权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Isolation and characterization of aluminum stress resistant genes using activati on tagging lines of Arabidopsis thaliana

使用拟南芥标签系激活分离和表征铝胁迫抗性基因

基本信息

批准号：
13660066
负责人：
EZAKI Bunichi
金额：
$ 2.18万
依托单位：
Okayama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
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/grant/KAKENHI-PROJECT-13660066/
关键词：
Resistant mechanism for Al stress Activation tagging line Al stress resistant gene Root hair Al toxicity Arabidopsis thaliana Root hair mutant Transgenic plant 形質転換体植物ストレス耐性 Aluminum(アルミニウム)ストレス酸化ストレス耐性遺伝子遺伝子操作 Arabidopsis(

项目摘要

To isolate new Al resistant genes, we performed a screening of resistant plants from a pool of Arabidopsis enhancer tagging lines. From approximately 16,000 lines, 20 clones were isolated as Al resistant candidates. Southern hybridization indicated that one line, #355-2, only carries single copy of the enhancer tag in chromosome DNA. A TAIL-PCR was performed for #355-2, and the result indicated that the tag is inserted in chromosome I (very closed to up-stream region of the F9E10.5 gene). RT-PCR was also carried out and we found that a higher gene expression of F9E10.5 and F9E10.6 was specifically detected in #355-2 line (approximately 8-and 7-folds, respectively) than control line (Col-0). DNA database analysts indicated that F9E10.6 encodes a DNA glycosylase, but the function of F9E10.5 is unknown.To characterize the A resistant mechanisms in #355-2 line, morphological observation of Col-0 and #355-2 was performed and we could see a clear difference in root-hair region. The length of root-hair of #355-2 was much shorter (approximately 1/3) than that of Col-o. Al treatment (300-500 μM, 2d) for these two lines indicated that their root-hairs were completely damaged by Al toxicity. Moreover, accumulation of Al in root region of #355-2 was smaller than that of Col-0. From these results, we speculated that root hair is one of the targets for Al attack and that the lower accumulation of Al ions in root region may be one of the Al resistant mechanisms in #355-2. These results also suggested that the Al influx is probably occurred not only in root-tip, but also in root-hair.We constructed two types of transgenic lines, over-expressing and suppressing of the F9E10.5 gene and determined their Al sensitivity and root-hair formation, but we suggested that F9E10.5 may not relate to these phenotype.

为了分离新的抗铝基因，我们从拟南芥增强子标签系中筛选抗性植物。从大约16，000个株系中，分离出20个克隆作为Al抗性候选物。Southern杂交结果表明，#355 - 2株系在染色体DNA中仅携带一个增强子标签拷贝。对#355 - 2进行TAIL-PCR，结果表明标签插入到染色体I中（非常接近F9E10.5基因的上游区域）。还进行了RT-PCR，我们发现在#355 - 2系中特异性地检测到F9E10.5和F9E10.6的基因表达（分别约为对照系（Col-0）的8倍和7倍）。DNA数据库分析表明，F9E10.6编码一种DNA糖基化酶，但F9E10.5的功能尚不清楚。#355 - 2的根毛长度比Col-o短得多（约1/3）。铝处理（300 - 500 μ M，2d）表明，这两个品系的根毛已被铝毒完全破坏。此外，355 - 2号根系中Al的积累量小于Col-0。根据这些结果，我们推测，根毛是铝攻击的目标之一，根区铝离子的低积累可能是#355 - 2的抗铝机制之一。我们构建了F9E10.5基因过表达和抑制两种类型的转基因株系，并对它们的铝敏感性和根毛形成进行了测定，但我们认为F9E10.5基因可能与这些表型无关。