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The Overproduction of Indole-3-Acetic Acid (IAA) in Endophytes Upregulates Nitrogen Fixation in Both Bacterial Cultures and Inoculated Rice Plants

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Abstract

Endophytic bacteria from roots and leaves of rice plants were isolated and identified in order to select the diazotrophs and improve their nitrogen-fixing abilities. The nitrogen-fixing endophytes were identified by PCR amplification of the nifH gene fragment. For this purpose, two isolates, Enterobacter cloacae RCA25 and Klebsiella variicola RCA26, and two model bacteria (Herbaspirillum seropedicae z67 and Sinorhizobium fredii NGR234) were transformed to increase the biosynthesis of the main plant auxin indole-3-acetic acid (IAA). A significant increase in the production of IAA was observed for all strains. When the expression of nifH gene and the activity of the nitrogenase enzyme were analyzed in liquid cultures, we found that they were positively affected in the IAA-overproducing endophytes as compared to the wild-type ones. Rice plants inoculated with these modified strains showed a significant upregulation of the nitrogenase activity when plants infected with the wild-type strains were used as reference. Similar results were obtained too with common bean plants infected with the S. fredii NGR234 strain. These findings suggest that IAA overproduction improves nitrogen-fixing apparatus of endophytic bacteria both in liquid cultures and in inoculated host plants. The present study highlights new perspectives to enhance nitrogen-fixing ability in non-legume crops. These strains could be used as bioinoculants to improve the growth and the yield of agricultural crops, offering an alternative to the use of chemical nitrogen fertilizers.

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Acknowledgments

We are grateful to Silvia Ardissone (Department of Microbiology and Molecular Medicine, Geneva) for kindly supplying the NGR234 strain. We are also grateful to Franco Nulli for providing us the rice plants used for the isolation of endophytes. We thank Mr. Rubino Stefano for technical assistance. This work was partially supported by a dedicated grant from the Italian Ministry of Economy and Finance to the National Research Council for the project CISIA “Innovazione e Sviluppo del Mezzogiorno—Conoscenze Integrate per Sostenibilità ed Innovazione del Made in Italy Agroalimentare—Legge no. 191/2009.” This work was also partially supported by the European Commission for funding the ABSTRESS project (FP7 KBBE-2011-289562).

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  1. Institute of Biosciences and Bioresources, Via P. Castellino 111, 80131, Naples, Italy

    Roberto Defez, Anna Andreozzi & Carmen Bianco

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  1. Roberto Defez
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  2. Anna Andreozzi
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Correspondence to Carmen Bianco.

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The nucleotide sequence data reported in this paper have been deposited in the National Center for Biotechnology Information database, and they are available using the supplied GenBank accession numbers.

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Defez, R., Andreozzi, A. & Bianco, C. The Overproduction of Indole-3-Acetic Acid (IAA) in Endophytes Upregulates Nitrogen Fixation in Both Bacterial Cultures and Inoculated Rice Plants. Microb Ecol 74, 441–452 (2017). https://doi.org/10.1007/s00248-017-0948-4

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