Morris Wayne L, Ducreux Laurence J M, Hedden Peter, Millam Steve, Taylor Mark A
Quality, Health and Nutrition, Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, UK.
J Exp Bot. 2006;57(12):3007-18. doi: 10.1093/jxb/erl061. Epub 2006 Jul 26.
Potato tubers were engineered to express a bacterial gene encoding 1-deoxy-D-xylulose 5-phosphate synthase (DXS) in order to investigate the effects of perturbation of isoprenoid biosynthesis. Twenty-four independent transgenic lines out of 38 generated produced tubers with significantly elongated shape that also exhibited an early tuber sprouting phenotype. Expression analysis of nine transgenic lines (four exhibiting the phenotype and five showing a wild-type phenotype) demonstrated that the phenotype was strongly associated with dxs expression. At harvest, apical bud growth had already commenced in dxs-expressing tubers whereas in control lines no bud growth was evident until dormancy was released after 56-70 d of storage. The initial phase of bud growth in dxs tubers was followed by a lag period of approximately 56 d, before further elongation of the developing sprouts could be detected. Thus dxs expression results in the separation of distinct phases in the dormancy and sprouting processes. In order to account for the sprouting phenotype, the levels of plastid-derived isoprenoid growth regulators were measured in transgenic and control tubers. The major difference measured was an increase in the level of trans-zeatin riboside in tubers at harvest expressing dxs. Additionally, compared with controls, in some dxs-expressing lines, tuber carotenoid content increased approximately 2-fold, with most of the increase accounted for by a 6-7-fold increase in phytoene.
为了研究类异戊二烯生物合成紊乱的影响,对马铃薯块茎进行基因工程改造,使其表达编码1-脱氧-D-木酮糖5-磷酸合酶(DXS)的细菌基因。在产生的38个独立转基因株系中,有24个产生了形状明显拉长的块茎,这些块茎还表现出早期块茎发芽的表型。对9个转基因株系(4个表现出该表型,5个表现出野生型表型)的表达分析表明,该表型与dxs表达密切相关。收获时,表达dxs的块茎顶端芽已经开始生长,而在对照株系中,直到储存56 - 70天后休眠解除,芽才明显生长。dxs块茎芽生长的初始阶段之后是大约56天的滞后期,之后才能检测到发育中的芽进一步伸长。因此,dxs表达导致休眠和发芽过程中出现不同阶段的分离。为了解释发芽表型,在转基因和对照块茎中测量了质体衍生的类异戊二烯生长调节剂的水平。测量的主要差异是收获时表达dxs的块茎中反式玉米素核苷水平增加。此外,与对照相比,在一些表达dxs的株系中,块茎类胡萝卜素含量增加了约2倍,其中大部分增加是由于八氢番茄红素增加了6 - 7倍。