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

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。

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.