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Rouws, L.* ; Leite, J.* ; Feitosa de Matos, G.* ; Zilli, J.E.* ; Coelho, M.R.* ; Xavier, G.R.* ; Fischer, D. ; Hartmann, A. ; Reis, V.M.* ; Baldani, J.I.*

Endophytic Bradyrhizobium spp. isolates from sugarcane obtained through different culture strategies.

Environ. Microbiol. Rep. 6, 354-363 (2014)
DOI
Open Access Gold as soon as Publ. Version/Full Text is submitted to ZB.
Brazilian sugarcane has been shown to obtain part of its nitrogen via biological nitrogen fixation (BNF). Recent reports, based on the culture independent sequencing of bacterial nifH complementary DNA (cDNA) from sugarcane tissues, have suggested that members of the Bradyrhizobium genus could play a role in sugarcane-associated BNF. Here we report on the isolation of Bradyrhizobium spp. isolates and a few other species from roots of sugarcane cultivar RB867515 by two cultivation strategies: direct isolation on culture media and capture of Bradyrhizobium spp. using the promiscuous legume Vigna unguiculata as trap-plant. Both strategies permitted the isolation of genetically diverse Bradyrhizobium spp. isolates, as concluded from enterobacterial repetitive intergenic consensus polymerase chain reaction (PCR) fingerprinting and 16S ribosomal RNA, nifH and nodC sequence analyses. Several isolates presented nifH phylotypes highly similar to nifH cDNA phylotypes detected in field-grown sugarcane by a culture-independent approach. Four isolates obtained by direct plate cultivation were unable to nodulate V. unguiculata and, based on PCR analysis, lacked a nodC gene homologue. Acetylene reduction assay showed in vitro nitrogenase activity for some Bradyrhizobium spp. isolates, suggesting that these bacteria do not require a nodule environment for BNF. Therefore, this study brings further evidence that Bradyrhizobium spp. may play a role in sugarcane-associated BNF under field conditions.
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Publication type Article: Journal article
Document type Scientific Article
Keywords Nitrogen-fixation; Photosynthetic Bradyrhizobia; Sp-nov.; Diversity; Sequences; Bacteria; Strains; Quantification; Symbionts; Rhizobia
ISSN (print) / ISBN 1758-2229
e-ISSN 1758-2229
Journal Environmental Microbiology Reports
Quellenangaben Volume: 6, Issue: 4, Pages: 354-363 Article Number: , Supplement: ,
Publisher Wiley
Publishing Place Hoboken
Reviewing status Peer reviewed
Institute(s) Research Unit Microbe-Plant Interactions (AMP)
Institute of Biochemical Plant Pathology (BIOP)
Research Unit Comparative Microbiome Analysis (COMI)