RESUMO
Ralstonia solanacearum species complex (RSSC) is a phytopathogenic bacterial group that causes bacterial wilt in several crops, being potato (Solanum tuberosum) one of the most important hosts. The relationship between the potato plant ionome (mineral and trace elements composition) and the resistance levels to this pathogen has not been addressed until now. Mineral content of xylem sap, roots, stems and leaves of potato genotypes with different levels of resistance to bacterial wilt was assessed in this work, revealing a positive correlation between divalent calcium (Ca) cation concentrations and genotype resistance. The aim of this study was to investigate the effect of Ca on bacterial wilt resistance, and on the growth and virulence of RSSC. Ca supplementation significantly decreased the growth rate of Ralstonia pseudosolanacearum GMI1000 in minimal medium and affected several virulence traits such as biofilm formation and twitching motility. We also incorporate for the first time the use of microfluidic chambers to follow the pathogen growth and biofilm formation in conditions mimicking the plant vascular system. By using this approach, a reduction in biofilm formation was observed when both, rich and minimal media, were supplemented with Ca. Assessment of the effect of Ca amendments on bacterial wilt progress in potato genotypes revealed a significant delay in disease progress, or a complete absence of wilting symptoms in the case of partially resistant genotypes. This work contributes to the understanding of Ca effect on virulence of this important pathogen and provides new strategies for an integrated control of bacterial wilt on potato. IMPORTANCE: Ralstonia solanacearum species complex (RSSC) includes a diverse group of bacterial strains that cause bacterial wilt. This disease is difficult to control due to pathogen aggressiveness, persistence, wide range of hosts, and wide geographic distribution in tropical, subtropical, and temperate regions. RSSC causes considerable losses depending on the pathogen strain, host, soil type, environmental conditions, and cultural practices. In potato, losses of $19 billion per year have been estimated for this pathogen worldwide. In this study, we report for the first time the mineral composition found in xylem sap and plant tissues of potato germplasm with different levels of resistance to bacterial wilt. This study underscores the crucial role of calcium (Ca) concentration in the xylem sap and stem in relation to the resistance of different genotypes. Our in vitro experiments provide evidence of Ca's inhibitory effect on the growth, biofilm formation, and twitching movement of the model RSSC strain R. pseudosolanacearum GMI1000. This study introduces a novel element, the Ca concentration, which should be included into the integrated disease control management strategies for bacterial wilt in potatoes.
Assuntos
Cálcio , Doenças das Plantas , Ralstonia solanacearum , Solanum tuberosum , Solanum tuberosum/microbiologia , Doenças das Plantas/microbiologia , Cálcio/metabolismo , Ralstonia solanacearum/fisiologia , Ralstonia solanacearum/genética , Ralstonia solanacearum/patogenicidade , Ralstonia solanacearum/crescimento & desenvolvimento , Virulência , Biofilmes/crescimento & desenvolvimento , Ralstonia/genética , Ralstonia/fisiologia , Raízes de Plantas/microbiologia , Xilema/microbiologiaRESUMO
Ralstonia pseudosolanacearum GMI1000 (Rpso GMI1000) is a soil-borne vascular phytopathogen that infects host plants through the root system causing wilting disease in a wide range of agro-economic interest crops, producing economical losses. Several features contribute to the full bacterial virulence. In this work we study the participation of light, an important environmental factor, in the regulation of the physiological attributes and infectivity of Rpso GMI1000. In silico analysis of the Rpso genome revealed the presence of a Rsp0254 gene, which encodes a putative blue light LOV-type photoreceptor. We constructed a mutant strain of Rpso lacking the LOV protein and found that the loss of this protein and light, influenced characteristics involved in the pathogenicity process such as motility, adhesion and the biofilms development, which allows the successful host plant colonization, rendering bacterial wilt. This protein could be involved in the adaptive responses to environmental changes. We demonstrated that light sensing and the LOV protein, would be used as a location signal in the host plant, to regulate the expression of several virulence factors, in a time and tissue dependent way. Consequently, bacteria could use an external signal and Rpsolov gene to know their location within plant tissue during the colonization process.
Assuntos
Proteínas de Bactérias/genética , Interações Hospedeiro-Patógeno/fisiologia , Ralstonia/fisiologia , Solanum lycopersicum/microbiologia , Aderência Bacteriana/fisiologia , Proteínas de Bactérias/metabolismo , Biofilmes/crescimento & desenvolvimento , Regulação Bacteriana da Expressão Gênica , Luz , Polissacarídeos Bacterianos/genética , Polissacarídeos Bacterianos/metabolismo , Ralstonia/patogenicidadeRESUMO
Fixed-bed reactors are usually designed for wastewater biotreatments, where the biofilm establishment and maintenance play the most important roles. Biofilm development strictly relies on different types of bacterial motility: swimming, swarming, and chemotaxis, which can be altered by the microenvironment conditions. The aim of this work is to do an integrated study on the effects of Cu(II), Cd(II), Zn(II) and Cr(VI) on swimming, swarming and chemotaxis of Pseudomonas veronii 2E, Delftia acidovorans AR and Ralstonia taiwanensis M2 to improve biofilm development and maintenance for metal loaded wastewater biotreatment in fixed-bed bioreactors. Swimming, swarming and chemotactic response evaluation experiments were carried out at different metal concentrations. P. veronii 2E motility was not affected by metal presence, being this strain optimal for fixed-bed reactors. D. acidovorans AR swarming was inhibited by Cd and Zn. Although R. taiwanensis M2 showed high resistance to Cu, Cd, Cr and Zn, motility was definitively altered, so further studies on R. taiwanensis M2 resistance mechanisms would be particularly interesting.