College of Life Science, Luoyang Normal University, Luoyang, Henan, 471934, China.
BMC Genomics. 2024 Sep 14;25(1):861. doi: 10.1186/s12864-024-10784-3.
Black spot disease in tree peony caused by the fungal necrotroph A. alternata, is a primary limiting factor in the production of the tree peony. The intricate molecular mechanisms underlying the tree peony resistance to A. alternata have not been thoroughly investigated.
The present study utilized high-throughput RNA sequencing (RNA-seq) technology to conduct global expression profiling, revealing an intricate network of genes implicated in the interaction between tree peony and A. alternata. RNA-Seq libraries were constructed from leaf samples and high-throughput sequenced using the BGISEQ-500 sequencing platform. Six distinct libraries were characterized. M1, M2 and M3 were derived from leaves that had undergone mock inoculation, while I1, I2 and I3 originated from leaves that had been inoculated with the pathogen. A range of 10.22-11.80 gigabases (Gb) of clean bases were generated, comprising 68,131,232 - 78,633,602 clean bases and 56,677 - 68,996 Unigenes. A grand total of 99,721 Unigenes were acquired, boasting a mean length of 1,266 base pairs. All these 99,721 Unigenes were annotated in various databases, including NR (Non-Redundant, 61.99%), NT (Nucleotide, 45.50%), SwissProt (46.32%), KEGG (Kyoto Encyclopedia of Genes and Genomes, 49.33%), KOG (clusters of euKaryotic Orthologous Groups, 50.18%), Pfam (Protein family, 47.16%), and GO (Gene Ontology, 34.86%). In total, 66,641 (66.83%) Unigenes had matches in at least one database. By conducting a comparative transcriptome analysis of the mock- and A. alternata-infected sample libraries, we found differentially expressed genes (DEGs) that are related to phytohormone signalling, pathogen recognition, active oxygen generation, and circadian rhythm regulation. Furthermore, multiple different kinds of transcription factors were identified. The expression levels of 10 selected genes were validated employing qRT-PCR (quantitative real-time PCR) to confirm RNA-Seq data.
A multitude of transcriptome sequences have been generated, thus offering a valuable genetic repository for further scholarly exploration on the immune mechanisms underlying the tree peony infected by A. alternata. While the expression of most DEGs increased, a few DEGs showed decreased expression.
芍药炭疽病是由真菌坏死营养型Alternaria alternata引起的，是芍药生产的主要限制因素。芍药对 A. alternata 抗性的复杂分子机制尚未得到深入研究。
本研究利用高通量 RNA 测序（RNA-seq）技术进行了全局表达谱分析，揭示了与芍药与 A. alternata 相互作用相关的基因的复杂网络。从叶片样本构建 RNA-seq 文库，并使用 BGISEQ-500 测序平台进行高通量测序。共构建了 6 个不同的文库。M1、M2 和 M3 来自未接种的叶片，而 I1、I2 和 I3 来自接种病原菌的叶片。产生了 10.22-11.80 千兆碱基（Gb）的清洁碱基，包括 68,131,232-78,633,602 个清洁碱基和 56,677-68,996 个 Unigenes。总共获得了 99,721 个 Unigenes，平均长度为 1,266 个碱基。所有这些 99,721 个 Unigenes均在各种数据库中进行了注释，包括 NR（非冗余，61.99%）、NT（核苷酸，45.50%）、SwissProt（46.32%）、KEGG（京都基因与基因组百科全书，49.33%）、KOG（真核同源群簇，50.18%）、Pfam（蛋白家族，47.16%）和 GO（基因本体论，34.86%）。总共有 66,641 个（66.83%）Unigenes在至少一个数据库中有匹配。通过对模拟和 A. alternata 感染的样本文库进行比较转录组分析，我们发现了与植物激素信号转导、病原体识别、活性氧生成和昼夜节律调节相关的差异表达基因（DEGs）。此外，还鉴定了多种不同类型的转录因子。通过 qRT-PCR（定量实时 PCR）验证了 10 个选定基因的表达水平，以确认 RNA-seq 数据。
生成了大量的转录组序列，为进一步研究芍药感染 A. alternata 的免疫机制提供了有价值的遗传资源。虽然大多数 DEGs 的表达水平增加，但少数 DEGs 的表达水平下降。
