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

小麦叶绿体DNA的全序列测定已经完成。根据小麦叶绿体测序数据，筛选出110个叶绿体基因。对这110个基因进行PCR扩增后，将这些基因以及25个与光合作用相关的核编码基因点在玻片上，组成叶绿体基因DNA微阵列。利用叶绿体基因芯片，监测了小麦生活史中13个不同组织中含有这种分化的叶绿体类型的全套叶绿体基因的稳态水平。为了评估小麦生活史中基因表达模式的全球变化，我们对普通小麦的表达序列标签(EST)进行了大规模的分析。总共从cDNA的两端获得了116232个序列。这些序列被分成25971个重叠群。为了研究幼穗和种子发育过程中的基因表达谱，利用YD阵列发现了3260个在幼穗中表达的EST，构建了包含2607个幼穗全长cDNA的YL阵列。最后，为了评估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个寡核苷酸基因芯片。通过这个项目，小麦研究进入了开展功能基因组学的时代。

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.