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Transcriptome Analysis of Tomato Leaf Spot Pathogen Fusarium proliferatum: De novo Assembly Expression Profiling and Identification of Candidate Effectors

机译:番茄叶斑病病原菌镰刀菌转录组的转录组分析:从头组装表达谱和候选效应子的鉴定。

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摘要

Leaf spot disease caused by the fungus Fusarium proliferatum (Matsushima) Nirenberg is a destructive disease of tomato plants in China. Typical symptoms of infected tomato plants are softened and wilted stems and leaves, leading to the eventual death of the entire plant. In this study, we resorted to transcriptional profile analysis to gain insight into the repertoire of effectors involved in F. proliferatum–tomato interactions. A total of 61,544,598 clean reads were de novo assembled to provide a F. proliferatum reference transcriptome. From these, 75,044 unigenes were obtained, with 19.46% of the unigenes being assigned to 276 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, with 22.3% having a homology with genes from F. fujikuroi. A total of 18,075 differentially expressed genes (DEGs) were identified, 720 of which were found to code for secreted proteins. Of these, 184 were identified as candidate effectors, while 79.89% had an upregulated expression. Moreover, 17 genes that were differentially expressed in RNA-seq studies were randomly selected for validation by quantitative real-time polymerase chain reaction (qRT–PCR). The study demonstrates that transcriptome analysis could be an effective method for identifying the repertoire of candidate effectors and may provide an invaluable resource for future functional analyses of F. proliferatum pathogenicity in F. proliferatum and tomato plant–host interactions.
机译:由真菌Fusarium proliferatum(Matsushima)Nirenberg引起的叶斑病是中国番茄植物的一种破坏性疾病。受感染的番茄植株的典型症状是茎叶变软和枯萎,导致整个植株最终死亡。在这项研究中,我们诉诸转录谱分析来深入了解参与F. proliferatum-tomato相互作用的效应子库。从头开始总共组装了61,544,598个干净读数,以提供F. proliferatum参考转录组。从这些中,获得了75,044个单基因,其中19.46%的单基因被分配给276个《京都基因与基因组百科全书》(KEGG)途径,其中22.3%的单基因与f.kujikuroi的基因同源。总共鉴定出18,075个差异表达基因(DEG),其中720个被编码为分泌蛋白。在这些蛋白中,有184个被鉴定为候选效应子,而79.89%的蛋白表达上调。此外,随机选择了17个在RNA-seq研究中差异表达的基因,通过定量实时聚合酶链反应(qRT-PCR)进行验证。该研究表明,转录组分析可能是一种识别候选效应子的有效方法,并且可能为今后在功能性拟南芥和番茄植物-宿主相互作用中功能性拟南芥致病性的功能分析提供宝贵的资源。

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