大螟水通道蛋白SiAQP基因的克隆与表达分析
Molecular cloning and expression analysis of the Sesamia inferens aquaporin (SiAQP) gene
肖天晶；汤小天；陆明星；马德英；杜予州
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DOI：10.7679/j.issn.2095-1353.2019.093
作者单位：新疆农业大学农学院&农林有害生物监测与安全防控重点实验室，乌鲁木齐 830001；扬州大学园艺与植物保护学院&扬州大学应用昆虫研究所，扬州 225009
中文关键词：大螟；组织；发育阶段；温度；湿度
英文关键词：Sesamia inferens; tissues; development; temperature; humidity
中文摘要：【目的】 水通道蛋白（AQP）是一种重要的膜结合蛋白，它可以在植物、细菌和动物的细胞膜上形成通道以促进水分和甘油等小分子的运输。本文研究了大螟水通道蛋白基因（SiAQP）的序列特征以及该基因的表达模式和功能。【方法】 运用RT-PCR和RACE技术克隆获取大螟水通道蛋白基因（SiAQP）；运用生物信息学软件分析该基因特征；用实时荧光定量PCR技术(qRTPCR)分析该基因在大螟的不同器官组织、不同发育阶段以及在不同温度和湿度条件下的表达模式。【结果】 SiAQP的开放阅读框（ORF）为831 bp，编码276个氨基酸，分子量为29.27 ku，等电点8.16。保守结构域分析发现其含有两个高度保守的水通道蛋白特征序列NPA（Asn-Pro-Ala）基序和6个跨膜结构域。qPCR结果显示，不同器官组织中，后肠的SiAQP表达量显著高于其它组织；不同发育阶段中成虫期的SiAQP表达量最高，卵和蛹期表达量相对较低；不同温度处理下SiAQP的表达量相对稳定，与27 ℃对比高温与低温处理都没有显著变化；随着湿度的上升大螟幼虫和雌雄蛹的SiAQP表达量都呈现出显著的先上升后下降趋势，表明高湿度和低湿度环境都会抑制SiAQP的表达。【结论】 SiAQP在大螟不同的器官组织表达具有明显的差异，温度对SiAQP的表达影响较小，而湿度的影响较大，成虫能较好的适应不同湿度环境。
英文摘要： [Objectives] To determine the sequence characteristics, expression patterns, and function, of the pink borer, Sesamia inferens (Walker) aquaporin gene (SiAQP), a membrane-bound protein gene that facilitates the movement of water, glycerol and other substances across cell membranes in plants, bacteria and animals via channels. [Methods] The aquaporin gene SiAQP was cloned from S. inferens by RT-PCR and RACE technology. The expression patterns of SiAQP in different tissues, developmental stages, temperatures and relative humidity was analyzed with Real time quantitative PCR. [Results] SiAQP contains a 831 bp open reading frame (ORF) and encodes 276 amino acids. It has a theoretical molecular weight of 29.27 ku and an isoelectric point of 8.16. SiAQP has six transmembrane domains and two highly conserved asparagine- proline-alanine motifs (NPA motifs). Real time quantitative PCR showed that expression in the hindgut was significantly higher than in other tissues. The expression of SiAQP was highest in adults and relatively low in eggs and pupae. The expression of SiAQP was relatively stable in different temperature treatments, and there was no significant difference between high and low temperature treatments relative to the 27 ℃ control. Expression in larvae and pupae first decreased, then increased, with increasing humidity, indicating that both high and low humidity can inhibit the expression of SiAQP. [Conclusion] The expression of SiAQP was significantly different in different tissues. Although the expression of SiAQP was not greatly affected by temperature, it was significantly affected by humidity. Adult S. inferens may be more tolerant to low or high humidity than larvae and pupae.