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Studies about the improvement of soybean storage lipids by using the combination of induced mutation and gene transfer techniques.

利用诱导突变和基因转移技术相结合改良大豆贮藏脂质的研究。

基本信息

批准号：
13460005
负责人：
TAKAGI Yutaka
金额：
$ 8.77万
依托单位：
Saga University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13460005/
关键词：
soybean storage lipid fatty acid desaturase mutant gene transfer 脂肪酸遺伝子組換え不飽和化酵素 DGAT

项目摘要

In this study, we have attempted to develop a novel soybean germplasm which equips with both modified fatty acid composition and high oil productivity, and have obtained following results by using the combination1. The corresponding genes were identified in several high oleic acid mutants, and two of them (KK21 and M23) were clarified that mutations occurred at GmFAD2-I locus which encodes a microsomal ω-6 fatty acid desaturase. In addition, J18 and M5 of low linolenic acid mutants have mutations in GmFAD3-1a, and the corresponding gene of M24 is also clarified that the mutation occurs at another GmFAD3-1b locus, which encode microsomal ω-3 fatty acid desaturase genes.2. We found that the addition of 20mM pyroglutamic acid to culture meda drastically improved the embryogenesis efficiency from soybean immature embryo and the addition of 0.2% polypeptone S was also effective for the embryo growth. Furthermore, we have developed three transgenic soybean lines of Japanese cultivar, Kyushu 126, by introducing EGFP gene under those conditions.3. We have successfully developed the transgenic rice with GmFAD3-1a gene which have improved the α- linolenic acid content of seeds over 10-fold higher than that of non-transgenic rice, have developed a novel RNAi vector for plant transformation, and it was clarified that these techniques were effective for the improvement of the soybean lipid using the transgenic technology.4. It has successfully cloned the genes (cDNA and genomic DNA) encoding diacylglycerol acyl transferase (DGAT) which could control the lipig composition (TAG/DAG ratio) and oil yield.

在本研究中，我们试图开发一种新的大豆种质，该种质具有改良的脂肪酸组成和高油产量，并通过使用组合1获得了以下结果。在几个高油酸突变体中鉴定了相应的基因，其中两个突变体（KK 21和M23）被证实发生在编码微粒体ω-6脂肪酸去饱和酶的GmFAD 2-I位点。此外，低亚麻酸突变体J18和M5的GmFAD 3 - 1a基因也发生了突变，M24的相应基因也明确了突变发生在另一个GmFAD 3 - 1b位点，该位点编码微粒体ω-3脂肪酸去饱和酶基因.在培养基中添加20mM焦谷氨酸可显著提高大豆幼胚的胚状体发生效率，添加0.2%的聚蛋白胨S对胚状体的生长也有促进作用。在此基础上，我们成功地将绿色荧光蛋白基因导入日本品种九州126中，获得了三个转基因大豆品系.我们成功培育了转GmFAD3 - 1a基因水稻，其种子中α-亚麻酸含量比未转基因水稻提高了10倍以上，并开发了一种新的RNAi植物转化载体，阐明了这些技术在利用转基因技术改良大豆油脂中的有效性.本研究成功克隆了控制脂肪组成（TAG/DAG）和油脂产量的二酰基甘油酰基转移酶（DGAT）基因（cDNA和基因组DNA）。