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水稻中脱落酸和赤霉素响应基因的转录谱分析:水稻与拟南芥的表型分析及比较

Transcriptional profiling of genes responsive to abscisic acid and gibberellin in rice: phenotyping and comparative analysis between rice and Arabidopsis.

作者信息

Yazaki Junshi, Shimatani Zenpei, Hashimoto Akiko, Nagata Yuko, Fujii Fumiko, Kojima Keiichi, Suzuki Kohji, Taya Toshiki, Tonouchi Mio, Nelson Charles, Nakagawa Allen, Otomo Yasuhiro, Murakami Kazuo, Matsubara Kenichi, Kawai Jun, Carninci Piero, Hayashizaki Yoshihide, Kikuchi Shoshi

机构信息

Department of Molecular Genetics, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan.

出版信息

Physiol Genomics. 2004 Apr 13;17(2):87-100. doi: 10.1152/physiolgenomics.00201.2003.

Abstract

We collected and completely sequenced 32,127 full-length complementary DNA clones from Oryza sativa L. ssp. japonica cv. "Nipponbare." Mapping of these clones to genomic DNA revealed approximately 20,500 transcriptional units (TUs) in the rice genome. For each TU, we selected 60-mers using an algorithm that took into account some DNA conditions such as base composition and sequence complexity. Using in situ synthesis technology, we constructed oligonucleotide arrays with these TUs on glass slides. We targeted RNAs prepared from normally grown rice callus and from callus treated with abscisic acid (ABA) or gibberellin (GA). We identified 200 ABA-responsive and 301 GA-responsive genes, many of which had never before been annotated as ABA or GA responsive in other expression analysis. Comparison of these genes revealed antagonistic regulation of almost all by both hormones; these had previously been annotated as being responsible for protein storage and defense against pathogens. Comparison of the cis-elements of genes responsive to one or antagonistic to both hormones revealed that the antagonistic genes had cis-elements related to ABA and GA responses. The genes responsive to only one hormone were rich in cis-elements that supported ABA and GA responses. In a search for the phenotypes of mutants in which a retrotransposon was inserted in these hormone-responsive genes, we identified phenotypes related to seed formation or plant height, including sterility, vivipary, and dwarfism. In comparison of cis-elements for hormone response genes between rice and Arabidopsis thaliana, we identified cis-elements for dehydration-stress response as Arabidopsis specific and for protein storage as rice specific.

摘要

我们从水稻(Oryza sativa L. ssp. japonica cv. “日本晴”)中收集了32,127个全长互补DNA克隆并进行了全序列测定。将这些克隆与基因组DNA进行比对,结果显示水稻基因组中大约有20,500个转录单元(TU)。对于每个转录单元,我们使用一种算法选择了60聚体,该算法考虑了一些DNA条件,如碱基组成和序列复杂性。利用原位合成技术,我们在载玻片上构建了含有这些转录单元的寡核苷酸阵列。我们将目标设定为从正常生长的水稻愈伤组织以及用脱落酸(ABA)或赤霉素(GA)处理过的愈伤组织中制备的RNA。我们鉴定出了200个对ABA有响应的基因和301个对GA有响应的基因,其中许多基因在其他表达分析中从未被注释为对ABA或GA有响应。对这些基因的比较显示,几乎所有基因都受到这两种激素的拮抗调控;这些基因先前被注释为负责蛋白质储存和抵御病原体。对响应一种激素或对两种激素都拮抗的基因的顺式元件进行比较,结果显示拮抗基因具有与ABA和GA响应相关的顺式元件。仅对一种激素有响应的基因富含支持ABA和GA响应的顺式元件。在寻找这些激素响应基因中插入了反转录转座子的突变体的表型时,我们鉴定出了与种子形成或株高相关的表型,包括不育、胎萌和矮化。在比较水稻和拟南芥中激素响应基因的顺式元件时,我们鉴定出脱水胁迫响应的顺式元件是拟南芥特有的,而蛋白质储存的顺式元件是水稻特有的。

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