Two-dimensional electrophoresis protein profile of the phytopathogenic fungus Botrytis cinerea

Fernández-Acero, Francisco Javier; Jorge, Inmaculada; Vallejo, Inmaculada; Carbú, María; Camafeita, Emilio; López, Juan Antonio; Cantoral, Jesús M.; Jorrı́n, Jesús

doi:10.1002/pmic.200500436

Cited by 75 publications

(68 citation statements)

References 44 publications

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“…Different procedures were used to optimize this protocol for P. chrysogenum proteins (see "Results"). The final optimal method (based on that described by Ferná ndez-Acero et al (27) for the fungus Botrytis cinerea) is detailed below and consisted of solubilizing proteins by phosphate buffer and precipitation with TCA/acetone.…”

Section: Methodsmentioning

confidence: 99%

Proteome Analysis of the Penicillin Producer Penicillium chrysogenum

Jami

Barreiro

Garcı́a-Estrada

et al. 2010

Molecular & Cellular Proteomics

119

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Proteomics is a powerful tool to understand the molecular mechanisms causing the production of high penicillin titers by industrial strains of the filamentous fungus Penicillium chrysogenum as the result of strain improvement programs. Penicillin biosynthesis is an excellent model system for many other bioactive microbial metabolites. The recent publication of the P. chrysogenum genome has established the basis to understand the molecular processes underlying penicillin overproduction. We report here the proteome reference map of P. chrysogenum Wisconsin 54-1255 (the genome project reference strain) together with an in-depth study of the changes produced in three different strains of this filamentous fungus during industrial strain improvement. Two-dimensional gel electrophoresis, peptide mass fingerprinting, and tandem mass spectrometry were used for protein identification. Around 1000 spots were visualized by "blue silver" colloidal Coomassie staining in a non-linear pI range from 3 to 10 with high resolution, which allowed the identification of 950 proteins (549 different proteins and isoforms). Comparison among the cytosolic proteomes of the wild-type NRRL 1951, Wisconsin 54-1255 (an improved, moderate penicillin producer), and AS-P-78 (a penicillin high producer) strains indicated that global metabolic reorganizations occurred during the strain improvement program. The main changes observed in the high producer strains were increases of cysteine biosynthesis (a penicillin precursor), enzymes of the pentose phosphate pathway, and stress response proteins together with a reduction in virulence and in the biosynthesis of other secondary metabolites different from penicillin (pigments and isoflavonoids). In the wild-type strain, we identified enzymes to utilize cellulose, sorbitol, and other carbon sources that have been lost in the high penicillin producer strains. Changes in the levels of a few specific proteins correlated well with the improved penicillin biosynthesis in the high producer strains. These results provide useful information to improve the production of many other bioactive secondary metabolites. Molecular & Cellular Proteomics 9:1182-1198, 2010.

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Section: Methodsmentioning

confidence: 99%

Proteome Analysis of the Penicillin Producer Penicillium chrysogenum

Jami

Barreiro

Garcı́a-Estrada

et al. 2010

Molecular & Cellular Proteomics

119

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“…Previously, such approaches have been used to study phytopathogenic fungi through protein profiling (2,5,12,42,44) and to identify host- (45) and morphogenesis-responsive (25) proteins. Recent sequencing of the genomes of the phytopathogenic fungi Magnaporthe grisea, Ustilago maydis, Fusarium graminearum, and Stagonospora nodorum (7,9,19,24) provides an opportunity for more thorough mass spectrometry (MS)-based proteomic analyses (2,42).…”

mentioning

confidence: 99%

A Signaling-Regulated, Short-Chain Dehydrogenase of Stagonospora nodorum Regulates Asexual Development

et al. 2008

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The fungus Stagonospora nodorum is a causal agent of leaf and glume blotch disease of wheat. It has been previously shown that inactivation of heterotrimeric G protein signaling in Stagonospora nodorum caused development defects and reduced pathogenicity [P. S. Solomon et al., Mol. Plant-Microbe Interact. 17: [456][457][458][459][460][461][462][463][464][465][466] 2004]. In this study, we sought to identify targets of the signaling pathway that may have contributed to phenotypic defects of the signaling mutants. A comparative analysis of Stagonospora nodorum wild-type and G␣-defective mutant (gna1) intracellular proteomes was performed via two-dimensional polyacrylamide gel electrophoresis. Several proteins showed significantly altered abundances when comparing the two strains. One such protein, the short-chain dehydrogenase Sch1, was 18-fold less abundant in the gna1 strain, implying that it is positively regulated by G␣ signaling. Gene expression and transcriptional enhanced green fluorescent protein fusion analyses of Sch1 indicates strong expression during asexual development. Mutant strains of Stagonospora nodorum lacking Sch1 demonstrated poor growth on minimal media and exhibited a significant reduction in asexual sporulation on all growth media examined. Detailed histological experiments on sch1 pycnidia revealed that the gene is required for the differentiation of the subparietal layers of asexual pycnidia resulting in a significant reduction in both pycnidiospore size and numbers.

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“…Mechanical cell disruption using glass beads in beating mills is, therefore, a preferred approach [15][16][17][18][19]. The mill can be also thermostatised at 4˚C avoiding increased temperatures by heat generation and protein lysis due to intracellular proteases or denaturation.…”

Section: Sample Homogenisationmentioning

confidence: 99%

A Review beyond the borders: Proteomics of microclonial black fungi and black yeasts

Marzban¹,

Tesei²,

Sterflinger³

2013

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Black microcolonial fungi and black yeasts are inhabitants of extreme environments like vulcanic, desert and polar regions, where they are exposed to enhanced temperature alterations and desiccation. They have developed, therefore, extraordinary biologic characteristics which are mainly based on the expression of proteins, however, these are rarely studied and known. The review article presented here is focused on the obstacles and solutions for the proteomic analyses of this very particular fungal species.

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Two-dimensional electrophoresis protein profile of the phytopathogenic fungus Botrytis cinerea

Cited by 75 publications

References 44 publications

Proteome Analysis of the Penicillin Producer Penicillium chrysogenum

Proteome Analysis of the Penicillin Producer Penicillium chrysogenum

A Signaling-Regulated, Short-Chain Dehydrogenase of Stagonospora nodorum Regulates Asexual Development

A Review beyond the borders: Proteomics of microclonial black fungi and black yeasts

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