Colletotrichum was isolated from coffee
berries (Coffea arabica) collected at two sites
in Chiang Mai, Thailand. Strains were isolated
from anthracnose lesions on infected berries
and from both the surface (epiphytes) and inner
masses (endophytes) of healthy berries. To
obtain isolates from fruit without visible
sporulation a modification of the procedure
described by Photita et al. (2005) and Than et
al. (2008a) was used. Three 5 × 5 mm pieces of
tissue were taken from the margin of infected
tissues, surface sterilized by dipping in 1%
sodium hypochlorite for 1 minute, immersed in
70% ethanol for 1 minute and rinsed three
times with sterilized water and finally dried in
sterilized tissue paper. Samples were placed on
water agar and incubated at room temperature
(28-30°C). The growing edges of any fungal
hyphae developing from the tissues were then
transferred aseptically to potato dextrose agar.
The fungi were identified following sporulation.
Single spore subcultures were obtained for
each Colletotrichum isolate using the
procedure described by Goh (1999). When
direct examination showed that the fungus was
sporulating on the berry, spore masses were
picked off with a sterilized wire loop and
streaked on the surface of water agar. After
incubation overnight (28-30°C), single germinated
spores were picked up with a sterilized
needle and transferred to PDA. Pure cultures
were stored in sterilized water in Eppendorf
tubes at 6°C (Abang, 2003). Cultures are
deposited in the culture collection of Mae Fah
Luang University, BIOTEC Culture Collection
(BCC), and CBS.
Morphological studies