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Molecular cloning and in silico analysis of potential Fusarium resistance genes in banana.doc

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MolecularcloningandinsilicoanalysisofpotentialFusariumresistancegenesinbananaMolecularcloningandinsilicoanalysisofpotentialFusariumresistancegenesinbanana

Molecular cloning and in silico analysis of potential Fusarium resistance genes in banana Abstract The fungus Fusarium oxysporum is responsible for causing a destructive wilting disease in numerous crops including banana. To date, only two non-TIR-NBS-LRR genes that confer resistance to F. oxysporum, the I2 and Fom-2 genes from tomato and melon, respectively, have been isolated. We previously identified two partial non-TIR-NBS sequences, designated RGC2 and RGC5, from FOC race 4 resistant banana, Musa acuminata ssp. malaccensis. In the present study, we have isolated and characterized the entire cDNA sequences of RGC2 and RGC5 from the root transcriptome of M. acuminata ssp. malaccensis. The open reading frames (ORFs) of both genes were predicted to encode the typical structure of non-TIR-NBS-LRR proteins. Similarity searches using the entire ORF of RGC2 and RGC5 revealed considerable sequence similarity to the I2 gene. Further, phylogenetic analyses showed that the banana RGC2 and RGC5 clustered within the ancient non-TIR clade N1.1. Interestingly, although phylogenetically distantly related, the known Fusarium R genes I2 and Fom-2 also grouped within the clade N1.1. The fact that the I2 and Fom-2 genes cluster in the same phylogenetic clade N1.1 opens the possibility that other unknown Fusarium R genes from different plant families may share the same clade. These findings may assist future attempts to identify Fusarium R genes in banana and other plants. Keywords Banana Disease Fusarium Non-TIR-NBS-LRR Resistance genes Introduction Banana (Musa spp.) is one of the most important fruit crops in many tropical and subtropical countries. Diseases are major constraints in the production of this crop with Panama disease considered one of the most devastating (Ploetz and Pegg 2000). This disease is caused by the soil-borne fungus Fusarium oxysporum forma specialis (f. sp.) cubense (FOC), which penetrates the roots of susceptible bananas causing a lethal vascular wi