黄曲条跳甲视觉基因的鉴定与表达分析
Identification and bioinformatic analysis of the Phyllotreta striolata visual gene
王 雪1** 肖 勇2 彭争科2 尹 飞2 王欢欢2 李振宇2 靖湘峰1***
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DOI：10.7679/j.issn.2095-1353.2025.090
作者单位：1. 西北农林科技大学植物保护学院，杨凌 712100；2. 广东省农业科学院植物保护研究所， 农业农村部华南果蔬绿色防控重点实验室，广东省植物保护新技术重点实验室，广州 510640
中文关键词：黄曲条跳甲；视蛋白；转录组；蛋白质结构；基因表达
英文关键词： Phyllotreta striolata; opsin; transcriptome; protein structure; gene expression
中文摘要：
【目的】 黄曲条跳甲Phyllotreta striolata作为十字花科蔬菜的重要害虫，其趋光性行为是防控策略的关键信息之一。本研究旨在通过转录组学、生物信息学分析和分子生物学技术解析黄曲条跳甲视觉基因特征，为揭示其趋光机制提供理论基础。【方法】 通过cDNA文库构建和PacBio测序对头部转录组进行分析和注释，使用BioEdit、ExPASY、TMHMM、AlphaFold等软件鉴定黄曲条跳甲视蛋白基因，并进行蛋白结构和功能位点分析，使用MEGA 7.0软件进行序列分析和进化树构建，最后利用实时荧光定量PCR分析4个视蛋白基因的时空表达特征及其在黄曲条跳甲雌雄虫中的表达差异。【结果】 通过黄曲条跳甲成虫头部转录组测序鉴定到4个黄曲条跳甲视蛋白基因，即长波敏感视蛋白PstrOpsin-LW、紫外光敏感视蛋白PstrOpsin-UV1和PstrOpsin-UV2以及和非视觉视蛋白Pstr-Pteropsin，且均属于G蛋白偶联受体的典型结构。其中，PstrOpsin-LW、PstrOpsin-UV1、PstrOpsin-UV2在黄曲条跳甲成虫期的表达水平均显著高于幼虫期和蛹期（P<0.05），PstrOpsin-LW在成虫期的表达量约为幼虫期的1 846倍，为蛹期表达量的46倍；PstrOpsin-UV1在成虫期的表达量约为幼虫期的80倍，为蛹期表达量的5倍；PstrOpsin-UV2在成虫期的表达量约为幼虫期的6倍，为蛹期表达量的3倍；Pstr-Pteropsin在不同龄期的表达量差异不显著（P>0.05）；这4个视觉基因在成虫头部表达量均显著高于成虫头部以外的虫体（P<0.05），其中PstrOpsin-LW的差异最大，在雌成虫头部的表达量约为头部以外虫体的1 314倍，在雄成虫头部的表达量约为头部以外虫体的2 291倍，Pstr-Pteropsin的表达差异最小，在雌成虫头部的表达量约为头部以外虫体的14倍，在雄成虫头部的表达量约为头部以外虫体的10倍。且PstrOpsin-UV1和PstrOpsin-UV2分别在雌雄成虫头部高表达，PstrOpsin-UV1在雌成虫头部的表达量显著高于雄成虫（P<0.05），为雄成虫头部表达量的1.58倍。PstrOpsin-UV2在雄成虫头部的表达量显著高于雌成虫（P<0.05），为雌成虫头部表达量的1.35倍。【结论】 鉴定到4个黄曲条跳甲视蛋白基因并明确了其分子特征，揭示了黄曲条跳甲视觉基因的时空表达规律，为研究黄曲条跳甲的趋光机制奠定了基础。
英文摘要：
[Aim] To identify and conduct a bioinformatic
analysis of the visual gene of Phyllotreta striolata and thereby provide
a basis for further study of phototaxis in this species. [Methods] The transcriptome was analyzed and annotated
by cDNA library construction and PacBio sequencing. BioEdit, ExPASY, TMHMM,
AlphaFold, and other software, were then used to analyze and identify the
physicochemical properties of the P. striolata opsin genes PstrOpsin-LW, PstrOpsin-UV1, PstrOpsin-UV2 and Pstr-Pteropsin, and predict
their protein structure and functional sites. Sequence analysis and
phylogenetic tree construction were performed using MEGA 7.0 software. Finally,
real-time fluorescent quantitative PCR was used to analyze the spatio-temporal
expression, and expression in each gender, of these 4 opsin genes. [Results] Four genes were identified by the
transcriptome sequencing, namely long-wave sensitive opsin PstrOpsin-LW,
ultraviolet light-sensitive opin PstrOpsin-UV1 and PstrOpsin-UV2,
and non-visual opin Pstr-Pteropsin, all of which belonged to the typical
structure of G-protein-coupled receptors. The expression levels of PstrOpsin-LW, PstrOpsin-UV1 and PstrOpsin-UV2 were significantly higher in
adults than in pupae and larvae (P<0.05), but there was no significant
difference in the expression of Pstr-Pteropsin in different instars (P>0.05). The
expression of the four genes in the head of adults was significantly higher
than in the body (P<0.05), and PstrOpsin-UV1 and Pstrosin-UV2 were highly expressed in the head of male and female
adults, respectively. [Conclusion] Four retinopsin genes were identified and
their molecular characteristics were clarified. Differences in the expression
of these genes during growth and development were revealed, which is useful
information for the prevention and control of P. striolata.