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

Márk Z. Németh, Alexandra Pintye, Áron N. Horváth, Pál Vági, G. Kovács, Markus Gorfer, L. Kiss

Research output: Contribution to journalArticle

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.

Original languageEnglish
Pages (from-to)1404-1416
Number of pages13
JournalPhytopathology
Volume109
Issue number8
DOIs
Publication statusPublished - Jan 1 2019

Fingerprint

Ampelomyces
green fluorescent protein
mildews
mycoparasites
powdery mildew
Agrobacterium radiobacter
hyphae
environmental fate
phyllosphere
saprophytes
optics
crossover interference
fluorescence microscopy
plant pathogens
molecular genetics
biological control agents
leaves
niches
host plants
soil sampling

Keywords

  • Tritrophic interactions

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Plant Science

Cite this

Green fluorescent protein transformation sheds more light on a widespread mycoparasitic interaction. / Németh, Márk Z.; Pintye, Alexandra; Horváth, Áron N.; Vági, Pál; Kovács, G.; Gorfer, Markus; Kiss, L.

In: Phytopathology, Vol. 109, No. 8, 01.01.2019, p. 1404-1416.

Research output: Contribution to journalArticle

Németh, Márk Z. ; Pintye, Alexandra ; Horváth, Áron N. ; Vági, Pál ; Kovács, G. ; Gorfer, Markus ; Kiss, L. / Green fluorescent protein transformation sheds more light on a widespread mycoparasitic interaction. In: Phytopathology. 2019 ; Vol. 109, No. 8. pp. 1404-1416.
@article{453e8c344c464ceeab7de47042777596,
title = "Green fluorescent protein transformation sheds more light on a widespread mycoparasitic interaction",
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.",
keywords = "Tritrophic interactions",
author = "N{\'e}meth, {M{\'a}rk Z.} and Alexandra Pintye and Horv{\'a}th, {{\'A}ron N.} and P{\'a}l V{\'a}gi and G. Kov{\'a}cs and Markus Gorfer and L. Kiss",
year = "2019",
month = "1",
day = "1",
doi = "10.1094/PHYTO-01-19-0013-R",
language = "English",
volume = "109",
pages = "1404--1416",
journal = "Phytopathology",
issn = "0031-949X",
publisher = "American Phytopathological Society",
number = "8",

}

TY - JOUR

T1 - Green fluorescent protein transformation sheds more light on a widespread mycoparasitic interaction

AU - Németh, Márk Z.

AU - Pintye, Alexandra

AU - Horváth, Áron N.

AU - Vági, Pál

AU - Kovács, G.

AU - Gorfer, Markus

AU - Kiss, L.

PY - 2019/1/1

Y1 - 2019/1/1

N2 - 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.

AB - 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.

KW - Tritrophic interactions

UR - http://www.scopus.com/inward/record.url?scp=85070367900&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85070367900&partnerID=8YFLogxK

U2 - 10.1094/PHYTO-01-19-0013-R

DO - 10.1094/PHYTO-01-19-0013-R

M3 - Article

C2 - 30900938

AN - SCOPUS:85070367900

VL - 109

SP - 1404

EP - 1416

JO - Phytopathology

JF - Phytopathology

SN - 0031-949X

IS - 8

ER -