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ABSTRACT: As it is extremely difficult to make DNA transformation for the obligate fungus, Blumeria graminis f. sp. tritici (Bgt), we developed a heterologous expression system for characterization of a Bgt gene, CYP51, which encodes 14α-demethylase. The CYP51 gene from Bgt was transformed into the necrotrophic fungus, Botrytis cinerea. Reverse transcription polymerase chain reaction showed that the Bgt CYP51 was transcribed in B. cinerea. Green fluorescence was observed in the transformants of B. cinerea carrying the Bgt CYP51-GFP fusion cassette, suggesting that its translation was successful. Fungicide sensitivity tests revealed that B. cinerea transformed with Bgt CYP51 showed reduced sensitivity to a sterol demethylation inhibitor triadimefon, but not to a benzimidazole fungicide carbendazim. These results indicated that this heterologous expression system can be used for functional analysis of other Bgt genes.
Journal of Eukaryotic Microbiology 09/2011; 59(1):88-92. · 2.66 Impact Factor
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ABSTRACT: The Magnaporthe oryzae genome contains two homologous CYP51 genes, MoCYP51A and MoCYP51B, that putatively encode sterol 14α-demethylase enzymes. Targeted gene deletion mutants of MoCYP51A were morphologically indistinguishable from the isogenic wild type M. oryzae strain Guy11 in vegetative culture, but were impaired in both conidiation and virulence. Deletion of MoCYP51B did not result in any obvious phenotypic changes compared with Guy11. The Δmocyp51A mutants were also highly sensitive to sterol demethylation inhibitor (DMI) fungicides, while Δmocyp51B mutants were unchanged in their sensitivity to these fungicides. Expression of both MoCYP51A and MoCYP51B was significantly induced by exposure to DMI fungicides. Analysis of intracellular localization of MoCyp51A showed that MoCyp51A was mainly localized to the cytoplasm of hyphae and conidia. Taken together, our results indicate that MoCYP51A is required for efficient conidiogenesis, full virulence and for mediating DMI sensitivity by the rice blast fungus.
Fungal Genetics and Biology 09/2010; 48(2):144-53. · 3.74 Impact Factor
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ABSTRACT: A specific and rapid real-time PCR assay for detecting Ralstonia solanacearum in the horticultural soil and plant tissues was developed in this study. The specific primers RSF/RSR were designed based on the upstream region of UDP-3-O-acyl-GlcNAc deacetylase gene from R. solanacearum, and a PCR product of 159 bp was amplified specifically from 28 strains of R. solanacearum, which represent all genetically diverse AluI types and all 6 biovars, but not from any other nontarget species. The detection limit of 102 CFU/g tomato stem and horticultural soil was achieved in this real-time PCR assay. The high sensitivity and specificity observed with filed samples as well as with artificially infected samples suggested that this method might be a useful tool for detection and quantification of R. solanacearum in precise forecast and diagnosis.
Journal of Microbiology and Biotechnology 01/2010; 20(1):193-201. · 1.38 Impact Factor
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ABSTRACT: Aflatoxins produced primarily by two closely related fungi, Aspergillus flavus and Aspergillus parasiticus, are mutagenic and carcinogenic in animals and humans. Of many approaches investigated to manage aflatoxin contamination, biological control method has shown great promise. Numerous organisms, including bacteria, yeasts and nontoxigenic fungal strains of A. flavus and A. parasiticus, have been tested for their ability in controlling aflatoxin contamination. Great successes in reducing aflatoxin contamination have been achieved by application of nontoxigenic strains of A. flavus and A. parasiticus in fields of cotton, peanut, maize and pistachio. The nontoxigenic strains applied to soil occupy the same niches as the natural occurring toxigenic strains. They, therefore, are capable of competing and displacing toxigenic strains. In this paper, we review recent development in biological control of aflatoxin contamination.
Journal of Zhejiang University SCIENCE B 11/2008; 9(10):787-92. · 1.10 Impact Factor
