中国水稻科学 ›› 2020, Vol. 34 ›› Issue (1): 37-45.DOI: 10.16819/j.1001-7216.2020.9034

• 研究报告 • 上一篇    下一篇

褐飞虱小分子量热激蛋白基因表达特性和功能

潘磊1,2, 王利华2, 朱凤3, 韩阳春2, 王培2, 方继朝1,2,4,*()   

  1. 1南京农业大学 植物保护学院, 南京 210095
    2江苏省农业科学院 植物保护研究所, 南京 210014
    3江苏省植物保护植物检疫站, 南京 210036
    4江苏省区域现代农业与环境保护协同创新中心, 江苏 淮安 223300
  • 收稿日期:2019-03-27 修回日期:2019-06-14 出版日期:2020-01-10 发布日期:2020-01-10
  • 通讯作者: 方继朝
  • 作者简介:

    作者简介:#共同第一作者

  • 基金资助:
    国家重点研发计划资助项目(2016YFD0300706);国家自然科学基金资助项目(31572004);江苏省自然科学基金资助项目(BK20170072);江苏省重点研发计划资助项目(BE2017366)

Expression Profiles and Functions of Small Heat Shock Proteins in Nilaparvata lugens

Lei PAN1,2, Lihua WANG2, Feng ZHU3, Yangchun HAN2, Pei WANG2, Jichao FANG1,2,4,*()   

  1. 1College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China
    2Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China
    3Jiangsu Plant Protection and Plant Quarantine Station, Nanjing 210036, China
    4Jiangsu Collaborative Innovation Center for Regional Modern Agriculture & Environmental Protection, Huaiyin Normal University, Huai’an 223300, China;
  • Received:2019-03-27 Revised:2019-06-14 Online:2020-01-10 Published:2020-01-10
  • Contact: Jichao FANG
  • About author:

    About author:#These authors contributed equally to this work

摘要:

【目的】研究褐飞虱小分子量热激蛋白的表达特性和功能,明确其在褐飞虱温度胁迫适应中的作用。【方法】采用BLAST从转录组数据库中筛选褐飞虱小分子量热激蛋白基因序列;利用Bioedit、Mega等分子生物学软件进行序列分析;利用qPCR技术分析目的基因在不同处理下的表达特性;利用原核表达技术研究其功能。【结果】筛选到6个含有α-晶体结构的小分子量热激蛋白基因NlHsp20.9NlHsp21.6NlHsp21.9NlHsp22.4NlHsp23.1NlHsp28.7,其ORF长度依次为561、531、570、570、588和735 bp,理论等电点为5.96、5.77、6.32、5.01、5.74和7.74。NlHsp28.7在3龄若虫中的表达量最高,而NlHsp21.9NlHsp23.1在雌成虫中的表达量最高。雌虫在低温胁迫后所有小分量热激蛋白基因的表达量均下降,高温胁迫后除NlHsp22.4外的其他5个基因表达量不同程度上调;3龄若虫在低温胁迫后一半sHsps表达量下降,另一半上升,高温胁迫后全部上调。转化褐飞虱sHsps的重组大肠杆菌热激存活率显著上升。【结论】褐飞虱小分子量热激蛋白具有龄期和诱导表达特性及热胁迫保护功能,可能在其高温胁迫应激中具有重要作用,在低温胁迫应激中的作用与虫态有关。

关键词: 褐飞虱, 小分子量热激蛋白, 温度, 表达

Abstract:

【Objective】To explore the adaptation mechanisms of brown planthoppers(BPH) to temperature stress, the expression profiles and functions of small heat shock proteins (sHSPs) in Nilaparvata lugens was studied. 【Method】The nucleotide sequence of small heat shock proteins in N. lugens was screened from transcriptome database by BLAST; The sequence analysis was carried out by molecular biological software such as Bioedit and Mega; the expression profiles were analyzed by qPCR; and the prokaryotic expression technology was used for functional research. 【Result】Six sHsps (NlHsp20.9, NlHsp21.6, NlHsp21.9, NlHsp22.4, NlHsp23.1 and NlHsp28.7) with conserved alpha crystal structure were screened. The ORF length of these genes was 561, 531, 570, 570, 588 and 735 bp, and the theoretical isoelectric point was 5.96, 5.77, 6.32, 5.01, 5.74 and 7.74, respectively. The peak expression of NlHsp28.7 was found in the third larvae and the peak of NlHsp21.9 and NlHsp23.1 in females. The expression of these genes decreased in females after exposure to low temperature, while increased after exposure to high temperature except NlHsp22.4. To the 3rd instar nymphs, the expression of about half of sHsps decreased after low temperature stress and half of sHsps increased, while all increased after high temperature stress. Moreover, the survival of transformed BL21 (DE3) was significantly increased after heat treatment. 【Conclusion】The small heat shock protein of N. lugens has stage-specific and inducible expression characteristics and show protection against heat stress. It may play an important role in response to high temperature stress, but its role in response to low temperature stress is related to insect developmental stages.

Key words: Nilaparvata lugens, small heat shock proteins, temperature, expression

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