Abstract

BERTANI, Romina P. et al. Genetic transformation of Acidovorax avenae subsp. avenae, the causal agent of Red stripe in sugarcane. Rev. ind. agric. Tucumán [online]. 2020, vol.97, n.2, pp.35-37. ISSN 1851-3018.

Red stripe (causal agent Acidovorax avenae subsp. avenae) is one of the main diseases that affect the sugarcane crop in Tucumán. At the moment, the information on sugarcane colonization and movement of A. avenae subsp. avenae during infections is limited, so the aim of this study was to develop tools to monitor the bacteria during the disease cycle. Three transformation methods were tested: chemical, electroporation and conjugation using plasmids containing different fluorescent proteins and selective gen markers. Mutant A. avenae subsp. avenae strains were evaluated for antibiotic resistance and expression of fluorescence signals. In addition, their identity was confirmed by PCR specie-specific. In this study, only transformation by conjugation presented stable fluorescent expression in the bacterium. The Aa::pHC60-gfp showed the highest expression of the green fluorescent protein gfp. In vitro assays showed not difference between the growth of wild type strain and the mutant one in liquid media. Plants inoculated either with A. avenae subsp. avenae or Aa::pHC60-gfp presented comparable incidence and severity 15 days after inoculation. This is the first report of an A. avenae subsp. avenae mutant strain expressing a heterologous gene and represents a valuable tool to study the biology of the bacterium and its colonization in sugarcane

Keywords : gfp; conjugation; disease cycle.

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