Isolation of endohyphal bacteria from foliar Ascomycota and In Vitro establishment of their symbiotic associations

Kayla R. Arendt, Kevin L. Hockett, Sarah J. Araldi-Brondolo, David A Baltrus, Anne E Arnold

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Endohyphal bacteria (EHB) can influence fungal phenotypes and shape the outcomes of plant-fungal interactions. Previous work has suggested that EHB form facultative associations with many foliar fungi in the Ascomycota. These bacteria can be isolated in culture, and fungi can be cured of EHB using antibiotics. Here, we present methods for successfully introducing EHB into axenic mycelia of strains representing two classes of Ascomycota. We first establish in vitro conditions favoring reintroduction of two strains of EHB (Luteibacter sp.) into axenic cultures of their original fungal hosts, focusing on fungi isolated from healthy plant tissue as endophytes: Microdiplodia sp. (Dothideomycetes) and Pestalotiopsis sp. (Sordariomycetes). We then demonstrate that these EHB can be introduced into a novel fungal host under the same conditions, successfully transferring EHB between fungi representing different classes. Finally, we manipulate conditions to optimize reintroduction in a focal EHB-fungal association. We show that EHB infections were initiated and maintained more often under low-nutrient culture conditions and when EHB and fungal hyphae were washed with MgCl2 prior to reassociation. Our study provides new methods for experimental assessment of the effects of EHB on fungal phenotypes and shows how the identity of the fungal host and growth conditions can define the establishment of these widespread and important symbioses.

Original languageEnglish (US)
Pages (from-to)2943-2949
Number of pages7
JournalApplied and Environmental Microbiology
Volume82
Issue number10
DOIs
StatePublished - May 1 2016

Fingerprint

Ascomycota
Bacteria
bacterium
bacteria
Fungi
fungus
fungi
reintroduction
In Vitro Techniques
phenotype
Sordariomycetes
Axenic Culture
Dothideomycetes
Pestalotiopsis
Endophytes
Phenotype
Magnesium Chloride
axenic culture
Symbiosis
Hyphae

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Food Science
  • Biotechnology
  • Ecology

Cite this

Isolation of endohyphal bacteria from foliar Ascomycota and In Vitro establishment of their symbiotic associations. / Arendt, Kayla R.; Hockett, Kevin L.; Araldi-Brondolo, Sarah J.; Baltrus, David A; Arnold, Anne E.

In: Applied and Environmental Microbiology, Vol. 82, No. 10, 01.05.2016, p. 2943-2949.

Research output: Contribution to journalArticle

@article{17929af6b7fd46fdafa226a92e4276f8,
title = "Isolation of endohyphal bacteria from foliar Ascomycota and In Vitro establishment of their symbiotic associations",
abstract = "Endohyphal bacteria (EHB) can influence fungal phenotypes and shape the outcomes of plant-fungal interactions. Previous work has suggested that EHB form facultative associations with many foliar fungi in the Ascomycota. These bacteria can be isolated in culture, and fungi can be cured of EHB using antibiotics. Here, we present methods for successfully introducing EHB into axenic mycelia of strains representing two classes of Ascomycota. We first establish in vitro conditions favoring reintroduction of two strains of EHB (Luteibacter sp.) into axenic cultures of their original fungal hosts, focusing on fungi isolated from healthy plant tissue as endophytes: Microdiplodia sp. (Dothideomycetes) and Pestalotiopsis sp. (Sordariomycetes). We then demonstrate that these EHB can be introduced into a novel fungal host under the same conditions, successfully transferring EHB between fungi representing different classes. Finally, we manipulate conditions to optimize reintroduction in a focal EHB-fungal association. We show that EHB infections were initiated and maintained more often under low-nutrient culture conditions and when EHB and fungal hyphae were washed with MgCl2 prior to reassociation. Our study provides new methods for experimental assessment of the effects of EHB on fungal phenotypes and shows how the identity of the fungal host and growth conditions can define the establishment of these widespread and important symbioses.",
author = "Arendt, {Kayla R.} and Hockett, {Kevin L.} and Araldi-Brondolo, {Sarah J.} and Baltrus, {David A} and Arnold, {Anne E}",
year = "2016",
month = "5",
day = "1",
doi = "10.1128/AEM.00452-16",
language = "English (US)",
volume = "82",
pages = "2943--2949",
journal = "Applied and Environmental Microbiology",
issn = "0099-2240",
publisher = "American Society for Microbiology",
number = "10",

}

TY - JOUR

T1 - Isolation of endohyphal bacteria from foliar Ascomycota and In Vitro establishment of their symbiotic associations

AU - Arendt, Kayla R.

AU - Hockett, Kevin L.

AU - Araldi-Brondolo, Sarah J.

AU - Baltrus, David A

AU - Arnold, Anne E

PY - 2016/5/1

Y1 - 2016/5/1

N2 - Endohyphal bacteria (EHB) can influence fungal phenotypes and shape the outcomes of plant-fungal interactions. Previous work has suggested that EHB form facultative associations with many foliar fungi in the Ascomycota. These bacteria can be isolated in culture, and fungi can be cured of EHB using antibiotics. Here, we present methods for successfully introducing EHB into axenic mycelia of strains representing two classes of Ascomycota. We first establish in vitro conditions favoring reintroduction of two strains of EHB (Luteibacter sp.) into axenic cultures of their original fungal hosts, focusing on fungi isolated from healthy plant tissue as endophytes: Microdiplodia sp. (Dothideomycetes) and Pestalotiopsis sp. (Sordariomycetes). We then demonstrate that these EHB can be introduced into a novel fungal host under the same conditions, successfully transferring EHB between fungi representing different classes. Finally, we manipulate conditions to optimize reintroduction in a focal EHB-fungal association. We show that EHB infections were initiated and maintained more often under low-nutrient culture conditions and when EHB and fungal hyphae were washed with MgCl2 prior to reassociation. Our study provides new methods for experimental assessment of the effects of EHB on fungal phenotypes and shows how the identity of the fungal host and growth conditions can define the establishment of these widespread and important symbioses.

AB - Endohyphal bacteria (EHB) can influence fungal phenotypes and shape the outcomes of plant-fungal interactions. Previous work has suggested that EHB form facultative associations with many foliar fungi in the Ascomycota. These bacteria can be isolated in culture, and fungi can be cured of EHB using antibiotics. Here, we present methods for successfully introducing EHB into axenic mycelia of strains representing two classes of Ascomycota. We first establish in vitro conditions favoring reintroduction of two strains of EHB (Luteibacter sp.) into axenic cultures of their original fungal hosts, focusing on fungi isolated from healthy plant tissue as endophytes: Microdiplodia sp. (Dothideomycetes) and Pestalotiopsis sp. (Sordariomycetes). We then demonstrate that these EHB can be introduced into a novel fungal host under the same conditions, successfully transferring EHB between fungi representing different classes. Finally, we manipulate conditions to optimize reintroduction in a focal EHB-fungal association. We show that EHB infections were initiated and maintained more often under low-nutrient culture conditions and when EHB and fungal hyphae were washed with MgCl2 prior to reassociation. Our study provides new methods for experimental assessment of the effects of EHB on fungal phenotypes and shows how the identity of the fungal host and growth conditions can define the establishment of these widespread and important symbioses.

UR - http://www.scopus.com/inward/record.url?scp=84966525933&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84966525933&partnerID=8YFLogxK

U2 - 10.1128/AEM.00452-16

DO - 10.1128/AEM.00452-16

M3 - Article

VL - 82

SP - 2943

EP - 2949

JO - Applied and Environmental Microbiology

JF - Applied and Environmental Microbiology

SN - 0099-2240

IS - 10

ER -