Development of breeding system with functional genomics of wheat by using cDNA microarray

利用cDNA微阵列开发小麦功能基因组育种系统

• 批准号: 13356001
• 负责人: OGIHARA Yasunari
• 金额: $ 26.37万
• 依托单位: Kihara Institute for Biological Research, Yokohama City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 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-13356001/
• 关键词: Common wheat; cDNA library; Large scale analysis of ESTs; Clustering of ESTs; Body map; Selection of clones; Construction of cDNA microarray; Functional genomics for breeding; ボディーマップ; EST; cDNAマイクロアレー; トランスクリプトーム; 種子形成過程; 遺伝子発現

Entire sequencing of wheat chloroplast DNA had been completed. Based on the sequencing data of wheat plastome, 110 chloroplast genes were selected. After PCR amplification of these 110 genes, these genes as well as 25 nuclear encoded photosynthesis-related genes were spotted onto the slide glass to make up chloroplast gene DNA microarray. By using the chloroplast gene microarray, steady-state levels of the whole set of plastid genes from 13 different tissues in the wheat life cycle containing such differentiated plastid types werre monitored.To assess global changes in gene expression patterns in the wheat life cycle, we conducted large-scale analysis of expressed sequence tags (ESTs) in common wheat. In total, 116232 sequences were obtained from both ends of cDNAs. Those sequences were groupted into 25971 contigs. To examine gene expression profiles in the young spikelets and developing seeds, yd array spotted the 3260 ESTs expressed in the young spikelets, yl array containing the 2607 full length cDNAs from the young spikelets were constructed. Additionally, the cDNA maicroarray spotting 3364 cDNAs from developing seeds had been also prepared.Finally, to assess global changes in gene expression patterns of wheat, 22000 oligonucleotide cDNA microarray had been successfully constructed in collaboration with Agilent Technology Co..By this project, wheat research now comes of age to carry out functional genomics.

完成了小麦叶绿体DNA的全序列测定。根据小麦质体基因组测序数据，筛选出110个叶绿体基因。将这110个基因进行PCR扩增后，与25个核编码的光合作用相关基因一起点样到载玻片上，制成叶绿体基因DNA微阵列。利用叶绿体基因微阵列技术，对小麦生活史中13个不同组织的质体基因进行了稳态水平的检测，并对普通小麦的表达序列标签（EST）进行了大规模分析，以评估小麦生活史中基因表达模式的总体变化。从cDNA的两端共获得116232个序列。这些序列被分组为25971个重叠群。为了研究基因在幼穗和发育种子中的表达谱，yd阵列标记了幼穗中表达的3260个EST，构建了包含2607个全长cDNA的yl阵列。最后，为了研究小麦基因表达模式的全球变化，与安捷伦科技公司合作，成功构建了22000个寡核苷酸cDNA微阵列。通过这个项目，小麦研究现在已经成熟，开展功能基因组学。

项目成果

Y.Ogihara, K.Mochida, Y.Nemoto, K.Murai, Y.Yamazaki, T.Shin-I, Y.Kohara: "Correlated clustering and virtual display of gene expression patterns in the wheat life cycle by large-scale statistical analyses of expressed sequence tags"The Plant Journal. 33(6)

Y.Ogihara、K.Mochida、Y.Nemoto、K.Murai、Y.Yamazaki、T.Shin-I、Y.Kohara：“通过大规模统计对小麦生命周期中基因表达模式进行相关聚类和虚拟显示

Y.Ogihara, K.Mochida, Y.Nemoto, K.Murai, Y.Yamazaki, T.Shin-I, Y.Kohara: "Correlated clustering and virtual display of gene expression patterns in the wheat life cycle by large-scale statistical analyses of expressed sequence tags"Plant Journal. 33. 1001-

Y.Ogihara、K.Mochida、Y.Nemoto、K.Murai、Y.Yamazaki、T.Shin-I、Y.Kohara：“通过大规模统计对小麦生命周期中基因表达模式进行相关聚类和虚拟显示

Y.Ogihara, K.Isono, T.Kojima, A.Endo et al.: "Structural features of wheat plastome as revealed by complete sequencing of chloroplast DNA"Molecular and General Genetics. 266. 740-746 (2002)

Y.Ogihara、K.Isono、T.Kojima、A.Endo 等人：“通过叶绿体 DNA 的完整测序揭示小麦质体的结构特征”分子和普通遗传学。

K.Mochida, Y.Yamazaki, Y.Ogihara: "Discernment of homoeologous gene expression in hexaploid wheat by SNP analysis of contigs grouped from a large number of expressed sequence tags."Molecular Genetics and Genomics. 270. 371-377 (2003)

K.Mochida、Y.Yamazaki、Y.Ogihara：“通过对从大量表达序列标签中分组的重叠群进行 S​​NP 分析来辨别六倍体小麦中的同源基因表达。”分子遗传学和基因组学。

Y.Ogihara, K.Mochida, K.Kawaura, et al.: "Construction of a full-length cDNA library from young spikelets of common wheat and its characterization by large scale sequencing of expressed sequence tags."DNA Research. (submitted).

Y.Ogihara、K.Mochida、K.Kawaura 等人：“从普通小麦幼穗中构建全长 cDNA 文库，并通过表达序列标签的大规模测序来表征其特征。”DNA 研究。