Search

The discovery of RNA interference (RNAi) has been the major recent breakthrough in biology. Only a few years after its discovery, RNAi has rapidly become a powerful reverse genetic tool, especially in organisms where gene targeting is inefficient and/or time-consuming. In filamentous fungi, RNAi is not currently used as widely as is gene targeting by homologous recombination that works with practical efficiencies in most model fungal species. However, to explore gene function in filamentous fungi, RNAi has the potential to offer new, efficient tools that gene disruption methods cannot provide. In this review, possible advantages and disadvantages of RNAi for fungal biology in the postgenomics era will be discussed. In addition, we will briefly review recent discoveries on RNAi-related biological phenomena (RNA silencing) in fungi. [Copyright &y& Elsevier]

Dicer is a ribonuclease III-like enzyme playing a key role in the RNA silencing pathway. Genome sequencing projects have demonstrated that eukaryotic genomes vary in the numbers of Dicer-like (DCL) proteins from one (human) to four (Arabidopsis). Two DCL genes, MDL-1 and -2 (Magnaporthe Dicer-like-l and -2) have been identified in the genome of the filamentous fungus Magnaporthe oryzae. Here we show that the knockout of MDL-2 drastically impaired gene silencing of enhanced green fluorescence protein by hairpin RNA and reduced related small interfering RNA (siRNA) accumulation to nondetectable levels. In contrast, mutating the other DCL, MDL-1, exhibited a gene silencing frequency similar to wild type and accumulated siRNA normally. The silencing-deficient phenotype and loss of siRNA accumulation in the mdl-2 mutant was restored by genetic complementation with the wild-type MDL-2 allele. These results indicate that only MDL-2 is responsible for siRNA production, and no functional redundancy exists between MDL-1 and MDL-2 in the RNA silencing pathway in M. oryzae. Our findings contrast with a recent report in the filamentous fungus Neurospora crassa, where two DCL proteins are redundantly involved in the RNA silencing pathway, but are similar to the resuits obtained in a more distantly related organism, Drosophila melanogaster, where an individual DCL protein has a distinct role in the siRNA/micro-RNA pathways. [ABSTRACT FROM AUTHOR]

Abstract: Pyricularia isolates from various host plants were subjected to a multilocus phylogenetic analysis based on rDNA-ITS, actin, β-tubulin, and calmodulin loci. A combined gene tree resolved seven groups with 100% BS support, suggesting that they are monophyletic groups supported concordantly by all four loci. By incorporating biological and morphological species criteria, each of the seven groups was considered to be a current species. However, phylogenetic relationships among these species were unresolved in the single-gene trees and in the combined tree. Furthermore, the transition from concordance to conflict occurred more than once in the combined gene tree. They were interpreted by assuming that Pyricularia has evolved through repeated species radiation. The transition point other than the current species limit was considered to be the limit of the former species. [Copyright &y& Elsevier]