Goldsbrough A., Belzile F., Yoder J. I.
Department of Vegetable Crops, University of California, Davis, Davis, California 95616.
Plant Physiol. 1994 Jun;105(2):491-496. doi: 10.1104/pp.105.2.491.
We isolated the dihydroflavonol 4-reductase (DFR) gene from tomato (Lycopersicon esculentum) using a previously characterized cDNA as probe. Earlier studies had indicated that the DFR gene is present in tomato as a single gene located on chromosome 2 near the locus anthocyanin without (aw). Mutant alleles of the aw locus result in the complete absence of anthocyanin pigmentation throughout all stages of plant development. When the genomic DFR clone was introduced by Agrobacterium-mediated transformation into plants bearing the aw mutation, primary transgenic seedlings accumulated anthocyanins that could be observed while the plants were still in tissue culture and which continued to be observed as the plants matured. Progeny of self pollinated and backcrossed transgenic plants segregated for anthocyanin pigmentation, and Southern hybridization analyses indicated the presence of the DFR transgene exclusively in those plants with pigmentation. These data indicate that the aw locus likely corresponds to the structural gene for DFR and that DFR can be used as a visual, nondestructive, plant-derived marker gene for tomato.
我们以一个先前已鉴定的cDNA为探针,从番茄(Lycopersicon esculentum)中分离出二氢黄酮醇4-还原酶(DFR)基因。早期研究表明,DFR基因在番茄中以单基因形式存在,位于2号染色体上靠近无花青素(aw)位点处。aw位点的突变等位基因导致植物发育的所有阶段完全没有花青素色素沉着。当通过农杆菌介导的转化将基因组DFR克隆导入携带aw突变的植物中时,初代转基因幼苗积累了花青素,在植物仍处于组织培养时就可以观察到,并且在植物成熟时仍可继续观察到。自花授粉和回交转基因植物的后代出现了花青素色素沉着的分离,Southern杂交分析表明DFR转基因仅存在于那些有色素沉着的植物中。这些数据表明,aw位点可能对应于DFR的结构基因,并且DFR可以用作番茄的一种可视化、非破坏性的植物源标记基因。