Liu Bo, Wang Haode, Ma Zhoujie, Gai Xiaotong, Sun Yanqiu, He Shidao, Liu Xian, Wang Yanfeng, Xuan Yuanhu, Gao Zenggui
Institute of Plant Immunology, Shenyang Agricultural University, Shenyang, Liaoning, China.
College of Life Sciences, Yan'an University, Yan'an, Shaanxi, China.
PeerJ. 2018 Jun 18;6:e5103. doi: 10.7717/peerj.5103. eCollection 2018.
AG1 IA is a soil-borne fungal phytopathogen that can significantly harm crops resulting in economic loss. This species overwinters in grass roots and diseased plants, and produces sclerotia that infect future crops. AG1 IA does not produce spores; therefore, understanding the molecular mechanism of sclerotia formation is important for crop disease control. To identify the genes involved in this process for the development of disease control targets, the transcriptomes of this species were determined at three important developmental stages (mycelium, sclerotial initiation, and sclerotial maturation) using an RNA-sequencing approach. A total of 5,016, 6,433, and 5,004 differentially expressed genes (DEGs) were identified in the sclerotial initiation vs. mycelial, sclerotial maturation vs. mycelial, and sclerotial maturation vs. sclerotial initiation stages, respectively. Moreover, gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) analyses showed that these DEGs were enriched in diverse categories, including oxidoreductase activity, carbohydrate metabolic process, and oxidation-reduction processes. A total of 12 DEGs were further verified using reverse transcription quantitative PCR. Among the genes examined, NADPH oxidase 1 () and superoxide dismutase () were highly induced in the stages of sclerotial initiation and maturation. In addition, the highest reactive oxygen species (ROS) production levels were detected during sclerotial initiation, and enzyme activities of NOX1, SOD, and catalase (CAT) matched with the gene expression profiles. To further evaluate the role of ROS in sclerotial formation, AG1 IA was treated with the CAT inhibitor aminotriazole and HO, resulting in the early differentiation of sclerotia. Taken together, this study provides useful information toward understanding the molecular basis of AG1 IA sclerotial formation and maturation, and identified the important role of ROS in these processes.
AG1 IA是一种土壤传播的真菌植物病原体，会对作物造成严重损害，导致经济损失。该物种在草根和患病植物中越冬，并产生菌核感染未来的作物。AG1 IA不产生孢子；因此，了解菌核形成的分子机制对于作物病害控制很重要。为了确定参与这一过程的基因以开发病害控制靶点，使用RNA测序方法在该物种的三个重要发育阶段（菌丝体、菌核起始和菌核成熟）测定了转录组。在菌核起始与菌丝体、菌核成熟与菌丝体以及菌核成熟与菌核起始阶段，分别鉴定出5016、6433和5004个差异表达基因（DEG）。此外，基因本体（GO）和京都基因与基因组百科全书（KEGG）分析表明，这些DEG富集在不同类别中，包括氧化还原酶活性、碳水化合物代谢过程和氧化还原过程。使用逆转录定量PCR进一步验证了总共12个DEG。在所检测的基因中，NADPH氧化酶1（）和超氧化物歧化酶（）在菌核起始和成熟阶段高度诱导。此外，在菌核起始期间检测到最高的活性氧（ROS）产生水平，并且NOX1、SOD和过氧化氢酶（CAT）的酶活性与基因表达谱相匹配。为了进一步评估ROS在菌核形成中的作用，用CAT抑制剂氨基三唑和HO处理AG1 IA，导致菌核早期分化。综上所述，本研究为理解AG1 IA菌核形成和成熟的分子基础提供了有用信息，并确定了ROS在这些过程中的重要作用。
