Identification of virulence genes in the corn pathogen Colletotrichum graminicola by Agrobacterium tumefaciens-mediated transformation
Muench S, Ludwig N, Floss DS, Sugui JA, Koszucka AM, Voll L, Sonnewald U, Deising HB (2011)
Publication Status: Published
Publication Type: Journal article
Publication year: 2011
Journal
Publisher: WILEY-BLACKWELL
Book Volume: 12
Pages Range: 43-55
Journal Issue: 1
DOI: 10.1111/j.1364-3703.2010.00651.x
Abstract
P>A previously developed Agrobacterium tumefaciens-mediated transformation (ATMT) protocol for the plant pathogenic fungus Colletotrichum graminicola led to high rates of tandem integration of the whole Ti-plasmid, and was therefore considered to be unsuitable for the identification of pathogenicity and virulence genes by insertional mutagenesis in this pathogen. We used a modified ATMT protocol with acetosyringone present only during the co-cultivation of C. graminicola and A. tumefaciens. Analysis of 105 single-spore isolates randomly chosen from a collection of approximately 2000 transformants, indicated that almost 70% of the transformants had single T-DNA integrations. Of 500 independent transformants tested, 10 exhibited attenuated virulence in infection assays on whole plants. Microscopic analyses primarily revealed defects at different pre-penetration stages of infection-related morphogenesis. Three transformants were characterized in detail. The identification of the T-DNA integration sites was performed by amplification of genomic DNA ends after endonuclease digestion and polynucleotide tailing. In one transformant, the T-DNA had integrated into the 5'-flank of a gene with similarity to allantoicase genes of other Ascomycota. In the second and third transformants, the T-DNA had integrated into an open reading frame (ORF) and into the 5'-flank of an ORF. In both cases, the ORFs have unknown function.
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Germany (DE)How to cite
APA:
Muench, S., Ludwig, N., Floss, D.S., Sugui, J.A., Koszucka, A.M., Voll, L.,... Deising, H.B. (2011). Identification of virulence genes in the corn pathogen Colletotrichum graminicola by Agrobacterium tumefaciens-mediated transformation. Molecular Plant Pathology, 12(1), 43-55. https://dx.doi.org/10.1111/j.1364-3703.2010.00651.x
MLA:
Muench, Steffen, et al. "Identification of virulence genes in the corn pathogen Colletotrichum graminicola by Agrobacterium tumefaciens-mediated transformation." Molecular Plant Pathology 12.1 (2011): 43-55.
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