Fig. 1 Heterologous expressions of eGFP and PlCYP6∷eGFP in P. lilacinum A：Observation of the eGFP green fluorescence signal of P. lilacinum（WT）,strains Pl-G（heterologous expression of eGFP）and Pl-GCYP6（heterologous expression of PlCYP6∷eGFP）by confocal microscope. B：Amplification of eGFP（about 1 000 bp）and PlCYP6∷eGFP（about 3 000 bp）by PCR,lane M is a DNA marker;lane 1 is WT;lane 2 and lane 3 are strain Pl-G;lane 4 and lane 5 are the strain Pl-GCYP6. C：Detection of eGFP or PlCYP6∷eGFP in the total protein of P. lilacinum by Western blot. Among them,using eGFP-anti as the primary antibody and goat anti-mouse IgG as the secondary antibody;lane M is Precision Plus ProteinTM Standards,lane 1 is the total protein of WT;lane 2 and lane 3 are the total protein of strain Pl-G;lane 4 and lane 5 are the total protein of strain Pl-GCYP6

Fig. 2 Immunoprecipitation to catch interacting proteins of PlCYP6 A：Detection of proteins by SDS-PAGE gel electrophoresis;M is Precision Plus ProteinTM Standards;lane 1 and lane 2 are the extracted total protein of Pl-G strain（heterologous expression of eGFP）and Pl-GCYP6 strain（heterologous expression of PlCYP6∷eGFP）after treatment with 1 mol/L NaCl for 0,15,30 and 60 min;lane 3 and lane 4 are the eluted products of total protein after immunoprecipitation;lane 5 and lane 6 are supernatant that washed for the first time when non-specific proteins were removed after immunoprecipitation. B：Western blot was used to hybridize the eluted product after immunoprecipitation;Input was the total protein solution;eGFP-anti as the primary antibody and goat anti-mouse IgG as the secondary antibody;“+” indicates containing and “-” indicates not containing

Fig. 3 Distribution of differential proteins Control group：Pl-G strain（heterologous expression of eGFP）. Treatment group：Pl-GCYP6 strain（heterologous expression of PlCYP6∷eGFP）. The protein solution after immunoprecipitation in the control group and the treatment group was analyzed by LC-MS/MS to identify the number and types of proteins respectively. The databases are UniProt and the protein databases based on genome annotation. Compared with the control group,the specific proteins in the treatment group were selected as candidate interacting proteins（482）

Fig. 4 GO function annotation and KEGG pathway analysis of candidate interacting proteins Based on the GO database and the KEGG pathway database,the candidate interacting proteins（482）are annotated with protein function. The abscissa is the number of proteins,and the ordinate is the classification item

Fig. 5 Interaction relationship between PlCYP6 and ADH1 protein by yeast two-hybrid A：Principle diagram of truncation analysis based on PlCYP6 protein domain, WD40 repeat and Cyclophilin-like are the predicted protein domains of PlCYP6. B：Yeast two-hybrid self-activation verification. C：Yeast twohybrid results between PlCYP6 and different segmented regions with ADH1. BD：PGBKT7;AD：PGADT7-7;“+” refers to AD or BD empty carrier. If yeast grew on DDO medium（two-deficient medium）,the target vectors have been successfully transformed into the yeast. If yeast grew on QDO/X/A medium（four-deficient medium）,there will be direct interaction relationship between target proteins

Fig. 6 Analysis of the gene expression levels of PlCYP6 and ADH1 under salt stress The wild-type P.lilacinum was treated with sterile water（Mock）and 1 mol/L NaCl for 60 min,and then the RNA was extracted. Actin was used as an internal control to measure the gene expression levels of PlCYP6 and ADH1. The t test was used to determine significant differences between samples. “***” refers to P<0.001