| 于苗苗 1, 2, 张思宇 2, 宋国锋 2, 陈巨莲 2, 解海翠 1*, 范 佳 2*.荻草谷网蚜ABCG1基因的克隆及表达模式分析[J].植物保护,2024,50(1):256-263. |
| 荻草谷网蚜ABCG1基因的克隆及表达模式分析 |
| Cloning and expression profiling of ATP-binding cassette transporter G1 gene in grain aphid Sitobion miscanthi |
| 投稿时间：2022-10-19 修订日期：2022-12-08 |
| DOI：10.16688/j.zwbh.2022648 |
| 中文关键词: 荻草谷网蚜 ABC转运蛋白 蜕皮激素 基因克隆 表达谱 |
| 英文关键词:Sitobion miscanthi ATP-binding cassette transporter ecdysone (20E) gene cloning expression profile |
| 基金项目:国家自然科学基金 (31871966);河北省高等学校科学技术研究项目(BJ2020049);省属高校基本科研业务费(2020JK001) |
| 作者 | 单位 | E-mail | | 于苗苗 1, 2, 张思宇 2, 宋国锋 2, 陈巨莲 2, 解海翠 1*, 范 佳 2* | 1. 河北科技师范学院, 河北省作物逆境生物学重点实验室(筹), 秦皇岛 066600
2. 中国农业科学院植物保护研究所, 植物病虫害综合治理全国重点实验室, 北京 100193 | 解海翠hcxie2008@126.com;范佳jfan@ippcaas.cn |
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| 中文摘要: |
| 荻草谷网蚜Sitobion miscanthi是严重威胁我国小麦生产安全的迁飞性害虫。蜕皮激素是参与蚜虫翅型分化调控的内激素, 在有翅成蚜体内保持高滴度, 且诱导后代产生更高比例的无翅蚜, 其进出靶细胞需要经过细胞膜上特定蛋白的转运。ATP结合盒转运蛋白家族G亚家族(ATP-binding cassette transporter G, ABCG)中的 ABCG1是通过跨膜转运昆虫类固醇、对蜕皮激素信号进行负调控的功能蛋白之一, 在蚜虫中尚未见报道。本文克隆了荻草谷网蚜ABCG1(SmisABCG1)基因, 并进行了序列比对、系统进化分析以及不同组织部位和发育时期表达模式分析。结果显示, SmisABCG1基因的开放阅读框全长为1 851 bp, 编码616个氨基酸, 含7个跨膜结构域, 符合ABCG蛋白家族典型结构特性, 基因登录ID:OP626323。昆虫间ABCG1较保守, 该蛋白系统进化关系与各自物种间亲缘关系的远近保持一致。其中, SmisABCG1与来自豌豆蚜、禾谷缢管蚜、棉蚜、花生蚜和雪松长足大蚜等的ABCG1氨基酸序列高度一致(>87%), 以上蚜虫聚为一支。与SmisABCG1亲缘关系最近的是豌豆蚜的ABCG1, 其次是半翅目的褐飞虱、白背飞虱和灰飞虱, 与膜翅目的新疆菜叶蜂、阿里山潜蝇茧蜂以及鞘翅目的赤拟谷盗、蜂箱小甲虫亲缘关系较远。该基因在伪胚胎和成蚜阶段高表达。包含伪胚胎的有翅、无翅成蚜整蚜SmisABCG1的转录水平无显著差异, 但其在来自有翅成蚜的伪胚胎中的转录水平高于无翅成蚜伪胚胎, 证实无翅成蚜自身的转录水平较高, 而有翅成蚜较低。进一步分析显示这一差异主要是无翅蚜胸部显著高表达所导致。基于该蛋白对蜕皮激素负调控, 与有翅成蚜转录水平低, 但蜕皮激素水平更高相符合。 |
| 英文摘要: |
| Grain aphid Sitobion miscanthi is a migratory pest as well as the dominant pest on wheat in China. Molting hormone, ecdysone (20E), as one of the internal hormone, involved in the regulation of wing differentiation of aphids, which induces a higher proportion of wingless aphid in the offspring and maintains a high titer in winged adults. To enter and exit the target cells, 20E requires the transport of specific membrane transporters. ABCG1, as one of 20E membrane transporters in insects, negatively regulates ecdysone signaling by transmembrane transport of insect ecdysone, which has not been reported in aphids yet. In the present study, SmisABCG1 was cloned and aligned with its orthologs, and their phylogenetic relationship was analyzed among insects. Expression patterns among organs or tissues of both winged and wingless adults, pseudo embryos, first to fourth instar nymphs were also analyzed. The results showed that the complete open reading frame of SmisABCG1 was 1 851 bp, encoding 616 amino acids, with seven trans-membrane regions, which was consistent with the traits of ABCG protein family member. The gene accession ID was OP626323. ABCG1 is conserved among insects, and the phylogenetic relationship of this protein confirmed that there is a close genetic relationship among species. SmisABCG1 shared high consistency (>87%) of sequence with ABCG1s in other insects such as Acyrthosiphon pisum, Rhopalosiphum padi, Aphis gossypii, A.craccivora, Cinara cedri, and these aphids clustered into a branch. The closest neighbor of SmisABCG1 is ApisABCG1 in A.pisum, followed by white-backed planthopper (Sogatella furcifera), plant hopper (Nilaparvata lugens), small brown planthopper (Laodelphax striatellus), sawfly (Athalia rosae), alishan dive fly cocoon bee (Fopius arisanus), red flour beetle (Tribolium castaneum) and Aethina tumida. RT-qPCR results showed that SmisABCG1 is mainly expressed in pseudo embryo as well as adults. Significantly high level of pseudo embryo in winged adult and equal level of winged and wingless adults with embryo indicated that wingless aphid expressed higher ABCG1 than winged aphid. Subsequent analysis found that this was due to a significantly higher expression in the thorax of wingless adulT. All other organs or tissues showed no significant differences between winged and wingless phenotype. Based on the negative regulation of ecdysone by this protein, it consistent with low transcript levels but high ecdysone levels in winged adult aphids. |
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