Methodology for sporulation and production of monoconidial isolates of Sphaeropsis sapinea

Abstract

Sphaeropsis sapinea is one of main pathogens that causes losses in commercial plantations of Pinus
spp. Studies for control based on genetic resistance can be done by inoculating different monosporic isolates on
pine populations and selectioning the resistant individuals. This work describes a methodology for pycnidia production in vitro and S. sapinea monoconidial cultures, using an agar-water medium with sterile pine needles. Four S. sapinea isolates were tested e pycnidia and mature conidia were observed after 14 days of incubation, under continuous fluorescent light, in BOD, at 25 °C. Pycnidia were triturated in sterile water and conidia germinated after two hours of incubation in water-agar, in BOD, at 25 °C. Germinated conidia formed colonies which were purified in malt extract-agar and maintained in test tubes with the same medium, covered with sterile mineral oil for preservation. This methodology revealed to be  adequated for production of monoconidial colonies
of S. sapinea