Abstract
Main conclusion
Rhizobium etli CE3-DsRed pMP604 drives infection 12-24 h earlier than R. etli CE3-DsRed and it is an excellent tool in live-cell imaging studies of IT developement in P. vulgaris roots.
Abstract
The study of the cellular dynamics of nodulation has frequently been limited by the difficulty of performing live-cell imaging in nodule primordia and legume roots, which are constituted by multiple cell layers, such is the case of Phaseolus vulgaris. Seeking conditions to reduce the time it takes for rhizobia to infect P. vulgaris root, we decided to explore the nodulation properties of Rhizobium etli CE3 pMP604, a strain that constitutively produces Nod factors through a flavonoids-independent transcriptional activation which is often used to purify Nod factors. Even though the strain infects 12–24 h earlier than the parental R. etli CE3 strain, infection thread (IT) formation, nodule organogenesis processes and N2-fixation activity are similar for both strains. Additionally, we have confirmed that R. etli CE3-DsRed pMP604 is an excellent tool to trace IT development in P. vulgaris roots.
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Abbreviations
- dpi:
-
Days post-inoculation
- hpi:
-
Hours post-inoculation
- IC:
-
Infection chamber
- IT:
-
Infection thread
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Acknowledgements
We thank M.Sc. Andrés M. Saralegui Amaro and Dr. J. Arturo Pimentel Cabrera (Laboratorio Nacional de Microscopía Avanzada, Instituto de Biotecnología, UNAM) for technical assistance in confocal microscopy and Alfonso Leija Salas (Centro de Ciencias Genómicas, UNAM) for acetylene reduction assays. We also thank Shirley E. Ainsworth for library services, Juan Manuel Hurtado, Roberto Rodríguez, Omar Arriaga, and Arturo Ocádiz for computer services. We are grateful to Drs. Claudia L. Treviño and Julio C. Chavez for their technical advice, Enrique Merino (Instituto de Biotecnología, UNAM) and Susana Brom (Centro de Ciencias Genómicas, UNAM) for critical reading of the manuscript. This work was partially supported by DGAPA-UNAM (IN207215 and IN206118 to RSL) and by Consejo Nacional de Ciencia y Tecnología (CONACyT) with PhD fellowships to RDD (288601), LABV (384814) and EMM (595064).
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Supplementary file1 R. etli CE3 and R. etli CE3 pMP604 growth curves. Bacteria were grown in 180 µl/well of liquid PY medium in a 96-well plate at 30 °C for 50 h with shaking. Optical density of seven replicates were collected every hour and mean values of each time point were plotted with GraphPad Prism version 6. Error bars represent ± SD. There was no difference between R. etli CE3 and R. etli CE3 pMP604 growth rates (unpaired Student-t two-tailed test P < .05) (PDF 75 kb)
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Dávila-Delgado, R., Bañuelos-Vazquez, L.A., Monroy-Morales, E. et al. Rhizobium etli CE3-DsRed pMP604: a useful biological tool to study initial infection steps in Phaseolus vulgaris nodulation. Planta 252, 69 (2020). https://doi.org/10.1007/s00425-020-03479-y
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DOI: https://doi.org/10.1007/s00425-020-03479-y