Abstract
We have developed an unsophisticated, non-disruptive and accurate method for evaluation of pathogen colonization in planta. In this protocol we use a Ralstonia solanacearum (R. solanacearum) UY031 strain genetically modified to constitutively generate light from a synthetic luxCDABE operon stably inserted in its chromosome. This system allows bacterial quantification in a high-throughput manner, avoiding time-consuming and tedious bacterial dilution plating and colony counting. In addition, this system could be especially useful in plant breeding programs to detect bacterial latent growth in symptomless parental lines before their inclusion in long-term disease resistance breeding programs.
Materials and Reagents
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Acknowledgments
This work was supported by grants SGR0052 and CONES2010-0030 from Comissionat per Universitats i Recerca of the Catalan Government (Generalitat de Catalunya), AGL2010-21870 from the Ministerio de Economía of the Spanish Government, and FMV_2009_1_3045 from the National Research Council in Uruguay (ANII). We thank I. van DijK and F. Monteiro for constructing original plasmids (pG-Pps, pRCG-GWY and pRCGent-Peplux) used as sources of inserts for cloning, F. Vilaró and M. González for providing the S. commersonii genotypes used in this study and S. Balcells for lending us a luminometer.
References
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