Miersch O, Wasternack C
Leibniz Institute of Plant Biochemistry, Halle, Germany.
Biol Chem. 2000 Aug;381(8):715-22. doi: 10.1515/BC.2000.092.
Jasmonates and their precursors, the octadecanoids, are signals in stress-induced alteration of gene expression. Several mRNAs coding for enzymes of jasmonic acid (JA) biosynthesis are up-regulated upon JA treatment or endogenous increase of the JA level. Here we investigated the positive feedback of endogenous JA on JA formation, as well as its beta-oxidation steps. JA-responsive gene expression was recorded in terms of proteinase inhibitor2 (pin2) mRNA accumulation. JA formed upon treatment of tomato (Lycopersicon esculentum cv. Moneymaker) leaves with JA derivatives carrying different lengths of the carboxylic acid side chain was quantified by gas chromatography-mass spectrometry (GC-MS). The data revealed that beta-oxidation of the side chain occurs up to a butyric acid moiety. The amount of JA formed from side-chain modified JA derivatives correlated with pin2-mRNA accumulation. JA derivatives with a carboxylic side chain of 3, 5 or 7 carbon atoms were unable to form JA and to express on pin2, whereas even-numbered derivatives were active. After treatment of tomato leaves with (10-(2)H)-(-)-12-oxophytoenoic acid, (4-(2)H)-(-)-JA and its methyl ester were formed and could be quantified separately from the endogenously nonlabeled JA pool by GC-MS analysis via isotopic discrimination. The level of 8 nmol per g fresh weight JA and its methyl ester originated exclusively from labeled 12-oxophytoenic acid. This and further data indicate that endogenous synthesis of the JA precursor 12-oxophytodienoic acid, as well as of JA and its methyl ester, are not induced in tomato leaves, suggesting that positive feedback in JA biosynthesis does not function in vivo.
茉莉酸及其前体十八烷类化合物是应激诱导基因表达改变中的信号分子。几种编码茉莉酸(JA)生物合成酶的mRNA在JA处理或JA水平内源性增加后会上调。在此,我们研究了内源性JA对JA形成及其β-氧化步骤的正反馈作用。以蛋白酶抑制剂2(pin2)mRNA积累来记录JA响应基因的表达。用带有不同长度羧酸侧链的JA衍生物处理番茄(Lycopersicon esculentum cv. Moneymaker)叶片后形成的JA,通过气相色谱-质谱联用(GC-MS)进行定量分析。数据显示,侧链的β-氧化会一直进行到丁酸部分。由侧链修饰的JA衍生物形成的JA量与pin2-mRNA积累相关。具有3、5或7个碳原子羧酸侧链的JA衍生物无法形成JA,也不能使pin2表达,而偶数碳侧链的衍生物则具有活性。用(10-(2)H)-(-)-12-氧代植物二烯酸处理番茄叶片后,形成了(4-(2)H)-(-)-JA及其甲酯,通过同位素鉴别,利用GC-MS分析可将它们与内源性未标记的JA库分开定量。每克鲜重8 nmol的JA及其甲酯水平完全源自标记的12-氧代植物二烯酸。这些以及其他数据表明,番茄叶片中不会诱导JA前体12-氧代植物二烯酸以及JA及其甲酯的内源性合成,这表明JA生物合成中的正反馈在体内不起作用。