title: A rapid and simple PCR method for identifying isolates of the genus Azospirillum within populations of rhizosphere bacteria.
abstract: OBJECTIVE
  To develop a rapid and simple genus-specific polymerase chain reaction (PCR) method for detecting and identifying isolates of the genus Azospirillum which is well-recognized as plant growth-promoting rhizobacterium.
  
  
  RESULTS
  Nine pairs of PCR primers were designed based on the Azospirillum 16S rRNA, ipdC, nifA and nifH genes to assess their genus specificity by testing against 12 Azospirillum (from seven species) and 15 non-Azospirillum reference strains, as compared with the fAZO/rAZO pair reported by Baudoin et al. (J Appl Microbiol, 108, 2010, 25). Among the primer pairs assessed, the Az16S-A pair designed on the 16S rRNA gene sequence showed the highest genus specificity: it successfully yielded a single amplicon of the expected size in all the 12 Azospirillum strains and for a close relative, Rhodocista centenaria. The PCR with the Az16S-A primers generated a detectable amount of the amplicon from ≥10³ CFU ml⁻¹ of Azospirillum cell suspensions even in the presence of contaminants and accurately discriminated Azospirillum and non-Azospirillum species in both 35 Azospirillum-like and 70 unknown isolates from plant roots and rhizosphere soils.
  
  
  CONCLUSIONS
  We developed a rapid and simple PCR method for detecting and identifying Azospirillum isolates within populations of rhizosphere bacteria.
  
  
  CONCLUSIONS
  The method developed would serve as a useful tool for isolating a variety of indigenous Azospirillum bacteria from agricultural samples.
authors.affiliation: National Agricultural Research Center for Hokkaido Region (NARCH), Hitsujigaoka 1, Toyohira-ku, Sapporo, Japan.
authors.forename: A
authors.initials: A
authors.lastname: Shime-Hattori
authors.forename: S
authors.initials: S
authors.lastname: Kobayashi
authors.forename: S
authors.initials: S
authors.lastname: Ikeda
authors.forename: R
authors.initials: R
authors.lastname: Asano
authors.forename: H
authors.initials: H
authors.lastname: Shime
authors.forename: T
authors.initials: T
authors.lastname: Shinano
mesh_terms: D016949:Azospirillum / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D005798:Genes, Bacterial; D057230:Limit of Detection; D018517:Plant Roots / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D058441:Rhizosphere; D017422:Sequence Analysis, DNA; D012988:Soil Microbiology; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S