The Chloroplast Function Database: a large-scale collection of Arabidopsis Ds/Spm- or T-DNA-tagged homozygous lines for nuclear-encoded chloroplast proteins, and their systematic phenotype analysis
The Chloroplast Function Database: a large-scale collection of Arabidopsis Ds/Spm- or T-DNA-tagged homozygous lines for nuclear-encoded chloroplast proteins, and their systematic phenotype analysis
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DOI：
10.1111/j.1365-313x.2009.04074.x
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发表时间：
2010-02-01
期刊：
影响因子：
7.2
通讯作者：
Shinozaki, Kazuo
中科院分区：
文献类型：
--
作者：
Myouga, Fumiyoshi;Akiyama, Kenji;Shinozaki, Kazuo
叶绿体的大部分蛋白质由核基因组编码，并在翻译后靶向叶绿体。从国际种子库存中心和我们自己的库存的标记插入品系数据库中，我们选择了 3246 个 Ds/Spm（解离子/抑制突变子）转座子或 T-DNA 标记的拟南芥品系，用于编码 1369 个叶绿体蛋白（约占 2090 个预测叶绿体蛋白的 66%）的基因，其中插入破坏了蛋白编码区。我们系统地观察了琼脂平板上生长的 3 周龄幼苗，鉴定了具有异常表型的突变体，并收集了具有野生型表型的纯合品系。我们还鉴定了未获得纯合植物的插入系。迄今为止，我们已经鉴定出 111 个具有可再现幼苗表型的品系，122 个我们无法获得纯合子的品系，以及 1290 个没有可见表型的纯合品系。叶绿体功能数据库提供了从该资源获得的分子和表型信息。该数据库提供了使用拟南芥基因组计划 (AGI) 基因座编号、标记品系编号和表型搜索突变品系的工具，并提供对标记品系资源详细信息的快速访问。此外，我们收集的插入纯合子提供了一个强大的工具来加速拟南芥中核编码叶绿体蛋白的功能分析。叶绿体功能数据库可在 http://rarge.psc.riken.jp/chromoplast/ 上免费获取。该项目中产生的纯合品系也可从各个拟南芥库存中心获得。我们已将插入纯合子捐赠给其原始种子储存中心。
A majority of the proteins of the chloroplast are encoded by the nuclear genome, and are post-translationally targeted to the chloroplast. From databases of tagged insertion lines at international seed stock centers and our own stock, we selected 3246 Ds/Spm (dissociator/suppressor-mutator) transposon- or T-DNA-tagged Arabidopsis lines for genes encoding 1369 chloroplast proteins (about 66% of the 2090 predicted chloroplast proteins) in which insertions disrupt the protein-coding regions. We systematically observed 3-week-old seedlings grown on agar plates, identified mutants with abnormal phenotypes and collected homozygous lines with wild-type phenotypes. We also identified insertion lines for which no homozygous plants were obtained. To date, we have identified 111 lines with reproducible seedling phenotypes, 122 lines for which we could not obtain homozygotes and 1290 homozygous lines without a visible phenotype. The Chloroplast Function Database presents the molecular and phenotypic information obtained from this resource. The database provides tools for searching for mutant lines using Arabidopsis Genome Initiative (AGI) locus numbers, tagged line numbers and phenotypes, and provides rapid access to detailed information on the tagged line resources. Moreover, our collection of insertion homozygotes provides a powerful tool to accelerate the functional analysis of nuclear-encoded chloroplast proteins in Arabidopsis. The Chloroplast Function Database is freely available at http://rarge.psc.riken.jp/chloroplast/. The homozygous lines generated in this project are also available from the various Arabidopsis stock centers. We have donated the insertion homozygotes to their originating seed stock centers.