| 谢金婷，李俊珏，周乐明，刘 雅，宾贵辉，杨振德，周 静,,桉树枝瘿姬小蜂气味结合蛋白基因OBPs的克隆及表达分析[J].环境昆虫学报,(): |
| 桉树枝瘿姬小蜂气味结合蛋白基因OBPs的克隆及表达分析 |
| Cloning and expression analysis of seven odour binding protein genes from Leptocybe invasa |
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| DOI： |
| 中文关键词: 桉树枝瘿姬小蜂 气味结合蛋白 植物挥发物 基因克隆 寄主识别 |
| 英文关键词:Leptocybe invasa odor binding protein plant volatiles gene cloning recognition of host |
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| 中文摘要: |
| 为明确桉树枝瘿姬小蜂Leptocybe invasa的嗅觉感受机制，本研究利用克隆技术从桉树枝瘿姬小蜂中成功克隆7个LinvOBPs基因：LinvOBP2、LinvOBP5、LinvOBP7、LinvOBP9、LinvOBP11、LinvOBP18、LinvOBP23。利用生物信息学软件分析7个基因序列编码蛋白的理化特性和结构特征；构建进化树分析桉树枝瘿姬小蜂OBPs及其它昆虫OBPs的进化关系；通过实时荧光定量PCR技术分析在不同挥发物处理下OBPs基因表达情况。结果表明，7个LinvOBPs基因长度约400 bp，含有6个保守的半胱氨酸，序列N端具有信号肽切割位点，在二级结构预测中均含有大量的α-螺旋结构，有利于疏水结合腔的形成，能够促进其与气味分子的结合。桉树枝瘿姬小蜂的LinvOBPs与白蛾周氏啮小蜂Chouioia cunea的CcOBPs聚在同一分支，亲缘性很高，推测二者可能具有相似的功能。不同LinvOBPs对于不同挥发物的响应存在显著差异，特定挥发物对LinvOBPs表达的调控作用，尤其是对伞花烃、桉叶油醇、右旋萜二烯能够上调多个LinvOBPs的表达量。7个LinvOBPs基因均在识别寄主桉树过程中发挥重要作用，其中，LinvOBPs对对伞花烃、桉叶油醇、右旋萜二烯的识别有利于揭示桉树枝瘿姬小蜂特异性识别寄主桉树的分子机制。 |
| 英文摘要: |
| In order to understand the olfactory perception mechanism of Leptocybe invasa, seven odour binding protein genes, named LinvOBP2, LinvOBP5, LinvOBP7, LinvOBP9, LinvOBP11, LinvOBP18, LinvOBP23 were successfully cloned from L. invasa. The bioinformatics software was used to analyze the physical and chemical characteristics and structural characteristics of the proteins encoded by the seven genes, and the phylogenetic relationships of OBPs and other insect OBPs were analyzed. Real-time quantitative PCR was used to analyze the expression of OBPs gene under different volatiles treatment. The results showed that the 7 LinvOBPs genes were about 400 bp in length and contained 6 conserved L-Cysteine. The N-terminal of the 7 LinvOBPs contained signal peptide cleavage sites and a large number of α-helices in secondary structure prediction, it was beneficial to the formation of hydrophobic binding cavity, and can promote its binding with odor molecules. The LinvOBPs of L. invasa and the CcOBPs of Chouioia cunea were clustered in the same branch and had high affinity. There were significant differences in the response of different LinvOBPs to different volatiles. The regulation of certain volatiles on LinvOBPs expression, especially for para-cymene, eucalyptol and d-terpenediene, could up-regulate the expression of multiple LinvOBPs. All the seven LinvOBPs genes play an important role in the recognition of host eucalyptus. Among them, the recognition of para-cymene, eucalyptol and d-terpendiene by LinvOBPs is helpful to reveal the molecular mechanism of the specific recognition of host eucalyptus by L. invasa. |
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