希金斯炭疽菌自噬相关基因<i>ChAtg3</i>的功能研究

段灵涛; 何九卿; 王莉; 陈炜伦; 祝一鸣; 周而勋

doi:10.7671/j.issn.1001-411X.202404046

希金斯炭疽菌自噬相关基因ChAtg3的功能研究

Study on the functions of the autophagy related gene ChAtg3 in Colletotrichum higginsianum

摘要

摘要:
目的研究希金斯炭疽病菌Colletotrichum higginsianum中自噬相关基因ChAtg3的作用。
方法对ChAtg3进行序列、系统发育和表达模式分析，并观察其在C. higginsianum侵染拟南芥后的表达变化。通过根癌农杆菌介导的遗传转化技术，进行ChAtg3基因的敲除和回补。
结果拟南芥接种C. higginsianum 8 ~ 40 h内ChAtg3基因表达量显著上升。敲除该基因后的突变体(ΔChAtg3)自噬过程受抑制，其黑色素含量、产孢量、附着胞形成率和致病力均显著下降，但与野生型相比，菌丝生长和孢子萌发率并无显著变化。而通过基因回补，突变体CΔChAtg3在生物学表型和致病性上得以恢复。
结论结果揭示了ChAtg3基因在希金斯炭疽病菌自噬过程、孢子产生和附着胞形成中的关键作用，是该病原菌致病力的关键调节因子。

Abstract:
Objective To study the role of ChAtg3, an autophagy-related gene in Colletotrichum higginsianum.
Method The sequence, phylogeny and expression pattern of ChAtg3 were analyzed, and the expression changes of ChAtg3 were observed in Arabidopsis thaliana after infection with C. higginsianum. An Agrobacterium tumefaciens-mediated genetic transformation system was employed to perform specific gene knockouts and their subsequent complementation.
Result The expression changes of ChAtg3 were observed after C. higginsianum infected A. thaliana for 8 to 40 h. The mutant ΔChAtg3, created through targeted ChAtg3 gene knockout, exhibited impaired autophagy, with markedly reduced melanin production, conidiation, appressorium formation, and pathogenic potential, although mycelial growth and conidial germination rate remained comparatively unaffected. Gene complementation in the CΔChAtg3 strain effectively reinstated its biological phenotype and pathogenicity.
Conclusion The results reveal that ChAtg3 play a key role in autophagy, conidiation, and appressorium formation of C. higginsianum, confirming it as a critical determinant of pathogenic capacity of C. higginsianum.

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