Multidrug-resistant bacteria arise mostly by the accumulation of plasmids and chromosomal mutations. Typically, these resistant determinants are costly to the bacterial cell. Yet, recently, it has been found that, in Escherichia coli bacterial cells, a mutation conferring resistance to an antibiotic can be advantageous to the bacterial cell if another antibiotic-resistance mutation is already present, a phenomenon called sign epistasis. Here we study the interaction between antibiotic-resistance chromosomal mutations and conjugative (i.e., self-transmissible) plasmids and find many cases of sign epistasis (40%)—including one of reciprocal sign epistasis where the strain carrying both resistance determinants is fitter than the two strains carrying only one of the determinants. This implies that the acquisition of an additional resistance plasmid or of a resistance mutation often increases the fitness of a bacterial strain already resistant to antibiotics. We further show that there is an overall antagonistic interaction between mutations and plasmids (52%). These results further complicate expectations of resistance reversal by interdiction of antibiotic use.

Summary 1.Plants can host microbial communities whose integrated functions are often responsible for their success. Understanding mechanisms regulating such functions is thus a major goal in ecology. 2. Fungal endophytes of grasses, particularly of the genus Neotyphodium , have been reported to reduce colonization of their host plant by arbuscular mycorrhizal (AM) fungi. However, it is unclear which mechanism(s) may explain the effects produced by the endophyte and whether these effects are present in nature. 3. We used Schedonorus phoenix (syn. Lolium arundinaceum ) plants that were endophyte-free or infected with one of two strains of Neotyphodium coenophialum known to produce different putative allelochemicals to test the hypotheses that (i) allelopathic effects of the endophyte reduce AM fungal spore germination; and (ii) the allelochemical compound(s) are leached into the soil even after the death of S. phoenix , where they reduce AM fungal colonization of other plants. 4. In a first experiment, aqueous extracts from the shoots of S. phoenix were applied onto spores of the AM fungus Glomus intraradices to test germination effects. Both endophyte strains reduced spore germination by approximately 10% relative to endophyte-free controls. 5. In a second experiment, we placed dried shoot material ('thatch') on the soil surface of pots containing Bromus inermis , which were either inoculated with G. intraradices or not. We watered the plants through the thatch, relying upon leaching to translocate potential allelochemicals to the soil. AM fungal colonization of B. inermis was significantly reduced when thatch was infected with the common strain, but not with AR542, compared to the endophyte-free thatch. Furthermore, the arbuscule : vesicle ratio was 11-fold smaller when thatch was infected with the common strain compared to endophyte-free thatch, suggesting that G. intraradices was stressed by the presence of common strain-leachate. 6. We observed situations whereby two ecologically widespread plant-microbe symbioses interact. Potential mechanisms may include allelopathic effects, although other factors are also possible, and leaching is a mode of entry of putative endophyte-induced AM fungal inhibitors in soil. Understanding these processes is important as they affect AM fungal communities which contribute to plant success and, consequentially, grassland ecosystem dynamics.

The factors which may influence temporal and spatial variation in plant arbuscular mycorrhizal (AM) colonization and propagule occurrence were evaluated in a Portuguese salt marsh poor in plant diversity. Two distinct sites were studied: a more-flooded (low marsh) and a less-flooded zone (high marsh). AM root colonization, AM fungal spore number and inoculum potential, soil edaphic parameters and tidal flooding time periods were analysed. Levels of AM colonization were considerable in Aster tripolium and Inula crithmoides but very low in Puccinellia maritima and non-existent in Spartina maritima, Halimione portulacoides, Arthrocnemum fruticosum and Arthrocnemum perenne. Fungal diversity was very low, with Glomus geosporum dominant at both marsh zones. Colonization showed no spatial variation within marsh zones but temporal variation was observed in the high marsh, dependent on plant phenological phases. In the low marsh, no significantly seasonal variation was observed. Apparently, plant phenological events were diluted by stressful conditions (e.g. flooding, salinity). Spore density was significantly different between marsh zones and showed temporal variation in both zones. This study showed that distribution of mycorrhizas in salt marsh is more dependent on host plant species than on environmental stresses.

Bacteria sometimes cooperate with co-inhabiting cells. Pathogenic bacteria, for example, often produce and excrete virulence factors, eventually benefitting both producer and non-producer cells. The role of social interactions involving antibiotic resistance, however, has been more elusive. Enzymes that inactivate β-lactam antibiotics such as ampicillin or penicillin (β-lactamases) are good candidates as public goods. Nonetheless, it has been claimed that bacteria harbouring plasmids of natural origin coding for β-lactamase almost do not protect sensitive bacteria. This does not fit with the fact that ampicillin-sensitive bacteria can be isolated from subjects undergoing ampicillin treatment. We hypothesised that there are two non-exclusive explanations for the discrepancy between previous works: (1) the range of values of demographic conditions (such as initial strain frequency, initial total cell density or habitat structure) has not been broad enough to include most scenarios, or (2) there are interactions between some of these factors. We performed experiments with Escherichia coli bacterial cells to measure the degree of protection of sensitive cells when co-cultured with cells harbouring RP4, R16a or the R1 plasmids, all of natural origin and coding for β-lactamases, and in presence of ampicillin. In these co-cultures, performed in structured and non-structured environments, both the initial total cell density and the initial frequency of sensitive cells spanned four orders of magnitude. We found protection of sensitive cells in 63% of tested conditions. All factors (plasmid, structure, frequency and density) significantly affect levels of protection. Moreover, all factors interact, with interactions revealing large or very large effect sizes.