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

基于基因组分析的基因鉴定新方法及其在植物分子育种中的应用。

• 批准号: 10554048
• 负责人: IIDA Shigeru
• 金额: $ 8.51万
• 依托单位: National Institute for Basic Biology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10554048/
• 关键词: anthocyanin biosynthesis gene; carnation; floricultural plants; gene identification; genome analysis; molecular breeding; the common morning glory; the Japanese morning glory

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.

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。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。转座元件与这些基因的整合导致了色素沉着表型的改变。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。

项目成果

Y.Yoshimoto: "Isolation and Characterization of a cDNA for phenylalanine ammonia-lyase (PAL) from Dianthus caryophyllus (carnation)"Plant Biotech.. 17. 325 (2000)

Y.Yoshimoto：“石竹（康乃馨）苯丙氨酸解氨酶 (PAL) cDNA 的分离和表征”植物生物技术.. 17. 325 (2000)

S. Takahashi: "Capturing of a genomic HMG domain sequence by an En/Spm related transposable element Tpn1 in the Japanese morning glory"Mol. Gen. Genet.. 261. 447-451 (1999)

S. Takahashi：“通过日本牵牛花中的 En/Spm 相关转座元件 Tpn1 捕获基因组 HMG 结构域序列”Mol。

H.Fujiwara: "cDNA cloning,gene expression and subcellular localization of anthocyanin 5-aromatic acyltransferase from Gentiana triflora" Plant J.16. 421-431 (1998)

H.Fujiwara：“三花龙胆花青素5-芳香酰基转移酶的cDNA克隆、基因表达和亚细胞定位”Plant J.16。

Y.Yoshimoto: "Isolation and Characterization of a cDNA for phenylalanine ammonia-lyase (PAL) from Dianthus caryophyllus (carnation)"Plant Biotech.. 17. 325-329 (2000)

Y.Yoshimoto：“石竹（康乃馨）中苯丙氨酸解氨酶（PAL）的 cDNA 的分离和表征”植物生物技术.. 17. 325-329 (2000)

S.Fukada-Tanaka, et al.: "Colour-enhancing protein in blue petals."Nature. 407. 581 (2000)

S.Fukada-Tanaka 等人：“蓝色花瓣中的增色蛋白”。《自然》。