Wageningen University, Postbus 386, 6700 AJ Wageningen, The Netherlands.
J Plant Physiol. 2013 Sep 15;170(14):1228-34. doi: 10.1016/j.jplph.2013.04.003. Epub 2013 May 16.
GA biosynthesis and catabolism has been shown to play an important role in regulating tuberization in potato. Active GAs are inactivated in the stolon tips shortly after induction to tuberization. Overexpression of a GA inactivation gene results in an earlier tuberization phenotype, while reducing expression of the same gene results in delayed tuberization. In addition, overexpression of genes involved in GA biosynthesis results in delayed tuberization, while decreased expression of those genes results in earlied tuberization. The final step in GA biosynthesis is catalysed by StGA3ox1 and StGA3ox2 activity, that convert inactive forms of GA into active GA1 and GA4. In this study we cloned StGA3ox2 gene in an RNAi construct and used this construct to transform potato plants. The StGA3ox2 silenced plants were smaller and had shorter internodes. In addition, we assayed the concentrations of various GAs in the transgenic plants and showed an altered GA content. No difference was observed on the time point of tuber initiation. However, the transgenic clones had increased number of tubers with the same yield, resulting in smaller average tuber weight. In addition, we cloned the promoter of StGA3ox2 to direct expression of the GUS reporter gene to visualize the sites of GA biosynthesis in the potato plant. Finally, we discuss how changes of several GA levels can have an impact on shoot, stolon and tuber development, as well as the possible mechanisms that mediate feed-forward and feed-back regulation loops in the GA biosynthetic pathway in potato.
GA 生物合成和分解代谢被证明在调节马铃薯块茎形成中起着重要作用。在诱导块茎形成后不久,活跃的 GA 在匍匐茎尖端失活。GA 失活基因的过表达导致更早的块茎形成表型,而相同基因的表达减少导致块茎形成延迟。此外,GA 生物合成相关基因的过表达导致块茎形成延迟,而这些基因的表达减少导致块茎形成提前。GA 生物合成的最后一步由 StGA3ox1 和 StGA3ox2 活性催化,将 GA 的非活性形式转化为活性 GA1 和 GA4。在这项研究中,我们在 RNAi 构建体中克隆了 StGA3ox2 基因,并使用该构建体转化了马铃薯植株。StGA3ox2 沉默的植株较小,节间较短。此外,我们检测了转基因植物中各种 GA 的浓度,并显示出 GA 含量的改变。在块茎起始的时间点没有观察到差异。然而,转基因克隆的块茎数量增加,产量相同,导致平均块茎重量减小。此外,我们克隆了 StGA3ox2 的启动子,以指导 GUS 报告基因的表达,从而可视化马铃薯植株中 GA 生物合成的部位。最后,我们讨论了几种 GA 水平的变化如何影响地上部、匍匐茎和块茎的发育,以及介导马铃薯中 GA 生物合成途径中前馈和反馈调节环的可能机制。