Control of citrus blue and green molds by Actinomycin X2 and its possible antifungal mechanism

Citrus blue and green molds caused by Penicillium digitatum, P. italicum, and P. polonicum, are the major postharvest diseases of citrus fruit. In the present study, Actinomycin X-2 (Act-X-2), a naturally occurring antibiotic produced by Streptomyces species, was found to show excellent antifungal effect against these three pathogens with a minimum inhibitory concentration (MIC) value of 62.5 mu g/mL for them all, which was better than the positive control thiophanate-methyl. Act-X-2 significantly reduced the percentage of spore germination, and highly inhibited the mycelial growth of P. italicum, P. digitatum, and P. polonicum with EC50 values being 34.34, 13.76, and 37.48 mu g/mL, respectively. In addition, Act-X-2 greatly decreased the intracellular protein content while increasing the reactive oxygen species (ROS) level and superoxide anion (O-2(-)) content in the mycelia of pathogens. In vivo test indicated that Act-X-2 strongly inhibited the infection of navel oranges by these three Penicillium species, with an inhibition percentage of >50% for them all at the concentration of 10 MIC. Transcriptome analysis suggested that Act-X-2 might highly influence the ribosomal functions of P. polonicum, which was supported as well by the molecular docking analysis of Act-X-2 with some key functional proteins and RNAs of the ribosome. Furthermore, Act-X-2 significantly reduced the decay percentage and improved the firmness, color, and sugar-acid ratio of navel oranges spray-inoculated with P. polonicum during the postharvest storage at 4 degrees C for 60 d.