New methods for gene identification based on genomic analysis and their application for molecular breeding in plants.

开蓝色花朵的日本牵牛花 (Ipomoea nil) 有着悠久的遗传和生理研究历史。许多与花和叶的颜色和形状相关的自发突变体已经被分离出来。多项证据表明，En/Spm 相关转座元件 Tpn1 及其相关元件是这些自发突变的主要来源。我们开发了一种新方法来识别通过插入 Tpn1 及其亲属标记的基因，我们将其命名为简化转座子展示 (STD)。随后，我们成功地应用了紫色可变等位基因，赋予带有蓝色斑点和扇区的紫色花朵。 STD 鉴定出的紫色基因被发现编码液泡 Na /H 交换剂，该交换剂负责增加液泡 pH 值以实现蓝色花的着色。我们还对康乃馨品种进行了表征，并成功地在 CHI 和 DFR 序列中识别出新的 Ac/Ds 相关转座元件，用于花青素生物合成。转座元件整合到这些基因中会导致色素沉着表型的改变。我们还能够通过引入 CHS 或 DFR cDNA 序列来生成显示浅色花朵的新蓝猪草品种。

The Japanese morning glory (Ipomoea nil) displaying blue flowers has an extensive history of genetic and physiological studies. A number of its spontaneous mutants related to the colors and shapes of the flowers and leaves have been isolated. Several lines of evidence indicate that an En/Spm-related transposable element Tpn1 and its relatives are major sources of these spontaneous mutations. We have developed a new method to identify a gene tagged by insertion of Tpn1 and its relatives, which we named Simplified Transposon Display (STD). Subsequently, we have successfully applied a purple-mutable allele conferring purple flowers with blue spots and sectors. The purple gene identified by STD was found to encode a vacuolar Na+/H+ exchanger that is responsible for increase vacuolar pH for blue flower coloration. We have also characterized carnation varieties and succeeded to identify new Ac/Ds-related transposable elements within the CHI and DFR senes for anthocyanin pigments biosynthesis. The integration of the transposable elements into these genes result in the altered pigmentation phenotypes. We were also able to generate new torenia varieties displaying pale flowers by introducing the CHS or DFR cDNA sequences.