Jan Asad, Nakamura Hidemitsu, Handa Hirokazu, Ichikawa Hiroaki, Matsumoto Hiroshi, Komatsu Setsuko
National Institute of Agrobiological Sciences, Tsukuba, 305-8602 Japan.
Plant Cell Physiol. 2006 Feb;47(2):244-53. doi: 10.1093/pcp/pci241. Epub 2005 Dec 13.
Pyruvate dehydrogenase kinase (PDK) is a negative regulator of the mitochondrial pyruvate dehydrogenase complex (mtPDC) that plays a key role in intermediary metabolism. OsPDK1 was identified as a gibberellin-up-regulated gene using a cDNA microarray. The full-length cDNA for OsPDK1 was 1498 bp and encoded a predicted polypeptide of 363 amino acids. Genomic DNA analysis showed the presence of another isoform of PDK, OsPDK2, in rice. Reverse transcriptase-PCR analysis revealed differential expression of the two isoforms. OsPDK1 was expressed in leaf blade and leaf sheath but not in callus and root, while OsPDK2 was expressed constitutively in all tissues examined. Maximum expression of OsPDK1 in leaf sheath was detected by Northern blot analysis when seedlings were treated with 5 microM GA3 for 24 h. OsPDK1 expression was up-regulated by GA3, and there was little effect of other plant hormones. Mitochondrial pyruvate dehydrogenase (PDH) activity was reduced compared with control plants in 2-week-old seedlings treated with GA3. The beta-glucuronidase (GUS) reporter gene, driven by a 2,067 bp OsPDK1 promoter region fragment, was mainly expressed in the aleurone layer of germinating seed and leaf sheath. Transgenic rice expressing PDK1 RNAi had altered vegetative growth with reduced accumulation of vegetative tissues. These results suggest that gibberellin modulates the activity of mtPDC by regulating OsPDK1 expression and subsequently controlling plant growth and development.
丙酮酸脱氢酶激酶(PDK)是线粒体丙酮酸脱氢酶复合体(mtPDC)的负调控因子,在中间代谢中起关键作用。利用cDNA微阵列,OsPDK1被鉴定为一个赤霉素上调基因。OsPDK1的全长cDNA为1498 bp,编码一个预测的363个氨基酸的多肽。基因组DNA分析表明,水稻中存在PDK的另一种同工型OsPDK2。逆转录聚合酶链反应(RT-PCR)分析揭示了这两种同工型的差异表达。OsPDK1在叶片和叶鞘中表达,但在愈伤组织和根中不表达,而OsPDK2在所有检测的组织中组成型表达。当用5 microM GA3处理幼苗24小时时,通过Northern印迹分析检测到叶鞘中OsPDK1的表达量最高。OsPDK1的表达受GA3上调,而其他植物激素的影响很小。在用GA3处理的2周龄幼苗中,线粒体丙酮酸脱氢酶(PDH)活性与对照植株相比有所降低。由2067 bp的OsPDK1启动子区域片段驱动的β-葡萄糖醛酸酶(GUS)报告基因主要在萌发种子的糊粉层和叶鞘中表达。表达PDK1 RNA干扰的转基因水稻改变了营养生长,营养组织的积累减少。这些结果表明,赤霉素通过调节OsPDK1的表达,进而控制植物生长发育,从而调节mtPDC的活性。