title: CmPEX6, a gene involved in peroxisome biogenesis, is essential for parasitism and conidiation by the sclerotial parasite Coniothyrium minitans.
abstract: Coniothyrium minitans is a sclerotial parasite of the plant-pathogenic fungus Sclerotinia sclerotiorum, and conidial production and parasitism are two important aspects for commercialization of this biological control agent. To understand the mechanism of conidiation and parasitism at the molecular level, we constructed a transfer DNA (tDNA) insertional library with the wild-type strain ZS-1. A conidiation-deficient mutant, ZS-1TN22803, was uncovered through screening of this library. This mutant could produce pycnidia on potato dextrose agar (PDA), but most were immature and did not bear conidia. Moreover, this mutant lost the ability to parasitize or rot the sclerotia of S. sclerotiorum. Analysis of the tDNA flanking sequences revealed that a peroxisome biogenesis factor 6 (PEX6) homolog of Saccharomyces cerevisiae, named CmPEX6, was disrupted by the tDNA insertion in this mutant. Targeted gene replacement and gene complementation tests confirmed that a null mutation of CmPEX6 was responsible for the phenotype of ZS-1TN22803. Further analysis showed that both ZS-1TN22803 and the targeted replacement mutants could not grow on PDA medium containing oleic acid, and they produced much less nitric oxide (NO) and hydrogen peroxide (H2O2) than wild-type strain ZS-1. The conidiation of ZS-1TN22803 was partially restored by adding acetyl-CoA or glyoxylic acid to the growth media. Our results suggest that fatty acid β-oxidation, reactive oxygen and nitrogen species, and possibly other unknown pathways in peroxisomes are involved in conidiation and parasitism by C. minitans.
authors.affiliation: State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, Hubei Province, People's Republic of China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wei
authors.forename: Wenjun
authors.initials: W
authors.lastname: Zhu
authors.forename: Jiasen
authors.initials: J
authors.lastname: Cheng
authors.forename: Jiatao
authors.initials: J
authors.lastname: Xie
authors.forename: Bo
authors.initials: B
authors.lastname: Li
authors.forename: Daohong
authors.initials: D
authors.lastname: Jiang
authors.forename: Guoqing
authors.initials: G
authors.lastname: Li
authors.forename: Xianhong
authors.initials: X
authors.lastname: Yi
authors.forename: Yanping
authors.initials: Y
authors.lastname: Fu
mesh_terms: D000105:Acetyl Coenzyme A; D000704:Analysis of Variance; D001203:Ascomycota / Q000235:genetics*; D001483:Base Sequence; D061046:Biological Control Agents; D015139:Blotting, Southern; D003470:Culture Media / Q000737:chemistry; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D015723:Gene Library; D005816:Genetic Complementation Test; D006038:Glyoxylates; D054884:Host-Pathogen Interactions; D006861:Hydrogen Peroxide / Q000378:metabolism; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009569:Nitric Oxide / Q000378:metabolism; D020675:Peroxisomes / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013172:Spores, Fungal / Q000235:genetics / Q000502:physiology*
chemical_list: D061046:Biological Control Agents; D003470:Culture Media; D004269:DNA, Bacterial; D006038:Glyoxylates; C036483:T-DNA; D009569:Nitric Oxide; D000105:Acetyl Coenzyme A; D006861:Hydrogen Peroxide; C031150:glyoxylic acid
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