Flower color modification of gentian plants by RNAi-mediated gene silencing
Flower color modification of gentian plants by RNAi-mediated gene silencing
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DOI：
10.5511/plantbiotechnology.25.61
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发表时间：
2008-01-01
影响因子：
1.6
通讯作者：
Nishihara, Masahiro
中科院分区：
文献类型：
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作者：
Nakatsuka, Takashi;Mishibaa, Kei-Ichiro;Nishihara, Masahiro
与反义和有义抑制相比，RNA 干扰 (RNAi) 是一种高效且强大的基因沉默技术。在此，我们报告了采用 RNAi 技术来改变龙胆花颜色的技术，其目标是抑制三种花青素生物合成基因；查耳酮合酶 (CHS)、花青素合酶 (ANS) 和类黄酮 3',5'-羟化酶 (F3'5'H)。 CHS基因被抑制的转基因龙胆植物的花瓣呈现纯白色至淡蓝色，而ANS基因被抑制的转基因龙胆植物的花瓣仅呈现淡蓝色。 F3'5'H基因的抑制减少了飞燕草素衍生物并增加了花青素衍生物，并导致洋红色的花色。 Northern印迹分析证实，所有表现出典型表型的转基因龙胆植物都强烈抑制了目标基因的转录，这与花瓣中花青素积累和组成的变化相对应。一些 rolCpro-CHSir 转基因龙胆表现出双色表型，沿维管束的花青素积累减少。这些数据表明，RNAi对花青素生物合成基因的抑制已成功应用于龙胆植物以改变花色，这可用于设计新颖的花色和图案。本研究产生的转基因龙胆植物可能在不久的将来作为龙胆植物育种的优良材料。
RNA interference (RNAi) is an efficient and powerful technique for gene silencing compared with antisense and sense suppression. Here we report adaptation of RNAi technology to modify flower colors in gentian, targeted for suppression of three anthocyanin biosynthetic genes; chalcone synthase (CHS), anthocyanidin synthase (ANS) and flavonoid 3',5'-hydroxylase (F3'5'H). The petals of transgenic gentian plants with a suppressed CHS gene exhibited pure white to pale-blue color, while those with a suppressed ANS gene showed only pale-blue. The suppression of the F3'5'H gene decreased delphinidin derivatives and increased cyanidin derivatives, and led to magenta flower colors. Northern blot analyses confirmed that all transgenic gentian plants showing typical phenotypes had strongly suppressed transcriptions of the targeted genes, corresponding with a change in anthocyanin accumulation and composition in their petals. Some rolCpro-CHSir transgenic gentians exhibited bicolor phenotypes with reduced anthocyanin accumulation along the vascular bundles. These data demonstrated that the suppression of anthocyanin biosynthetic genes by RNAi was successfully applied to gentian plants to change flower color, and this could be useful for designing novel flower color and patterns. Transgenic gentian plants produced in this study might be utilized as elite materials in the breeding of gentian plants in the near future.