丝状真菌Podospora anserina中光敏色素基因的鉴定及功能分析

Identification and functional characterization of phytochrome genes in the filamentous fungus Podospora anserina

摘要:

光敏色素在细菌和植物发育中起着关键作用，但它们在真菌中的生物学功能尚不完全清楚。【目的】探究光敏色素基因PaPhy1和PaPhy2在Podospora anserina有性生殖和无性发育中的作用及其调控机制。【方法】利用同源重组方法对P.anserina中2个光敏色素基因PaPhy1和PaPhy2进行定点敲除，获得光敏色素基因缺失菌株ΔPaPhy1和ΔPaPhy2，并通过遗传杂交构建双重突变体ΔPaPhy1ΔPaPhy2；分析突变型菌株和野生型菌株在不同光照下有性生殖、无性发育、生长速率和活性氧代谢等方面的差异，明确光敏色素基因在P.anserina中的主要功能。【结果】白光和蓝光诱导P.anserina子实体的形成，ΔPaPhy在光照下产生子实体的数量减少，ΔPaPhy的生命周期延长。【结论】光敏色素基因与P.anserina有性生殖密切相关；ΔPaPhy的衰老延迟和活性氧代谢有关。本研究的结果为进一步探索光照对丝状真菌繁殖调控机制以及抗衰老研究提供了新的思路。

Abstract:

Phytochromes play a key role in bacterial and plant development, while their roles in fungi are not fully understood. [Objective] To explore the roles of phytochrome genes PaPhy1 and PaPhy2 in the sexual reproduction and asexual development of Podospora anserina and decipher the regulatory mechanisms. [Methods] The homologous recombination method was employed to knock out the two phytochrome genes, and the resulting mutants, ΔPaPhy1 and ΔPaPhy2, were obtained. A double mutant, ΔPaPhy1ΔPaPhy2, was constructed via genetic cross. We compared the sexual production, asexual development, growth rate, and reactive oxygen metabolism between the mutants and wild-type strains under different light conditions to reveal the main roles of the phytochrome genes in P. anserina. [Results] Blue light and white light induced the formation of P. anserina ascospore shells. ΔPaPhy produced fewer ascospores in the light and showed prolonged life cycle. [Conclusion] Phytochrome genes are associated with the sexual reproduction of P. anserina. The aging delay of ΔPaPhy is related to reactive oxygen metabolism. The results of this study provide new ideas for further exploring the mechanism of light in regulating the reproduction of filamentous fungi as well as anti-aging studies.