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Study on enhancement of wide-range stress tolerance by rhizosphere control on moleculartolerance by rhizosphere control on molecular base.

根际分子调控增强大范围胁迫耐受性的研究

基本信息

批准号：
13460029
负责人：
OSAKI Mitsuru
金额：
$ 8万
依托单位：
Hokkaido 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 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13460029/
关键词：
rhizosphere control low pH A1 tolerance low P tolerance root exudation mycorrhiza haypae 広域ストレス根圏環境制御 PEPC 酸性フォスファターゼフィターゼ形質転換プロモーター酸性フォスターゼ

项目摘要

Acid and alkaline soils occupy over half of arable land in the world. Plant growth is inhibited in these soils because of the insolubilization of nutrients and toxicity of inorganic compounds such as A1 and heavy metals. Recently, it is indicated that the plant roots have some strategy to improve the rhizosphere environment (about several μm from root surface). We have studied to increase the capacity of function for rhizosphere regulation. The results of this study are summarized as follows;1) A cDNA for LASAP2 encoding acid phosphatase secreted from white lupin roots was transformed into tobacco plants. Genomic DNA containing promoter region was transformed into rice plants. These plants were able to secrete the acid phosphatase which had mostly same properties with native mature protein of white lupin roots.2) To introduce the PEPC (phosphoenolpyruvate carboxylase) gene into rice plants, some gene constructs for transformation using Agrobacterium were designed. For the efficient expression of PEPC, a transgenic rice line containing PEPC gene under regulation of actin promoter was selected. Novel gene for PEPC was isolated from rice plant, and the function was analyzed. The novel PEPC was C3 type and specifically induced in leaves. The identity was about 80% with root type PEPC. The A1 and low P tolerances were improved in PEPC transgenic rice line because overexpression of mRNA and protein for PEPC causes the increase of oxalate secretion from roots.3) Effect of root exudates of P-deficient onion on hyphal growth of arbuscular mycorrhizal fungi were studied. A1 fraction (Rf:0.47) was isolated by TLC from ethyl acetate fraction of onion root exudates. Spores of Gigaspora margarita were incubated in this fraction and deionized water (control) for 7 days. Length of external hyphae and number of 2nd or higher order branches were higher in A1 fraction than in control.

酸性和碱性土壤占据了世界上一半以上的可耕地。植物生长受到抑制，在这些土壤中，因为养分的不溶性和毒性的无机化合物，如铝和重金属。近年来的研究表明，植物根系具有改善根际环境（距根表几μ m）的策略。我们研究了提高根际调节功能的能力。本研究的主要结果如下：1）将编码白色羽扇豆根分泌酸性磷酸酶的LASAP 2基因的cDNA转化烟草植株。将含有启动子区的基因组DNA转化到水稻植株中。2）为了将PEPC（phosphoenolpyruvate carboxylase，磷酸烯醇式丙酮酸羧化酶）基因导入水稻，设计了几种适合农杆菌介导的基因转化载体。为了使PEPC基因在水稻中得到高效表达，我们筛选了一个含有肌动蛋白启动子调控的PEPC基因的转基因水稻品系。从水稻植株中克隆了一个新的PEPC基因，并对其功能进行了分析。新的PEPC为C3型，在叶片中特异诱导。与根型PEPC的同源性约为80%。转PEPC基因水稻对A1和低磷的耐受性得到提高，这是由于PEPC基因mRNA和蛋白的过量表达导致根系草酸分泌增加。3）研究了缺磷洋葱根系分泌物对丛枝菌根真菌菌丝生长的影响。洋葱根分泌物乙酸乙酯部分经薄层层析分离得到A1组分（Rf：0.47）。将Gigaspora margarita的孢子在该级分和去离子水（对照）中孵育7天。A1组的菌丝长度和二级以上分枝数均高于对照组。