Antioxidant treatments counteract the non-culturability of bacterial endophytes isolated from legume nodules

In many wild legumes, attempts to cultivate nodule bacteria fail. We hypothesized that the limited culturability could be related to injury from oxidative stress caused by disruption of plant tissues during isolation. To test that, we isolated bacteria from nodules of Hedysarum spinosissimum and Tet...

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Bibliographic Details
Published in:Archives of microbiology 2013-06, Vol.195 (6), p.385-391
Main Authors: Muresu, Rosella, Tondello, Alessandra, Polone, Elisa, Sulas, Leonardo, Baldan, Barbara, Squartini, Andrea
Format: Article
Language:English
Subjects:
Antioxidants
Antioxidants - metabolism
Bacteria
Biochemistry
Biomedical and Life Sciences
Biotechnology
Buffers
Cell Biology
Culture Media - chemistry
Ecology
Endophytes
Endophytes - growth & development
Endophytes - isolation & purification
Fabaceae - microbiology
Hedysarum
Hydrogen peroxide
Hydrogen Peroxide - metabolism
Legumes
Life Sciences
Microbial Ecology
Microbiology
Nitrogen
Original Paper
Oxidative Stress
Oxidoreductases - metabolism
Pathogens
Plant tissues
Rhizobiaceae
Rhizobiaceae - growth & development
Rhizobiaceae - isolation & purification
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Summary:In many wild legumes, attempts to cultivate nodule bacteria fail. We hypothesized that the limited culturability could be related to injury from oxidative stress caused by disruption of plant tissues during isolation. To test that, we isolated bacteria from nodules of Hedysarum spinosissimum and Tetragonolobus purpureus using buffers supplemented with scavenging systems to prevent damage from reactive oxygen species (ROS). Treatments included the following: antioxidants (glutathione, ascorbate, EDTA) or enzymes (catalase, peroxidase, superoxide dismutase), tested either as modified squashing buffers or added in plates. Some combinations yielded dramatic increases of culturability. Different endophytes were found, including additional Rhizobiaceae that were not the primary symbiont and were unable to nodulate. Their H 2 O 2 tolerance in broth culture showed differences consistent with the unequal culturability observed. In wild legumes species, ROS generation during extraction appears to be a major factor limiting microbiota isolation, and protocols presented here significantly improve the recovery of culturable bacterial endophytes from plants.
ISSN:0302-8933
1432-072X
DOI:10.1007/s00203-013-0886-6