Verwaaijen Bart, Wibberg Daniel, Kröber Magdalena, Winkler Anika, Zrenner Rita, Bednarz Hanna, Niehaus Karsten, Grosch Rita, Pühler Alfred, Schlüter Andreas
Center for Biotechnology, Bielefeld University, Bielefeld, Germany.
Leibniz-Institute of Vegetable and Ornamental Crops (IGZ), Großbeeren, Germany.
PLoS One. 2017 May 9;12(5):e0177278. doi: 10.1371/journal.pone.0177278. eCollection 2017.
The necrotrophic pathogen Rhizoctonia solani is one of the most economically important soil-borne pathogens of crop plants. Isolates of R. solani AG1-IB are the major pathogens responsible for bottom-rot of lettuce (Lactuca sativa L.) and are also responsible for diseases in other plant species. Currently, there is lack of information regarding the molecular responses in R. solani during the pathogenic interaction between the necrotrophic soil-borne pathogen and its host plant. The genome of R. solani AG1-IB (isolate 7/3/14) was recently established to obtain insights into its putative pathogenicity determinants. In this study, the transcriptional activity of R. solani AG1-IB was followed during the course of its pathogenic interaction with the host plant lettuce under controlled conditions. Based on visual observations, three distinct pathogen-host interaction zones on lettuce leaves were defined which covered different phases of disease progression on tissue inoculated with the AG1-IB (isolate 7/3/14). The zones were defined as: Zone 1-symptomless, Zone 2-light brown discoloration, and Zone 3-dark brown, necrotic lesions. Differences in R. solani hyphae structure in these three zones were investigated by microscopic observation. Transcriptional activity within these three interaction zones was used to represent the course of R. solani disease progression applying high-throughput RNA sequencing (RNA-Seq) analysis of samples collected from each Zone. The resulting three transcriptome data sets were analyzed for their highest expressed genes and for differentially transcribed genes between the respective interaction zones. Among the highest expressed genes was a group of not previously described genes which were transcribed exclusively during early stages of interaction, in Zones 1 and 2. Previously described importance of up-regulation in R. solani agglutinin genes during disease progression could be further confirmed; here, the corresponding genes exhibited extremely high transcription levels. Most differentially higher expressed transcripts were found within Zone 2. In Zone 3, the zone with the strongest degree of interaction, gene transcripts indicative of apoptotic activity were highly abundant. The transcriptome data presented in this work support previous models of the disease and interaction cycle of R. solani and lettuce and may influence effective techniques for control of this pathogen.
坏死营养型病原菌立枯丝核菌是农作物中最具经济重要性的土传病原菌之一。立枯丝核菌AG1-IB分离株是导致生菜(Lactuca sativa L.)底部腐烂的主要病原菌,也会引发其他植物物种的病害。目前,关于这种坏死营养型土传病原菌与其寄主植物之间致病相互作用过程中立枯丝核菌的分子反应,我们还缺乏相关信息。立枯丝核菌AG1-IB(分离株7/3/14)的基因组最近已被确定,以便深入了解其假定的致病决定因素。在本研究中,在可控条件下,观察了立枯丝核菌AG1-IB与寄主植物生菜致病相互作用过程中的转录活性。基于肉眼观察,在生菜叶片上定义了三个不同的病原菌-寄主相互作用区域,这些区域涵盖了接种AG1-IB(分离株7/3/14)的组织上不同的病害进展阶段。这些区域被定义为:区域1无症状,区域2浅褐色变色,区域3深褐色坏死病斑。通过显微镜观察研究了这三个区域中立枯丝核菌菌丝结构的差异。利用高通量RNA测序(RNA-Seq)分析从每个区域收集的样本,以这三个相互作用区域内的转录活性来代表立枯丝核菌病害进展过程。对得到的三个转录组数据集分析了其表达量最高的基因以及各个相互作用区域之间差异转录的基因。在表达量最高的基因中,有一组此前未被描述的基因,它们仅在相互作用的早期阶段,即在区域1和区域2中转录。此前描述的立枯丝核菌凝集素基因在病害进展过程中上调的重要性在此得到了进一步证实;在这里,相应基因表现出极高的转录水平。大多数差异表达较高的转录本出现在区域2中。在相互作用程度最强的区域3中,指示凋亡活性的基因转录本高度丰富。本研究中呈现的转录组数据支持了立枯丝核菌与生菜病害及相互作用周期的先前模型,并可能影响控制这种病原菌的有效技术。