Vibrio cholerae vexH Encodes a Multiple Drug Efflux Pump That Contributes to the Production of Cholera Toxin and the Toxin Co-Regulated Pilus

The resistance-nodulation-division (RND) efflux systems are ubiquitous transporters that function in antimicrobial resistance. Recent studies showed that RND systems were required for virulence factor production in Vibrio cholerae. The V. cholerae genome encodes six RND efflux systems. Three of the RND systems (VexB, VexD, and VexK) were previously shown to be redundant for in vitro resistance to bile acids and detergents. A mutant lacking the VexB, VexD, and VexK RND pumps produced wild-type levels of cholera toxin (CT) and the toxin co-regulated pilus (TCP) and was moderately attenuated for intestinal colonization. In contrast, a RND negative mutant produced significantly reduced amounts of CT and TCP and displayed a severe colonization defect. This suggested that one or more of the three uncharacterized RND efflux systems (i.e. VexF, VexH, and VexM) were required for pathogenesis. In this study, a genetic approach was used to generate a panel of V. cholerae RND efflux pump mutants in order to determine the function of VexH in antimicrobial resistance, virulence factor production, and intestinal colonization. VexH contributed to in vitro antimicrobial resistance and exhibited a broad substrate specificity that was redundant with the VexB, VexD, and VexK RND efflux pumps. These four efflux pumps were responsible for in vitro antimicrobial resistance and were required for virulence factor production and intestinal colonization. Mutation of the VexF and/or VexM efflux pumps did not affect in vitro antimicrobial resistance, but did negatively affect CT and TCP production. Collectively, our results demonstrate that the V. cholerae RND efflux pumps have redundant functions in antimicrobial resistance and virulence factor production. This suggests that the RND efflux systems contribute to V. cholerae pathogenesis by providing the bacterium with protection against antimicrobial compounds that are present in the host and by contributing to the regulated expression of virulence factors

In vitro propagation and germplasm conservation of wild orchids from South America

Orchids are an important part of plant biodiversity on this planet due to their high variability among species and their habitats. South America represents more than thirty percent of all known orchid species, Colombia, Ecuador, Brazil, Peru, and Bolivia being among the richest countries in the world in terms of orchid biodiversity. Nevertheless, concerning the orchid conservation status, in Colombia precisely orchids occupy the unlucky first place as the plant family with the highest number of threatened species. There is a similar situation in the rest of the South American countries. The two main threats to orchid survival are both anthropogenic: the first one is deforestation, and the second largest threat to orchids is collection from the wild. One desirable action to safeguard these endangered species is to develop procedures that make possible their massive propagation, which would provide material for both environmental restoration and commercial purposes avoiding extractions from nature. Likewise, the development of systems that allow the ex situ conservation of orchid germplasm is imperative. This chapter reviews the progresses of different in vitro approaches for orchid propagation and germplasm conservation, safeguarding the genetic biodiversity of these species. Several study cases are presented and described to exemplify the protocols developed in the Botanical Institute of Northeast (UNNE-CONICET) for propagating and long-term storing the germplasm of wild orchids from Argentina (Cattleya lundii, Cohniella cepula, C. jonesiana, Gomesa bifolia, Aa achalensis, Cyrtopodium brandonianum, C. hatschbachii, Habenaria bractescens). Moreover, it has been attempted to put together most of the available literature on in vitro propagation and germplasm conservation for South American orchids using different explants and procedures. There are researches of good scientific quality that even cover critical insights into the physiology and factors affecting growth and development as well as storage of several orchid materials. Moreover, studies are still necessary to cover a major number of South American species as well as the use of selected material (clonal) for both propagation and conservation approaches.Fil: Dolce, Natalia Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: Medina, Ricardo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: Terada, Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: González Arnao, María Teresa. Universidad Veracruzana; MéxicoFil: Flachsland, Eduardo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; Argentin