Flexible gateway constructs for functional analyses of genes in plant pathogenic fungi

Rahim Mehrabi*, Amir Mirzadi Gohari, Gilvan Ferreira da Silva, Gero Steinberg, Gert H.J. Kema, Pierre J.G.M. de Wit

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

9 Citations (Scopus)

Abstract

Genetic manipulation of fungi requires quick, low-cost, efficient, high-throughput and molecular tools. In this paper, we report 22 entry constructs as new molecular tools based on the Gateway technology facilitating rapid construction of binary vectors that can be used for functional analysis of genes in fungi. The entry vectors for single, double or triple gene-deletion mutants were developed using hygromycin, geneticin and nourseothricin resistance genes as selection markers. Furthermore, entry vectors containing green fluorescent (GFP) or red fluorescent (RFP) in combination with hygromycin, geneticin or nourseothricin selection markers were generated. The latter vectors provide the possibility of gene deletion and simultaneous labelling of the fungal transformants with GFP or RFP reporter genes. The applicability of a number of entry vectors was validated in Zymoseptoria tritici, an important fungal wheat pathogen.

Original languageEnglish
Pages (from-to)186-192
JournalFungal Genetics and Biology
Volume79
DOIs
Publication statusPublished - 2015

Fingerprint

Streptothricins
Plant Genes
Gene Deletion
Fungi
Reporter Genes
Genes
Triticum
Technology
Costs and Cost Analysis
antibiotic G 418
hygromycin A

Keywords

  • Binary vectors
  • Gateway technology
  • Gene deletion
  • GFP
  • RFP
  • Zymoseptoria tritici

Cite this

Mehrabi, Rahim ; Mirzadi Gohari, Amir ; da Silva, Gilvan Ferreira ; Steinberg, Gero ; Kema, Gert H.J. ; de Wit, Pierre J.G.M. / Flexible gateway constructs for functional analyses of genes in plant pathogenic fungi. In: Fungal Genetics and Biology. 2015 ; Vol. 79. pp. 186-192.
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Flexible gateway constructs for functional analyses of genes in plant pathogenic fungi. / Mehrabi, Rahim; Mirzadi Gohari, Amir; da Silva, Gilvan Ferreira; Steinberg, Gero; Kema, Gert H.J.; de Wit, Pierre J.G.M.

In: Fungal Genetics and Biology, Vol. 79, 2015, p. 186-192.

Research output: Contribution to journalArticleAcademicpeer-review

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AU - Mehrabi, Rahim

AU - Mirzadi Gohari, Amir

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AU - Kema, Gert H.J.

AU - de Wit, Pierre J.G.M.

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AB - Genetic manipulation of fungi requires quick, low-cost, efficient, high-throughput and molecular tools. In this paper, we report 22 entry constructs as new molecular tools based on the Gateway technology facilitating rapid construction of binary vectors that can be used for functional analysis of genes in fungi. The entry vectors for single, double or triple gene-deletion mutants were developed using hygromycin, geneticin and nourseothricin resistance genes as selection markers. Furthermore, entry vectors containing green fluorescent (GFP) or red fluorescent (RFP) in combination with hygromycin, geneticin or nourseothricin selection markers were generated. The latter vectors provide the possibility of gene deletion and simultaneous labelling of the fungal transformants with GFP or RFP reporter genes. The applicability of a number of entry vectors was validated in Zymoseptoria tritici, an important fungal wheat pathogen.

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