Narita J O, Gruissem W
Department of Botany, University of California, Berkeley 94720.
Plant Cell. 1989 Feb;1(2):181-90. doi: 10.1105/tpc.1.2.181.
The activity of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR) and the level of its mRNA have been determined at various stages of tomato fruit development. The HMGR reaction makes mevalonate, a necessary component in the synthesis of all isoprene containing compounds, such as sterols and carotenoids. A cDNA clone encoding the active site region of HMGR has been isolated from a tomato library derived from young-fruit mRNA. The clone hybridizes to a one- or two-copy fragment in high-stringency DNA gel blot analyses and detects an mRNA of approximately 3.0 kb. Both HMGR activity and mRNA levels are high in early stages of tomato fruit development, when rapid cell division occurs, as well as in the subsequent early stages of cellular expansion. In contrast, ripening fruit have very low levels of reductase activity and mRNA, even though large amounts of the carotenoid lycopene are synthesized during this period. Furthermore, in vivo inhibition of HMGR during early fruit stages disrupts subsequent development, whereas inhibition during later stages of fruit expansion has no apparent effect on ripening. We conclude that the pool of mevalonate responsible for the synthesis of phytosterols is synthesized primarily during the first half of tomato fruit development. In addition, the final period of fruit expansion and ripening is not dependent upon HMGR activity, but instead utilizes a preexisting pool of pathway intermediates or requires the use of salvage pathways in the cell.
已在番茄果实发育的各个阶段测定了3-羟基-3-甲基戊二酰辅酶A还原酶(HMGR)的活性及其mRNA水平。HMGR反应生成甲羟戊酸,甲羟戊酸是合成所有含异戊二烯化合物(如甾醇和类胡萝卜素)的必需成分。已从一个源自幼果mRNA的番茄文库中分离出一个编码HMGR活性位点区域的cDNA克隆。在高严谨度DNA凝胶印迹分析中,该克隆与一个或两个拷贝的片段杂交,并检测到一条约3.0 kb的mRNA。在番茄果实发育早期,即细胞快速分裂发生时,以及随后的细胞膨大早期,HMGR活性和mRNA水平都很高。相比之下,成熟果实的还原酶活性和mRNA水平非常低,尽管在此期间大量合成了类胡萝卜素番茄红素。此外,在果实发育早期对HMGR进行体内抑制会扰乱后续发育,而在果实膨大后期进行抑制对成熟没有明显影响。我们得出结论,负责植物甾醇合成的甲羟戊酸池主要在番茄果实发育的前半期合成。此外,果实膨大与成熟的最后阶段并不依赖于HMGR活性,而是利用预先存在的途径中间产物池,或者需要利用细胞中的补救途径。