Green fluorescent protein transformation sheds more light on a widespread mycoparasitic interaction

Nemeth, Mark Z. and Pintye, Alexandra and Horvath, Aron N. and Vagi, Pal and Kovacs, Gabor M. and Gorfer, Markus and Kiss, Levente (2019) Green fluorescent protein transformation sheds more light on a widespread mycoparasitic interaction. Phytopathology: International Journal of the American Phytopathological Society, 109 (8). pp. 1404-1416. ISSN 0031-949X


Abstract

Powdery mildews, ubiquitous obligate biotrophic plant pathogens, are often attacked in the field by mycoparasitic fungi belonging to the genus Ampelomyces. Some Ampelomyces strains are commercialized biocontrol agents of crop pathogenic powdery mildews. Using Agrobacterium tumefaciens-mediated transformation (ATMT), we produced stable Ampelomyces transformants that constitutively expressed green fluorescent protein (GFP) to (i) improve the visualization of the mildew–Ampelomyces interaction and (ii) decipher the environmental fate of Ampelomyces fungi before and after acting as a mycoparasite. Detection of Ampelomyces structures, and especially hyphae, was greatly enhanced when diverse powdery mildew, leaf, and soil samples containing GFP transformants were examined with fluorescence microscopy compared with brightfield and differential interference contrast optics. We showed for the first time, to our knowledge, that Ampelomyces strains can persist up to 21 days on mildew-free host plant surfaces, where they can attack powdery mildew structures as soon as these appear after this period. As saprobes in decomposing, powdery mildew-infected leaves on the ground and also in autoclaved soil, Ampelomyces strains developed new hyphae but did not sporulate. These results indicate that Ampelomyces strains occupy a niche in the phyllosphere where they act primarily as mycoparasites of powdery mildews. Our work has established a framework for a molecular genetic toolbox for the genus Ampelomyces using ATMT.


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Item Type: Article (Commonwealth Reporting Category C)
Refereed: Yes
Item Status: Live Archive
Additional Information: Published version not displayed due to copyright restrictions.
Faculty/School / Institute/Centre: Current - Institute for Life Sciences and the Environment - Centre for Crop Health (24 Mar 2014 -)
Faculty/School / Institute/Centre: Current - Institute for Life Sciences and the Environment - Centre for Crop Health (24 Mar 2014 -)
Date Deposited: 09 Aug 2019 03:54
Last Modified: 18 Sep 2019 06:02
Uncontrolled Keywords: tritrophic interactions; GFP transformation; powdery mildew; plant pathology
Fields of Research : 06 Biological Sciences > 0605 Microbiology > 060505 Mycology
Identification Number or DOI: 10.1094/PHYTO-01-19-0013-R
URI: http://eprints.usq.edu.au/id/eprint/36866

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