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

开蓝色花的日本牵牛花(Ipomoea Nil)具有广泛的遗传和生理研究历史。已分离出一些与花和叶的颜色和形状有关的自发突变体。一些证据表明，与EN/SPM相关的转座元件Tpn1及其近亲是这些自发突变的主要来源。我们发展了一种新的方法来鉴定通过插入Tpn1及其近亲标记的基因，我们称之为简化转座子显示(STD)。随后，我们成功地利用了一个紫色突变等位基因，该等位基因赋予紫色花朵蓝色的斑点和扇形。经STD鉴定的紫色基因编码液泡Na/H交换器，导致液泡pH升高，导致蓝色花朵着色。我们还鉴定了康乃馨品种，并成功地在CHI和DFR SENE中发现了与Ac/DS相关的转座元件，用于花青素的生物合成。转座元件整合到这些基因中，导致色素沉着表型改变。我们还能够通过引入CHS或DFR的cDNA序列来培育出显示淡色花朵的新的香菇品种。

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.