Endophyte isolation and characterization: Current methods, practical insights, and challenges in medicinal plant research

Endophytes in medicinal plants represent a versatile and largely untapped source of structurally diverse secondary metabolites, many of which exhibit potent and selective bioactivities. To fully harness this potential, optimized protocols for the isolation and characterization of endophytic communities are essential. The diversity of culturable microorganisms directly impacts the likelihood of discovering novel bioactive compounds, making methodological precision critical. To support this endeavor, the article offers practical guidance on current methodologies for isolating and characterizing endophytic fungi and bacteria from medicinal plants, emphasizing key protocols such as sample collection, surface sterilization, culture-dependent and culture-independent techniques, media optimization, growth conditions, and accurate identification methods. It also addresses common challenges, including the effectiveness of surface sterilization, the unculturable nature of some endophytes, and issues related to misidentification. By addressing these challenges and refining existing protocols, researchers can harness the full potential of endophytes, enhancing their role in drug discovery and other biotechnological applications.