Center for Plant Biotechnology and Genomics, Campus de Montegancedo, Universidad Politecnica de Madrid, 28223 Pozuelo de Alarcon, Madrid, Spain.
Plant Physiol. 2014 Feb;164(2):978-91. doi: 10.1104/pp.113.225730. Epub 2013 Dec 4.
Climate-driven heat stress is a key factor affecting forest plantation yields. While its effects are expected to worsen during this century, breeding more tolerant genotypes has proven elusive. We report here a substantial and durable increase in the thermotolerance of hybrid poplar (Populus tremula×Populus alba) through overexpression of a major small heat shock protein (sHSP) with convenient features. Experimental evidence was obtained linking protective effects in the transgenic events with the unique chaperone activity of sHSPs. In addition, significant positive correlations were observed between phenotype strength and heterologous sHSP accumulation. The remarkable baseline levels of transgene product (up to 1.8% of total leaf protein) have not been reported in analogous studies with herbaceous species. As judged by protein analyses, such an accumulation is not matched either by endogenous sHSPs in both heat-stressed poplar plants and field-grown adult trees. Quantitative real time-polymerase chain reaction analyses supported these observations and allowed us to identify the poplar members most responsive to heat stress. Interestingly, sHSP overaccumulation was not associated with pleiotropic effects that might decrease yields. The poplar lines developed here also outperformed controls under in vitro and ex vitro culture conditions (callus biomass, shoot production, and ex vitro survival), even in the absence of thermal stress. These results reinforce the feasibility of improving valuable genotypes for plantation forestry, a field where in vitro recalcitrance, long breeding cycles, and other practical factors constrain conventional genetic approaches. They also provide new insights into the biological functions of the least understood family of heat shock protein chaperones.
气候驱动的热应激是影响人工林产量的一个关键因素。尽管预计在本世纪内其影响会加剧,但培育更具耐受性的基因型一直难以实现。我们在这里报告了通过过量表达具有便利特性的主要小热休克蛋白 (sHSP) ,显著且持久地提高了杂种杨树(Populus tremula×Populus alba)的耐热性。实验证据表明,转基因事件中的保护作用与 sHSP 的独特伴侣活性有关。此外,还观察到表型强度与异源 sHSP 积累之间存在显著的正相关。在与草本物种类似的研究中,尚未报道过类似的转基因产物(高达总叶片蛋白的 1.8%)的显著基线水平。根据蛋白质分析,在热胁迫的杨树植物和田间生长的成年树木中,这种积累与内源性 sHSP 并不匹配。定量实时聚合酶链反应分析支持了这些观察结果,并使我们能够鉴定对热应激最敏感的杨树成员。有趣的是,sHSP 的过度积累与可能降低产量的多效性效应无关。与对照相比,这里开发的杨树系在体外和离体培养条件下(愈伤组织生物量、芽产量和离体存活率)表现更好,即使在没有热应激的情况下也是如此。这些结果增强了为人工林林业改良有价值基因型的可行性,在该领域,体外抗性、长育种周期和其他实际因素限制了常规遗传方法的应用。它们还为最不了解的热休克蛋白伴侣家族的生物学功能提供了新的见解。