Diversity of Micromonospora strains isolated from nitrogen fixing nodules and rhizosphere of Pisum sativum analyzed by multilocus sequence analysis

It was recently reported that Micromonospora inhabits the intracellular tissues of nitrogen fixing nodules of the wild legume Lupinus angustifolius. To determine if Micromonospora populations are also present in nitrogen fixing nodules of cultivated legumes such as Pisum sativum, we carried out the...

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Bibliographic Details
Published in:Systematic and applied microbiology 2012-03, Vol.35 (2), p.73-80
Main Authors: Carro, Lorena, Spröer, Cathrin, Alonso, Pilar, Trujillo, Martha E.
Format: Article
Language:English
Subjects:
Bacterial Typing Techniques
Base Sequence
Biological and medical sciences
DNA Gyrase - genetics
DNA, Bacterial - genetics
DNA, Ribosomal - genetics
Fundamental and applied biological sciences. Psychology
Genetic Variation
Microbiology
Micromonospora
Micromonospora - classification
Micromonospora - genetics
Micromonospora - isolation & purification
MLSA
Molecular Sequence Data
Multilocus Sequence Typing
Nitrogen Fixation
Nodule
Phylogeny
Pisum sativum
Pisum sativum - microbiology
Rec A Recombinases - genetics
Restriction Mapping
Rhizosphere
RNA, Bacterial - genetics
RNA, Ribosomal, 16S - analysis
RNA, Ribosomal, 16S - genetics
Root Nodules, Plant - microbiology
Sequence Analysis, DNA
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Summary:It was recently reported that Micromonospora inhabits the intracellular tissues of nitrogen fixing nodules of the wild legume Lupinus angustifolius. To determine if Micromonospora populations are also present in nitrogen fixing nodules of cultivated legumes such as Pisum sativum, we carried out the isolation of this actinobacterium from P. sativum plants collected in two man-managed fields in the region of Castilla and León (Spain). In this work, we describe the isolation of 93 Micromonospora strains recovered from nitrogen fixing nodules and the rhizosphere of P. sativum. The genomic diversity of the strains was analyzed by amplified ribosomal DNA restriction analysis (ARDRA). Forty-six isolates and 34 reference strains were further analyzed using a multilocus sequence analysis scheme developed to address the phylogeny of the genus Micromonospora and to evaluate the species distribution in the two studied habitats. The MLSA results were evaluated by DNA–DNA hybridization to determine their usefulness for the delineation of Micromonospora at the species level. In most cases, DDH values below 70% were obtained with strains that shared a sequence similarity of 98.5% or less. Thus, MLSA studies clearly supported the established taxonomy of the genus Micromonospora and indicated that genomic species could be delineated as groups of strains that share >98.5% sequence similarity based on the 5 genes selected. The species diversity of the strains isolated from both the rhizosphere and nodules was very high and in many cases the new strains could not be related to any of the currently described species.
ISSN:0723-2020
1618-0984
DOI:10.1016/j.syapm.2011.11.003