title: The ipdC, hisC1 and hisC2 genes involved in indole-3-acetic production used as alternative phylogenetic markers in Azospirillum brasilense.
abstract: Plant growth-promoting bacteria of the genus Azospirillum are present in the rhizosphere and as endophytes of many crops. In this research we studied 40 Azospirillum strains isolated from different plants and geographic regions. They were first characterized by 16S rDNA restriction analysis, and their phylogenetic position was established by sequencing the genes 16S rDNA, ipdC, hisC1, and hisC2. The latter three genes are involved in the indole-3-pyruvic acid (IPyA) biosynthesis pathway of indole-3-acetic acid (IAA). Furthermore, the suitability of the 16S-23S rDNA intergenic spacer sequence (IGS) for the differentiation of closely related Azospirillum taxa and development of PCR protocols allows for specific detection of strains. The IGS-RFLP analysis enabled intraspecies differentiation, particularly of Azospirillum brasilense and Azospirillum lipoferum strains. Results demonstrated that the ipdC, hisC1, and hisC2 genes are highly conserved in all the assessed A. brasilense isolates, suggesting that these genes can be used as an alternative phylogenetic marker. In addition, IAA production determined by HPLC ranged from 0.17 to 98.2 μg mg(-1) protein. Southern hybridization with the A. brasilense ipdC gene probe did not show, a hybridization signal with A. lipoferum, Azospirillum amazonense, Azospirillum halopreferans and Azospirillum irakense genomic DNA. This suggests that these species produce IAA by other pathways. Because IAA is mainly synthesized via the IPyA pathway in A. brasilense strains, a species that is used worldwide in agriculture, the identification of ipdC, hisC1, and hisC2 genes by PCR may be suitable for selecting exploitable strains.
authors.affiliation: Centro de Investigaciones en Ciencias Microbiológicas, Benemérita Universidad Autónoma de Puebla (BUAP), 72000, Puebla, Puebla, Mexico.
authors.forename: Saúl
authors.initials: S
authors.lastname: Jijón-Moreno
authors.forename: Cynthia
authors.initials: C
authors.lastname: Marcos-Jiménez
authors.forename: Raúl O
authors.initials: RO
authors.lastname: Pedraza
authors.forename: Alberto
authors.initials: A
authors.lastname: Ramírez-Mata
authors.forename: I García
authors.initials: IG
authors.lastname: de Salamone
authors.forename: Ana
authors.initials: A
authors.lastname: Fernández-Scavino
authors.forename: Claudia A
authors.initials: CA
authors.lastname: Vásquez-Hernández
authors.forename: Lucia
authors.initials: L
authors.lastname: Soto-Urzúa
authors.forename: Beatriz E
authors.initials: BE
authors.lastname: Baca
mesh_terms: D015806:Azospirillum brasilense / Q000145:classification* / Q000235:genetics* / Q000378:metabolism; D053898:Biosynthetic Pathways / Q000235:genetics*; D015139:Blotting, Southern; D002851:Chromatography, High Pressure Liquid; D016000:Cluster Analysis; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D021903:DNA, Ribosomal Spacer; D005798:Genes, Bacterial; D007210:Indoleacetic Acids / Q000378:metabolism*; D008969:Molecular Sequence Data; D010802:Phylogeny; D010944:Plants / Q000382:microbiology; D012150:Polymorphism, Restriction Fragment Length; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012338:RNA, Ribosomal, 23S / Q000235:genetics; D017422:Sequence Analysis, DNA
chemical_list: D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D021903:DNA, Ribosomal Spacer; D007210:Indoleacetic Acids; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S; C030737:indoleacetic acid