CSIRO Plant Industry, 306 Carmody Rd, St Lucia, Qld 4067, Australia.
J Exp Bot. 2014 Feb;65(2):539-57. doi: 10.1093/jxb/ert399. Epub 2013 Dec 9.
Heat shock factors (Hsfs) play a central regulatory role in acquired thermotolerance. To understand the role of the major molecular players in wheat adaptation to heat stress, the Hsf family was investigated in Triticum aestivum. Bioinformatic and phylogenetic analyses identified 56 TaHsf members, which are classified into A, B, and C classes. Many TaHsfs were constitutively expressed. Subclass A6 members were predominantly expressed in the endosperm under non-stress conditions. Upon heat stress, the transcript levels of A2 and A6 members became the dominant Hsfs, suggesting an important regulatory role during heat stress. Many TaHsfA members as well as B1, C1, and C2 members were also up-regulated during drought and salt stresses. The heat-induced expression profiles of many heat shock protein (Hsp) genes were paralleled by those of A2 and A6 members. Transactivation analysis revealed that in addition to TaHsfA members (A2b and A4e), overexpression of TaHsfC2a activated expression of TaHsp promoter-driven reporter genes under non-stress conditions, while TaHsfB1b and TaHsfC1b did not. Functional heat shock elements (HSEs) interacting with TaHsfA2b were identified in four TaHsp promoters. Promoter mutagenesis analysis demonstrated that an atypical HSE (GAACATTTTGGAA) in the TaHsp17 promoter is functional for heat-inducible expression and transactivation by Hsf proteins. The transactivation of Hsp promoter-driven reporter genes by TaHsfC2a also relied on the presence of HSE. An activation motif in the C-terminal domain of TaHsfC2a was identified by amino residue substitution analysis. These data demonstrate the role of HsfA and HsfC2 in regulation of Hsp genes in wheat.
热休克因子 (Hsfs) 在获得性耐热性中发挥着核心调节作用。为了了解主要分子在小麦适应热胁迫中的作用,研究了小麦中的 Hsf 家族。生物信息学和系统发育分析鉴定了 56 个 TaHsf 成员,它们分为 A、B 和 C 类。许多 TaHsfs 组成型表达。亚类 A6 成员在非胁迫条件下主要在胚乳中表达。在热胁迫下,A2 和 A6 成员的转录水平成为主要的 Hsfs,表明其在热胁迫期间具有重要的调节作用。许多 TaHsfA 成员以及 B1、C1 和 C2 成员在干旱和盐胁迫下也被上调。许多热休克蛋白 (Hsp) 基因的热诱导表达谱与 A2 和 A6 成员的表达谱相似。转激活分析表明,除了 TaHsfA 成员 (A2b 和 A4e) 之外,TaHsfC2a 的过表达在非胁迫条件下激活 TaHsp 启动子驱动的报告基因的表达,而 TaHsfB1b 和 TaHsfC1b 则没有。在四个 TaHsp 启动子中鉴定出与 TaHsfA2b 相互作用的功能热休克元件 (HSE)。启动子诱变分析表明,TaHsp17 启动子中典型的 HSE (GAACATTTTGGAA) 对于 Hsf 蛋白的热诱导表达和转激活是功能性的。TaHsfC2a 对 Hsp 启动子驱动的报告基因的转激活也依赖于 HSE 的存在。通过氨基酸残基取代分析鉴定了 TaHsfC2a 中 C 端结构域的激活基序。这些数据表明 HsfA 和 HsfC2 在调节小麦中 Hsp 基因方面的作用。