Agrobacterium tumefaciens-mediated transformation of the plant pathogenic fungus, Magnaporthe grisea.

H. S. Rho, Seogchan Kang, Y. H. Lee

Research output: Contribution to journalArticle

167 Citations (Scopus)

Abstract

An effective way to study the infection mechanisms of fungal pathogens is to disrupt their genes via transformation in both targeted and random manners. This isolates the mutants that exhibit altered virulence. In this paper, we report the successful transformation of Magnaporthe grisea, the causal agent for rice blast, that is mediated by Agrobacterium tumefaciens. Employing the binary vector pBHt2, which carries the bacterial hygromycin B phosphotransferase gene under the control of the Aspergillus nidulans trpC promoter as a selectable marker, led to the production of 500 to > 1,000 hygromycin B-resistant transformants per 1 x 10(6) conidia of M. grisea. The transformation efficiency is correlated with the number of A. tumefaciens cells used, pre-treating bacterial cells with acetosyringone prior to co-cultivation with fungal spores, and the duration of co-cultivation. All of the transformants tested remained mitotically stable, maintaining their hygromycin B resistance after several generations of growth in the absence of hygromycin B. A genomic Southern blot analysis showed that over 60% of the transformants contained a single T-DNA insert on their genome. Considering the efficiency and flexibility of A. tumefaciens-mediated transformation (ATMT), this technique offers highly efficient means for characterizing the genes that are important for the pathogenicity of M. grisea.

Original languageEnglish (US)
Pages (from-to)407-411
Number of pages5
JournalMolecules and cells
Volume12
Issue number3
StatePublished - Jan 1 2001

Fingerprint

Hygromycin B
Magnaporthe
Agrobacterium tumefaciens
Fungi
hygromycin-B kinase
Fungal Spores
Virulence
Genes
Aspergillus nidulans
Mycoses
Southern Blotting
Genome
Growth

All Science Journal Classification (ASJC) codes

  • Medicine(all)

Cite this

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title = "Agrobacterium tumefaciens-mediated transformation of the plant pathogenic fungus, Magnaporthe grisea.",
abstract = "An effective way to study the infection mechanisms of fungal pathogens is to disrupt their genes via transformation in both targeted and random manners. This isolates the mutants that exhibit altered virulence. In this paper, we report the successful transformation of Magnaporthe grisea, the causal agent for rice blast, that is mediated by Agrobacterium tumefaciens. Employing the binary vector pBHt2, which carries the bacterial hygromycin B phosphotransferase gene under the control of the Aspergillus nidulans trpC promoter as a selectable marker, led to the production of 500 to > 1,000 hygromycin B-resistant transformants per 1 x 10(6) conidia of M. grisea. The transformation efficiency is correlated with the number of A. tumefaciens cells used, pre-treating bacterial cells with acetosyringone prior to co-cultivation with fungal spores, and the duration of co-cultivation. All of the transformants tested remained mitotically stable, maintaining their hygromycin B resistance after several generations of growth in the absence of hygromycin B. A genomic Southern blot analysis showed that over 60{\%} of the transformants contained a single T-DNA insert on their genome. Considering the efficiency and flexibility of A. tumefaciens-mediated transformation (ATMT), this technique offers highly efficient means for characterizing the genes that are important for the pathogenicity of M. grisea.",
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Agrobacterium tumefaciens-mediated transformation of the plant pathogenic fungus, Magnaporthe grisea. / Rho, H. S.; Kang, Seogchan; Lee, Y. H.

In: Molecules and cells, Vol. 12, No. 3, 01.01.2001, p. 407-411.

Research output: Contribution to journalArticle

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