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烟粉虱(MEAM 1)中热休克蛋白基因的差异表达模式与热响应

Differential expression patterns among heat-shock protein genes and thermal responses in the whitefly Bemisia tabaci (MEAM 1).

作者信息

Díaz Fernando, Orobio Rony F, Chavarriaga Paul, Toro-Perea Nelson

机构信息

Department of Biology, Universidad del Valle, Calle 13 No. 100-00, Cali, Colombia.

Department of Biology, Universidad del Valle, Calle 13 No. 100-00, Cali, Colombia.

出版信息

J Therm Biol. 2015 Aug;52:199-207. doi: 10.1016/j.jtherbio.2015.07.004. Epub 2015 Jul 16.

Abstract

There is convincing evidence that heat-shock proteins (HSP) are upregulated by stress conditions in insects; however, the relative contribution of each HSP gene to the heat-shock response remains unclear. Here we considered the whitefly Bemisia tabaci (MEAM 1), a phloem feeder and invasive species whose molecular stress response is an important mechanism for overcoming heat stress. We assessed the expression of the hsp23, 40, 70 and 90 genes at the mRNA level when submitted to heat shocks of 40 and 44°C/1h (control at 25°C). For this, we evaluated a set of available and suitable reference genes in order to perform data normalization using the real-time polymerase chain reaction (qRT-PCR) technique, and then confirmed the production of HSP70 protein based on Western blot. Results were compared with the hardening capacity of B. tabaci, measured by fitness components as a response to heat shocks, using 40°C as the induction temperature. Three of the four genes (hsp23, 70 and 90) were upregulated by heat stress at mRNA, showing differential expression patterns. Hsp70 expression was confirmed at the protein level. Hardening significantly increased fitness following heat stress, suggesting that HSPs may contribute to hardening capacity in B. tabaci. Potential role of each gene in the heat-shock response for whiteflies is discussed.

摘要

有令人信服的证据表明,昆虫在应激条件下热休克蛋白(HSP)会上调;然而,每个HSP基因对热休克反应的相对贡献仍不清楚。在这里,我们以烟粉虱(MEAM 1)为研究对象,它是一种吸食韧皮部汁液的入侵物种,其分子应激反应是克服热应激的重要机制。我们在40和44°C/1小时热休克处理(25°C为对照)下,评估了hsp23、40、70和90基因在mRNA水平的表达。为此,我们评估了一组可用且合适的内参基因,以便使用实时聚合酶链反应(qRT-PCR)技术进行数据标准化,然后基于蛋白质印迹法确认HSP70蛋白的产生。将结果与烟粉虱的硬化能力进行比较,以40°C作为诱导温度,通过适合度成分来衡量其对热休克的反应。四个基因中的三个(hsp23、70和90)在mRNA水平上受热应激上调,呈现出不同的表达模式。在蛋白质水平上证实了Hsp70的表达。硬化显著提高了热应激后的适合度,表明HSPs可能有助于烟粉虱的硬化能力。本文还讨论了每个基因在粉虱热休克反应中的潜在作用。

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