Home⇛NRCP Research Journal⇛vol. 24 no. 2 (2025)

Effectiveness of Different Surface Sterilization Protocols for the Isolation of Endolichenic Fungi from Ramalina

Thomas Edison E. Dela Cruz | Ghimel P. Espinosa | John Joshua T. Bellen | Sittie Aisha B. Macabago | Melfei E. Bungihan

Discipline: molecular biology, biophysics and biochemistry

 

Abstract:

Endolichenic fungi (ELF) thrives within the thallus of lichens. Its isolation remains challenging given the indeterminate number of microorganisms cohabiting within the lichen thallus. This study determines the most effective surface sterilization method from the different protocols and their modifications to isolate ELF from the fruticose lichen Ramalina. Our findings showed that the protocol as described by Maduranga et al. (2018) with the treatment of the lichen thalli with 70% ethanol for 10 secs followed by 0.5% commercial bleach solution for 3 mins, and 1 min washing of sterile distilled water for 3 consecutive times remained the most ideal for isolation. All lichen thallus explants treated with this surface-sterilization method yielded 100% isolation rate while the culture plate for the tissue prints exhibited no fungal growth (100% effectivity rate). Modifications of this method resulted in 95–100% isolation rates, but lower effectivity rates of 80–86%. In comparison with the other tested surface-sterilization protocols and their modifications, their effectivity rates vary from 57-95%, albeit with isolation rates between 95-100%. All nine surface-sterilization protocols tested in this study resulted in the isolation of 156 ELF from a single lichen host. Our study highlighted Philippine lichens as ideal hosts for a diverse assemblage of endolichenic fungi.



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