Molecular cloning, characterization and expression analysis of a pacC homolog in the mycoparasite Coniothyrium minitans

Han, Yongchao; Li, Guoqing; Yang, Long

doi:10.1007/s11274-010-0469-5

Cited by 15 publications

(16 citation statements)

References 37 publications

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“…Oxalic acid, a pathogenicity factor of S. sclerotiorum, is likely to be a signal for this interaction. C. minitans produces antifungal substances with increasing activity at low pH caused by oxalic acid and produces beta-1,3-glucanase, an fCWDE, at higher pH after oxalic acid is degraded (15)(16)(17). Theoretically, the interactions between C. minitans and S. sclerotiorum are similar to that between the plant pathogen and host plants, and many similar pathways are likely to be involved in this hyperparasitism.…”

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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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“…The mechanism is that the activated PacC induces transcription of fungal genes preferentially expressed under alkaline conditions and simultaneously represses transcription of genes preferentially expressed under acidic conditions (Tilburn et al ., ; Díez et al ., ). Han and colleagues () identified a pacC homologue, cmpacC , in C. minitans . Han and colleagues () also found that transcription of cmpacC as well as the two mycoparasitism‐associated genes cmch1 and cmg1 in C. minitans was increased in parallel when ambient pH increased from 3 to 8.…”

Section: Discussionmentioning

confidence: 99%

“…Han and colleagues () identified a pacC homologue, cmpacC , in C. minitans . Han and colleagues () also found that transcription of cmpacC as well as the two mycoparasitism‐associated genes cmch1 and cmg1 in C. minitans was increased in parallel when ambient pH increased from 3 to 8. Therefore, cmpacC may positively regulate transcription of Cmch1 and Cmg1 , but negatively regulate transcription of Cmoxdc1 , in C. minitans .…”

Section: Discussionmentioning

confidence: 99%

“…The resulting two DNA fragments were gel-purified and cloned separately into the pMD18-T vector (TaKaRa Biotechnology, Dalian, China), and sequenced. Then, the 3′-and 5′-terminal regions flanking the cloned DNA sequences were obtained using the 3′RACE and 5′RACE techniques respectively (Han et al, 2011). The putative promoter region for each gene was obtained using the inverse PCR technique (Mäkelä et al, 2009).…”

Section: Cloning Of Cmoxdc1 and Cmoxdc2mentioning

confidence: 99%

“…The gene structure of Cmoxdc2 was deduced on the basis of the full-length DNA sequence using the Softberry program (http://www.softberry.com), as we failed to obtain the full-length cDNA sequence for that gene using RT-PCR. The amino acid sequences encoded by Cmoxdc1 and Cmoxdc2 were deduced using the DNAman software (version 5.2.2, Lynnon, Vaudreuil, Quebec, Canada) (Han et al, 2011). The molecular weight and the pI for CMOXDC1 and CMOXDC2 were inferred using the ProtParam tool (http://us.expasy.org/tools/protparam.html) (Swiss Institute of Bioinformatics, Lausanne, Switzerland).…”

Section: Sequence Analysis Of Cmoxdc1 and Cmoxdc2mentioning

confidence: 99%

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Degradation of oxalic acid by the mycoparasite Coniothyrium minitans plays an important role in interacting with Sclerotinia sclerotiorum

Zeng

Zhang

Han

et al. 2014

Environmental Microbiology

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Coniothyrium minitans (Cm) is a mycoparasite of the phytopathogenic fungus Sclerotinia sclerotiorum (Ss). Ss produces a virulence factor oxalic acid (OA) which is toxic to plants and also to Cm, and Cm detoxifies OA by degradation. In this study, two oxalate decarboxylase genes, Cmoxdc1 and Cmoxdc2, were cloned from Cm strain Chy-1. OA and low pH induced expression of Cmoxdc1, but not Cmoxdc2. Cmoxdc1 was partially responsible for OA degradation, whereas Cmoxdc2 had no effect on OA degradation. Disruption of Cmoxdc1 in Cm reduced its ability to infect Ss in dual cultures where OA accumulated. Compared with Chy-1, the Cmoxdc1-disrupted mutants had reduced expression levels of two mycoparasitism-related genes chitinase (Cmch1) and β-1,3-glucanase (Cmg1), and had no detectable activity of extracellular proteases in the presence of OA. On the other hand, the cultural filtrates of the Cmoxdc1-disrupted mutants in OA-amended media showed enhanced antifungal activity, possibly because of increased production of antifungal substances under acidic pH condition resulted from reduced Cmoxdc1-mediated OA degradation. This study provides direct genetic evidence of OA degradation regulating mycoparasitism and antibiosis of Cm against Ss, and sheds light on the sophisticated strategies of Cm in interacting with metabolically active mycelia and dormant sclerotia of Ss.

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Fungal Genes and Metabolites Associated with the Biocontrol of Soil-borne Plant Pathogenic Fungi

Daguerre¹,

Edel-Hermann²,

Steinberg³

2016

Fungal Metabolites

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Molecular cloning, characterization and expression analysis of a pacC homolog in the mycoparasite Coniothyrium minitans

Cited by 15 publications

References 37 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

Degradation of oxalic acid by the mycoparasite Coniothyrium minitans plays an important role in interacting with Sclerotinia sclerotiorum

Fungal Genes and Metabolites Associated with the Biocontrol of Soil-borne Plant Pathogenic Fungi

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