THE USE OF SPECIES-SPECIFIC DNA PROBES FOR THE IDENTIFICATION OF MYCOSPHAERELLA FIJIENSIS AND M-MUSICOLA, THE CAUSAL AGENTS OF SIGATOKA DISEASE OF BANANA
A. Johanson et al., THE USE OF SPECIES-SPECIFIC DNA PROBES FOR THE IDENTIFICATION OF MYCOSPHAERELLA FIJIENSIS AND M-MUSICOLA, THE CAUSAL AGENTS OF SIGATOKA DISEASE OF BANANA, Plant Pathology, 43(4), 1994, pp. 701-707
The polymerase chain reaction (PCR)-based technique of random amplific
ation of polymorphic DNA (RAPD) was used to differentiate DNA from spe
cies of the genus Mycosphaerella. DNA from two pathogens which cause S
igatoka leafspot diseases of banana, M. fijiensis and M. musicola, and
two other Mycosphaerella species which are commonly found on banana,
M. musae and M. minima, gave distinct RAPD banding patterns with all P
CR primers tested. PCR, using primer RC07, amplified a 1250 bp RAPD fr
agment from all isolates of M. fijiensis obtained from 11 geographical
origins. This fragment was absent from the other species of Mycosphae
rella. In Southern blots of genomic DNA, this band hybridized exclusiv
ely to DNA from M. fijiensis, and the pattern of hybridization suggest
ed that it was binding to repeated DNA. A RAPD band amplified with pri
mer PM06 obtained from M. musicola was also found to be species-specif
ic. Southern analysis suggested that the fragment hybridized to a sing
le-copy sequence in the M. musicola genome. Total genomic DNA from M.
musicola was found to be a species-specific hybridization probe. Dot-b
lots confirmed the specificity of these probes, and could be used to i
dentify isolates of Mycosphaerella which cause Sigatoka disease of ban
ana in south-east Asia.