Colonization of Madagascar periwinkle (Catharanthus roseus), by endophytes encoding gfp marker
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This study reports the introduction of gfp marker in two endophytic bacterial strains (Pantoea agglomerans C33.1, isolated from cocoa, and Enterobacter cloacae PR2/7, isolated from citrus) to monitor the colonization in Madagascar perinwinkle (Catharanthus roseus). Stability of the plasmid encoding gfp was confirmed in vitro for at least 72 h of bacterial growth and after the colonization of tissues, under non-selective conditions. The colonization was observed using fluorescence microscopy and enumeration of culturable endophytes in inoculated perinwinkle plants that grew for 10 and 20 days. Gfp-expressing strains were re-isolated from the inner tissues of surface-sterilized roots and stems of inoculated plants, and the survival of the P. agglomerans C33:1gfp in plants 20 days after inoculation, even in the absence of selective pressure, suggests that is good colonizer. These results indicated that both gfp-tagged strains, especially P. agglomerans C33.1, may be useful tools to deliver enzymes or other proteins in plant.
KeywordsMadagascar periwinkle Endophytes Green fluorescent protein
Funding for this research was provided by FAPESP. A. R. Torres and M. Mondin are CAPES fellows. P. Rossetto and W. L. Araújo are FAPESP fellows. L. Cursino, M. Hungria, and J. L. Azevedo are CNPq fellows. The authors thank M. C. Quecine for help in statistical analysis and Drs. Glaciela Kaschuk and Paulo Lacava for suggestions on the manuscript.
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