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An ipdC gene knock-out of Azospirillum brasilense strain SM and its implications on indole-3-acetic acid biosynthesis and plant growth promotion

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Abstract

The indole-3-pyruvate decarboxylase gene (ipdC), coding for a key enzyme of the indole-3-pyruvic acid pathway of IAA biosynthesis in Azospirillum brasilense SM was functionally disrupted in a site-specific manner. This disruption was brought about by group II intron-based Targetron gene knock-out system as other conventional methods were unsuccessful in generating an IAA-attenuated mutant. Intron insertion was targeted to position 568 on the sense strand of ipdC, resulting in the knock-out strain, SMIT568s10 which showed a significant (∼50%) decrease in the levels of indole-3-acetic acid, indole-3-acetaldehyde and tryptophol compared to the wild type strain SM. In addition, a significant decrease in indole-3-pyruvate decarboxylase enzyme activity by ∼50% was identified confirming a functional knock-out. Consequently, a reduction in the plant growth promoting response of strain SMIT568s10 was observed in terms of root length and lateral root proliferation as well as the total dry weight of the treated plants. Residual indole-3-pyruvate decarboxylase enzyme activity, and indole-3-acetic acid, tryptophol and indole-3-acetaldehyde formed along with the plant growth promoting response by strain SMIT568s10 in comparison with an untreated set suggest the presence of more than one copy of ipdC in the A. brasilense SM genome.

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Acknowledgements

Authors thank Prof. Dieter Haas (Universite de Lausanne, Switzerland) for the gift of pME3468. M.M. acknowledges financial support by Council of Scientific and Industrial Research, India. The authors also acknowledge financial assistance by Department of Biotechnology, Govt. of India to S.S. and facilities supported by University Grants Commission under the SAP program and Department of Science and Technology under the FIST program in the Department of Genetics, UDSC, New Delhi.

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  1. Department of Genetics, University of Delhi South Campus, Benito Juarez Road, New Delhi, 110021, India

    Mandira Malhotra & Sheela Srivastava

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  1. Mandira Malhotra
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  2. Sheela Srivastava
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Correspondence to Sheela Srivastava.

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Malhotra, M., Srivastava, S. An ipdC gene knock-out of Azospirillum brasilense strain SM and its implications on indole-3-acetic acid biosynthesis and plant growth promotion . Antonie van Leeuwenhoek 93, 425–433 (2008). https://doi.org/10.1007/s10482-007-9207-x

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