Departamento de Biología, Facultad de Ciencias, Universidad de Chile, Santiago, Chile.
PLoS One. 2013;8(3):e58144. doi: 10.1371/journal.pone.0058144. Epub 2013 Mar 29.
Plant carotenoids are synthesized and accumulated in plastids through a highly regulated pathway. Lycopene β-cyclase (LCYB) is a key enzyme involved directly in the synthesis of α-carotene and β-carotene through the cyclization of lycopene. Carotenoids are produced in both carrot (Daucus carota) leaves and reserve roots, and high amounts of α-carotene and β-carotene accumulate in the latter. In some plant models, the presence of different isoforms of carotenogenic genes is associated with an organ-specific function. D. carota harbors two Lcyb genes, of which DcLcyb1 is expressed in leaves and storage roots during carrot development, correlating with an increase in carotenoid levels. In this work, we show that DcLCYB1 is localized in the plastid and that it is a functional enzyme, as demonstrated by heterologous complementation in Escherichia coli and over expression and post transcriptional gene silencing in carrot. Transgenic plants with higher or reduced levels of DcLcyb1 had incremented or reduced levels of chlorophyll, total carotenoids and β-carotene in leaves and in the storage roots, respectively. In addition, changes in the expression of DcLcyb1 are accompanied by a modulation in the expression of key endogenous carotenogenic genes. Our results indicate that DcLcyb1 does not possess an organ specific function and modulate carotenoid gene expression and accumulation in carrot leaves and storage roots.
植物类胡萝卜素是通过高度调控的途径在质体中合成和积累的。番茄红素 β-环化酶(LCYB)是一种直接参与α-胡萝卜素和β-胡萝卜素合成的关键酶,通过番茄红素的环化作用。类胡萝卜素在胡萝卜(Daucus carota)叶片和储备根中都有产生,并且在后一种中积累了大量的α-胡萝卜素和β-胡萝卜素。在一些植物模型中,不同同工型的类胡萝卜素生物合成基因的存在与器官特异性功能有关。D. carota 含有两个 Lcyb 基因,其中 DcLcyb1 在胡萝卜发育过程中在叶片和贮藏根中表达,与类胡萝卜素水平的增加相关。在这项工作中,我们表明 DcLCYB1 定位于质体中,并且是一种功能性酶,这通过在大肠杆菌中的异源互补以及在胡萝卜中的过表达和转录后基因沉默得到证实。具有较高或较低 DcLcyb1 水平的转基因植物在叶片和贮藏根中分别具有增加或减少的叶绿素、总类胡萝卜素和β-胡萝卜素水平。此外,DcLcyb1 的表达变化伴随着关键内源性类胡萝卜素生物合成基因的表达调节。我们的结果表明,DcLcyb1 不具有器官特异性功能,并且调节胡萝卜叶片和贮藏根中的类胡萝卜素基因表达和积累。