Development of a Rapid Molecular Detection Marker for Colletotrichum species with AFLP

Abstract

Sweet persimmons have been increasingly cultivated in the southern part of Korea. However, anthracnose disease caused by Colletotrichum species is one of the major hindrances in cultivation and productions. In this study, we used polymerase chain reaction (PCR) to detect Colletotrichum species with the AFLP (amplified fragment length polymorphism) method. In AFLP, we used E3 (′5-GACTGCGTACCAATTCTA-3′) and Ml (5′-GATGAGTCCTGAGTAACAG-3′) primer combination and, as a result, 262 bp segment was observed in Colletotrichum species only. Specific PCR primers were designed from the sequence data and used to detect the presence of the fungus in genomic DNA isolated from symptomless sweet persimmon plants. Based on sequence data for specific segments, Co.Bl (5′-GAGAGAGTAGAATTGCGCTG-3′) and Co.B2 (5′-CTAC-CATTCTTCTA GGTGGG-3′) were designed to detect Colletotrichum species. The 220 bp segment was observed in Colletotrichum species only, but not in other fungal and bacterial isolates.

Key Words:

• AFLP
• Anthracnose
• Colletotrichum
• species
• Sweet persimmons