Department of Molecular Microbiology, VIB, Leuven, Belgium.
Plant Physiol. 2013 Mar;161(3):1158-71. doi: 10.1104/pp.112.211391. Epub 2013 Jan 22.
Introduction of microbial trehalose biosynthesis enzymes has been reported to enhance abiotic stress resistance in plants but also resulted in undesirable traits. Here, we present an approach for engineering drought stress tolerance by modifying the endogenous trehalase activity in Arabidopsis (Arabidopsis thaliana). AtTRE1 encodes the Arabidopsis trehalase, the only enzyme known in this species to specifically hydrolyze trehalose into glucose. AtTRE1-overexpressing and Attre1 mutant lines were constructed and tested for their performance in drought stress assays. AtTRE1-overexpressing plants had decreased trehalose levels and recovered better after drought stress, whereas Attre1 mutants had elevated trehalose contents and exhibited a drought-susceptible phenotype. Leaf detachment assays showed that Attre1 mutants lose water faster than wild-type plants, whereas AtTRE1-overexpressing plants have a better water-retaining capacity. In vitro studies revealed that abscisic acid-mediated closure of stomata is impaired in Attre1 lines, whereas the AtTRE1 overexpressors are more sensitive toward abscisic acid-dependent stomatal closure. This observation is further supported by the altered leaf temperatures seen in trehalase-modified plantlets during in vivo drought stress studies. Our results show that overexpression of plant trehalase improves drought stress tolerance in Arabidopsis and that trehalase plays a role in the regulation of stomatal closure in the plant drought stress response.
已有研究报道,引入微生物海藻糖生物合成酶可以增强植物的非生物胁迫抗性,但也会导致不良性状。在这里,我们提出了一种通过修饰拟南芥(Arabidopsis thaliana)内源海藻糖酶活性来工程抗旱性的方法。AtTRE1 编码拟南芥海藻糖酶,这是该物种中唯一已知能够特异性地将海藻糖水解为葡萄糖的酶。构建了 AtTRE1 过表达和 Attre1 突变体系,并对它们在干旱胁迫测定中的表现进行了测试。AtTRE1 过表达植株的海藻糖水平降低,干旱胁迫后恢复更好,而 Attre1 突变体的海藻糖含量升高,表现出抗旱性表型。叶片脱落测定表明,Attre1 突变体比野生型植物更快失去水分,而 AtTRE1 过表达植株具有更好的保水能力。体外研究表明,ABA 介导的气孔关闭在 Attre1 系中受损,而 AtTRE1 过表达植株对 ABA 依赖性气孔关闭更敏感。在体内干旱胁迫研究中,观察到海藻糖酶修饰的幼苗叶片温度的变化进一步支持了这一观察结果。我们的结果表明,过表达植物海藻糖酶可以提高拟南芥的抗旱性,并且海藻糖酶在植物干旱胁迫反应中调节气孔关闭中起作用。