Table 1

List of primers"

Fig.1

Phylogenetic analysis of Exo70 homologs from different organisms"

Fig.2

Targeted gene deletion and complementation A: Construction strategy of mutants and complementary strain; B: Specific primers were used to verify the transformed strains by PCR.Primer 1:External primer for target gene External- Exo70for/External-Exo70rev, M: 1 000 marker; Primer 2:Target gene internal primer Internal-Exo70for/Internal-Exo70rev, M: 250 marker."

Fig.3

Radial growth in CcExo70 deletion mutant A: Colony morphologies of the wild-type, mutant, and complemented strains after 2 and 7 days of growth on PDA plates; B: The mycelia morphology of the strains after 7 d of growth in liquid PDB; C.Statistical analysis of colony diameters of the indicated strains after 2 d on media.Error bars are standard deviation and different lowercases represent significant difference at P < 0.05."

Fig.4

Mycelium morphology of CcExo70 deletion mutant A: The strain was cultured at 25℃ on a slide containing PDA ramp for 3 days, then the mycelia morphology was observed under microscope; B: The number of single mycelium branches was counted under microscope.Error bars are standard deviation and different lowercases represent significant difference at P < 0.05."

Fig.5

Measurement of oxidative stress in CcExo70 deletion mutants A: Growth status of wild-type, mutant and complementary strains grown on PDA plates with or without drip addition (CK) of 2 mol·L-1 and 3.5 mol·L-1 H2O2.All strains were incubated at 25℃ for 3 d and photographed.B: The inhibition diameters of different strains were quantitatively analyzed.Error bars are standard deviation and different lowercases represent significant difference at P < 0.05."

Fig.6

Pathogenicity determination of CcExo70 deletion mutant A: Infection symptoms on detached poplar twigs inoculated with the wild-type, mutant, and complemented strains after 3 d and 7 d.The area of lesions is outlined with a dotted line.B: Area of lesions produced by the different strains on twigs.Error bars are standard deviation and different lowercases represent significant difference at P < 0.05."

Fig.7

Detection of superoxide production A: Detection of superoxide production by DAB staining.Dark DAB dye in wild type strain suggested a high ROS level, while the mutant showed less color, indicating a reduced ROS accumulation.The reconstituted strain exhibited a darker color similar to wild type.B: Accumulation of ROS in single hyphae after two days of DAB staining.All strains were cultured on PDA plate at 25℃ for 2 days, then observed under light microscope after DAB staining."

Fig.8

Distribution of chitin in the CcExo70 deletion mutant A: Mycelium of WT, ΔCcExo70 and the ΔCcExo70/C strains were stained by calcofluor white (CFW) to observe chitin accumulation.B: Chitin deposition at the tip of mycelia.Compared with the wild type and complementary strains, there was no obvious chitin deposition at the tip of the mutant hyphae.C: After CFW staining, the septal distribution of wild type and complementary strains was significantly less than that of mutant strains under fluorescence microscope.D: The number of septa in mycelium cytoplasm of wild type, ΔCcExo70 and complementary strains.Error bars are standard deviation and different lowercases represent significant difference at P < 0.05."

Fig.9

Distribution of vesicles in the CcExo70 deletion mutant The mycelium were stained with 5 μmol·L-1 FM4-64 for confocal imaging after cultured for 3 days."

Table 2

List of proteins with abundances fall by over half in ΔCcExo70-F compared to WT-F"