In Vitro Antifungal Activity Of Synthetic Dsrna Molecules Against Two Pathogens Of Banana, Fusarium Oxysporum F. Sp. Cubense And Mycosphaerella Fijiensis
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Date
2013-01-03
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Publisher
Pest Management Science
Abstract
BACKGROUND: A key challenge for designing RNAi-based crop protection strategies is the identification of effective target genes in the pathogenic organism. In this study, in vitro antifungal activities of a set of synthetic double-stranded RNA molecules on spore germination of two major pathogenic fungi of banana, Fusarium oxysporum Schlecht f. sp. cubense WC Snyder & HN Hans (Foc) and Mycosphaerella fijiensis Morelet (Mf) were evaluated.
RESULTS: All the tested synthetic dsRNAs successfully triggered the silencing of target genes and displayed varying degrees of potential to inhibit spore germination of both tested banana pathogens. When Foc dsRNAs were applied to Foc spores, inhibition ranged from 79.8 to 93.0%, and from 19.9 to 57.8% when Foc dsRNAs were applied to Mf spores. However, when Mf dsRNAs were applied on Mf spores, inhibition ranged from 34.4 to 72.3%, and from 89.7 to 95.9% when Mf dsRNAs were applied to Foc spores.
CONCLUSION: The dsRNAs for adenylate cyclase, DNA polymerase alpha subunit and DNA polymerase delta subunit showed high levels of spore germination inhibition during both self- and cross-species tests, making them the most promising targets for RNA-mediated resistance in banana against these fungal pathogens.
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Keywords
double-stranded RNA, genes, RNA interference, spore 1 INTRODUCTION Two major fungal diseases, black Sigatoka caused by Mycosphaerella fijiensis Morelet (Mf) and Panama disease or Fusarium wilt caused by Fusarium oxysporum Schlecht f. sp. cubense WC Snyder & HN Hans (Foc), threaten banana production worldwide. They are reported to cause a yield loss of over 38% and up to 50% respectively in some banana-producing regions.1, 2 Host resistance has long been identified as the most feasible and sustainable way of managing diseases in crops, including banana. This is achievable using either conventional cross-breeding and/or genetic modification (GM) approaches. However, owing to the paucity of disease-resistant germplasms and the sterility of most edible banana cultivars, genetic engineering is now regarded as the best way to introduce resistance genes into this hard- to-breed crop. Significant successes in the genetic engineering of banana have been obtained as far as agrobacterium- mediated transformation and microprojectile bombardment of cell suspension are concerned.3–6 Transformation systems using apical meristems7, 8 as well as intercalary meristematic tissues9 have also been reported. These achievements have enabled the experimentation of a number of technology platforms that can confer an acceptable level of resistance to banana pests and pathogens. Strategies rely mainly on the expression of germination inhibition