Biological control potential of a soil fungus Trichoderma asperellum K1-02 against Neoscytalidium dimidiatum causing stem canker of dragon fruit

Abstract

Stem canker on dragon fruit caused by the fungus Neoscytalidium dimidiatum negatively impact dragon fruit cultivation and production. The use of Trichoderma species, the biological control agent against stem canker disease is an alternative way instead of use of synthetic fungicides. This study aimed to test the antifungal activities of soil fungi Trichoderma against N. dimidiatum. Ten isolates of Trichoderma species were tested against N. dimidiatum via dual culture assay, and the results showed that percentages of inhibition ranged from 72.75 to 84.45%. Trichoderma species K1-02 was found to be the most effective strain against N. dimidiatum (84.45%). Trichoderma K1-02 was identified based on a morphology and molecular study of internal transcribed spacer (ITS), the second largest subunit of RNA polymerase II (rbp2), and the translation elongation factor 1-alpha (tef1-α) as Trichoderma asperellum K1-02. The pre-colonized plate method demonstrated that T. asperellum K1-02 covered the tested plates at a rate of 60%, which was significantly higher than N. dimidiatum (40%), and T. asperellum K1-02 aggressively covered the mycelia of N. dimidiatum as observed via stereomicroscopy. An enzyme assay was conducted to test the activities of cell wall-degrading enzymes (CWDEs) in cell-free culture filtrate (CF) released by T. asperellum K1-02, and the results showed that the activities of chitinase and β-1,3-glucanase were significantly higher than those of the control (potato dextrose broth alone). Scanning electron microscopy of N. dimidiatum mycelia incubated with cell free CF of T. asperellum K1-02 showed abnormal morphology. Furthermore, an in vivo test was conducted on dragon fruit cladodes to test the ability of Trichoderma with respect to the reduction of canker lesions. The results revealed that the application of 106 spore/mL T. asperellum K1-02 spore suspension suppressed disease development via the reduction of canker areas (0.09 cm2) on dragon fruit cladodes at a significantly higher level than those inoculated by pathogens alone (0.49 cm2). Thus, T. asperellum K1-02 demonstrated antifungal abilities against N. dimidiatum and has the potential to control stem canker on dragon fruit.