The efficient genetic transformation of Cordyceps militaris by using mononuclear protoplasts

Lou, Haiwei; Ye, Zhi‐Wei; Yu, Yinghao; Lin, Jizhen; Guo, Li‐Qiong; Chen, Bai‐Xiong; Tang, Hong-Biao; Wei, Tao; Chen, Letao; Fan, Yun

doi:10.1016/j.scienta.2018.08.043

Cited by 20 publications

(13 citation statements)

References 31 publications

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“…Most of them focused on developing suitable methodologies and interventions to avoid (or minimize) culture degeneration [21][22][23][24][25][26][27][28][29][30][31], on improving ascomata and mycelia production [12,15,[32][33][34][35][36], and/or on enhancing cordycepin output by adopting various approaches and the use of both solid and liquid substrates [12,15,23,32,[37][38][39][40][41][42][43][44][45][46][47]. In addition, the application of genetic engineering breakthrough technologies has resulted in a considerable improvement in C. militaris biomass content in various metabolites, including cordycepin [37,[48][49][50][51].…”

Section: Introductionmentioning

confidence: 99%

Biomass and Cordycepin Production by the Medicinal Mushroom Cordyceps militaris—A Review of Various Aspects and Recent Trends towards the Exploitation of a Valuable Fungus

Kontogiannatos

Koutrotsios

Xekalaki

et al. 2021

JoF

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Cordyceps militaris is an entomopathogenic ascomycete with similar pharmacological importance to that of the wild caterpillar fungus Ophiocordyceps sinensis. C. militaris has attracted significant research and commercial interest due to its content in bioactive compounds beneficial to human health and the relative ease of cultivation under laboratory conditions. However, room for improvement exists in the commercial-scale cultivation of C. militaris and concerns issues principally related to appropriate strain selection, genetic degeneration of cultures, and substrate optimization. In particular, culture degeneration—usually expressed by abnormal fruit body formation and reduced sporulation—results in important economic losses and is holding back investors and potential growers (mainly in Western countries) from further developing this highly promising sector. In the present review, the main factors that influence the generation of biomass and metabolites (with emphasis on cordycepin biosynthesis) by C. militaris are presented and evaluated in conjunction with the use of a wide range of supplements or additives towards the enhancement of fungal productivity in large-scale cultivation processes. Moreover, physiological and genetic factors that increase or reduce the manifestation of strain degeneration in C. militaris are outlined. Finally, methodologies for developing protocols to be used in C. militaris functional biology studies are discussed.

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Section: Introductionmentioning

confidence: 99%

Biomass and Cordycepin Production by the Medicinal Mushroom Cordyceps militaris—A Review of Various Aspects and Recent Trends towards the Exploitation of a Valuable Fungus

Kontogiannatos

Koutrotsios

Xekalaki

et al. 2021

JoF

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“…In our case the combined activity of Yatalase and Glucanex was necessary for efficient digestion of T. thlaspeos cell wall, although they appear to have overlapping enzymatic properties. Similar additive effects have recently been shown for the ascomycete Hirsutella sinensis [48], while in Cordyceps militaris the enzymes mix is less active than Glucanex alone [49]. Hence, during the establishment of conditions for protoplasting, various lytic enzymes and combinations should be tested to reach an optimal cell wall degradation [30].…”

Section: Discussionmentioning

confidence: 64%

“…Cordyceps militaris the enzymes mix is less active than Glucanex alone [49]. Hence, during the establishment of conditions for protoplasting, various lytic enzymes and combinations should be tested to reach an optimal cell wall degradation [30].…”

Section: Discussionmentioning

confidence: 99%

Genetic manipulation of the Brassicaceae smut fungus Thecaphora thlaspeos

Plücker

Bösch

Geißl

et al. 2020

Preprint

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Investigation of plant-microbe interactions greatly benefits from genetically tractable partners to address molecularly the virulence and defense mechanisms. The smut fungus Ustilago maydis is a model pathogen in that sense: efficient homologous recombination and a small genome allow targeted modification. On the host side, maize is limiting with regard to rapid genetic alterations. By contrast, the model plant Arabidopsis thaliana is an excellent model with a vast amount of information and techniques as well as genetic resources. Here, we present a transformation protocol for the Brassicaceae smut fungus Thecaphora thlaspeos. Using the well-established transformation of protoplasts, we generated the first reporter strains expressing fluorescent proteins to follow mating. As a proof-of-principle for homologous recombination, we deleted the pheromone receptor pra1. As expected, this mutant cannot mate. Further analysis will contribute to our understanding of the role of mating for infection biology in this novel model fungus. From now on, the genetic manipulation of T. thlaspeos, which is able to colonize the model plant A. thaliana, provides us with a pathosystem, in which both partners are genetically amenable to study smut infection biology.

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“…Similar additive effects have recently been shown for the ascomycete Hirsutella sinensis [ 49 ] and Ag. aegerita [ 31 ]; while in Cordyceps militaris, the enzymes mix is less active than Glucanex alone [ 50 ]. Hence, during the establishment of conditions for protoplasting, various lytic enzymes and combinations should be tested to reach optimal cell wall degradation [ 30 ].…”

Section: Discussionmentioning

confidence: 99%

Genetic Manipulation of the Brassicaceae Smut Fungus Thecaphora thlaspeos

Plücker

Bösch

Geißl

et al. 2021

JoF

View full text Add to dashboard Cite

Investigation of plant–microbe interactions greatly benefit from genetically tractable partners to address, molecularly, the virulence and defense mechanisms. The smut fungus Ustilago maydis is a model pathogen in that sense: efficient homologous recombination and a small genome allow targeted modification. On the host side, maize is limiting with regard to rapid genetic alterations. By contrast, the model plant Arabidopsis thaliana is an excellent model with a vast amount of information and techniques as well as genetic resources. Here, we present a transformation protocol for the Brassicaceae smut fungus Thecaphora thlaspeos. Using the well-established methodology of protoplast transformation, we generated the first reporter strains expressing fluorescent proteins to follow mating. As a proof-of-principle for homologous recombination, we deleted the pheromone receptor pra1. As expected, this mutant cannot mate. Further analysis will contribute to our understanding of the role of mating for infection biology in this novel model fungus. From now on, the genetic manipulation of T. thlaspeos, which is able to colonize the model plant A. thaliana, provides us with a pathosystem in which both partners are genetically amenable to study smut infection biology.

show abstract

The efficient genetic transformation of Cordyceps militaris by using mononuclear protoplasts

Cited by 20 publications

References 31 publications

Biomass and Cordycepin Production by the Medicinal Mushroom Cordyceps militaris—A Review of Various Aspects and Recent Trends towards the Exploitation of a Valuable Fungus

Biomass and Cordycepin Production by the Medicinal Mushroom Cordyceps militaris—A Review of Various Aspects and Recent Trends towards the Exploitation of a Valuable Fungus

Genetic manipulation of the Brassicaceae smut fungus Thecaphora thlaspeos

Genetic Manipulation of the Brassicaceae Smut Fungus Thecaphora thlaspeos

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