Unité Mixte de Recherche 1136 'Interactions Arbres/Microorganismes', INRA (Institut National de la Recherche Agronomique)/Université de Lorraine, Centre INRA de Nancy, Champenoux, France.
PLoS One. 2012;7(8):e44408. doi: 10.1371/journal.pone.0044408. Epub 2012 Aug 30.
Biotroph pathogens establish intimate interactions with their hosts that are conditioned by the successful secretion of effectors in infected tissues and subsequent manipulation of host physiology. The identification of early-expressed pathogen effectors and early-modulated host functions is currently a major goal to understand the molecular basis of biotrophy. Here, we report the 454-pyrosequencing transcriptome analysis of early stages of poplar leaf colonization by the rust fungus Melampsora larici-populina. Among the 841,301 reads considered for analysis, 616,879 and 649 were successfully mapped to Populus trichocarpa and M. larici-populina genome sequences, respectively. From a methodological aspect, these results indicate that this single approach is not appropriate to saturate poplar transcriptome and to follow transcript accumulation of the pathogen. We identified 19 pathogen transcripts encoding early-expressed small-secreted proteins representing candidate effectors of interest for forthcoming studies. Poplar RNA-Seq data were validated by oligoarrays and quantitatively analysed, which revealed a highly stable transcriptome with a single transcript encoding a sulfate transporter (herein named PtSultr3;5, POPTR_0006s16150) showing a dramatic increase upon colonization by either virulent or avirulent M. larici-populina strains. Perspectives connecting host sulfate transport and biotrophic lifestyle are discussed.
生物寄生菌与宿主建立密切的相互作用,这取决于在感染组织中成功分泌效应子,以及随后对宿主生理机能的操纵。鉴定早期表达的病原体效应子和早期调节的宿主功能,是理解生物寄生现象分子基础的主要目标。在这里,我们报告了杨树叶被锈菌 Melampsora larici-populina 早期定殖的 454 焦磷酸测序转录组分析。在考虑进行分析的 841,301 个读段中,分别有 616,879 和 649 个被成功映射到杨属基因组序列和 M. larici-populina 基因组序列。从方法学的角度来看,这些结果表明,这种单一方法并不适合饱和杨属转录组,也不适合跟踪病原体转录物的积累。我们鉴定了 19 个病原体转录本,这些转录本编码早期表达的小分泌蛋白,代表了即将进行的研究中感兴趣的候选效应子。通过寡核苷酸微阵列对杨树 RNA-Seq 数据进行了验证和定量分析,结果显示转录组高度稳定,只有一个编码硫酸盐转运蛋白(命名为 PtSultr3;5,POPTR_0006s16150)的转录本在受到强毒或弱毒 M. larici-populina 菌株侵染时显著增加。本文还讨论了连接宿主硫酸盐转运和生物寄生生活方式的观点。