胶孢炭疽菌自噬诱导和MDC染色方法的建立和条件优化
Establishment and Condition Optimization of Autophagy Induction and MDC Staining Methods for Colletotrichum gloeosporioides
廖至雯 1黄志睿 1罗红丽1
作者信息
- 1. 海南大学热带作物学院,海口,570228
- 折叠
摘要
自噬(Autophagy)是真核细胞中普遍存在的生理代谢过程,对植物病原真菌的致病性具有决定性的作用.MDC(Monodansylcadaverine)染色是检测自噬体形成的常用方法,具有简单快速的优点.本研究通过优化培养基种类、MDC染色浓度和染色时间、氮饥饿诱导时间、PMSF使用浓度等影响橡胶树胶孢炭疽菌自噬诱导和观测的主要因素,确立适合于橡胶树胶孢炭疽菌的自噬诱导和MDC染色条件.结果表明,在基础培养基的基础上,在4 mmol/L PMSF存在的条件下氮饥饿3 h能够有效诱导胶孢炭疽菌产生自噬;当用100 μmol/LMDC溶液对菌丝体染色30 min时染色效果最佳.本研究所建立的自噬诱导和检测的方法技术体系为研究胶孢炭疽菌的自噬机理提供理论基础.
Abstract
Autophagy is a physiological metabolic process in eukaryotic cells and plays a decisive role in the pathogenicity of plant pathogenic fungi.MDC(Monodansylcadaverine)staining is a common method to detect au-tophagosome formation,which is simple and rapid.In this study,the appropriate conditions for autophagy induc-tion and MDC staining of Colletotrichum gloeosporioides were established and modified,including the type of medium,the concentration and time of MDC staining,the induction time of nitrogen starvation,and the concentra-tion of PMSF.The results showed that autophagy was effectively induced by nitrogen starvation for 3 hours in the presence of 4 mmol/L PMSF in the basal medium MM,and the best staining effect was achieved when the myceli-um was stained with 100 μmol/L MDC solution for 30 min.The autophagy induction and detection method laid a foundation for studying the autophagy mechanism of C.gloeosporioides.
关键词
胶孢炭疽菌/自噬诱导/MDC染色Key words
Colletotrichum gloeosporioides/Autophagy induction/MDC staining引用本文复制引用
基金项目
国家自然科学基金项目(32260716)
国家自然科学基金项目(32060591)
出版年
2024
NETL
NSTL
万方数据