| 姜万千 1, 杨 磊 2, 孟建玉3, 杨昌利 1, 张长禹 1*.桃蚜组织蛋白酶基因CTSB-16D和CTSB-348的克隆、表达谱及对不同环境胁迫的响应[J].植物保护,2024,50(1):137-145. |
| 桃蚜组织蛋白酶基因CTSB-16D和CTSB-348的克隆、表达谱及对不同环境胁迫的响应 |
| Cloning and expression profiling of the cathepsin genes CTSB-16D and CTSB-348 and their response to different environmental stresses in Myzus persicae (Hemiptera: Aphididae) |
| 投稿时间：2022-12-27 修订日期：2023-02-11 |
| DOI：10.16688/j.zwbh.2022811 |
| 中文关键词: 桃蚜 组织蛋白酶 基因克隆 表达分析 |
| 英文关键词:Myzus persicae cathepsin gene cloning expression analysis |
| 基金项目:贵州省教育厅特色领域项目(黔教合KY〔2021〕057)；贵州省烟草公司贵阳市公司科技项目(2022-07)； 中国烟草总公司贵州省公司项目（2021XM09） |
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| 中文摘要: |
| 为了探索组织蛋白酶基因CTSB-16D和CTSB-348在桃蚜Myzus persicae(Sulzer)响应高低温和UV-B胁迫中的作用, 通过RT-PCR技术克隆了桃蚜CTSB-16D和CTSB-348基因, 利用生物信息学方法分析了它们的序列特征;采用实时荧光定量PCR(quantitative real-time PCR, RT-qPCR)技术检测了2个基因在桃蚜不同发育阶段的相对表达量及经高温、低温和UV-B胁迫不同时长的无翅成虫中的相对表达量。克隆获得了桃蚜CTSB-16D(GenBank登录号:MZ962352)和CTSB-348(GenBank登录号:MZ962353)基因, 序列长度分别为1 096 bp和1 101 bp, 开放阅读框(ORF)分别为1 017 bp和1 023 bp, 分别编码338个和340个氨基酸, 蛋白相对分子量为38.14 kD和37.52 kD, 等电点(pI)为5.46和5.99。系统发育分析表明, 桃蚜CTSB-16D和CTSB-348均与豌豆蚜Acyrthosiphon pisum (XP_003246386.1、NP_001119608.1) CTSB亲缘关系最近。RT-qPCR分析表明, CTSB-16D和CTSB-348在桃蚜不同发育阶段均有表达, 分别在1龄若虫和无翅成虫中的表达量最高。高、低温和UV-B胁迫对CTSB-16D和CTSB-348基因的表达具有明显的诱导作用, 且表达量均随时间的延长呈先上升后下降的趋势;4℃胁迫下, CTSB-16D和CTSB-348表达量均在90 min时达到峰值;36℃胁迫下, CTSB-16D和CTSB-348表达量均在30 min时达到峰值;UV-B胁迫下, CTSB-16D和CTSB-348表达量分别在60 min和120 min时达峰值。桃蚜CTSB-16D和CTSB-348基因在不同发育阶段差异表达, 且响应温度和UV-B的胁迫, 推测两个基因参与桃蚜的生长发育并在桃蚜响应环境胁迫中发挥了重要作用。 |
| 英文摘要: |
| To explore the role of CTSB-16D and CTSB-348 of Myzus persicae (Sulzer) in response to high and low temperature and UV-B stresses, they were cloned by RT-PCR and their sequence characteristics were analyzed using bioinformatics approaches. The relative expression levels of the two genes in aphids at different developmental stages and in wingless adults exposed to high temperature, low temperature and UV-B stresses for different durations were detected by quantitative real-time PCR (RT-qPCR). The sequences of CTSB-16D and CTSB-348 (GenBank acc. nos. MZ962352 and MZ962353) were 1 096 bp and 1 101 bp in length, with an open reading frame of 1 017 bp and 1 023 bp, encoding 338 and 340 amino acids, respectively. The relative molecular weight of the proteins are 38.14 kD and 37.52 kD, with an isoelectric point of 5.46 and 5.99, respectively. Phylogenetic analysis showed that both CTSB-16D and CTSB-348 had the closest relationship with CTSB of Acyrthosiphon pisum (XP_003246386.1, NP_001119608.1). RT-qPCR analysis showed that the two genes were expressed at different developmental stages, with the highest expression levels of CTSB-16D and CTSB-348 in the 1st-instar nymph and wingless adults, respectively. Low and high temperature and UV-B stresses obviously induced the expression of CTSB-16D and CTSB-348 genes, and the expression increasing first and then decreased with increasing exposure time. Under 4℃, the expression levels of CTSB-16D and CTSB-348 reached the peak after 90 min exposure, respectively, but under 36℃, their expression reached the peak after 30 min exposure. Under UV-B stress, the expression levels of CTSB-16D and CTSB-348 peaked after 60 min and 120 min exposure, respectively. The different expressions of CTSB-16D and CTSB-348 at different developmental stages and their different responses to temperature and UV-B stresses suggested that the two genes were involved in the growth and development of M. persicae in response to environmental stresses. |
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