Lee D, Ellard M, Wanner L A, Davis K R, Douglas C J
Department of Botany, University of British Columbia, Vancouver, Canada.
Plant Mol Biol. 1995 Aug;28(5):871-84. doi: 10.1007/BF00042072.
An Arabidopsis cDNA clone encoding 4-coumarate:CoA ligase (4CL), a key enzyme of phenylpropanoid metabolism, was identified and sequenced. The predicted amino acid sequence is similar to those of other cloned 4CL genes. Southern blot analysis indicated that 4CL is single-copy gene in Arabidopsis. Northern blots showed that 4CL expression was activated early during seedling development. The onset of 4CL expression was correlated with the onset of lignin deposition in cotyledons and roots 2-3 days after germination. The timing of the expression of a parsley 4CL1-GUS fusion in transgenic Arabidopsis seedlings was examined in parallel and was very similar to that of endogenous 4CL. In mature plants, highest 4CL expression was observed in bolting stems, where relatively large amounts of lignin accumulate. Both 4CL and 4CL1-GUS mRNA accumulation was strongly and transiently activated by wounding of mature Arabidopsis leaves. 4CL expression was specifically activated within 6 h after infiltration of Arabidopsis ecotype Columbia leaves with a Pseudomonas syringae pv. maculicola strain harboring the bacterial avirulence gene avrB, which causes in incompatible interaction. The timing of 4CL activation was identical to the previously observed activation of PAL gene expression in this interaction. No activation of 4CL expression was observed in a compatible interaction caused by a Pseudomonas syringae pv. maculicola strain without avrB.
一个编码4-香豆酸:辅酶A连接酶(4CL)的拟南芥cDNA克隆被鉴定并测序,4CL是苯丙烷代谢的关键酶。预测的氨基酸序列与其他已克隆的4CL基因相似。Southern杂交分析表明4CL在拟南芥中是单拷贝基因。Northern杂交显示4CL表达在幼苗发育早期被激活。4CL表达的开始与萌发后2 - 3天子叶和根中木质素沉积的开始相关。同时检测了转基因拟南芥幼苗中一个欧芹4CL1 - GUS融合基因的表达时间,其与内源4CL的表达时间非常相似。在成熟植株中,4CL在抽薹茎中表达最高,那里积累了相对大量的木质素。成熟拟南芥叶片受伤后,4CL和4CL1 - GUS的mRNA积累都被强烈且短暂地激活。用携带细菌无毒基因avrB的丁香假单胞菌番茄致病变种菌株浸润拟南芥生态型哥伦比亚叶片后6小时内,4CL表达被特异性激活,这种相互作用是不亲和的。4CL激活的时间与之前在这种相互作用中观察到的苯丙氨酸解氨酶(PAL)基因表达的激活时间相同。在由不含avrB的丁香假单胞菌番茄致病变种菌株引起 的亲和性相互作用中未观察到4CL表达的激活。