Abstract
Problem statement: Morphological characterization of Fusarium species which emphasize on microscopic and cultural characteristics are not sufficient to characterize Fusarium Oxysporum F. sp. Cubense (FOC) from banana as these characteristics could easily influence by environmental factors. As an alternative molecular methods were used to characterize and to assess genetic variation of FOC from different banana cultivars. Knowledge on the genetic variation is important to determine the genetic relationship between FOC isolates from different banana cultivars. Approach: Two PCR-based methods, Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR) and restriction analysis of the Internal Transcribed Spacer and 5.8S regions (ITS+5.8S regions) were used to characterize Fusarium Oxysporum F. sp. Cubense (FOC) isolates from different banana cultivars. The genetic relationship of the FOC isolates were analyzed using Un-weighted Pair-Group Method with Arithmetic Averages (UPGMA) cluster analysis based on Jaccard Similarity Coefficient. Results: Restriction patterns of the ITS+5.8S regions using nine restriction enzymes namely, Alu I, Eco RI, Eco 88I, Bsu RI, Bsu 15I, Hin fI, Hin 6I, Msp I and Taq I and ERIC-PCR showed low variation among the FOC isolates studied, indicating close relationship among the isolates. Un-weighted Pair-group Method with Arithmetic Averages (UPGMA) cluster analysis based on Jaccard Similarity Coefficient showed that the FOC isolates were grouped into two main clusters with similarity value of 41.4-100% in PCR-RFLP of ITS + 5.8S and 45-100% similarity based on ERIC-PCR analysis, respectively. Cluster analysis of the combined data also showed that the FOC isolates were grouped into two clusters, sharing 42.9-100% similarity. Conclusion/Recommendations: The results of the present study indicate that the FOC isolates were closely related regardless of banana cultivars and location.
Highlights
The Internal Transcribed Spacer (ITS) + 5.8S of all the FOC isolates could be digested using Taq I, Msp I, Alu I, Bsu 15I, Bsu RI, Hin fI, Hin 6I and Eco RI indicating that the ITS + 5.8S regions of the FOC isolates contain recognition sites for these enzymes
The results of the present study showed combined data, separated the FOC isolates into two that the FOC isolates were closely related regardless of main clusters, showing 42.9-100% similarity (Fig. 5). banana cultivars and locations
ERIC-PCR is faster and less tedious compared to PCR-RFLP of ITS + 5.8S and a large number of samples could be analyzed in a short time[6]
Summary
Cubense (FOC) is the causal agent of banana wilt or Panama disease. It is a cosmopolitan soil-borne fungus which colonizes the vascular system of the host plant. Nangka Awak Masam used in characterization of Fusarium species. These techniques served as supportive means which provide. Awak Awak genetic insight of Fusarium species In this preliminary study, PCR-RFLP of Restriction analysis: About 5-10 μL of PCR products. ITS+5.8S regions and ERIC-PCR were conducted to were digested using nine restriction enzymes, namely characterize and assess genetic variation among FOC Alu I, Eco RI, Eco 88I, Bsu RI, Bsu 15I, Hin fI, Hin 6I, isolates from different banana cultivars in Malaysia.
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