Transformation of the cultivated mushroom,Agaricus bisporus, to hygromycin B resistance

Rhee, M. D. van de; Graça, Paula; Huizing, H. J.; Mooibroek, H.

doi:10.1007/bf02174382

Cited by 58 publications

(45 citation statements)

References 26 publications

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“…As the dominant selectable marker gene hph has previously been successfully employed to develop hygromycin resistancebased transformation systems for basidiomycete species, such as the edible mushrooms A. bisporus and P. ostreatus and the model systems C. cinerea and S. commune (10,27,35,42,55), we used the same gene to establish transformation systems of C. passeckerianus. We were successful in generating hygromycin-resistant transformants by using both Agrobacterium-and PEG-mediated methods.…”

Section: Vol 75 2009 Genetic Transformation Of Clitopilus Passeckermentioning

confidence: 99%

“…Vectors containing the Escherichia coli hph gene (for hygromycin resistance) coupled with PEG-mediated and/or Agrobacterium-mediated transformation systems have previously been developed for a few basidiomycete species, mainly model systems such as Coprinopsis cinerea (10) and Schizophyllum commune (35,51), or edible fungi, including Agaricus bisporus (55), Lentinula edodes (50), and Pleurotus ostreatus (42). Phleomycin resistance vectors were also employed to transform C. cinerea (27), S. commune (52), and Trametes versicolor (1).…”

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confidence: 99%

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Establishing Molecular Tools for Genetic Manipulation of the Pleuromutilin-Producing FungusClitopilus passeckerianus

Kilaru

Collins

Hartley

et al. 2009

Appl Environ Microbiol

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We describe efficient polyethylene glycol (PEG)-mediated and Agrobacterium-mediated transformation systems for a pharmaceutically important basidiomycete fungus, Clitopilus passeckerianus, which produces pleuromutilin, a diterpene antibiotic. Three dominant selectable marker systems based on hygromycin, phleomycin, and carboxin selection were used to study the feasibility of PEG-mediated transformation of C. passeckerianus. The PEG-mediated transformation of C. passeckerianus protoplasts was successful and generated hygromycinresistant transformants more efficiently than either phleomycin or carboxin resistance. Agrobacterium-mediated transformation with plasmid pBGgHg containing hph gene under the control of the Agaricus bisporus gpdII promoter led to hygromycin-resistant colonies and was successful when homogenized mycelium and fruiting body gill tissue were used as starting material. Southern blot analysis of transformants revealed the apparently random integration of the transforming DNA to be predominantly multiple copies for the PEG-mediated system and a single copy for the Agrobacterium-mediated system within the genome. C. passeckerianus actin and tubulin promoters were amplified from genomic DNA and proved successful in driving green fluorescent protein and DsRed expression in C. passeckerianus, but only when constructs contained a 5 intron, demonstrating that the presence of an intron is prerequisite for efficient transgene expression. The feasibility of RNA interference-mediated gene silencing was investigated using gfp as a target gene easily scored in C. passeckerianus. Upon transformation of gfp antisense constructs into a highly fluorescent strain, transformants were recovered that exhibited either reduced or undetectable fluorescence. This was confirmed by Northern blotting showing depletion of the target mRNA levels. This demonstrated that gene silencing is a suitable tool for modulating gene expression in C. passeckerianus. The molecular tools developed in this study should facilitate studies aimed at gene isolation or characterization in this pharmaceutically important species.

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Section: Vol 75 2009 Genetic Transformation Of Clitopilus Passeckermentioning

confidence: 99%

mentioning

confidence: 99%

Establishing Molecular Tools for Genetic Manipulation of the Pleuromutilin-Producing FungusClitopilus passeckerianus

Kilaru

Collins

Hartley

et al. 2009

Appl Environ Microbiol

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“…Pleurotus ostreatus [5][6][7] and Agaricus bisporus 8) have been transformed by PEG/CaCl 2 . Protoplasts of Agrocybe aegerita prototrophs have been transformed to prototrophy by electroporation with transformation frequencies ranging from 1 to 26 transformants/mg of DNA.…”

Section: )mentioning

confidence: 99%

Transformation of the Mycorrhizal Basidiomycetes,Suillus grevilleiandS. bovinus, by Particle Bombardment

Sunagawa

Murata

Miyazaki

et al. 2007

Bioscience, Biotechnology, and Biochemistry

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Mycorrhizal basidiomycetes, Suillus grevillei and S. bovinus, were transformed by particle bombardment. We isolated eight and four transformants from S. grevillei and S. bovinus respectively under three conditions differing in terms of target distance and helium pressure. The transformation frequencies were 1.2 and 0.6 transformants/g DNA. Plasmid DNAs were integrated into the genomic DNA of the transformants and stably maintained. This is the first description of the transformation of S. grevillei and S. bovinus by particle bombardment.

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“…In U. maydis, the linearization of the vector pHL1 increased the transformation frequency by about 20 times (34), and studies into other fungi, such as A. bisporus, M. grisea, and S. cerevisae have reported the same observation (14,24,31). Whitehead et al (35) showed that the use of the linearized vector pSTA10, which carries the A. niger niaD gene, increased the transformation frequency from 6.2 to 13.3 transformants/μg DNA.…”

Section: Transformationmentioning

confidence: 93%

Development of a transformation system for Penicillium brevicompactum based on the Fusarium oxysporum nitrate reductase gene

Varavallo¹,

Queiroz²,

Pereira³

et al. 2005

Braz. J. Microbiol.

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Penicillium brevicompactum is a filamentous fungus that presents a potential for industrial use due its efficient pectinase production. A heterologous transformation system was developed for P. brevicompactum based on the complementation of a nitrate reductase mutant. Nitrate reductase mutants were obtained by resistance to chlorate in a rate of 23.24% when compared to other mutations that lead to the chlorate resistance. One mutant named 4457-18X was chosen for the transformation experiments with the pNH24 vector containing de Fusarium oxysporum nitrate reductase gene. A frequency of approximately 3 transformants/µg DNA was obtained using the circular vector pNH24. This frequency was multiplied about 10 fold using the linearized vector with the Xba I restriction enzyme. Southern analysis of the transformants showed a tendency of the linearized vector to diminish the number of integrations compared to the use of the circular vector. The integration was random and stable in the analyzed transformants. The establishment of a transformation system for P. brevicompactum is fundamental for genetic manipulation of this microorganism.

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Transformation of the cultivated mushroom,Agaricus bisporus, to hygromycin B resistance

Cited by 58 publications

References 26 publications

Establishing Molecular Tools for Genetic Manipulation of the Pleuromutilin-Producing FungusClitopilus passeckerianus

Establishing Molecular Tools for Genetic Manipulation of the Pleuromutilin-Producing FungusClitopilus passeckerianus

Transformation of the Mycorrhizal Basidiomycetes,Suillus grevilleiandS. bovinus, by Particle Bombardment

Development of a transformation system for Penicillium brevicompactum based on the Fusarium oxysporum nitrate reductase gene

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