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Tools to evaluate Herbaspirillum seropedicae abundance and nifH and rpoC expression in inoculated maize seedlings grown in vitro and in soil

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Abstract

The plant growth promoting bacteria Herbaspirillum seropedicae is an important model to study biological nitrogen fixation and it is proposed as crop inoculants for grasses. In this study we developed new tools to evaluate the abundance of H. seropedicae SmR1 and its expression in planta and investigated the association of the plant growth promoting bacteria H. seropedicae with maize grown in sterile and nonsterile conditions. Maize seedlings (P30F53) were inoculated with H. seropedicae SmR1 and grown in vitro and in soil. The plants were sampled at 4, 7 and 10 (in vitro) or 14, 21 and 28 days after inoculation (soil). Using qPCR we quantified H. seropedicae DNA and measured nifH, rpoC and hrcN levels of bacterial transcripts. In vitro assay inoculated plants presented highest amount of DNA and transcript contents, not detected in control plants. nifH and rpoC gene expression were detected on roots of inoculated maize cultivated in both growth conditions. However, it was not possible to detect hrcN gene expression in maize roots cultivated in soil. TaqMan assay is species-specific for H. seropedicae DNA and nifH and rpoC transcript levels could be used to monitor H. seropedicae gene expression in planta in sterile and nonsterile growth conditions. We developed specific, reliable and efficient tools to monitor H. seropedicae DNA abundance and H. seropedicae nifH and rpoC expression in planta.

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Acknowledgements

This work was financially supported by the National Institute of Science and Technology-Biological Nitrogen Fixation (INCT-FBN), National Counsel of Technological and Scientific Development (CNPq), Ministry of Science and Technology, Brazil. Pâmela Dall Asta, Tomás Pellizzaro Pereira and Fernanda Plucani Amaral were recipients of PhD fellowships from Coordination of Personnel Improvement of Higher Education (CAPES), Ministry of Education, Brazil and Ana Carolina Maisonnave Arisi is recipient of research fellowship (PQ2) from CNPq.

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  1. CAL, CCA, UFSC, Food Science and Technology Department, Federal University of Santa Catarina, Rod. Admar Gonzaga, 1346, 88034-001, Florianópolis, SC, Brazil

    Pâmela Dall’Asta, Tomás Pellizzaro Pereira, Fernanda Plucani do Amaral & Ana Carolina Maisonnave Arisi

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  1. Pâmela Dall’Asta
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  2. Tomás Pellizzaro Pereira
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  3. Fernanda Plucani do Amaral
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Correspondence to Ana Carolina Maisonnave Arisi.

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Dall’Asta, P., Pereira, T.P., do Amaral, F.P. et al. Tools to evaluate Herbaspirillum seropedicae abundance and nifH and rpoC expression in inoculated maize seedlings grown in vitro and in soil. Plant Growth Regul 83, 397–408 (2017). https://doi.org/10.1007/s10725-017-0306-z

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