Comments on abstract, title, references

The MS entitled ‘Rapid adaptation of signaling networks in the fungal pathogen Magnaporthe oryzae’ by Bohnert etc. described experimental assay as well as DNA and RNA sequencing of ‘adapted’ mutations of Loss of function (LOF) mutants of some high osmolarity glycerol (HOG) signaling pathway components in the filamentous fungus Magnaporthe oryzae. However, the title failed to point out what 'signaling network' it is.

Comments on introduction/background

The authors first introduced molecular mechanisms underlie rapid adaptation of microbes to stress environments. Then they gave very detailed, published studies to summarize functional roles of lof mutants used in this study. Of note, they used a nice schematic map showing pathways of lof mutants, which I think greatly improved readibility of this paper.

Comments on methodology

The paper has given very detailed descriptions on how exprements are performed. And listed availability of NGS data. For some online resources, website links have been provided. Given this study have executed many bioinformatic analyses, I suggest that the authors could further improve reliablity and reproducibility of this study via providing detailed bioinformatic pipelines/scripts used in their study.

Comments on data and results

For Results part, I think this study should also tested LOF mutants growing on normal conditions. It may better if the control that grow in normal conditions is included in the comparison. For bar plot, such as Fig. 3, 4 and 6, although error bar has been given, statistical significance between different groups should also be presented. Different panels in figures should be labeled as A, B, not 1 or 2.

Comments on discussion and conclusions

In this paper, the author used stress-inducing to get ‘adapted’ mutants. While many other studies in filamentous fungus (such as Yang Li 2017 Plos genetics, Poonguzhali Selvaraj 2017 Molecular microbiology, Qiurong Xie 2019 Current genetics etc.) found spontaneous suppressor mutations under normal growth conditions also result in same phenotypic recovery. I speculate normal growth conditions may enough to give ‘adapted’ mutants for these LOF mutants. This possibility should also be discussed. Some statements are overstated, and need more evidence to support.