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Analysis of Function and Agricultural Application of Plant Aldehyde oxidase using Transgenic Plants

利用转基因植物分析植物醛氧化酶的功能及农业应用

基本信息

批准号：
10559017
负责人：
KOSHIDA Tomokazu
金额：
$ 1.47万
依托单位：
TOKYO METOROPOLITAN UNIVERSITY
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 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10559017/
关键词：
abscisic acid auxin aldehyde oxidase Arabidopsic thaliana cDNA cloning transgenic plant 遺伝子組み換え植物

项目摘要

Aldehyde oxidase (AO) has been extensively investigated in animals and microorganisms. In plants, however, there was only a limited information on this enzyme. A great interest in this enzyme has been in its possible involvement in indole-3-acetic acid (IAA) and abscisic acid (ABA) biosynthesis. In the present study, we cloned four full-length AO cDNAs from Arabidopsis thaliana and investigate their physiological functions using transgenic plants with these genes. We also tried to develop these results to agricultural application.(1) Four different putative AO cDNA clones were obtained from A. thaliana. Their products were analyzed by using antibodies each specific to four AOs in A. thaliana.(2) The four AO cDNAs were introduced into yeast Pichia pastoris and the products were functionally expressed in the yeast cells. The result revealed that two AO genes encoded three AOs in A. thaliana seedlings and another AO gene produced a leaf AO. Biochemical studies using AOs expressed in yeast indicated that one of seedling AO might be involved in IAA biosynthesis and a leaf AO is a most possible candidate of an AO that catalyzes oxidation of abscisic aldehyde to form ABA.(3) The four AO cDNA clones were introduced into A. thaliana and some over expressed and suppressed lines were obtained for each DNA construct.(4) We also succeeded screening several knockout lines with these genes.

醛氧化酶（AO）在动物和微生物中已被广泛研究。然而，在植物中，关于这种酶的信息非常有限。对这种酶的极大兴趣在于其可能参与吲哚-3-乙酸（IAA）和脱落酸（阿坝）的生物合成。在本研究中，我们从拟南芥中克隆了四个全长AO cDNA，并利用这些基因的转基因植物研究其生理功能。我们还尝试将这些结果推广到农业应用中。(1)从A. thaliana.通过使用各自特异于A. thaliana. (2)将这四个AO cDNA导入毕赤酵母中，并在酵母细胞中进行功能性表达。结果表明，两个AO基因编码A.拟南芥幼苗和另一个AO基因产生了叶片AO。利用在酵母中表达的AO进行的生化研究表明，幼苗AO可能参与IAA的生物合成，叶片AO是催化脱落醛氧化形成阿坝的AO的最有可能的候选者。(3)将这4个AO cDNA克隆导入A.和一些过表达和抑制的株系。(4)我们还成功地筛选出了几个具有这些基因的敲除系。