温度和毒死蜱对灰飞虱雌成虫热激蛋白70和90基因的诱导表达特性研究
Induced expression of the heat shock protein 70 and 90 genes by exposure to high temperature and chlorpyrifos in female ofLaodelphax striatellus
王利华** 单 丹 姚 静 方继朝***
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DOI：10.7679/j.issn.2095-1353.2015.097
作者单位：江苏省农业科学院植物保护研究所， 南京 210014
中文关键词：灰飞虱，高温，毒死蜱，Lhsp90-1，表达特性，适应性
英文关键词：Laodelphax striatellus, high temperature, chlorpyrifos, Lhsp90-1, expression patterns, tolerance
中文摘要：
【目的】 为了明确灰飞虱Laodelphax striatellus对温度和杀虫剂等环境胁迫因子的适应性进化机制，本文研究了高温和毒死蜱对该害虫热激蛋白70和90基因的诱导表达特性。【方法】 利用生物测定法研究了灰飞虱的热激存活率和对毒死蜱的敏感性，采用实时荧光定量PCR技术测定了不同诱导温度、LC25剂量的毒死蜱诱导不同时间以及不同品系的Lhsp70-1、Lhsp70-2、Lhsp90-1和Lhsp90-2 4个基因表达量的变化。【结果】 在27℃和31℃条件下，耐高温品系对毒死蜱的敏感性比对照品系分别降低1.39倍和1.95倍，42℃热激后毒死蜱抗性品系的存活率比敏感品系高23.58%。经过30、34、38和42℃热激1 h后，灰飞虱雌成虫Lhsp70-2、Lhsp90-1和Lhsp90-2的表达量分别上升1.4~2.5、1.7~3.3和1.1~2.0倍，Lhsp70-1表达量下降1.0~1.7倍；LC25剂量的毒死蜱处理0.5、8和12 h后，以上4个基因的表达量变化不显著或呈下降趋势。然而毒死蜱长期筛选后Lhsp70-2、Lhsp90-1和Lhsp90-2表达量分别上升2.32、1.53和2.28倍，高温长期筛选后Lhsp90-1表达量上升1.58倍。【结论】 Lhsp90-1表达量增加很可能是灰飞虱对高温和毒死蜱产生交互适应性的重要原因。
英文摘要：
[Objectives] To explore the mechanisms used by Laodelphax striatellus to cope with environmental stress; specifically, the expression patterns of the heat shock protein 70 and 90 genes induced by exposure to high temperature and chlorpyrifos. [Methods] A bioassay was performed to test the sensitivity of female L. striatellus to chlorpyrifos and heat, and real-time quantitative PCR used to analyze the expression levels of the Lhsp70-1, Lhsp70-2, Lhsp90-1 and Lhsp90-2 genes in thermo-tolerant and chlorpyrifos-resistant L. striatellus strains. Changes in the expression of these genes were measured after exposure to 1 hour at 10-42℃ or LC25 dose of chlorpyrifos for 0.5-12 hours. [Results] The sensitivity of the thermo-tolerant strain to chlorpyrifos at 27 and 31℃ was 1.39 and 1.95 times, respectively, less than that of the control strain. Survival of the chlorpyrifos-resistant strain increased by 23.58% after exposure to 42℃. After 1 hour at either 30, 34, 38 or 42℃, the expression of the Lhsp70-2, Lhsp90-1 and Lhsp90-2 genes increased by 1.4-2.5, 1.7-3.3 and 1.1-2.0 times, respectively, and the expression of the Lhsp70-1 gene decreased 1.0-1.7 times. The expression of the above four heat shock protein genes either did not change much, or tended to decrease, following exposure to an LC25 dose of chlorpyrifos for 0.5, 8 and 24 h. However, the expression of the Lhsp70-2, Lhsp90-1 and Lhsp90-2 genes increased by 2.32, 1.53 and 2.28 times, respectively following long-term selection for chlorpyrifos resistance, and Lhsp90-1 expression increased by 1.58 times after long-term selection for tolerance to sub-lethal high temperatures. [Conclusion] These results suggested that the increased expression of Lhsp90-1 may promote the tolerance of L. striatellus to both high temperatures and chlorpyrifos.