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Article(id=1277293368128434633, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1277293236137890180, articleNumber=null, orderNo=null, doi=10.3969/j.issn.1000-2561.2024.11.016, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1715875200000, receivedDateStr=2024-05-17, revisedDate=1716307200000, revisedDateStr=2024-05-22, acceptedDate=null, acceptedDateStr=null, onlineDate=1782460069924, onlineDateStr=2026-06-26, pubDate=1732464000000, pubDateStr=2024-11-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1782460069924, onlineIssueDateStr=2026-06-26, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1782460069924, creator=13701087609, updateTime=1782460069924, updator=13701087609, issue=Issue{id=1277293236137890180, tenantId=1146029695717560320, journalId=1235980609244409860, year='2024', volume='45', issue='11', pageStart='2243', pageEnd='2486', issueExtLink='null', onlineDate='null', pubDate='1732464000000', pubDateStr='2024-11-25', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1782460038455, creator='13701087609', updateTime=1782815269280, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1278783182988358204, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1277293236137890180, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1278783182988358205, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1277293236137890180, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=2380, endPage=2389, ext={EN=ArticleExt(id=1277293368459784651, articleId=1277293368128434633, tenantId=1146029695717560320, journalId=1235980609244409860, language=EN, title=Molecular Docking of Five Chemosensory Proteins in Xyleborus affinis with Semiochemicals, columnId=1236292524264968282, journalTitle=Chinese Journal of Tropical Crops, columnName=Plant Protection & Bio-safety, runingTitle=null, highlight=null, articleAbstract=

Xyleborus affinis, a main pest of rubber trees, is effectively monitored and controlled using semiochemicals, a key method in rubber pest management. Chemosensory proteins (CSPs) are believed to play an important role in the process of binding and transporting odorants in insect. To elucidate the role of chemosensory proteins in semiochemicals recognizing by X. affinis, the XaffCSP1~XaffCSP5 genes were cloned and identified in this study. The characteristics of nucleotide and deduced amino acid sequences of the genes were analyzed using bioinformatics methods. The binding mode and ability with 15 different ligands (14 host plant volatiles and 1 aggregation pheromone) was analyzed using molecular docking technology. Results revealed that the length of the open reading frame (ORF) of the five XaffCSPs genes was 366-417 bp, encoding 121-138 amino acids. The predicted protein molecular weight was 13.95-16.08 kDa and all five XaffCSPs contained signal peptide composed of 17-20 amino acid residues at N-terminal. The secondary structure of the proteins was mainly composed of α-helix. All five XaffCSPs possessed four conserved cysteine residues, conforming to the typical characteristics of insect chemosensory proteins. Phylogenetic tree anaylsis indicated that except for XaffCSP5, XaffCSP1 to XaffCSP4 were clustered with CSPs of other bark beetle species. The three-dimensional models of the the five XaffCSPs were with good model quality according to model assessment results, and the models of the five XaffCSPs were composed of six α-helices. Molecular docking results demonstrated all five XaffCSPs had a strong binding affinity with (-)-α-copaene and (+)-longifolene. Additionally, XaffCSP4 showed strong binding to terpinolene and limonene, while XaffCSP5 showed strong binding to camphene, α-pinene, and β-pinene. The binding ability of the five XaffCSPs to (S)-cis-verbenol was relatively weak. The results demonstrated that the five XaffCSPs had good affinity with host volatiles and may be participated in the process of recognizing host plant volatiles by X. affinis. This study would provide a theoretical basis for elucidating the molecular mechanism of X. affinis in recognizing semiochemicals and is of great significance for the control of X. affinis.

, authors=null, authorsList=Kai ZHANG, Jixing GUO, Qian WANG, Qi WU, Zheyuan XU, Linan DENG, Xiang ZHOU, authorCompany=null, correspAuthors=Xiang ZHOU, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1277293372029137372, articleId=1277293368128434633, tenantId=1146029695717560320, journalId=1235980609244409860, language=CN, title=橡胶材小蠹5个化学感受蛋白与信息化合物的分子对接, columnId=1236292524520820846, journalTitle=热带作物学报, columnName=植物保护与生物安全, runingTitle=null, highlight=null, articleAbstract=

橡胶材小蠹（Xyleborus affinis）是橡胶树的主要害虫之一，利用信息化合物对其进行监测或诱集防控已成为橡胶害虫防治的关键手段。在昆虫识别信息化合物的过程中，化学感受蛋白（chemosensory proteins, CSPs）发挥着重要作用。为了探究化学感受蛋白在橡胶材小蠹识别信息化合物中的作用，本研究克隆并鉴定了5个橡胶材小蠹化学感受蛋白XaffCSP1~XaffCSP5基因。利用生物信息学方法分析了XaffCSPs核酸及氨基酸序列特征，利用分子对接技术解析了XaffCSPs与橡胶材小蠹15种信息化合物的结合能力及结合模式，其中包括14种寄主植物挥发物和1种聚集信息素。结果表明：橡胶材小蠹5个XaffCSPs基因开放阅读框（open reading frame, ORF）长度为366~417 bp，编码121~138个氨基酸，预测蛋白质分子量为13.95~16.08 kDa，氨基端含有17~20个氨基酸残基组成的信号肽，蛋白质结构二级结构均以α-螺旋为主，并且5个XaffCSPs均具有4个保守的半胱氨酸位点，符合昆虫化学感受蛋白的典型特征。系统发育树结果显示，除XaffCSP5单独聚为一支外，XaffCSP1~XaffCSP4分别与其他小蠹不同CSPs聚为一支。利用trRosetta平台对XaffCSPs进行模型构建，5个XaffCSPs的三维模型均由6个α-螺旋组成，模型评估结果表明模型质量良好。分子对接结果显示5个XaffCSPs对α-古巴烯和长叶烯均表现出很强的结合能力；此外，XaffCSP4对萜品油烯和柠檬烯结合较强，而XaffCSP5对莰烯、α-蒎烯和β-蒎烯有较强结合；5个XaffCSPs与顺式-马鞭草烯醇结合能力相对较弱。研究表明5个XaffCSPs与寄主植物挥发物有较好的亲和力，可能参与橡胶材小蠹对寄主植物挥发物的识别，研究结果为阐明橡胶材小蠹识别信息化合物的分子机制奠定理论基础，对橡胶材小蠹的防控具有重要意义。

, authors=

* 同等贡献作者

张凯（1999—），女，硕士研究生，研究方向：农业昆虫与害虫防治

郭技星（1989—），男，博士，讲师，研究方向：农业昆虫与害虫防治。

, authorsList=张凯, 郭技星, 王倩, 邬祺, 许哲源, 邓礼楠, 周祥, authorCompany=null, correspAuthors=周祥, authorNote=null, correspAuthorsNote=
** 周祥（ZHOU Xiang），E-mail：。
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张凯（1999—），女，硕士研究生，研究方向：农业昆虫与害虫防治

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张凯（1999—），女，硕士研究生，研究方向：农业昆虫与害虫防治

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郭技星（1989—），男，博士，讲师，研究方向：农业昆虫与害虫防治。

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Scientific Reports, 2016, 6(1): 33981., articleTitle=Predicted structure of a Minus-C OBP from Batocera horsfieldi (Hope) suggests an intermediate structure in evolution of OBPs, refAbstract=null)], funds=[Fund(id=1277293395953447464, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, awardId=32060644, language=CN, fundingSource=国家自然科学基金地区基金项目(32060644), fundOrder=null, country=null), Fund(id=1277293396301574697, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, awardId=322MS012, language=CN, fundingSource=海南省自然科学基金项目(322MS012), fundOrder=null, country=null), Fund(id=1277293396377072170, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, awardId=KJRC2023D01, language=CN, fundingSource=海南省科技人才创新项目(KJRC2023D01), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1277293372268212702, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, xref=null, ext=[AuthorCompanyExt(id=1277293372276601311, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, companyId=1277293372268212702, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=School of Tropical Agriculture and Forestry, Hainan University / Key Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests, Ministry of Education, Danzhou, Hainan 571737, China), AuthorCompanyExt(id=1277293372289184224, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, companyId=1277293372268212702, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=海南大学热带农林学院/热带农林生物灾害绿色防控教育部重点实验室，海南儋州　571737)])], figs=[ArticleFig(id=1277293386767921680, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, language=EN, label=Fig. 1, caption=PCR amplification of XaffCSPs ORF

M: DL2000 DNA marker; 1-5: PCR product of XaffCSP1-XaffCSP5.

, figureFileSmall=BEsKH25HRARKYldQOLwmsA==, figureFileBig=gp17Q0h8wNvwx1CHLmu5xg==, tableContent=null), ArticleFig(id=1277293387145409041, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, language=CN, label=图1, caption=XaffCSPs基因ORF的PCR扩增

M：DL2000 DNA marker；1~5：XaffCSP1~XaffCSP5 PCR产物。

, figureFileSmall=BEsKH25HRARKYldQOLwmsA==, figureFileBig=gp17Q0h8wNvwx1CHLmu5xg==, tableContent=null), ArticleFig(id=1277293387925549586, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, language=EN, label=Fig. 2, caption=Conserved motifs prediction of XaffCSPs protein, figureFileSmall=XDCFPCOp08WFqEopjLmxhw==, figureFileBig=Au9M6hgPyngc4veDD6SmRQ==, tableContent=null), ArticleFig(id=1277293389175452179, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, language=CN, label=图2, caption=XaffCSPs蛋白的保守基序分析, figureFileSmall=XDCFPCOp08WFqEopjLmxhw==, figureFileBig=Au9M6hgPyngc4veDD6SmRQ==, tableContent=null), ArticleFig(id=1277293389607465492, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, language=EN, label=Fig. 3, caption=Sequence alignment of XaffCSPs

The red background represents conserved amino acids, the blue box represents identical or similar amino acids, and the green numbers represent disulfide bonds formed by cysteine.

, figureFileSmall=8xSGp1HGpFA88gUG4MNdUw==, figureFileBig=TeBhp13xNChilFtdmulQtA==, tableContent=null), ArticleFig(id=1277293390031090197, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, language=CN, label=图3, caption=XaffCSPs的序列比对

红色背景为保守氨基酸，蓝色框为相同或相似的氨基酸，绿色数字为半胱氨酸形成的二硫键。

, figureFileSmall=8xSGp1HGpFA88gUG4MNdUw==, figureFileBig=TeBhp13xNChilFtdmulQtA==, tableContent=null), ArticleFig(id=1277293390437937686, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, language=EN, label=Fig. 4, caption=Phylogenetic analysis of amino acid sequence of XaffCSPs and bark beetles CSPs, figureFileSmall=AO5yeI45dhVm22qjv4Basg==, figureFileBig=RMK5spXoBQbjCdTzCPDMeg==, tableContent=null), ArticleFig(id=1277293390526018071, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, language=CN, label=图4, caption=XaffCSPs与其他小蠹虫CSPs的系统发育树, figureFileSmall=AO5yeI45dhVm22qjv4Basg==, figureFileBig=RMK5spXoBQbjCdTzCPDMeg==, tableContent=null), ArticleFig(id=1277293390907699736, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, language=EN, label=Fig. 5, caption=3D model of XaffCSPs

A-E are 3D structures of XaffCSP1-XaffCSP5; Nt and Ct represent the amino terminal and the carboxyl terminal of the protein, respectively; α indicates α-helix of the protein.

, figureFileSmall=6NmPsmohIX07qefA3X9cOw==, figureFileBig=2XMwG/0Qp4BButi3VCfrJQ==, tableContent=null), ArticleFig(id=1277293391260021273, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, language=CN, label=图5, caption=XaffCSPs的三维模型

A~E为XaffCSP1~XaffCSP5的三维结构；Nt和Ct分别代表蛋白质的氨基端和羧基端；α表示蛋白质的α-螺旋。

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Red: Most favored regions, including A, B and L areas; Yellow: Additional allowed regions, including a, b, l, p areas; Pale yellow: Generously allowed regions, including ~a, ~b, ~l, ~p areas; White: Disallowed regions.

, figureFileSmall=fq8Xsd740CoTLzJpcqt1Wg==, figureFileBig=uyzH6Bn/xeTNLPgBKUUQdQ==, tableContent=null), ArticleFig(id=1277293391729783323, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, language=CN, label=图6, caption=XaffCSPs模型的拉式构象图

红色：最佳区域，包括A、B、L区域；黄色：许可区域，包括a、b、l、p区域；浅黄色：较合理区域，包括~a、~b、~l、~p区域；白色：不合理区域。

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A: XaffCSP1; B: XaffCSP2; C: XaffCSP3; D: XaffCSP4; E: XaffCSP5. The same color represents the same ligand.

, figureFileSmall=8Y8XP8nd2ocIhfeqOnwg+A==, figureFileBig=OWSXhUseK4oZveULDv88GQ==, tableContent=null), ArticleFig(id=1277293392182768157, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, language=CN, label=图7, caption=XaffCSPs的分子对接图

A：XaffCSP1；B：XaffCSP2；C：XaffCSP3；D：XaffCSP4；E：XaffCSP5。相同颜色代表同一配体。

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Primers used in this study

, figureFileSmall=null, figureFileBig=null, tableContent=
引物Primer序列（5′-3′）Sequence (5′-3′)
XaffCSP1-FATGAAAAATCTAATCGCTGCC
XaffCSP1-RGTTGCCTAGAAACTGCCTA
XaffCSP2-FATGTGGAAGTTGACCCTACTT
XaffCSP2-RCTGAGCATCGACAGGCTTC
XaffCSP3-FATGGTTTTGTTTACTTCGTTACT
XaffCSP3-RCTTATCTACAAGTATTTTACTC
XaffCSP4-FATGAAGCTTTTGGTAATCGC
XaffCSP4-RCAGATTGATTCCATCCTTCTTC
XaffCSP5-FATGAGAAGCATAGTGGTTGT
XaffCSP5-RAATGTCATACTCTGCCTTCC
), ArticleFig(id=1277293394204422687, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, language=CN, label=表1, caption=

本研究所用引物

, figureFileSmall=null, figureFileBig=null, tableContent=
引物Primer序列（5′-3′）Sequence (5′-3′)
XaffCSP1-FATGAAAAATCTAATCGCTGCC
XaffCSP1-RGTTGCCTAGAAACTGCCTA
XaffCSP2-FATGTGGAAGTTGACCCTACTT
XaffCSP2-RCTGAGCATCGACAGGCTTC
XaffCSP3-FATGGTTTTGTTTACTTCGTTACT
XaffCSP3-RCTTATCTACAAGTATTTTACTC
XaffCSP4-FATGAAGCTTTTGGTAATCGC
XaffCSP4-RCAGATTGATTCCATCCTTCTTC
XaffCSP5-FATGAGAAGCATAGTGGTTGT
XaffCSP5-RAATGTCATACTCTGCCTTCC
), ArticleFig(id=1277293394292503072, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, language=EN, label=Tab. 2, caption=

Physicochemical properties of putative XaffCSPs protein

, figureFileSmall=null, figureFileBig=null, tableContent=
蛋白名称Protein name全长Full length/bp开放阅读框ORF size/bp氨基酸数量Number of amino acids信号肽Signal peptide分子量Molecular weight/kDa等电点pI
XaffCSP15743661211~1813.959.36
XaffCSP28474171381~2016.085.45
XaffCSP316044051341~1815.419.05
XaffCSP415393841271~1714.638.79
XaffCSP59793901291~2015.305.07
), ArticleFig(id=1277293394623853089, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, language=CN, label=表2, caption=

XaffCSPs蛋白的理化性质

, figureFileSmall=null, figureFileBig=null, tableContent=
蛋白名称Protein name全长Full length/bp开放阅读框ORF size/bp氨基酸数量Number of amino acids信号肽Signal peptide分子量Molecular weight/kDa等电点pI
XaffCSP15743661211~1813.959.36
XaffCSP28474171381~2016.085.45
XaffCSP316044051341~1815.419.05
XaffCSP415393841271~1714.638.79
XaffCSP59793901291~2015.305.07
), ArticleFig(id=1277293394695156258, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, language=EN, label=Tab. 3, caption=

XaffCSPs protein secondary structure prediction %

, figureFileSmall=null, figureFileBig=null, tableContent=
蛋白名称Protein nameα-螺旋α-helixβ-转角β-turn延伸链Extended strand无规则卷曲Random coil
XaffCSP162.814.963.3128.93
XaffCSP252.905.806.5234.78
XaffCSP364.186.722.9926.12
XaffCSP461.428.664.7225.20
XaffCSP565.894.651.5527.91
), ArticleFig(id=1277293395064255011, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, language=CN, label=表3, caption=

XaffCSPs蛋白二级结构预测

, figureFileSmall=null, figureFileBig=null, tableContent=
蛋白名称Protein nameα-螺旋α-helixβ-转角β-turn延伸链Extended strand无规则卷曲Random coil
XaffCSP162.814.963.3128.93
XaffCSP252.905.806.5234.78
XaffCSP364.186.722.9926.12
XaffCSP461.428.664.7225.20
XaffCSP565.894.651.5527.91
), ArticleFig(id=1277293395152335396, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, language=EN, label=Tab. 4, caption=

Homology analysis of XaffCSPs gene

, figureFileSmall=null, figureFileBig=null, tableContent=
基因Gene物种名称Species name同源基因Homologous gene登录号Accession No.相似度Identity/%
XaffCSP1华山松大小蠹CSP2AXF53965.168.07
XaffCSP2云南切梢小蠹CSP1AMP19496.179.03
XaffCSP3枣飞象CSP9WJJ63294.176.98
XaffCSP4星天牛CSPAUF72999.188.04
XaffCSP5黄粉虫CSP9AJO62215.160.95
), ArticleFig(id=1277293395471102501, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, language=CN, label=表4, caption=

XaffCSPs基因的同源性分析

, figureFileSmall=null, figureFileBig=null, tableContent=
基因Gene物种名称Species name同源基因Homologous gene登录号Accession No.相似度Identity/%
XaffCSP1华山松大小蠹CSP2AXF53965.168.07
XaffCSP2云南切梢小蠹CSP1AMP19496.179.03
XaffCSP3枣飞象CSP9WJJ63294.176.98
XaffCSP4星天牛CSPAUF72999.188.04
XaffCSP5黄粉虫CSP9AJO62215.160.95
), ArticleFig(id=1277293395559182886, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, language=EN, label=Tab. 5, caption=

Molecular docking of XaffCSPs with 15 ligands

, figureFileSmall=null, figureFileBig=null, tableContent=
编号No.配体物质Ligand substanceCAS号CAS No.分子式Formula形状（环状/链状）Shape (cyclic/chain)结合能/(kJ·mol-1)
CSP1CSP2CSP3CSP4CSP5
1棕榈酸甲酯methyl palmitate112-39-0C17H34O2链状-4.95-5.01-4.82-5.80-4.78
2月桂烯myrcene123-35-3C10H16链状-4.36-4.35-4.46-5.18-4.85
33-蒈烯3-carene13466-78-9C10H16环状-5.24-5.21-5.20-5.51-5.66
4β-蒎烯(-)-beta-pinene18172-67-3C10H16环状-5.54-5.79-5.52-5.79-6.17
5顺式-马鞭草烯醇(S)-cis-verbenol18881-04-4C10H16O环状-5.56-5.80-5.42-5.61-5.81
6桧烯sabinen3387-41-5C10H16环状-5.17-5.30-5.19-5.73-5.60
7α-古巴烯(-)-alpha-copaene3856-25-5C15H24环状-7.71-7.26-7.46-7.85-7.59
8长叶烯(+)-longifolene475-20-7C15H24环状-5.36-6.27-7.31-6.33-7.24
9间伞花烃m-cymene535-77-3C10H14环状-5.40-5.11-4.97-5.78-5.29
10萜品油烯terpinolene586-62-9C10H16环状-5.57-5.57-5.51-6.32-5.83
11柠檬烯(-)-limonene5989-54-8C10H16环状-5.45-5.43-5.53-6.21-5.61
122-苯基-2-丙醇2-phenyl-2-propanol617-94-7C9H12O环状-4.48-5.01-4.51-5.53-4.82
13十四烷tetradecane629-59-4C14H30链状-4.83-4.47-4.42-5.86-4.83
14莰烯camphene79-92-5C10H16环状-5.60-5.69-5.42-5.72-6.25
15α-蒎烯alpha-piene80-56-8C10H16环状-4.42-5.70-5.51-5.76-6.22
), ArticleFig(id=1277293395634680359, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1277293368128434633, language=CN, label=表5, caption=

XaffCSPs与15种配体的分子对接

, figureFileSmall=null, figureFileBig=null, tableContent=
编号No.配体物质Ligand substanceCAS号CAS No.分子式Formula形状（环状/链状）Shape (cyclic/chain)结合能/(kJ·mol-1)
CSP1CSP2CSP3CSP4CSP5
1棕榈酸甲酯methyl palmitate112-39-0C17H34O2链状-4.95-5.01-4.82-5.80-4.78
2月桂烯myrcene123-35-3C10H16链状-4.36-4.35-4.46-5.18-4.85
33-蒈烯3-carene13466-78-9C10H16环状-5.24-5.21-5.20-5.51-5.66
4β-蒎烯(-)-beta-pinene18172-67-3C10H16环状-5.54-5.79-5.52-5.79-6.17
5顺式-马鞭草烯醇(S)-cis-verbenol18881-04-4C10H16O环状-5.56-5.80-5.42-5.61-5.81
6桧烯sabinen3387-41-5C10H16环状-5.17-5.30-5.19-5.73-5.60
7α-古巴烯(-)-alpha-copaene3856-25-5C15H24环状-7.71-7.26-7.46-7.85-7.59
8长叶烯(+)-longifolene475-20-7C15H24环状-5.36-6.27-7.31-6.33-7.24
9间伞花烃m-cymene535-77-3C10H14环状-5.40-5.11-4.97-5.78-5.29
10萜品油烯terpinolene586-62-9C10H16环状-5.57-5.57-5.51-6.32-5.83
11柠檬烯(-)-limonene5989-54-8C10H16环状-5.45-5.43-5.53-6.21-5.61
122-苯基-2-丙醇2-phenyl-2-propanol617-94-7C9H12O环状-4.48-5.01-4.51-5.53-4.82
13十四烷tetradecane629-59-4C14H30链状-4.83-4.47-4.42-5.86-4.83
14莰烯camphene79-92-5C10H16环状-5.60-5.69-5.42-5.72-6.25
15α-蒎烯alpha-piene80-56-8C10H16环状-4.42-5.70-5.51-5.76-6.22
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橡胶材小蠹5个化学感受蛋白与信息化合物的分子对接
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张凯 , 郭技星 , 王倩 , 邬祺 , 许哲源 , 邓礼楠 , 周祥 **
热带作物学报 | 植物保护与生物安全 2024,45(11): 2380-2389
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热带作物学报 |植物保护与生物安全 2024 , 45 (11) : 2380 -2389
橡胶材小蠹5个化学感受蛋白与信息化合物的分子对接
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张凯, 郭技星, 王倩, 邬祺, 许哲源, 邓礼楠, 周祥**
作者信息
  • 海南大学热带农林学院/热带农林生物灾害绿色防控教育部重点实验室，海南儋州　571737
通讯作者:
** 周祥（ZHOU Xiang），E-mail：。
Molecular Docking of Five Chemosensory Proteins in Xyleborus affinis with Semiochemicals
Kai ZHANG, Jixing GUO, Qian WANG, Qi WU, Zheyuan XU, Linan DENG, Xiang ZHOU**
Affiliations
  • School of Tropical Agriculture and Forestry, Hainan University / Key Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests, Ministry of Education, Danzhou, Hainan 571737, China
出版时间: 2024-11-25 doi: 10.3969/j.issn.1000-2561.2024.11.016
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摘要
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橡胶材小蠹（Xyleborus affinis）是橡胶树的主要害虫之一，利用信息化合物对其进行监测或诱集防控已成为橡胶害虫防治的关键手段。在昆虫识别信息化合物的过程中，化学感受蛋白（chemosensory proteins, CSPs）发挥着重要作用。为了探究化学感受蛋白在橡胶材小蠹识别信息化合物中的作用，本研究克隆并鉴定了5个橡胶材小蠹化学感受蛋白XaffCSP1~XaffCSP5基因。利用生物信息学方法分析了XaffCSPs核酸及氨基酸序列特征，利用分子对接技术解析了XaffCSPs与橡胶材小蠹15种信息化合物的结合能力及结合模式，其中包括14种寄主植物挥发物和1种聚集信息素。结果表明：橡胶材小蠹5个XaffCSPs基因开放阅读框（open reading frame, ORF）长度为366~417 bp，编码121~138个氨基酸，预测蛋白质分子量为13.95~16.08 kDa，氨基端含有17~20个氨基酸残基组成的信号肽，蛋白质结构二级结构均以α-螺旋为主，并且5个XaffCSPs均具有4个保守的半胱氨酸位点，符合昆虫化学感受蛋白的典型特征。系统发育树结果显示，除XaffCSP5单独聚为一支外，XaffCSP1~XaffCSP4分别与其他小蠹不同CSPs聚为一支。利用trRosetta平台对XaffCSPs进行模型构建，5个XaffCSPs的三维模型均由6个α-螺旋组成，模型评估结果表明模型质量良好。分子对接结果显示5个XaffCSPs对α-古巴烯和长叶烯均表现出很强的结合能力；此外，XaffCSP4对萜品油烯和柠檬烯结合较强，而XaffCSP5对莰烯、α-蒎烯和β-蒎烯有较强结合；5个XaffCSPs与顺式-马鞭草烯醇结合能力相对较弱。研究表明5个XaffCSPs与寄主植物挥发物有较好的亲和力，可能参与橡胶材小蠹对寄主植物挥发物的识别，研究结果为阐明橡胶材小蠹识别信息化合物的分子机制奠定理论基础，对橡胶材小蠹的防控具有重要意义。

橡胶材小蠹  /  化学感受蛋白  /  序列分析  /  信息化合物  /  分子对接

Xyleborus affinis, a main pest of rubber trees, is effectively monitored and controlled using semiochemicals, a key method in rubber pest management. Chemosensory proteins (CSPs) are believed to play an important role in the process of binding and transporting odorants in insect. To elucidate the role of chemosensory proteins in semiochemicals recognizing by X. affinis, the XaffCSP1~XaffCSP5 genes were cloned and identified in this study. The characteristics of nucleotide and deduced amino acid sequences of the genes were analyzed using bioinformatics methods. The binding mode and ability with 15 different ligands (14 host plant volatiles and 1 aggregation pheromone) was analyzed using molecular docking technology. Results revealed that the length of the open reading frame (ORF) of the five XaffCSPs genes was 366-417 bp, encoding 121-138 amino acids. The predicted protein molecular weight was 13.95-16.08 kDa and all five XaffCSPs contained signal peptide composed of 17-20 amino acid residues at N-terminal. The secondary structure of the proteins was mainly composed of α-helix. All five XaffCSPs possessed four conserved cysteine residues, conforming to the typical characteristics of insect chemosensory proteins. Phylogenetic tree anaylsis indicated that except for XaffCSP5, XaffCSP1 to XaffCSP4 were clustered with CSPs of other bark beetle species. The three-dimensional models of the the five XaffCSPs were with good model quality according to model assessment results, and the models of the five XaffCSPs were composed of six α-helices. Molecular docking results demonstrated all five XaffCSPs had a strong binding affinity with (-)-α-copaene and (+)-longifolene. Additionally, XaffCSP4 showed strong binding to terpinolene and limonene, while XaffCSP5 showed strong binding to camphene, α-pinene, and β-pinene. The binding ability of the five XaffCSPs to (S)-cis-verbenol was relatively weak. The results demonstrated that the five XaffCSPs had good affinity with host volatiles and may be participated in the process of recognizing host plant volatiles by X. affinis. This study would provide a theoretical basis for elucidating the molecular mechanism of X. affinis in recognizing semiochemicals and is of great significance for the control of X. affinis.

Xyleborus affinis  /  chemosensory proteins  /  sequence analysis  /  semichemicals  /  molecular docking
张凯, 郭技星, 王倩, 邬祺, 许哲源, 邓礼楠, 周祥. 橡胶材小蠹5个化学感受蛋白与信息化合物的分子对接. 热带作物学报, 2024 , 45 (11) : 2380 -2389 . DOI: 10.3969/j.issn.1000-2561.2024.11.016
Kai ZHANG, Jixing GUO, Qian WANG, Qi WU, Zheyuan XU, Linan DENG, Xiang ZHOU. Molecular Docking of Five Chemosensory Proteins in Xyleborus affinis with Semiochemicals[J]. Chinese Journal of Tropical Crops, 2024 , 45 (11) : 2380 -2389 . DOI: 10.3969/j.issn.1000-2561.2024.11.016
正文
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橡胶材小蠹（Xyleborus affinis）属鞘翅目（Coleoptera）象甲科（Curculionidae）小蠹亚科（Scolytinae），原产于美洲，现已广泛分布于全球热带和亚热带地区，且寄主范围广，是多种重要经济作物的害虫[1]。橡胶材小蠹是我国橡胶树害虫的优势种之一[2-3]，通常在极端天气或人为割胶导致树势衰弱后入侵树体，在木质部中构筑坑道为害[4-5]。橡胶材小蠹目前主要以农业防治和化学防治为主，但由于该害虫主要以蛀干为害，隐蔽性强、适应性广，且具有较强的繁殖能力，化学防治效果较差[3,6]。利用信息化合物对害虫进行监测或诱集防控，因其绿色、高效等特点而被广泛关注。已有研究利用乙醇及其与ɑ-蒎烯的混合物成功诱集到橡胶材小蠹[7]；干威[8]利用月桂烯、S-(-)-柠檬烯、顺式-马鞭草烯醇和莰烯在橡胶林中成功诱集到橡胶材小蠹；徐凡丁等[9]鉴定了橡胶树韧皮部挥发物成分，并通过室内行为测定试验筛选到长叶烯、十四烷、2-苯基-2-丙醇3种化合物对橡胶材小蠹具有行为活性。ROMERO等[1]测定了橡胶材小蠹对3种寄主植物挥发物的触角电生理反应，发现裂榄（Bursera simaruba）树皮挥发物能引起橡胶材小蠹较强的触角电位反应，其主要成分包括3-蒈烯、莰烯、ɑ-蒎烯、β-蒎烯、桧烯、伞花烃、柠檬烯、ɑ-古巴烯、萜品油烯等。
昆虫识别外界信息化合物需要多种蛋白质相互协作，其中化学感受蛋白（chemosensory proteins, CSPs）是嗅觉识别过程最早参与的蛋白质之一。CSPs由100~120个氨基酸组成，其典型特征为含有4个保守的半胱氨酸（cysteine, Cys），分别由2个二硫键（S-S键）连接[10-11]。该类蛋白质三维空间结构通常小而紧凑，其6个α-螺旋形成一个“疏水结合口袋”，用于结合信息化合物[12-13]。CSPs主要存在于化学感受器淋巴液中，负责结合气味或信息素分子，并传递给气味受体（odorant receptors, ORs），因此，CSPs在识别和运输气味分子中发挥着重要作用[14]。
昆虫化学感受研究中，同源建模和分子对接技术被广泛应用于CSPs、气味结合蛋白（odorant binding proteins, OBPs）及ORs的功能预测，这些技术可以方便快捷地研究蛋白质与小分子物质的结合模式及其结合力大小，从而了解它们之间的相互作用[15-17]。本研究从橡胶材小蠹转录组中鉴定到5个橡胶材小蠹化学感受蛋白XaffCSPs，通过构建XaffCSPs三维模型，利用分子对接技术解析XaffCSPs与信息化合物的结合能力及结合方式，为阐明橡胶材小蠹识别信息化合物的分子机制奠定理论基础。
1　材料与方法
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1.1　材料
供试虫源：橡胶材小蠹采集于海南省儋州市海南大学农科基地，使用锯木断收集和诱捕法2种方法采集。
1.2　方法
1.2.1　 XaffCSPs基因的克隆
从前期橡胶材小蠹转录组中筛选鉴定化学感受蛋白基因XaffCSPs序列[18]，使用Primer Premier 5.0软件设计特异性引物，由生工生物工程（上海）股份有限公司合成（表1）。
以橡胶材小蠹cDNA为模板进行PCR扩增，反应体系如下：Phanta®Max Super-Fidelity DNA Polymerase 0.4 μL、2×Phanta® Max Buffer 10 μL、dNTP Mix 0.4 μL、引物对各0.8 μL、cDNA模板0.2 μL、ddH2O 7.4 μL，总体系为20 μL。反应条件：95 ℃预变性3 min；95 ℃变性15 s，58 ℃退火15 s，72 ℃延伸30 s，反应35个循环；72 ℃延伸10 min。用1%琼脂糖凝胶电泳检测获得PCR产物，利用琼脂糖凝胶DNA回收试剂盒[天根生化科技（北京）有限公司]纯化回收目的条带，将纯化产物与pMD18-T载体连接，并转化至感受态细胞DH5α中。挑取单克隆菌落进行PCR验证，筛选阳性克隆提取质粒，送至生工生物工程（上海）股份有限公司进行测序。
1.2.2　XaffCSPs的序列分析
利用NCBI ORF Finder（https://www.ncbi.nlm.nih.gov/orffinder/）预测XaffCSPs基因的开放阅读框（open reading frame, ORF）；使用SignalP 4.1（https://services.healthtech.dtu.dk/services/SignalP-4.1/）预测XaffCSPs的信号肽位置；通过Expasy ProtParam（https://web.expasy.org/protparam）预测XaffCSPs的分子量和等电点。利用SOPMA（http://npsapbil.ibcp.fr/cgi-bin/npsa_automat.pl?page=npsa_sopma.html）对XaffCSPs氨基酸序列二级结构进行预测；使用NCBI BLASTx（https://blast.ncbi.nlm.nih.gov/Blast.cgi）分析XaffCSPs与其他鞘翅目昆虫化学感受蛋白序列的相似性；利用DNAman软件进行氨基酸序列比对。利用MEGA 11软件采用邻接法（neighbor-joining, NJ）构建XaffCSPs与其他昆虫CSPs的系统发育树，bootstrap为1000次。使用MEME（https://meme-suite.org/meme/index.html）设置最大Motif数量为5，对XaffCSPs的保守基序进行评估。
1.2.3　XaffCSPs的三维建模与模型评价
利用trRosetta平台[19]对蛋白质的三级结构进行预测。使用SAVES 6.0（https://saves.mbi.ucla.edu/）的ERRAT和ProCheck对XaffCSPs三维模型进行评价。
1.2.4　分子对接
以15种对橡胶材小蠹有行为或电生理活性的化合物作为配体，从PubChem数据库（https://pubchem.ncbi.nlm.nih.gov/）中下载配体PDB文件，通过AutoDock Tool 1.5.6对XaffCSPs蛋白质与配体的三维模型进行能量优化，并赋予原子电荷及力场。通过POCASA1.1（https://g6altair.sci.hokudai.ac.jp/g6/service/pocasa/）预测蛋白质结合口袋的位置，设置受体与配体的结合搜寻空间Grid Box，进行半柔性对接[20]。自由结合能越低且位点构象合理的物质，即为XaffCSPs的潜在结合物质。结合能越小，认为配体与受体的结合越好。
2　结果与分析
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2.1　5个XaffCSPs的序列分析
从前期转录组数据中鉴定到5个注释为化学感受蛋白的Unigene序列，分别命名为XaffCSP1~XaffCSP5。以橡胶材小蠹cDNA为模版对ORF进行PCR扩增，电泳结果显示5个XaffCSPs基因扩增条带均位于300~500 bp之间，条带大小与预期一致（图1）。获得测序结果后对转录组原始序列进行校正。
5个XaffCSPs基因均包含完整ORF，长度为366~417 bp，编码121~138个氨基酸，预测的蛋白质分子量为13.95~16.08 kDa，氨基端含有17~20个氨基酸残基组成的信号肽（表2）。二级结构预测结果表明，5个XaffCSPs蛋白质结构均以α-螺旋为主，其次为无规则卷曲（表3）。对XaffCSPs进行基序预测分析，5个XaffCSPs序列中发现5个保守基序（图2）。
2.2　 XaffCSPs的序列比对与系统发育分析
对XaffCSPs进行Blast比对分析表明，在NCBI数据库中与XaffCSP1~XaffCSP5相似性最高的分别是华山松大小蠹（Dendroctonus armandi）CSP2、云南切梢小蠹（Tomicus yunnanensis）CSP1、枣飞象（Pachyrhinus yasumatsui）CSP9、星天牛（Anoplophora chinensis）CSP和黄粉虫（Tenebrio molitor）CSP9基因（表4）。序列比对结果表明，XaffCSP1~XaffCSP5均具有4个保守半胱氨酸位点，符合C1-X6-C2-X18-C3-X2-C4的排列模式（图3）。构建XaffCSPs与其他5种小蠹CSPs的系统发育树，除XaffCSP5单独聚为一支外，XaffCSP1~XaffCSP4分散在不同的分支（图4），XaffCSP1与华山松大小蠹DarmCSP2、中欧山松大小蠹（D. ponderosae）DponCSP2和松圆头小蠹（D. adjunctus）DadjCSP2聚为一支，亲缘关系近；XaffCSP2与华山松大小蠹、中欧山松大小蠹、松圆头小蠹的CSP3及云南切梢小蠹TyunCSP1聚为一支；XaffCSP3与中欧山松大小蠹、松圆头小蠹及云杉八齿小蠹（Ips typographus）的CSP4及华山松大小蠹DarmCSP5聚为一支；XaffCSP4与华山松大小蠹、中欧山松大小蠹及松圆头小蠹的CSP8聚为一支。
2.3　XaffCSPs的三维建模构建及质量评估
通过trRosetta平台构建XaffCSPs的蛋白质模型，XaffCSP1~XaffCSP5的三维模型均由6个α-螺旋组成（图5）。XaffCSP1~XaffCSP5三维模型的ERRAT值分别为100%、98.14%、100%、100%和100%，均大于50%，表明非共价键相互作用合理。利用ProCheck程序对蛋白质三维结构进行评估，拉氏构象图表明XaffCSP1~XaffCSP15三维模型分别有95.70%、94.30%、94.20%、94.90%和93.90%的氨基酸残基位于最佳区域，分别有4.30%、5.70%、5.80%、5.10%和6.10%的氨基酸位于较合适区域，无氨基酸位于勉强许可区和不合理区域，表明通过trRosetta构建的XaffCSP1~XaffCSP15三维模型构象合理（图6）。
2.4　XaffCSPs与信息化合物的分子对接
将5个XaffCSPs与15种信息化合物进行分子对接，结果表明，XaffCSP1~XaffCSP5对α-古巴烯均表现出很强的结合能力，结合能分别为-7.71、-7.26、-7.46、-7.85、-7.59 kJ/mol；其次为长叶烯，XaffCSP2、XaffCSP3、XaffCSP4和XaffCSP5与该化合物的结合能分别为-6.27、-7.31、-6.33、-7.24 kJ/mol，XaffCSP1与长叶烯的结合能力相对较弱。此外，XaffCSP4对萜品油烯和柠檬烯也表现出较强的结合能力，结合能分别为-6.32、-6.21 kJ/mol，而XaffCSP5对莰烯、α-蒎烯和β-蒎烯表现出较强的结合能力，结合能分别为-6.25、-6.22、-6.17 kJ/mol。15个化合物中，5个XaffCSPs对十四烷和月桂烯均表现出较弱的结合能力。在与聚集信息素顺式马鞭草醇的结合中，与其他XaffCSPs相比，XaffCSP5表现出较强的结合能力，其结合能为-5.81 kJ/mol（表5）。
XaffCSP1主要通过范德华力和疏水作用与α-古巴烯结合，其中氨基酸残基Ile15、Leu22、Tyr25、Val26、Val29、Leu42、Phe64、Val65、Leu72形成疏水作用，氨基酸残基Ile12、Leu16、Gly39、Ile46、Gly68、Ala69、Trp80、Ile83形成范德华力。在与其他化合物的对接结果中，氨基酸残基Ile46、Val65、Ala69、Ile83参与了多数化合物的结合（图7A）。XaffCSP2与α-古巴烯的结合中，形成疏水作用的氨基酸残基包括Val15、Ile17、Ile20、Tyr30、Leu47、Lys65，形成范德华力的氨基酸残基有Tyr12、Asp13、Asn14、Asp43、Gln66、Gly69、Ser70；而在与长叶烯的结合中，氨基酸残基Leu34、Met51、Ile55、Met74、Leu77、Leu88、Tyr98主要形成疏水作用，Leu47、Ala54、Ser70、Met73、Trp85、Gln89形成范德华力（图7B）。α-古巴烯与长叶烯的结合位置均位于XaffCSP2的疏水性结合腔内，但位置略有差异，2个氨基酸残基Leu47和Ser70在2种配体结合中均发挥作用。
根据XaffCSP3与配体的对接结果，α-古巴烯和长叶烯位于XaffCSP3疏水性结合腔内相同的位置（图7C）。形成疏水作用的氨基酸残基Tyr8、Phe47、Ile50和形成范德华力的氨基酸残基Gln66和Thr70均在XaffCSP3与这2种信息化合物的结合中均发挥作用。
XaffCSP4主要通过范德华力和疏水作用与α-古巴烯、长叶烯、萜品油烯和柠檬烯结合，这4种配体位于XaffCSP4疏水性结合腔的不同位置（图7D）。在与α-古巴烯的结合中，XaffCSP4的氨基酸残基Arg68、Met71、Arg72、Ile75、Trp82、Tyr95、Tyr99、Leu103形成疏水性作用力，氨基酸残基Lys96、Glu102、Asp106形成范德华力。在与长叶烯的结合中，Leu31、Leu44、Leu48、Leu52、Ile70、Met71、Leu74、Leu85形成疏水作用，Met24、Typ82、Ser67形成范德华力。Tyr9、Val12、Leu14、Leu44、Ile70通过疏水作用作用于萜品油烯，Asp10、Thr62、Gln63、Gly66、Ser67通过范德华力与萜品油烯相互作用。与柠檬烯的结合中，Leu31、Leu48、Leu52、Leu74、Trp82、Leu85、Trp82形成疏水作用，Met71、Glu86、Tyr89、Asp90通过范德华力发挥作用。
XaffCSP5与α-古巴烯、长叶烯、莰烯、α-蒎烯和β-蒎烯的结合主要通过范德华力和疏水作用（图7E）。形成疏水作用的氨基酸残基Tyr9、Ile12、Tyr14、Leu44与形成范德华力的氨基酸残基Glu40、Glu43、Gln63在XaffCSP5与以上5种信息化合物的结合中发挥作用。XaffCSP5通过范德华力、疏水作用和氢键与顺式-马鞭草烯醇结合，其氨基酸残基Ile12、Tyr14、Leu44、Ile47、Val67通过疏水作用与顺式-马鞭草烯醇互作，氨基酸残基Asp10、Glu40、Glu43、Gln63通过范德华力与聚集信息素互作，Tyr9与顺式-马鞭草烯醇形成氢键。
3　讨论
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随着基因组与转录组学技术在昆虫化学感受研究领域的应用，许多昆虫的化学感受基因已经被鉴定，其中包括多种小蠹虫的CSPs基因。例如在中欧山松大小蠹D. ponderosae中，鉴定得到11个CSPs基因，在华山松大小蠹（D. armandi）的转录组中发现9个CSPs，在云南切梢小蠹（T. yunnanensis）头部转录组中得到8个CSPs，红脂大小蠹（D. valens）和云杉八齿小蠹（I. typographus）触角转录组中各发现6个CSPs[21-24]。本研究利用橡胶材小蠹转录组数据鉴定到5个XaffCSPs，其序列分析表明这些XaffCSPs的结构具有高度保守性，均包含4个半胱氨酸位点，其排列模式为C-X6-C-X18-C-X2-C，符合鞘翅目昆虫CSPs的普遍结构[25]。
鞘翅目昆虫中，CSPs在识别信息化合物过程中发挥着重要作用。一些CSPs能与寄主植物的多种挥发物紧密结合。如绿豆象（Callosobruchus chinensis）的CchiCSP1可以与绿豆的4种挥发物紧密结合[26]。在甘薯小象甲（Cylas formicarius）中发现有3个CSPs对17种植物挥发物具有较高的结合能力，通过RNAi干扰CforCSP1、CforCSP5和CforCSP6的表达后，甘薯小象甲丧失了对部分植物挥发物的嗅觉反应[27]。CSPs也参与了鞘翅目昆虫信息素的识别，如华山松大小蠹的DarmCSP2可能是多种信息素和寄主植物挥发物的载体[23]。本研究通过分子对接技术分析了5个XaffCSPs与橡胶材小蠹信息化合物的结合能力，发现XaffCSPs与α-古巴烯和长叶烯有较强的结合能力，而对聚集信息素结合能力相对较弱。因此，XaffCSPs可能在橡胶材小蠹识别寄主植物挥发物的过程中发挥重要作用。XaffCSPs与植物挥发物的结合主要是通过范德华力和疏水作用进行结合，在这些配体中，棕榈酸甲酯、十四烷和月桂烯的分子结构呈链状，而其余配体的分子结构为环状。根据分子对接的结果，发现5个XaffCSPs对环状配体的结合能力普遍强于对链状配体的结合能力。结合腔的形状及其与配体分子形状的契合度可能是影响CSPs结合能的重要因素，如果配基与结合腔能够进行较好的契合，配体与形成结合腔的不同氨基酸充分接触，通过更牢固作用力将配基固定在结合腔内[28]，也有研究表明配体的弹性及碳链长度也会影响其与蛋白质的结合[29-30]。
综上所述，本研究克隆并分析了5个橡胶材小蠹化学感受蛋白XaffCSPs基因，并利用分子对接技术分析了其与15种橡胶材小蠹信息化合物的结合模式，结果表明这5个XaffCSPs与α-古巴烯和长叶烯结合能力较强。本研究为阐明橡胶材小蠹识别信息化合物的分子机制提供了理论基础。由于昆虫的生活环境及生理状态具有复杂性和多样性，尽管分子对接技术可以模拟CSPs与配体的结合过程，但仍存在一定的局限性，还需通过荧光竞争结合试验和定点突变等技术验证蛋白质与配体的结合能力及关键氨基酸残基的作用，以及通过RNAi试验验证CSPs在橡胶材小蠹识别信息化合物中的作用。
基金
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  • 国家自然科学基金地区基金项目(32060644)
  • 海南省自然科学基金项目(322MS012)
  • 海南省科技人才创新项目(KJRC2023D01)
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doi: 10.3969/j.issn.1000-2561.2024.11.016
  • 接收时间：2024-05-17
  • 首发时间：2026-06-26
  • 出版时间：2024-11-25
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  • 收稿日期：2024-05-17
  • 修回日期：2024-05-22
基金
国家自然科学基金地区基金项目(32060644)
海南省自然科学基金项目(322MS012)
海南省科技人才创新项目(KJRC2023D01)
作者信息
    海南大学热带农林学院/热带农林生物灾害绿色防控教育部重点实验室，海南儋州　571737

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** 周祥（ZHOU Xiang），E-mail：。
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2种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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