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热应激诱导的 BBX18 负调控拟南芥的耐热性。

Heat stress-induced BBX18 negatively regulates the thermotolerance in Arabidopsis.

机构信息

State Key Laboratory Breeding Base for Crop Germplasm Innovation and Utilization, College of Biosciences and Biotechnology, Hunan Agricultural University, Changsha, 410128, China.

出版信息

Mol Biol Rep. 2013 Mar;40(3):2679-88. doi: 10.1007/s11033-012-2354-9. Epub 2012 Dec 14.

Abstract

There is increasing evidence for considerable interlinking between the responses to heat stress (HS) and light signaling. In the present work, we provide molecular evidence that BBX18, a negative regulator in photomorphogenesis belonging to the B-box zinc finger protein family in Arabidopsis thaliana, is involved in the regulation of thermotolerance. Using quantitative RT-PCR, GUS staining and immunoblot analysis, our results indicate that the expression of BBX18 was induced by HS. BBX18-RNAi and 35S::BBX18 transgenic Arabidopsis plants were obtained for functional analysis of BBX18. Under-expression of BBX18 displayed increased both basal and acquired thermotolerance in the transgenic plants, while over-expression of BBX18 reduced tolerance to HS in transgenic lines. Moreover, when wild-type, BBX18-RNAi and 35S::BBX18 transgenic plants were treated with HS, HR-related digalactosyldiacylglycerol synthase 1 (DGD1) was down-regulated by BBX18 in both normal and heat shock conditions. Besides, the expression levels of Hsp70, Hsp101 and APX2 were increased in BBX18-RNAi transgenic plants, but lower in 35S::BBX18 transgenic plants. However, the expression of HsfA2 was lower in BBX18-RNAi transgenic plants and higher in the 35S::BBX18 after high-temperature treatment. These results suggesting that, by modulated expression of a set of HS-responsive genes, BBX18 weakened tolerance to HS in Arabidopsis. So our data indicate that BBX18 plays a negative role in thermotolerance.

摘要

越来越多的证据表明,热应激(HS)和光信号之间存在着相当大的相互联系。在本工作中,我们提供了分子证据,表明 BBX18,一种拟南芥光形态建成中属于 B 盒锌指蛋白家族的负调控因子,参与了耐热性的调节。通过定量 RT-PCR、GUS 染色和免疫印迹分析,我们的结果表明 BBX18 的表达受 HS 诱导。为了对 BBX18 进行功能分析,获得了 BBX18-RNAi 和 35S::BBX18 转基因拟南芥植物。BBX18 的低表达在转基因植物中显示出了基础耐热性和获得性耐热性的增加,而 BBX18 的过表达则降低了转基因系对 HS 的耐受性。此外,当野生型、BBX18-RNAi 和 35S::BBX18 转基因植物受到 HS 处理时,HR 相关的二半乳糖基二酰基甘油合酶 1(DGD1)在正常和热休克条件下均受 BBX18 的下调。此外,在 BBX18-RNAi 转基因植物中,Hsp70、Hsp101 和 APX2 的表达水平升高,但在 35S::BBX18 转基因植物中表达水平降低。然而,在 BBX18-RNAi 转基因植物中 HsfA2 的表达水平较低,而在高温处理后 35S::BBX18 转基因植物中的表达水平较高。这些结果表明,通过调节一组 HS 响应基因的表达,BBX18 削弱了拟南芥对 HS 的耐受性。因此,我们的数据表明 BBX18 在耐热性中发挥负作用。

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