Department of Plant Physiology, Ruhr-Universität, 44801 Bochum, Germany.
Plant Physiol. 2010 May;153(1):114-27. doi: 10.1104/pp.110.155093. Epub 2010 Mar 26.
Lipases are involved in the generation of jasmonates, which regulate responses to biotic and abiotic stresses. Two sn-1-specific acyl hydrolases, DEFECTIVE IN ANTHER DEHISCENCE1 (DAD1) and DONGLE (DGL), have been reported to be localized in plastids and to be essential and sufficient for jasmonate biosynthesis in Arabidopsis (Arabidopsis thaliana) leaves. Here, we show that levels of 12-oxo-phytodienoic acid (OPDA) and jasmonic acid in three different DGL RNA interference lines and the dad1 mutant were similar to wild-type levels during the early wound response as well as after Pseudomonas infection. Due to the lack of sn-2 substrate specificity, synthesis of dinor OPDA was not expected and also not found to be affected in DGL knockdown and DGL-overexpressing lines. As reported, DAD1 participates in jasmonate formation only in the late wound response. In addition, DGL protein was found to be localized in lipid bodies and not in plastids. Furthermore, jasmonate levels in 16 additional mutants defective in the expression of lipases with predicted chloroplast localization did not show strong differences from wild-type levels after wounding, except for a phospholipase A (PLA) PLA-Igamma1 (At1g06800) mutant line that displayed diminished wound-induced dinor OPDA, OPDA, and jasmonic acid levels. A quadruple mutant defective in four DAD1-like lipases displayed similar jasmonate levels as the mutant line of PLA-Igamma1 after wounding. Hence, we identify PLA-Igamma1 as a novel target gene to manipulate jasmonate biosynthesis. Our results suggest that, in addition to DAD1 and PLA-Igamma1, still unidentified enzymes with sn-1 and sn-2 hydrolase activity are involved in wound- and pathogen-induced jasmonate formation, indicating functional redundancy within the lipase family.
脂肪酶参与茉莉酸的生成,茉莉酸调节生物和非生物胁迫的反应。已有报道称,两种 sn-1 特异性酰基水解酶——DEFECTIVE IN ANTHER DEHISCENCE1(DAD1)和 DONGLE(DGL)——定位于质体中,对于拟南芥(Arabidopsis thaliana)叶片中茉莉酸的生物合成是必需且充分的。在这里,我们发现,在早期创伤反应以及 Pseudomonas 感染后,三个不同的 DGL RNAi 系和 dad1 突变体中的 12-氧代-植二烯酸(OPDA)和茉莉酸水平与野生型水平相似。由于缺乏 sn-2 底物特异性,预计不会合成二降 OPDA,并且在 DGL 敲低和 DGL 过表达系中也未发现其受到影响。如前所述,DAD1 仅在晚期创伤反应中参与茉莉酸的形成。此外,发现 DGL 蛋白定位于脂滴而不是质体中。此外,在 16 个另外的脂酶表达缺陷突变体中,预测这些脂酶具有质体定位,在创伤后,除了磷脂酶 A(PLA)PLA-Igamma1(At1g06800)突变体外,其表现出创伤诱导的二降 OPDA、OPDA 和茉莉酸水平降低外,这些突变体的茉莉酸水平与野生型水平没有明显差异。在四个 DAD1 样脂酶缺陷的四重突变体中,创伤后其表现出与 PLA-Igamma1 突变体相似的茉莉酸水平。因此,我们将 PLA-Igamma1 鉴定为一种新的靶基因,用于操纵茉莉酸的生物合成。我们的结果表明,除了 DAD1 和 PLA-Igamma1 之外,仍然有具有 sn-1 和 sn-2 水解酶活性的未鉴定酶参与了创伤和病原体诱导的茉莉酸形成,这表明脂肪酶家族内存在功能冗余。