PCR-RFLP based genetic diversity of Colletotrichum Musae isolates of banana in Jaffna district

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.