in the gut and fat body tissues. We thus tested the effects of endectocyte insecticide abamectin （B 1 a and Blb avermectins） on BmorCSP gene expression. Quantitative real-time PCR experi- ments showed that a single brief exposure to insecticide abamectin increased dramatically CSP expression not only in the antennae but in most tissues, including gut and fat body. Furthermore, our study showed coordinate expression of CSPs and metabolic cytochrome P450 enzymes in a tissue-dependent manner in response to the insecticide. The function of CSPs remains unknown. Based on our results, we suggest a role in detecting xenobiotics that are then detoxified by cytochrome P450 anti-xenobiotic enzymes.
基金Supported by the National Natural Science Foundation of China (Grant No. 30270896)
文摘Chemosensory proteins (CSPs) are ubiquitous soluble small proteins isolated from sensory organs thought to be involved in chemical communication. Here we report the first cDNA of CSPs, called Ac-ASP3, cloned and characterized from antennas of adult worker bees in Chinese honeybee, Apis cerana cerana. The Ac-ASP3 cDNA comprises 2 exons, with an ORF of 393-bp encoding 130 aa. Protein signature analyses show that the protein consists of four conserved cysteines and a signal peptide with 19 aa in the N-terminal sequence. The deduced protein sequence shares high homology with Am-ASP3 of Apis mellifera and low similarity with other species of insects. Immunocytochemical localization shows that Ac-ASP3 is only specifically expressed on the antenna contact chemosensilla such as sensilla trichodea B and sensilla basiconica, whereas Ac-ASP3 is scarcely expressed on olfactory chemosensilla such as sensilla placodea. Real-time PCR of Ac-ASP3 transcripts shows that Ac-ASP3 is highly expressed on wings and legs, but expression is lower on antenna. Temporal expression patterns suggest that Ac-ASP3 is expressed during the period of pupa and adults from 1-d to 6-d stages when bees act as house bees, cleaning the comb and taking care of the queen and larvae in comb. The above evidence suggests that Ac-ASP3 is unique in species and is generally not involved in olfaction during searching for honey and pollen. Rather, the protein seems to function in recognition of chemosensory substances on bees' cuticle and mechanical movement of antenna.
文摘【目的】建立花椒窄吉丁Agrilus zanthoxylumi成虫触角转录组数据库,挖掘嗅觉相关基因,为今后研究其触角的化学感受机制及生物防控提供理论支撑。【方法】采用高通量测序平台Illumina NovaSeq 6000对花椒窄吉丁雌雄成虫触角进行转录组测序,用Trinity软件对获得的高质量reads进行序列拼接与组装;使用BLAST软件将触角转录组数据比对NR,NT,Swiss-Prot,GO,KEGG,BLASTX,eggNOG,Pfam,TmHMM,SignalP,KO,Map,BLASTP和RNAMMER公共数据库;基于初步筛选到的花椒窄吉丁候选气味结合蛋白(odorant binding protein,OBP)和化学感受蛋白(chemosensory protein,CSP)以及其他鞘翅目昆虫的同源蛋白的核苷酸序列,利用MEGA软件进行系统进化分析。运用RPKM(reads per kilobase per million mapped reads)值对嗅觉相关基因表达量进行分析。【结果】花椒窄吉丁雌雄成虫触角转录组测序共获得36209条基因和90982条转录本,其N50分别为2103和2523 bp,组装完整性较高。注释到NR数据库的基因最多(41.62%),其中与赤拟谷盗Tribolium castaneum相似基因所占比例最高(19%)。在GO数据库中比对到11614个基因,按功能分为细胞组分、分子功能与生物学进程三大类57个分支,其中分子功能大类中的结合(70.57%)与催化活性(45.51%)相关基因占比最多;KEGG代谢途径分析表明,7427条基因参与了5类代谢通路,其中涉及信号转导的基因最多,为815条;筛选到7个候选OBP基因和5个具有全长开放阅读框的CSP基因,其编码蛋白均具有化学感受蛋白家族的典型特征。系统进化分析表明,花椒窄吉丁OBPs和CSPs分别与苹果小吉丁A.mali的OBPs和CSPs氨基酸序列一致性最高。RPKM值表明,嗅觉相关基因AzanOBP1和AzanOBP2在雌成虫触角中不表达,在雄成虫触角中微量表达;AzanOBP3在雄成虫触角中高丰度表达。【结论】首次获得了花椒窄吉丁成虫触角转录组数据,筛选到了花椒窄吉丁OBP,CSP,Or,IR和SNMP等的嗅觉相关�
基金supported by the National 973 Program of China (2012CB114104)the National Natural Science Foundation of China (31171858)
文摘A chemosensory protein named HarmCSP5 in cotton bollworm Helicoverpa armigera （Hvbner） was obtained from antennal eDNA libraries and expressed in Escherichia coll. The real time quantitative PCR （RT-qPCR） results indicated that HarmCSP5 gene was mainly expressed in male and female antennae but also expressed in female legs and wings. Competitive binding assays were performed to test the binding affinity of recombinant HarmCSP5 to 60 odor molecules including some cotton volatiles. The resules showed that HarmCSP5 showed strong binding abilities to 4-ehtylbenzaldehyde and 3,4-dimethlbenz aldehyde, whereas methyl phenylacetate, 2-decanone, 1-pentanol, carvenol, isobomeol, nerolidol, 2- nonanone and ethyl heptanoate have relatively weak binding affinity. Moreover, the predicted 3D model of HarmCSP5 consists of six α-helices located among residues 33-38 （αl）, 40-48 （α2）, 62-72 （α3）, 80-96 （α4）, 98-108 （α5）, and 116-119 （α6）, two pairs of disulfide bridges Cys49-Cys55, Cys75-Cys78. The two amino acid residues, Ile94 and Trpl01, may play crucial roles in HarmCSP5 binding with ligands and need further study for confirmation.
基金supported by the National Natural Science Foundation of China (31772172)the earmarked fund for China Agriculture Research System (CARS25)the Beijing Key Laboratory for Pest Control and Sustainable Cultivation of Vegetables。
文摘Chemosensory proteins(CSPs) perform several functions in insects.This study performed the gene expression,ligand-binding,and molecular docking assays on the EforCSP3 identified in the parasitoid wasp Encarsia formosa,to determine whether EforCSP3 functions in olfaction,especially in host location and host preference.The results showed that EforCSP3 was highly expressed in the female head,and its relative expression was much higher in adults than in other developmental stages.The fluorescence binding assays suggested that the EforCSP3 exhibited high binding affinities to a wide range of host-related volatiles,among which dibutyl phthalate,1-octene,β-elemene,and tridecane had the strongest binding affinity with EforCSP3,besides α-humulene and β-myrcene,and should be assessed as potential attractants.Protein structure modeling and molecular docking predicted the amino acid residues of EforCSP3possibly involved in volatile binding.α-Humulene and β-myrcene attracted E.formosa in a previous study and exhibited strong binding affinities with EforCSP3 in the current study.In conclusion,EforCSP3 may be involved in semiochemical reception by E.formosa.
