Melan M A, Dong X, Endara M E, Davis K R, Ausubel F M, Peterman T K
Department of Biological Sciences, Wellesley College, Massachusetts 02181.
Plant Physiol. 1993 Feb;101(2):441-50. doi: 10.1104/pp.101.2.441.
We isolated and characterized a 2.8-kb, full-length, Arabidopsis thaliana cDNA clone encoding a lipoxygenase. DNA sequence analysis showed that the deduced amino acid sequence of the Arabidopsis protein is 72 to 78% similar to that of legume seed lipoxygenases. DNA blot analysis indicated that Arabidopsis contains a single gene, LOX1, with appreciable homology to the cDNA clone. RNA blot analysis showed that the LOX1 gene is expressed in Arabidopsis leaves, roots, inflorescences, and young seedlings. LOX1 expression levels were highest in roots and young seedlings. In mature plants, LOX1 mRNA levels increased upon treatment with the stress-related hormones abscisic acid and methyl jasmonate and remained high for at least 96 h. Expression of the LOX1 gene was examined following infiltration of leaves with virulent (Psm ES4326) and avirulent (Pst MM1065) strains of Pseudomonas syringae. LOX1 mRNA levels were induced approximately 6-fold by both virulent and avirulent strains; however, the response to avirulent strains was much more rapid. Infiltration of leaves with Pst MM1065 resulted in maximal induction within 12 h, whereas maximal induction by Psm ES4326 did not occur until 48 h. When a cloned avr gene, avrRpt2, was transferred to Psm ES4326, LOX1 mRNA accumulated in a pattern similar to that observed for the avirulent strain Pst MM1065.
我们分离并鉴定了一个编码脂氧合酶的2.8 kb拟南芥全长cDNA克隆。DNA序列分析表明,拟南芥蛋白推导的氨基酸序列与豆科种子脂氧合酶的序列相似度为72%至78%。DNA印迹分析表明,拟南芥含有一个与该cDNA克隆具有明显同源性的单基因LOX1。RNA印迹分析表明,LOX1基因在拟南芥的叶片、根、花序和幼苗中表达。LOX1的表达水平在根和幼苗中最高。在成熟植株中,用与胁迫相关的激素脱落酸和茉莉酸甲酯处理后,LOX1的mRNA水平升高,并至少持续96小时保持在高位。在用丁香假单胞菌的强毒株(Psm ES4326)和无毒株(Pst MM1065)浸润叶片后,检测了LOX1基因的表达。强毒株和无毒株均可使LOX1的mRNA水平诱导增加约6倍;然而,对无毒株的反应要快得多。用Pst MM1065浸润叶片在12小时内诱导达到最大值,而用Psm ES4326诱导的最大值直到48小时才出现。当将一个克隆的avr基因avrRpt2转入Psm ES4326时,LOX1 mRNA的积累模式与无毒株Pst MM1065相似。