Plant Biotechnology Institute, National Research Council of Canada, Saskatoon, Saskatchewan, Canada.
Plant Physiol. 2011 Dec;157(4):2108-19. doi: 10.1104/pp.111.182584. Epub 2011 Oct 5.
The phytohormone abscisic acid (ABA) regulates stress responses and controls numerous aspects of plant growth and development. Biosynthetic precursors and catabolites of ABA have been shown to trigger ABA responses in physiological assays, but it is not clear whether these are intrinsically active or whether they are converted into ABA in planta. In this study, we analyzed the effect of ABA precursors, conjugates, and catabolites on hormone signaling in Arabidopsis (Arabidopsis thaliana). The compounds were also tested in vitro for their ability to regulate the phosphatase moiety of ABA receptor complexes consisting of the protein phosphatase 2C ABI2 and the coreceptors RCAR1/PYL9, RCAR3/PYL8, and RCAR11/PYR1. Using mutants defective in ABA biosynthesis, we show that the physiological activity associated with ABA precursors derives predominantly from their bioconversion to ABA. The ABA glucose ester conjugate, which is the most widespread storage form of ABA, showed weak ABA-like activity in germination assays and in triggering ABA signaling in protoplasts. The ABA conjugate and precursors showed negligible activity as a regulatory ligand of the ABI2/RCAR receptor complexes. The majority of ABA catabolites were inactive in our assays. To analyze the chemically unstable 8'- and 9'-hydroxylated ABA catabolites, we used stable tetralone derivatives of these compounds, which did trigger selective ABA responses. ABA synthetic analogs exhibited differential activity as regulatory ligands of different ABA receptor complexes in vitro. The data show that ABA precursors, catabolites, and conjugates have limited intrinsic bioactivity and that both natural and synthetic ABA-related compounds can be used to probe the structural requirements of ABA ligand-receptor interactions.
植物激素脱落酸(ABA)调节应激反应,并控制植物生长和发育的许多方面。ABA 的生物合成前体和代谢物已被证明在生理测定中触发 ABA 反应,但尚不清楚这些物质是否具有内在活性,或者它们是否在植物体内转化为 ABA。在这项研究中,我们分析了 ABA 前体、缀合物和代谢物对拟南芥(Arabidopsis thaliana)激素信号的影响。还在体外测试了这些化合物调节由蛋白磷酸酶 2C ABI2 和核心受体 RCAR1/PYL9、RCAR3/PYL8 和 RCAR11/PYR1 组成的 ABA 受体复合物的磷酸酶部分的能力。使用 ABA 生物合成缺陷突变体,我们表明与 ABA 前体相关的生理活性主要源自它们转化为 ABA。ABA 葡萄糖酯缀合物是 ABA 最广泛的储存形式,在萌发测定中和在原生质体中触发 ABA 信号中表现出微弱的 ABA 样活性。ABA 缀合物和前体作为 ABI2/RCAR 受体复合物的调节配体几乎没有活性。在我们的测定中,大多数 ABA 代谢物没有活性。为了分析化学不稳定的 8'-和 9'-羟基化 ABA 代谢物,我们使用了这些化合物的稳定四氢酮衍生物,它们确实引发了选择性的 ABA 反应。ABA 合成类似物在体外作为不同 ABA 受体复合物的调节配体表现出不同的活性。这些数据表明,ABA 前体、代谢物和缀合物具有有限的内在生物活性,天然和合成的 ABA 相关化合物都可用于研究 ABA 配体-受体相互作用的结构要求。