Fungal

Ma, Xiao-Ya, Hyde, Kevin D., Phillips, Alan J. L., Kang, Ji-Chuan, Chomnunti, Putarak & Doilom, Mingkwan, 2021, Three new host records of endophytic Neofusicoccum species reported from Dendrobium orchids, Phytotaxa 494 (2), pp. 193-207 : 194

Fungal isolation and cultivation

Isolations were carried out within 48 hours after collection. Surface sterilization was conducted following the method of Nontachaiyapoom et al. (2010) and Doilom et al. (2017) with modifications. Materials were washed with tap water and immersed in a solution containing 3% (v/v) H 2 O 2 and 70% (v/v) ethanol for 5 mins. They were then rinsed with sterile distilled water for three times. The sterilized materials were cut into 2 mm 2 pieces with sterilized knife in laminar flowing workplace. And then kept them on potato dextrose agar (PDA) containing 50 μg/ml oxytetracycline, 50 μg/ml penicillin and 50 μg/ml streptomycin, which could reduce the contamination opportunity for the target pieces ( Otero et al. 2002). The efficiency of the method was checked by printing sterilized tissues on PDA as described in Petrini (1991). Plates were incubated at 28 °C in the dark. Cultures were observed every day. Vegetative mycelium was transferred to fresh PDA for purification. Pure cultures were deposited at Mae Fah Luang University Culture Collection (MFLUCC) and in the culture collection of Guizhou University (GZAC).