Disruption of the Coniothyrium minitans PIF1 DNA helicase gene impairs growth and capacity for sclerotial mycoparasitism

Rogers, Christopher W.; Challen, Michael P.; Muthumeenakshi, S.; Sreenivasaprasad, S.; Whipps, J. M.

doi:10.1099/mic.0.2008/017020-0

Cited by 11 publications

(7 citation statements)

References 40 publications

(51 reference statements)

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“…Several genes and pathways regulating C. minitans conidiation have recently been elucidated, including signaling mediated by nitric oxide, cGMP, cyclic AMP (cAMP), the mitogen-activated protein (MAP) kinase cascade, and the PIF1 DNA helicase gene, as well as by many cell walldegrading enzymes (fCWDE), such as beta-1,3-glucanase, chitinase, and so on (9)(10)(11)(12)(13)(14).…”

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confidence: 99%

CmPEX6 , a Gene Involved in Peroxisome Biogenesis, Is Essential for Parasitism and Conidiation by the Sclerotial Parasite Coniothyrium minitans

Wei

Zhu

Chéng

et al. 2013

Appl Environ Microbiol

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bConiothyrium 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 wildtype 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 (H 2 O 2 ) 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 ␤-oxidation, reactive oxygen and nitrogen species, and possibly other unknown pathways in peroxisomes are involved in conidiation and parasitism by C. minitans. Coniothyrium minitans, a mycoparasite of the plant fungal pathogen Sclerotinia sclerotiorum, can parasitize and decay both hyphae and sclerotia of Sclerotinia spp. It also reduces the germination of sclerotia and inhibits further infection by S. sclerotiorum hyphae; therefore, its existence in crop fields may play a very important role in suppressing Sclerotinia diseases (1-5). Its antagonistic properties have made C. minitans a well-known biological control microorganism, and several formulations based on C. minitans have been developed and registered commercially (4, 6).Coniothyrium minitans is a coelomycete fungus producing conidia in pycnidia. Understanding the conidiation of C. minitans at the molecular level may help to improve the efficiency of conidial production, which is important for the commercial use of biological control agents. Asexual reproduction of fungi which do not have a sexual stage in their life cycle is essential for survival and spread in nature; however, many previous studies on conidiation have focused on hyphomycete fungi (unenclosed conidia), such as Aspergillus nidulans, Neurospora crassa, and Magnaporthe oryzae, and little is known about conidiation in coelomycetes, except for the chestnut blight fungus Cryphonectria parasitica (7; for a review, see reference 8). Studies on the conidiation of C. minitans may improve our understanding of the general nature of asexual reproduction by coelomycetes.Conidiation by C. min...

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confidence: 99%

CmPEX6 , a Gene Involved in Peroxisome Biogenesis, Is Essential for Parasitism and Conidiation by the Sclerotial Parasite Coniothyrium minitans

Wei

Zhu

Chéng

et al. 2013

Appl Environ Microbiol

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“…These helicases are reportedly involved in various functions, such as replication, transcription, translation, recombination, DNA repair, and ribosomal biogenesis [22]. Abnormal helicase activity causes several developmental defects in fungi and plants, such as slow growth, premature aging and aberrant assembly of chromosomes [23], [24]. These observations suggest a close relationship between the abnormalities observed and this helicase.…”

Section: Resultsmentioning

confidence: 95%

Identification of Degenerate Nuclei and Development of a SCAR Marker for Flammulina velutipes

Kim

et al. 2014

PLoS ONE

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Flammulina velutipes is one of the major edible mushrooms in the world. Recently, abnormalities that have a negative impact on crop production have been reported in this mushroom. These symptoms include slow vegetative growth, a compact mycelial mat, and few or even no fruiting bodies. The morphologies and fruiting capabilities of monokaryons of wild-type and degenerate strains that arose through arthrospore formation were investigated through test crossing. Only one monokaryotic group of the degenerate strains and its hybrid strains showed abnormal phenotypes. Because the monokaryotic arthrospore has the same nucleus as the parent strain, these results indicated that only one aberrant nucleus of the two nuclei in the degenerate strain was responsible for the degeneracy. A sequence-characterized amplified region marker that is linked to the degenerate monokaryon was identified based on a polymorphic sequence that was generated using random primers. Comparative analyses revealed the presence of a degenerate-specific genomic region in a telomere, which arose via the transfer of a genomic fragment harboring a putative helicase gene. Our findings have narrowed down the potential molecular targets responsible for this phenotype for future studies and have provided a marker for the detection of degenerate strains.

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“…hPIF1 has therefore been proposed as a cancer therapy target [ 28 ]. The mechanism may be related to its effect on maintaining mitochondrial stability in response to reactive oxygen species (ROS) [ 37 ]. ROS are essential for the initiation, progression, and metastasis of cancer, and altered ROS levels in cancer cells are one of the reasons for recurrence/relapse [ 38 ].…”

Section: Discussionmentioning

confidence: 99%

Identifying PIF1 as a Potential Target of Wenxia Changfu Formula in Promoting Lung Cancer Cell Apoptosis: Bioinformatics Analysis and Biological Evidence

Yin

Kan

Ruan

et al. 2021

Evidence-Based Complementary and Alternative Medicine

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Lung cancer remains the leading cause of cancer-related deaths worldwide. Traditional Chinese medicine (TCM) is a valuable resource of active natural products and plays an important role in cancer treatment with the advantages of high efficiency and safety. Wenxia Changfu formula (WCF) is modified from Dahuang Fuzi decoction from Han Dynasty and has been used for treating lung cancer in China. Our previous research showed that WCF had an antitumor effect in vivo and in vitro, while the mechanism has not been well illustrated. In this study, the effect of WCF on the proliferative ability in three lung cancer cells and one noncancerous human cell line was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. WCF suppressed A549, H460, and PC-9 cell viability in a dose-dependent manner, with no inhibition of noncancerous MRC-5 cells after 48 h treatment with WCF (0–50 mg/mL). Furthermore, we screened for genes in A549 cells using four WCF-treated samples and four control samples on a gene expression profile microarray. 21 genes were significantly downregulated by WCF, which may potentially play an important role in the proliferation of A549 cells. High-content screening evaluated whether silencing the 21 genes affected A549 cell growth. The results showed that PIF1 knockdown exhibited the most potent inhibition of cell proliferation compared with the other genes. Downregulation of PIF1 increased A549 cell apoptosis and the activity of caspase 3/7. Besides, RT-PCR showed that the expression levels of PIF1 mRNA decreased significantly in A549, H460, and PC-9 cells after WCF treatment. In conclusion, the present observations indicate that WCF may inhibit lung cancer cell proliferation by promoting apoptosis via regulating the expression of PIF1.

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Disruption of the Coniothyrium minitans PIF1 DNA helicase gene impairs growth and capacity for sclerotial mycoparasitism

Cited by 11 publications

References 40 publications

CmPEX6 , a Gene Involved in Peroxisome Biogenesis, Is Essential for Parasitism and Conidiation by the Sclerotial Parasite Coniothyrium minitans

CmPEX6 , a Gene Involved in Peroxisome Biogenesis, Is Essential for Parasitism and Conidiation by the Sclerotial Parasite Coniothyrium minitans

Identification of Degenerate Nuclei and Development of a SCAR Marker for Flammulina velutipes

Identifying PIF1 as a Potential Target of Wenxia Changfu Formula in Promoting Lung Cancer Cell Apoptosis: Bioinformatics Analysis and Biological Evidence

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