Department of Plant Molecular Biology, University of Delhi, South Campus, Benito Juarez Road, Dhaula Kuan, New Delhi 110 021, India.
Plant Cell Environ. 2012 Nov;35(11):1912-31. doi: 10.1111/j.1365-3040.2012.02525.x. Epub 2012 May 22.
The nuclear-encoded chloroplast small heat shock proteins (sHSPs) are present in all plant species from algae to angiosperms. Expression analysis shows that the wheat chloroplastic sHSP (HSP26) is highly inducible by heat stress in almost all the vegetative and generative tissues and is also expressed constitutively in certain developmental growth stages. We characterize wheat chloroplastic sHSP 26 through transgenic approach using Arabidopsis and report cloning of the promoter and its characterization. Transgenic Arabidopsis plants were substantially tolerant under continuous high temperature regimen than wild-type plants, as measured by photosystem II (PSII) activity, accumulation of more photosynthetic pigments, higher biomass and seed yield. Transgenic plants produced bold seeds under high temperature, having higher germination potential than the wild-type plants. Further, antisense Arabidopsis plants showed negligible tolerance even for non-lethal heat shock, impaired in basal thermo-tolerance, and accumulated less biomass and seed yield under normal growth conditions. Promoter analysis revealed the presence of several heat and other abiotic stress responsive cis-acting elements along with developmental stage and tissue-specific elements. Analysis of promoter through GUS reporter system in both transgenic rice and Arabidopsis further confirms the role of chloroplastic sHsp26 in heat and other abiotic stresses as well as during seed maturation and germination. Genome-wide expression analysis of overexpression Arabidopsis plants revealed that the transcriptome remained unchanged in the transgenic plants and the tolerance was due to the overexpression of chloroplastic heat shock protein (HSP) only.
核编码的叶绿体小分子热休克蛋白(sHSPs)存在于所有植物物种中,从藻类到被子植物。表达分析表明,小麦叶绿体 sHSP(HSP26)在几乎所有营养和生殖组织中对热胁迫高度诱导,并且在某些发育生长阶段也持续表达。我们通过使用拟南芥的转基因方法对小麦叶绿体 sHSP26 进行了表征,并报告了其启动子的克隆及其特性。与野生型植物相比,转基因拟南芥植物在连续高温处理下具有更高的耐受性,表现在 PSII(光合系统 II)活性、更多光合色素的积累、更高的生物量和种子产量。转基因植物在高温下产生的种子更大胆,具有比野生型植物更高的发芽潜力。此外,反义拟南芥植物即使在非致死性热休克下也几乎没有耐受性,基础耐热性受损,在正常生长条件下积累的生物量和种子产量更少。启动子分析显示,存在多个热和其他非生物胁迫响应顺式作用元件,以及发育阶段和组织特异性元件。在转基因水稻和拟南芥中通过 GUS 报告基因系统对启动子进行分析进一步证实了叶绿体 sHsp26 在热胁迫和其他非生物胁迫以及种子成熟和萌发过程中的作用。过表达拟南芥植物的全基因组表达分析表明,转基因植物的转录组没有变化,并且耐受性仅归因于叶绿体热休克蛋白(HSP)的过表达。