利用黑斑病的高抗无性系美洲黑杨I-69及黑斑病的高感无性系欧美杨I-45为材料建立的2个cDNA文库,随机挑取cDNA克隆进行5'端EST序列测序,共获得有效序列20 023条.序列经聚类分析和拼接后,共获得10 816个Unigene,其中包括3 734个Contig,7 082个独立的Singleton.被注释的8 853个具有同源性匹配序列基因中,按照GO的分子功能、生物过程和细胞组分3个不同分类角度进行分类.在具有功能注释的8 853个Unigene中,选出330个与完成全基因组测序的毛果杨序列进行BLAST分析,结果发现有177个抗病相关候选基因出现在282个Unigene中,其中135个分布于杨树的18个不同连锁群上,其他42个基因位于还没定位的scaffolds上.所测定的这些EST序列为后期在基因组水平上研究杨树黑斑病的水平抗性遗传机制及进一步的相关基因发现奠定基础.%In an attempt to elucidate the molecular mechanism for resistance of black spot disease in poplar, gene expression profiles in leaves of Populus deltoides 'Lus' ( 1-69/55 ) and P.euramericana ' Ⅰ- 45/51 ' , which were inoculated with the pathogen Marssonina brunnea f.sp.brunnea, were analyzed based on expressed sequence tags (ESTs).A total of 20 023 valid ESTs from the 5' terminal ends derived from corresponding cDNA libraries of the two poplar species were sequenced.Cluster analysis of the 20 023 sequences yielded 10 816 tentative unigenes, including 3 734 contigs and 7 082 singletons.All tentative unigenes were classified by Gene Ontology functional categories.To find resistance-associated candidate genes and locate them on poplar genome, 330 unigenes was chosen from 8 853 annotated tentative unigenes, and their BLAST alignment was conducted with Populus trichocarpa assembly, 177 related candidate resistant genes were found, and they presented in 282 unigenes.Among these genes, there were 135 genes located on 18 different linkage groups of poplar genome, and 42 genes located on the different scaffolds.This study supplied a resource of candidate genes for further exploring the genetic mechanism for the host horizontal resistance to the pathogen Marssonina brunnea at the whole genome range, and provided important information for further gene discovery.
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