Building blocks method, efficient gene silencing of multiple genes

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In the post-genomics era, a major obstacle to functional genomics is functional redundancy among gene family members. To overcome this difficulty, we have developed an RNAi strategy designated building blocks method, in which a 40-60 bp sequence is chosen from a target gene, and such sequences from multiple gene family members were combined to make an artificial RNAi trigger by synthetic DNA. In this study, the building blocks method was applied to knock-down (KD) two gene families, xylanases and cellulases, to elucidate their roles in the pathogenicity of the blast fungus, Magnaporthe oryzae. In the resulting KD mutants, the transcript levels of all the target genes were significantly decreased. The number of lesions, rate of penetration, and extent of infected cells were all reduced in KD mutant-infected leaves. Thus, we concluded that xylanases and cellulases play significant roles in vertical penetration and horizontal expansion of M. oryzae in infected plants.