Abstract:
Anthracnose disease is a major post-harvest disease of banana in Sri Lanka. The present study was
conducted to determine the genomic variations among Colletotrichum musae isolates, causing
anthracnose disease of banana, by PCR-RFLP analysis. Morphologically-different six colletotrichum
musae isolates were isolated from 3 different banana local cultivars (‘Kathali’, ‘Kappal’, ‘Etharai’).
Genomic DNA was extracted from each purified isolate using a modified CTAB method. PCR
amplification was done with ITS1 and ITS 4 primers to amplify the 5.8S-ITS subunit. For fungi, the
internal transcribed spacer (ITS) region in the ribosomal RNA (rRNA) operon has been accepted as
the formal fungal barcode. A 590 bp PCR product were resulted by two Colletotrichum musae isolates
(Cm2 and Cm5). Colletotrichum musae isolates (Cm1, Cm2, Cm3, Cm4, Cm5 and Cm6) showed two
polymorphic groups based on PCR-RFLP by HaeIII. The same isolates showed no polymorphism based
on PCR-RFLP by RsaI and MspI. Amplification of 5.8S-ITS region and subsequent PCR-RFLP by HaeIII
is an effective and reliable method to determine genomic variation among Colletotrichum musae isolates
causing anthracnose disease in banana fruits.
