Fungal

Silva, Rejane M. F., De Mélo, Mayara A. C., Oliveira, Thays G. L., De Oliveira, Rafael J. V., Souza-Motta, Cristina M. & Silva, Gladstone A., 2024, Nigrospora solani (Amphisphaeriales, Apiosporaceae) a new endophytic species from Brazil, Phytotaxa 669 (3), pp. 273-284 : 274

Fungal isolation

Healthy S. lycopersicum var. cerasiforme leaves (27 sample) were collected from an organic farming system in the municipality of Lagoa de Itaenga (7°53’42.9”S, 35°16’52.5”W), Pernambuco, Brazil. The leaves were transported to the laboratory and processed within 24 h. The leaves were first washed with tap water to remove dust particles from the surface and were then cut into six 2.5 × 2.5 mm fragments (Gamboa et al. 2002). Subsequently, the fragments were disinfected with 70% alcohol for 30 s and 2% sodium hypochlorite (NaOCl) for 2 min and washed twice with sterile distilled water to remove the NaOCl ( Araújo et al. 2002; modified). After disinfection, the leaf fragments were distributed in Petri dishes containing Potato Dextrose Agar (PDA) culture medium with chloramphenicol (100 µg/mL), and incubated at room temperature (± 28°C) for seven days. To confirm the effectiveness of disinfection, 1 mL of the distilled water used for the last bath was also plated in Petri dishes with PDA medium. The isolate was cultured on PDA, and synthetic nutrient-poor agar (SNA). After two-week incubation, the colony diameters were measured, and their morphology was described. Colony colours on the surface and reverse were assessed according to the colour charts of Rayner (1970), and micromorphological descriptions were obtained from cultures grown for 14 days. Microscope slides were made in lactoglycerol and at least 10 conidiogenous cells and 30 conidia were measured. The reference strain was deposited in the URM culture collection (Micoteca URM Profª Maria Auxiliadora Cavalcanti), whereas prepared microscope slides were deposited in the HURM herbarium (Herbário HURM Padre Camille Torrend) at the Universidade Federal de Pernambuco Recife, Brazil.