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Investigation of genetically modified (GM) crops reducing nitrate content

减少硝酸盐含量的转基因 (GM) 作物的研究

基本信息

批准号：
12660009
负责人：
OGAWA Kyoko
金额：
$ 2.18万
依托单位：
JAPAN WOMEN'S UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12660009/
关键词：
Nitrate reductase Nitrite reductase Genetically modified crops Accumulation of nitrate Metohaemoglobinaemia

项目摘要

Nitrate is a major source of inorganic nitrogen utilized by higher plants, and vegetables accumulate high levels of nitrate. Nitrate is not toxic, but when reduced to nitrite during digestion, the ion can oxidize haemoglobin to methaemoglobin, causing methaemoglobinemia. This is important to us.The purpose of the investigation presented here is to introduce genes related to nitrate assimilation and to create a vegetable reducing nitrate content.(1) Analysis of transformants of nitrate reductase(NR)-deficient mutants introducing NR geneIn biochemical and molecular biological analysis of transformants, they showed overexpression of NR mRNA and high level of NR activity. Therefore, it would be expect to reduce nitrate content and to increase protein content.(2) Assay methods of concentration of nitrate and nitrite ionsWe examined a possible to utilize capillary electrophoresis for assay of nitrate and nitrite contents. Our results indicated that nitrate ion was able to utilize capillary electrophoresis, but nitrite ion was unable to utilize it. Accordingly nitrite content was determined by previous method.(3) Introduction of NR gene to lettuce and spinachIn lettuce, method of plant regeneration has already established. On the other hand in this study we could get regenerated plants of spinach. Transformation method employing Agrobacterium was used to transfer NR gene into lettuce and spinach. Nitrate and nitrite contents were assayed in transformants producing.

硝酸盐是高等植物利用无机氮的主要来源，蔬菜中硝酸盐含量较高。硝酸盐是无毒的，但在消化过程中被还原为亚硝酸盐时，离子会将血红蛋白氧化为高血红蛋白，引起高血红蛋白血症。这对我们很重要。本研究的目的是引入与硝酸盐同化有关的基因，并创造一种降低硝酸盐含量的蔬菜。(1)引入NR基因的硝酸还原酶（NR）缺陷突变体的转化子分析在转化子的生化和分子生物学分析中，它们表现出NR mRNA的过表达和高水平的NR活性。因此，有望降低硝酸盐含量，提高蛋白质含量。(2)硝酸盐和亚硝酸盐离子浓度的测定方法研究了毛细管电泳法测定硝酸盐和亚硝酸盐含量的可能性。结果表明，硝酸盐离子可以利用毛细管电泳，而亚硝酸盐离子不能利用毛细管电泳。据此，用上述方法测定亚硝酸盐含量。(3) NR基因导入莴苣和菠菜在莴苣中，已经建立了植株再生的方法。另一方面，在这项研究中，我们可以得到再生的菠菜植株。采用农杆菌转化的方法将NR基因转入生菜和菠菜中。测定了变压器生产过程中硝酸盐和亚硝酸盐的含量。