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Biotization of Arabidopsis thaliana with Pseudomonas putida and assessment of its positive effect on in vitro growth

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Abstract

Determination of the optimal inoculation method and concentration to use for plant-bacteria interaction studies is important in many cases, such as the phytoremediation of heavy metals and other toxic compounds in contaminated areas. The aim of this study was to compare different concentrations and times of inoculation of Pseudomonas putida with various growth stages of Arabidopsis thaliana in 14-d in vitro cultures. A significant beneficial impact of the bacterium was detected in the shoot length and root weight of seedlings. The highest shoot length and root fresh and dry weights were detected in 14-d and 2 × 103 cfu mL−1 inoculated samples. In addition, the increase in root weight could be visualized with crystal violet staining, as relatively more root hair and lateral root formation occurred in seedlings inoculated with moderate concentrations of bacteria, possibly due to the ability of P. putida to produce indole-acetic acid. Moreover, the highest photosynthetic pigment accumulation was obtained with the highest bacterial inoculum (2 × 106 cfu mL−1), which was tested in 0- or 3-d-old seedlings. Rhizospheric bacterial colonization was also visualized with GFP-labeled bacteria by confocal microscopy. These results showed that biotization of A. thaliana with P. putida KT2440 did not cause severe oxidative stress in seedlings, because H2O2 accumulation levels together with CAT and POX activities were not significantly induced. Therefore, this strain could be used for several applications based on plant-bacteria interactions.

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Acknowledgments

The authors would like to thank Prof. Dr. Victor de Lorenzo (Systems and Synthetic Biology Program, Centro Nacional de Biotecnología, Madrid, Spain) for the pSEVA237-G plasmid (http://seva.cnb.csic.es), Prof. Yelda Özden Çiftçi for her valuable comments, and Dr. Metin Çetin for his help for confocal microscopy.

Funding

This study was funded by a grant (2018-A105-41) from Gebze Technical University.

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Authors and Affiliations

  1. Department of Molecular Biology and Genetics, Gebze Technical University, 41400, Kocaeli, Turkey

    Ebru Arslan & Özlem Akkaya

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  1. Ebru Arslan
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  2. Özlem Akkaya
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Correspondence to Özlem Akkaya.

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Editor: Masaru Nakano

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Arslan, E., Akkaya, Ö. Biotization of Arabidopsis thaliana with Pseudomonas putida and assessment of its positive effect on in vitro growth. In Vitro Cell.Dev.Biol.-Plant 56, 184–192 (2020). https://doi.org/10.1007/s11627-019-10045-z

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