Ougham H J, Thomas A M, Thomas B J, Frick G A, Armstrong G A
Cell Biology Department, Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth, Ceredigion SY23 3EB, Wales, UK.
J Exp Bot. 2001 Jul;52(360):1447-54. doi: 10.1093/jexbot/52.360.1447.
The gibberellin-insensitive overgrowth mutant of barley, slender, exhibits altered expression of a number of nuclear genes in comparison with the wild type. There is a particularly marked reduction in slender seedlings of transcript encoding protochlorophyllide oxidoreductase (POR), the enzyme which catalyses the penultimate and only light-requiring step in chlorophyll biosynthesis. The expression of the two barley genes encoding light-dependent POR, PORA and PORB was investigated. Expression of both genes was found to be reduced in slender seedlings relative to the wild type, in both etiolated and light-grown leaf tissue; this was most marked in the zone of rapid cell extension. Western blot analysis showed that POR protein was also less abundant in etiolated and in light-grown slender than in the equivalent wild-type leaf tissue, although the effect was less pronounced than at the transcript level. Protochlorophyllide content in etiolated slender seedlings was reduced in comparison with wild-type seedlings, though chlorophyll content in light-grown leaf blades was unaffected. The reduction in POR expression in slender barley may reflect a novel response to the constitutive activation of gibberellin signalling in this mutant. Despite the consequences of the mutation for POR gene expression, slender seedlings develop apparently normal chloroplasts in the light, and etioplasts with well-defined prolamellar bodies when grown in continuous darkness. This suggests that the POR content of wild-type barley seedlings is well in excess of the minimum required for normal plastid development.
大麦的赤霉素不敏感过度生长突变体“细长型”与野生型相比,许多核基因的表达发生了改变。在细长型幼苗中,编码原叶绿素酸酯氧化还原酶(POR)的转录本显著减少,该酶催化叶绿素生物合成中倒数第二步且唯一需要光的步骤。对编码光依赖型POR的两个大麦基因PORA和PORB的表达进行了研究。结果发现,在黄化和光照生长的叶片组织中,相对于野生型,细长型幼苗中这两个基因的表达均降低;在快速细胞伸展区域最为明显。蛋白质免疫印迹分析表明,与野生型叶片组织相比,黄化和光照生长的细长型幼苗中POR蛋白含量也较少,尽管这种影响不如转录水平明显。与野生型幼苗相比,黄化的细长型幼苗中原叶绿素酸酯含量降低,不过光照生长叶片中的叶绿素含量未受影响。细长型大麦中POR表达的降低可能反映了对该突变体中赤霉素信号组成型激活的一种新反应。尽管该突变对POR基因表达有影响,但细长型幼苗在光照下能发育出明显正常的叶绿体,在持续黑暗中生长时能发育出具有明确原片层体的黄化质体。这表明野生型大麦幼苗的POR含量远超过正常质体发育所需的最低水平。