Blee Kristopher A, Choi Joon W, O'Connell Ann P, Schuch Wolfgang, Lewis Norman G, Bolwell G Paul
School of Biological Sciences, Royal Holloway, University of London, Egham, Surrey TW20 0EX, UK.
Phytochemistry. 2003 Sep;64(1):163-76. doi: 10.1016/s0031-9422(03)00212-7.
A tobacco peroxidase isoenzyme (TP60) was down-regulated in tobacco using an antisense strategy, this affording transformants with lignin reductions of up to 40-50% of wild type (control) plants. Significantly, both guaiacyl and syringyl levels decreased in essentially a linear manner with the reductions in lignin amounts, as determined by both thioacidolysis and nitrobenzene oxidative analyses. These data provisionally suggest that a feedback mechanism is operative in lignifying cells, which prevents build-up of monolignols should oxidative capacity for their subsequent metabolism be reduced. Prior to this study, the only known rate-limiting processes in the monolignol/lignin pathways involved that of Phe supply and the relative activities of cinnamate-4-hydroxylase/p-coumarate-3-hydroxylase, respectively. These transformants thus provide an additional experimental means in which to further dissect and delineate the factors involved in monolignol targeting to precise regions in the cell wall, and of subsequent lignin assembly. Interestingly, the lignin down-regulated tobacco phenotypes displayed no readily observable differences in overall growth and development profiles, although the vascular apparatus was modified.
利用反义策略使烟草中的一种烟草过氧化物酶同工酶(TP60)表达下调,这使得转基因植株的木质素含量比野生型(对照)植株降低了40%-50%。值得注意的是,通过硫代酸解和硝基苯氧化分析确定,随着木质素含量的降低,愈创木基和紫丁香基水平基本上呈线性下降。这些数据初步表明,在木质化细胞中存在一种反馈机制,当单木质醇后续代谢的氧化能力降低时,可防止单木质醇的积累。在本研究之前,单木质醇/木质素途径中唯一已知的限速过程分别涉及苯丙氨酸供应以及肉桂酸-4-羟化酶/对香豆酸-3-羟化酶的相对活性。因此,这些转基因植株提供了一种额外的实验手段,可进一步剖析和阐明参与单木质醇靶向细胞壁精确区域以及后续木质素组装的因素。有趣的是,尽管维管系统发生了改变,但木质素下调的烟草表型在整体生长和发育概况上没有明显的差异。