Genetic manipulation of condensed tannins in higher plants II. Analysis of birdsfoot trefoil plants harboring antisense dihydroflavonol reductase constructs
Genetic manipulation of condensed tannins in higher plants II. Analysis of birdsfoot trefoil plants harboring antisense dihydroflavonol reductase constructs
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DOI：
10.1104/pp.116.3.1133
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发表时间：
1998-03-01
期刊：
影响因子：
7.4
通讯作者：
Morris, P
中科院分区：
文献类型：
--
作者：
Robbins, MP;Bavage, AD;Morris, P
我们已经生产并分析了含有反义二氢黄酮醇还原酶（AS-DFR）序列的转基因鸟足三叶草（Lotus corniculatus L.）植物。在最初的实验中，评估了将三种不同的反义金鱼草L.DFR构建体引入单一受体基因型(S50)的效果。对植物生物量没有明显影响，但一些反义品系的叶、茎和根组织中缩合单宁的水平显示出统计上的减少。还发现了转化事件，导致缩合单宁水平增加。在随后的实验中，使用 pMAJ2（包含 A. majus cDNA 5' 一半的反义构建体）在基因型 S33 中进行了 AS-DFR 表型的详细研究。在这种情况下，叶和茎组织以及幼枝组织中的单宁水平降低。对单宁下调枝条组织中可溶性黄酮类化合物和异黄酮类化合物的分析表明，几乎没有明显的默认产物。当两个 S33 AS-DFR 品系异型杂交时，后代植物中转基因序列的代表性不足，并且没有观察到反义表型遗传的例子。据我们所知，这是对高等植物中缩合单宁生物合成进行基因操作的第一份报告。
We have produced and analyzed transgenic birdsfoot trefoil (Lotus corniculatus L.) plants harboring antisense dihydroflavonol reductase (AS-DFR) sequences. In initial experiments the effect of introducing three different antisense Antirrhinum majus L. DFR constructs into a single recipient genotype (S50) was assessed. There were no obvious effects on plant biomass, but levels of condensed tannins showed a statistical reduction in leaf, stem, and root tissues of some of the antisense lines. Transformation events were also found, which resulted in increased levels of condensed tannins. In subsequent experiments a detailed study of AS-DFR phenotypes was carried out in genotype S33 using pMAJ2 (an antisense construct comprising the 5' half of the A. majus cDNA). In this case, reduced tannin levels were found in leaf and stem tissues and in juvenile shoot tissues. Analysis of soluble flavonoids and isoflavonoids in tannin down-regulated shoot tissues indicated few obvious default products. When two S33 AS-DFR lines were outcrossed, there was an underrepresentation of transgene sequences in progeny plants and no examples of inheritance of an antisense phenotype were observed. To our knowledge, this is the first report of the genetic manipulation of condensed tannin biosynthesis in higher plants.