Cunillera N, Boronat A, Ferrer A
Departament de Bioquímica i Biologia Molecular, Facultat de Farmàcia, Universitat de Barcelona, Spain.
Plant Mol Biol. 2000 Dec;44(6):747-58. doi: 10.1023/a:1026588708849.
Farnesyl diphosphate synthase (FPS), the enzyme that catalyses the synthesis of farnesyl diphosphate (FPP) from isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP), is considered a regulatory enzyme of plant isoprenoid biosynthesis. The promoter regions of the FPS1 and FPS2 genes controlling the expression of isoforms FPS1S and FPS2, respectively, were fused to the beta-glucuronidase (GUS) reporter gene and introduced into Arabidopsis thaliana plants. The FPS1S:GUS gene is widely expressed in all plant tissues throughout development, thus supporting a role for FPS1S in the synthesis of isoprenoids serving basic plant cell functions. In contrast, the FPS2:GUS gene shows a pattern of expression restricted to specific organs at particular stages of development. The highest levels of GUS activity are detected in flowers, especially in pollen grains, from the early stages of flower development. After pollination, much lower levels of GUS activity are detected in the rest of floral organs, with the exception of the ovary valves, which remain unstained throughout flower development. GUS activity is also detected in developing and mature seeds. In roots, GUS expression is primarily detected at sites of lateral root initiation and in junctions between primary and secondary roots. No GUS activity is detected in root apical meristems. GUS expression is also observed in junctions between primary and secondary stems. Overall, the pattern of expression of FPS2:GUS suggests a role for FPS2 in the synthesis of particular isoprenoids with specialized functions. Functional FPS2 gene promoter deletion analysis in transfected protoplasts and transgenic A. thaliana plants indicate that all the cis-acting elements required to establish the full pattern of expression of the FPS2 gene are contained in a short region extending from positions -111 to +65. The potential regulatory role of specific sequences within this region is discussed.
法尼基二磷酸合酶(FPS)可催化由异戊烯基二磷酸(IPP)和二甲基烯丙基二磷酸(DMAPP)合成法尼基二磷酸(FPP),被认为是植物类异戊二烯生物合成的一种调控酶。分别控制FPS1S和FPS2亚型表达的FPS1和FPS2基因的启动子区域与β-葡萄糖醛酸酶(GUS)报告基因融合,并导入拟南芥植株中。FPS1S:GUS基因在植物发育过程中的所有组织中广泛表达,因此支持FPS1S在合成具有基本植物细胞功能的类异戊二烯中的作用。相比之下,FPS2:GUS基因的表达模式仅限于发育特定阶段的特定器官。在花发育早期,在花中,尤其是花粉粒中检测到最高水平的GUS活性。授粉后,在其余花器官中检测到的GUS活性水平低得多,但子房瓣膜除外,其在整个花发育过程中均未染色。在发育中的种子和成熟种子中也检测到GUS活性。在根中,GUS表达主要在侧根起始部位和主根与次生根的连接处检测到。在根尖分生组织中未检测到GUS活性。在主茎和次茎的连接处也观察到GUS表达。总体而言,FPS2:GUS的表达模式表明FPS2在合成具有特殊功能的特定类异戊二烯中起作用。在转染原生质体和转基因拟南芥植株中对功能性FPS2基因启动子进行缺失分析表明,建立FPS2基因完整表达模式所需的所有顺式作用元件都包含在从-111至+65位延伸的短区域内。讨论了该区域内特定序列的潜在调控作用。