Sacadura Nuno T, Saville Barry J
Department of Botany, University of Toronto at Mississauga, 3359 Mississauga Road North, Mississauga, Ont., Canada L5L 1C6.
Fungal Genet Biol. 2003 Oct;40(1):47-64. doi: 10.1016/s1087-1845(03)00078-1.
Ustilago maydis grows in its host Zea mays eliciting the formation of obvious tumors that are full of black teliospores. Teliospores are thick-walled, dormant, diploid cells that have evolved for dispersal and survival of the pathogen. Their germination leads to new rounds of infection and is temporally linked to meiosis. We are investigating gene expression during teliospore germination to gain insight into the control of this process. Here we identify genes expressed through creation of an expressed sequence tag (EST) library. We generated 2871 ESTs that are assembled into 1293 contiguous sequences. Based upon a blast search similarity cutoff of E < or =10(-5) 38% of all contigs were orphans while 62% showed similarity to sequences in the protein database. Analyses of blast searches were used to functionally classify genes. Northern hybridizations using specific cDNA clones reveal a relative level of expression consistent with the number of sequences per contig. Identified genes and expression information provide a base for genome annotation of U. maydis and further investigation of teliospore germination and pathogenesis.
玉蜀黍黑粉菌在其寄主玉米中生长,引发明显肿瘤的形成,这些肿瘤充满黑色冬孢子。冬孢子是厚壁、休眠的二倍体细胞,其进化目的是实现病原体的传播和存活。它们的萌发会引发新一轮感染,并且在时间上与减数分裂相关联。我们正在研究冬孢子萌发过程中的基因表达,以深入了解这一过程的调控机制。在此,我们通过创建一个表达序列标签(EST)文库来鉴定表达的基因。我们生成了2871个EST,这些EST被组装成1293个连续序列。基于E≤10^(-5)的Blast搜索相似性阈值,所有重叠群中有38%是孤儿序列,而62%与蛋白质数据库中的序列具有相似性。利用Blast搜索分析对基因进行功能分类。使用特定cDNA克隆进行的Northern杂交揭示了与每个重叠群中序列数量一致的相对表达水平。鉴定出的基因和表达信息为玉蜀黍黑粉菌的基因组注释以及冬孢子萌发和致病机制的进一步研究提供了基础。