GUS and GFP transformation of the biocontrol strain Clonostachys rosea IK726 and the use of these marker genes in ecological studies

Lübeck, Mette; Knudsen, Inge M.B.; Jensen, Birgit; Thrane, Ulf; Janvier, Céline; Jensen, Dan Funck

doi:10.1017/s095375620200607x

Cited by 60 publications

(31 citation statements)

References 53 publications

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“…In many cases, the heterologous expression of fungal enzymes previously reviewed [38] was controlled either by inducible promoters that were homologous to the expression host or by the constitutive A. nidulans glyceraldehyde-3-phosphate dehydrogenase (gpdA) promoter, which was reported to be functional in industrially used Penicillium and Aspergillus species [58] as well as other species, e.g., Trichoderma and Clonostachys [59,60]. The expression of the most prominent cellobiohydrolase, Cel7A (syn.…”

Section: Promoters For Driving the Heterologous Expression Of Cellobimentioning

confidence: 99%

Heterologous expression of cellobiohydrolases in filamentous fungi – An update on the current challenges, achievements and perspectives

Zoglowek

Lübeck

Ahring

et al. 2015

Process Biochemistry

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Section: Promoters For Driving the Heterologous Expression Of Cellobimentioning

confidence: 99%

Heterologous expression of cellobiohydrolases in filamentous fungi – An update on the current challenges, achievements and perspectives

Zoglowek

Lübeck

Ahring

et al. 2015

Process Biochemistry

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“…Since Ustilago maydis was successfully transformed with gfp [5], the exploitation of GFP technology has been adapted for use with other filamentous fungi (for reviews, see [6,7]). We have used GFP constitutively expressed in various fungi for studying fungal ecology in relation to biological control of plant diseases [8] and colonization of ryegrass by the endophyte Neotyphodium lolii [9]. However, there is still a need for developing gfp constructs for use in fungi as many of the present gene constructs have been designed for use with other organisms.…”

Section: Introductionmentioning

confidence: 99%

Expression of the red fluorescent protein DsRed-Express in filamentous ascomycete fungi

Mikkelsen¹,

Sarrocco²,

Lübeck³

et al. 2003

FEMS Microbiology Letters

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The recently reported red fluorescent protein DsRed from the reef coral Discosoma sp. represents a new marker that has been codonoptimized for high expression in mammalian cells. To facilitate expression of DsRed in ascomycete fungi, we used the clone pDsRedExpress (Clontech) for constructing a plasmid vector, pPgpd-DsRed, containing the constitutive Aspergillus nidulans glyceraldehyde 3-phosphate (gpd) promoter. This vector was used for co-transformation of Penicillium paxilli, Trichoderma harzianum and Trichoderma virens (syn. Gliocladium virens) together with either pAN7-1 or gGFP, both containing a gene for hygromycin resistance for transformant selection. In addition, gGFP contains a green fluorescent protein (GFP) gene for expression in Ascomycetes. Expression of DsRedExpress was obtained in all three fungi, indicating that DsRed can be used as a highly effective vital marker in Ascomycetes. Dual marked transformants expressed both DsRed-Express and GFP in the same mycelium and were used for non-quantitative comparison of the intensity of the fluorescence using confocal laser scanning microscopy.

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“…Unfortunately, this reporter gene, which needs a substrate to stain hyphae, is not easy to use in studies of root colonization in soil. Other reporter genes, such as the green fluorescent protein (GFP) gene or the DsRed2 gene, have also been used to transform strains of pathogenic fungi and antagonistic bacteria or fungi (4,10,14,16,22,23). When strains marked with different reporter genes are used simultaneously, both microorganisms can be observed simultaneously on the same root (5).…”

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Colonization of Tomato Root by Pathogenic and Nonpathogenic Fusarium oxysporum Strains Inoculated Together and Separately into the Soil

Olivain

Humbert

Nahálková

et al. 2006

Appl Environ Microbiol

135

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In soil, fungal colonization of plant roots has been traditionally studied by indirect methods such as microbial isolation that do not enable direct observation of infection sites or of interactions between fungal pathogens and their antagonists. Confocal laser scanning microscopy was used to visualize the colonization of tomato roots in heat-treated soil and to observe the interactions between a nonpathogenic strain, Fo47, and a pathogenic strain, Fol8, inoculated onto tomato roots in soil. When inoculated separately, both fungi colonized the entire root surface, with the exception of the apical zone. When both strains were introduced together, they both colonized the root surface and were observed at the same locations. When Fo47 was introduced at a higher concentration than Fol8, it colonized much of the root surface, but hyphae of Fol8 could still be observed at the same location on the root. There was no exclusion of the pathogenic strain by the presence of the nonpathogenic strain. These results are not consistent with the hypothesis that specific infection sites exist on the root for Fusarium oxysporum and instead support the hypothesis that competition occurs for nutrients rather than for infection sites.

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GUS and GFP transformation of the biocontrol strain Clonostachys rosea IK726 and the use of these marker genes in ecological studies

Cited by 60 publications

References 53 publications

Heterologous expression of cellobiohydrolases in filamentous fungi – An update on the current challenges, achievements and perspectives

Heterologous expression of cellobiohydrolases in filamentous fungi – An update on the current challenges, achievements and perspectives

Expression of the red fluorescent protein DsRed-Express in filamentous ascomycete fungi

Colonization of Tomato Root by Pathogenic and Nonpathogenic Fusarium oxysporum Strains Inoculated Together and Separately into the Soil

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