Abstract
Penicillium expansum is a destructive phytopathogen causing postharvest decay on many stored fruits. To develop effective and safe management strategies, it is important to investigate its pathogenicity-related mechanisms. In this study, a bioinformatic pipeline was constructed and 50 core effector genes were identified in P. expansum using multiple RNA-seq data sets and their putative functions were implicated by comparatively homologous analyses using pathogen–host interaction database. To functionally characterize P. expansum LysM domain proteins during infection, null mutants for the 15 uncharacterized putative LysM effectors were constructed and the fungal growth rate on either PDA or Cazpek medium or lesion expansion rate on the infected apple fruits was evaluated. The results showed the growth rate of knockout mutants from PeLysM5, PeLysM12 and PeLysM15 was retarded on PDA medium. No significant difference in growth rate was observed between wild type and all mutants on solid Cazpek medium. Nevertheless, the hypha of wild type displayed deeper yellow on the back of Cazpek medium than those of knockout mutants. On the infecting apples fruits, the knockout mutants from PeLysM5, PeLysM7, PeLysM8, PeLysM9, PeLysM10, PeLysM11, PeLysM14, PeLysM15, PeLysM16, PeLysM18 and PeLysM19 showed enhanced fungal virulence, with faster decaying on infected fruits than those from wild type. By contrast, the knockout mutation at PeLysM12 locus led to reduced lesion expansion rate on the infected apple fruits. In addition, P. expansum-apple interaction RNA-seq experiment was performed using apple fruit tissues infected by the wild type and knockout mutant ΔPeLysM15, respectively. Transcriptome analyses indicated that deletion of PeLysM15 could activate expression of several core effector genes, such as PEX2_055830, PEX2_036960 and PEX2_108150, and a chitin-binding protein, PEX2_064520. These results suggest PeLysM15 may play pivotal roles in fungal growth and development and involve pathogen–host interaction by modulating other effector genes’ expression. Our results could provide solid data reference and good candidates for further pathogen-related studies in P. expansum.
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Data availability
All the raw RNA-seq data analyzed in this study could be freely downloaded from SRA database in NCBI (https://www.ncbi.nlm.nih.gov/sra), with the accession number represented in our paper.
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Funding
This study was funded by National Natural Science Foundation of China (Nos. 31972474 &31461143008) and Israel Science Foundation (ISF1936/14), awarded to Prof. Samir Droby and Prof. Yongsheng Liu.
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Danyang Chen and Guangwei Li contribute equally to this work.
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Fig. S1 Construction of 16 PeLysM genes’ knockout mutants. (a) The schematic diagram of knockout mutants’ construction of PeLysM genes; (b) PCR verification of 16 PeLysM genes’ knockout mutants.
Fig. S2 Growth rates comparisons between wild type and PeLysMs’ null mutants on PDA medium. The morphology of wild type and PeLysMs’ null mutants on PDA medium at 144 hpi were shown. The significant difference was determined by Student's t-test (*, p value < 0.05). M: PeLysMs’ null mutant; W: wild type.
Fig. S3 Growth rates comparisons between wild type and PeLysMs’ null mutants on solid Cazpek medium. The morphology of wild type and PeLysMs’ null mutants on front and back of medium plates at 144 hpi were shown. M: PeLysMs’ null mutants; W: wild type.
Fig. S4 Decay development comparisons between wild type and PeLysMs’ null mutants in apple fruits. The effects of 16 PeLysM genes’ knockout on pathogenicity in apple fruits were studied in apple fruits. Progression of decay development was expressed as lesion diameter (mm) at different time of post inoculation. The pictures of infected apples represented the phenotype at 144 hpi. The significant difference was determined by Student's t-test (*, p value < 0.05). M: PeLysMs’ null mutant; W: wild type.
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Chen, D., Li, G., Liu, J. et al. Multiple transcriptomic analyses and characterization of pathogen-related core effectors and LysM family members reveal their differential roles in fungal growth and pathogenicity in Penicillium expansum. Mol Genet Genomics 295, 1415–1429 (2020). https://doi.org/10.1007/s00438-020-01710-9
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DOI: https://doi.org/10.1007/s00438-020-01710-9