Fungal Transformation: From Protoplasts to Targeted Recombination Systems

Martı́n, Juan F.

doi:10.1007/978-3-319-10142-2_1

Cited by 10 publications

(10 citation statements)

References 134 publications

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“…The range of inorganic salts, sugars and sugar alcohols that can be used as osmotic stabilisers for fungal protoplasts had been described previously . There has been no final conclusion for the superiority of one component over another in filamentous fungi . In this study, sucrose was found to be the best osmotic stabiliser for L. lecanii , a result that tends to contradict the previous assumption that inorganic salts are more effective with filamentous fungi .…”

Section: Discussioncontrasting

confidence: 67%

“…37 There has been no final conclusion for the superiority of one component over another in filamentous fungi. 24 In this study, sucrose was found to be the best osmotic stabiliser for L. lecanii, a result that tends to contradict the previous assumption that inorganic salts are more effective with filamentous fungi. 38 The enzymolysis time also significantly influenced the efficiency of fungal protoplast generation.…”

Section: Discussioncontrasting

confidence: 63%

“…Since the first report of a DNA-mediated transformation system for a fungus in 1973, 33 many fungal species have been transformed and a range of protocols now exist not only for PEG-mediated transformation of protoplasts but also for electroporation, biolistics and Agrobacterium-mediated transformation. 24 However, what is apparent from the literature is that a process for one fungus may not necessarily work for another, and each fungus will usually require its own specific conditions. In this paper, we studied the conditions of protoplast transformation for L. lecanii by taking a step-by-step approach in which most of the parameters described to be important in protoplast generation were systematically tested.…”

Section: Discussionmentioning

confidence: 99%

“…22 Various transformation systems have been developed for a large number of flamentous fungi, and the transformation of protoplasts mediated by PEG remains a dominant method. 23,24 Protoplast generation is affected by many factors (such as fungal materials used, types and concentrations of lyric enzymes and osmotic stabilisers), and the cell wall composition varies among fungal species. Thus, no recommendations for the optimised parameters about protoplast generation can be used in certain fungus species.…”

Section: -6mentioning

confidence: 99%

“…To date, only one transformation method using Agrobacterium tumefaciens has been reported for L. lecanii . Various transformation systems have been developed for a large number of flamentous fungi, and the transformation of protoplasts mediated by PEG remains a dominant method . Protoplast generation is affected by many factors (such as fungal materials used, types and concentrations of lyric enzymes and osmotic stabilisers), and the cell wall composition varies among fungal species.…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Establishment of polyethylene‐glycol‐mediated protoplast transformation for Lecanicillium lecanii and development of virulence‐enhanced strains against Aphis gossypii

Zhang

Xie

Zhang

et al. 2016

Pest Management Science

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Section: Discussioncontrasting

confidence: 67%

Section: Discussioncontrasting

confidence: 63%

Section: Discussionmentioning

confidence: 99%

Section: -6mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Establishment of polyethylene‐glycol‐mediated protoplast transformation for Lecanicillium lecanii and development of virulence‐enhanced strains against Aphis gossypii

Zhang

Xie

Zhang

et al. 2016

Pest Management Science

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Genetic Transformation of Filamentous Fungi: Achievements and Challenges

Lichius

Ruiz

Zeilinger

2020

Grand Challenges in Biology and Biotechnology

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A natural short pathway synthesizes roquefortine C but not meleagrin in three different Penicillium roqueforti strains

Kosalková

Domínguez-Santos

Coton

et al. 2015

Appl Microbiol Biotechnol

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The production of mycotoxins and other secondary metabolites in Penicillium roqueforti is of great interest because of its long history of use in blue-veined cheese manufacture. In this article, we report the cloning and characterization of the roquefortine gene cluster in three different P. roqueforti strains isolated from blue cheese in the USA (the type strain), France, and the UK (Cheshire cheese). All three strains showed an identical roquefortine gene cluster organization and almost identical (98-99%) gene nucleotide sequences in the entire 16.6-kb cluster region. When compared with the Penicillium chrysogenum roquefortine/meleagrin seven-gene cluster, the P. roqueforti roquefortine cluster contains only four genes (rds, rdh, rpt, and gmt) encoding the roquefortine dipeptide synthetase, roquefortine D dehydrogenase, roquefortine prenyltransferase, and a methyltransferase, respectively. Silencing of the rds or rpt genes by the RNAi strategy reduced roquefortine C production by 50% confirming the involvement of these two key genes in roquefortine biosynthesis. An additional putative gene, orthologous of the MFS transporter roqT, is rearranged in all three strains as a pseudogene. The same four genes and a complete (not rearranged) roqT, encoding a MFS transporter containing 12 TMS domains, occur in the seven-gene cluster in P. chrysogenum although organized differently. Interestingly, the two "late" genes of the P. chrysogenum roquefortine/meleagrin gene cluster that convert roquefortine C to glandicoline B and meleagrin are absent in the P. roqueforti four-gene cluster. No meleagrin production was detected in P. roqueforti cultures grown in YES medium, while P. chrysogenum produces meleagrin in these conditions. No orthologous genes of the two missing meleagrin synthesizing genes were found elsewhere in the recently released P. roqueforti genome. Our data suggest that during evolution, the seven-gene cluster present in P. chrysogenum, and probably also in other glandicoline/meleagrin producing fungi, has been trimmed down to a short cluster in P. roqueforti leading to the synthesis of roquefortine C rather than meleagrin as a final product.

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Fungal Transformation: From Protoplasts to Targeted Recombination Systems

Cited by 10 publications

References 134 publications

Establishment of polyethylene‐glycol‐mediated protoplast transformation for Lecanicillium lecanii and development of virulence‐enhanced strains against Aphis gossypii

Establishment of polyethylene‐glycol‐mediated protoplast transformation for Lecanicillium lecanii and development of virulence‐enhanced strains against Aphis gossypii

Genetic Transformation of Filamentous Fungi: Achievements and Challenges

A natural short pathway synthesizes roquefortine C but not meleagrin in three different Penicillium roqueforti strains

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