Optimization of surface sterilization techniques to enhance the diversity of leaf endophytic fungal isolates from bitter ginger

作者：Marlina Puspita Sari, Suryo Wiyono, Giyanto Giyanto, Awang Maharijaya, Dono Wahyuno · 发表于：IOP Conference Series Earth and Environmental Science · 年份：2025 · DOI：10.1088/1755-1315/1494/1/012022 · 被引用次数：3 · 研究领域：Plant Pathogens and Fungal Diseases、Plant-Microbe Interactions and Immunity、Plant tissue culture and regeneration

Abstract Surface sterilization techniques are crucial in isolating plant endophytic fungi. The method may vary depending on the plant tissue structure as well as the characteristics of the fungus. The appropriate technique can minimize the isolation of epiphytic fungi and contaminants, thereby increasing the diversity of isolated endophytic fungi. This study aims to determine the optimal concentration and exposure time of NaOCl to enhance the diversity of endophytic fungi isolated from the leaves of bitter ginger. The tested NaOCl concentrations were 1%, 3%, and 5%, with exposure times of 1, 3, and 5 minutes. The leaves were rinsed with running water, and under a laminar airflow, the leaf laminae were sterilized sequentially: first by immersing them in 70% ethanol for 1 minute, followed by treatment with NaOCl as specified, and finally in 70% ethanol for 30 seconds. After that, the leaves were rinsed three times with sterile distilled water, cut into 5x5 mm 2 pieces, and air-dried. The sterilized leaf pieces-print were planted onto 20% potato dextrose agar (PDA) medium enriched with 0.25% chloramphenicol and incubated at 20°C for 7-21 days. The higher the concentration and exposure time of NaOCl, the lower the colonization rate of endophytic fungi. However, this does not always correspond to the diversity of isolated endophytic fungi. At too low NaOCl concentrations, colonization rates are high, but diversity is narrow due to the dominance of certain fungi. Surface sterilizat...