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The ipdC, hisC1 and hisC2 genes involved in indole-3-acetic production used as alternative phylogenetic markers in Azospirillum brasilense

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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.

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Acknowledgments

We are grateful to Dr. C. Elmerich to the gift of Pseudomonas stutzeri A1501 strain, Noé Arroyo for technical assistance, and Ira Fogel for English editing. This work was supported by the Consejo Nacional de Ciencia y Tecnología) to B.E.B (CONACyT grant 49227-Z) and funding by a collaborative grant to B.E.B., R.P., I. G de S. and A.F.S. from Ciencia y Tecnología para el Desarrollo (CYTED). S. J. M., C. M. J. and L.S.U. are recipients CONACYT fellowships, respectively. Authors appreciate to Editor and Reviewers for constructive and valuable comments.

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Authors and Affiliations

  1. Centro de Investigaciones en Ciencias Microbiológicas, Benemérita Universidad Autónoma de Puebla (BUAP), 72000, Puebla, Puebla, Mexico

    Saúl Jijón-Moreno, Cynthia Marcos-Jiménez, Alberto Ramírez-Mata, Claudia A. Vásquez-Hernández, Lucia Soto-Urzúa & Beatriz E. Baca

  2. Facultad de Agronomía y Zootecnia, Universidad Nacional de Tucumán, San Miguel de Tucumán, Argentina

    Raúl O. Pedraza

  3. Facultad de Agronomía, Universidad de Buenos Aires, Buenos Aires, Argentina

    I. García de Salamone

  4. Facultad de Química Universidad de la República, 11100, Montevideo, Uruguay

    Ana Fernández-Scavino

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  1. Saúl Jijón-Moreno
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  9. Beatriz E. Baca
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Correspondence to Beatriz E. Baca.

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Jijón-Moreno, S., Marcos-Jiménez, C., Pedraza, R.O. et al. The ipdC, hisC1 and hisC2 genes involved in indole-3-acetic production used as alternative phylogenetic markers in Azospirillum brasilense . Antonie van Leeuwenhoek 107, 1501–1517 (2015). https://doi.org/10.1007/s10482-015-0444-0

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