Müssig Carsten, Fischer Sabine, Altmann Thomas
Max-Planck-Institut für Molekulare Pflanzenphysiologie, Department Willmitzer, Am Mühlenberg 1, 14476 Golm, Germany.
Plant Physiol. 2002 Jul;129(3):1241-51. doi: 10.1104/pp.011003.
Major brassinosteroid (BR) effects such as BR-induced growth are mediated through genomic pathways because RNA synthesis inhibitors and protein synthesis inhibitors interfere with these processes. A limited number of BR-regulated genes have been identified hitherto. The majority of genes (such as BRU1, CycD3, Lin6, OPR3, and TRIP-1) were identified by comparisons of BR-treated versus control-treated plants. However, altered transcript levels after BR application may not reflect normal physiological events. A complementary approach is the comparison of BR-deficient plants versus wild-type plants. No artificial treatments interfere with endogenous signaling pathways, but a subset of phenotypic alterations of phytohormone-deficient plants most probably is secondary. To identify genes that are subject to direct BR regulation, we analyzed CPD antisense and dwf1-6 (cbb1) mutant plants. Both show a mild phenotype in comparison with BR-deficient mutants such as cpd/cbb3, det2, and dwf4. Plants were grown under two different environments to filter out BR deficiency effects that occur only at certain environmental conditions. Finally, we established expression patterns after BR treatment of wild-type and dwf1-6 (cbb1) plants. Ideally, a BR-regulated gene displays a dose-response relationship in such a way that a gene with decreased transcript levels in BR-deficient plants is BR inducible and vice versa. Expression profile analysis of above ground part of plants was performed by means of Affymetrix Arabidopsis Genome Arrays.
主要的油菜素内酯(BR)效应,如BR诱导的生长,是通过基因组途径介导的,因为RNA合成抑制剂和蛋白质合成抑制剂会干扰这些过程。迄今为止,已鉴定出数量有限的BR调控基因。大多数基因(如BRU1、CycD3、Lin6、OPR3和TRIP-1)是通过比较BR处理的植物与对照处理的植物而鉴定出来的。然而,施用BR后转录水平的改变可能无法反映正常的生理事件。一种互补的方法是比较BR缺陷型植物与野生型植物。没有人工处理会干扰内源性信号通路,但植物激素缺陷型植物的一部分表型改变很可能是继发性的。为了鉴定受BR直接调控的基因,我们分析了CPD反义植株和dwf1-6(cbb1)突变体植株。与BR缺陷型突变体如cpd/cbb3、det2和dwf4相比,这两种植株都表现出轻微的表型。将植株种植在两种不同的环境中,以滤除仅在特定环境条件下出现的BR缺乏效应。最后,我们确定了野生型和dwf1-6(cbb1)植株经BR处理后的表达模式。理想情况下,一个受BR调控的基因呈现剂量反应关系,即一个在BR缺陷型植株中转录水平降低的基因是BR可诱导的,反之亦然。通过Affymetrix拟南芥基因组芯片对植株地上部分进行表达谱分析。