Use of nodulation pattern, stress tolerance, nodC gene amplification, RAPD-PCR and RFLP-16S rDNA analysis to discriminate genotypes of Rhizobium leguminosarum biovar viciae

6533b7d0fe1ef96bd125a457

RESEARCH PRODUCT

Use of nodulation pattern, stress tolerance, nodC gene amplification, RAPD-PCR and RFLP-16S rDNA analysis to discriminate genotypes of Rhizobium leguminosarum biovar viciae

Anna Lucia Peluso Roberto Defez Marilena Anastasio Olimpia Pepe Andrea Protopapa Giancarlo Moschetti

subject

DNA Bacterial Root nodule Genotype Stress tolerance Biology medicine.disease_cause N-Acetylglucosaminyltransferases Applied Microbiology and Biotechnology Microbiology DNA Ribosomal Polymerase Chain Reaction Mediterranean area Rhizobium leguminosarum Rhizobia Bacterial Proteins Rhizobium leguninosarumNodC geneStress toleranceWild legumesStrains diversityMediterranean areaSymbiotic characteristics RNA Ribosomal 16S medicine Symbiosis Ecology Evolution Behavior and Systematics Genetics Principal Component Analysis Rhizobium leguminosarum food and beverages Fabaceae Nucleic acid amplification technique NodC gene Hydrogen-Ion Concentration Rhizobium leguninosarum 16S ribosomal RNA biology.organism_classification Strains diversity Symbiotic characteristics RAPD Bacterial Typing Techniques Random Amplified Polymorphic DNA Technique Rhizobium Wild legume Restriction fragment length polymorphism Nucleic Acid Amplification Techniques Polymorphism Restriction Fragment Length Settore AGR/16 - Microbiologia Agraria

description

Twenty-seven new Rhizobium isolates were obtained from root nodules of wild and crop legumes belonging to the genera Vicia, Lathyrus and Pisum from different agroecological areas in central and southern Italy. A polyphasic approach including phenotypic and genotypic techniques was used to study their diversity and their relationships with other biovars and species of rhizobia. Analysis of symbiotic properties and stress tolerance tests revealed that wild isolates, showed a wide spectrum of nodulation and a marked variation in stress tolerance compared with reference strains tested in this study. All rhizobial isolates (except for the isolate CG4 from Galega officinalis) were presumptively identified as Rhizobium leguminosarum biovar viciae both by their symbiotic properties and the specific amplification of the nodC gene. In particular, we found that the nodC gene could be used as a diagnostic molecular marker for strains belonging to the bv. viciae. RFLP-PCR 16S rDNA analysis confirms these results, with the exception of two strains that showed different RFLP-genotypes from those of the reference strains of R. leguminosarum bv. viciae. Analysis of intraspecies relationship among strains by using the RAPD-PCR technique showed a high level of genetic polymorphism, grouping our isolates and reference strains into six different major clusters with a similarity level of 20%. Data from seven parameters of phenotypic and genotypic analyses were evaluated by using principal component analysis which indicated the differences among strains and allowed them to be divided into seven different groups. (c) 2005 Elsevier GmbH. All rights reserved.

year	journal	country	edition	language
2005-01-01

http://hdl.handle.net/11588/348562