GFP-tagged multimetal-tolerant bacteria and their detection in the rhizosphere of white mustard

The introduction of rhizobacteria that tolerate heavy metals is a promising approach to support plants involved in phytoextraction and phytostabilisation. In this study, soil of a metal-mine wasteland was analyzed for the presence of metal-tolerant bacterial isolates, and the tolerance patterns of the isolated strains for a number of heavy metals and antibiotics were compared. Several of the multimetal-tolerant strains were tagged with a broad host range reporter plasmid (i.e. pPROBE-NT) bearing a green fluorescent protein marker gene (gfp). Overall, the metal-tolerant isolates were predominately Gram-negative bacteria. Most of the strains showed a tolerance to five metals (Zn, Cu, Ni, Pb and Cd), but with differing tolerance patterns. From among the successfully tagged isolates, we used the transconjugant Pseudomonas putida G25 (pPROBE-NT) to inoculate white mustard seedlings. Despite a significant decrease in transconjugant abundance in the rhizosphere, the gfp-tagged cells survived on the root surfaces at a level previously reported for root colonisers

Characterization of the Metabolically Modified Heavy Metal-Resistant Cupriavidus metallidurans Strain MSR33 Generated for Mercury Bioremediation

BACKGROUND: Mercury-polluted environments are often contaminated with other heavy metals. Therefore, bacteria with resistance to several heavy metals may be useful for bioremediation. Cupriavidus metallidurans CH34 is a model heavy metal-resistant bacterium, but possesses a low resistance to mercury compounds. METHODOLOGY/PRINCIPAL FINDINGS: To improve inorganic and organic mercury resistance of strain CH34, the IncP-1β plasmid pTP6 that provides novel merB, merG genes and additional other mer genes was introduced into the bacterium by biparental mating. The transconjugant Cupriavidus metallidurans strain MSR33 was genetically and biochemically characterized. Strain MSR33 maintained stably the plasmid pTP6 over 70 generations under non-selective conditions. The organomercurial lyase protein MerB and the mercuric reductase MerA of strain MSR33 were synthesized in presence of Hg(2+). The minimum inhibitory concentrations (mM) for strain MSR33 were: Hg(2+), 0.12 and CH(3)Hg(+), 0.08. The addition of Hg(2+) (0.04 mM) at exponential phase had not an effect on the growth rate of strain MSR33. In contrast, after Hg(2+) addition at exponential phase the parental strain CH34 showed an immediate cessation of cell growth. During exposure to Hg(2+) no effects in the morphology of MSR33 cells were observed, whereas CH34 cells exposed to Hg(2+) showed a fuzzy outer membrane. Bioremediation with strain MSR33 of two mercury-contaminated aqueous solutions was evaluated. Hg(2+) (0.10 and 0.15 mM) was completely volatilized by strain MSR33 from the polluted waters in presence of thioglycolate (5 mM) after 2 h. CONCLUSIONS/SIGNIFICANCE: A broad-spectrum mercury-resistant strain MSR33 was generated by incorporation of plasmid pTP6 that was directly isolated from the environment into C. metallidurans CH34. Strain MSR33 is capable to remove mercury from polluted waters. This is the first study to use an IncP-1β plasmid directly isolated from the environment, to generate a novel and stable bacterial strain useful for mercury bioremediation

Effect of temperature on growth and fatty acids profile of the biodiesel producing microalga Scenedesmus acutus

Description of the subject. The present study examined the effect of temperature (15, 20, 25, 30, 35 and 40 °C) on biomass, esterified fatty acids content and fatty acid productivity of Scenedesmus acutus. Objectives. This work aimed to study the effect of variation in temperature on lipid productivity and fatty acid profiles of S. acutus as a feedstock for biodiesel production. Method. The alga was grown under different temperatures and its biomass, as well as fatty acid content and composition, were determined. Results. The maximum growth rate of S. acutus was achieved at 30 °C , but there was no significant difference in biomass productivity at 25 and 30 °C (0.41 and 0.42 g·l-1·d-1), respectively. The highest fatty acid content (104.1 mg·g-1 CDW) was recorded at low temperature (15 °C) and decreased with increasing temperature. As a result of high biomass production, fatty acids productivity showed the highest values (41.27 and 42.10 mg·l-1·d-1) at 25 and 30 °C, respectively. The proportion of saturated and mono-unsaturated fatty acids increased from 13.72 to 23.79% and from 11.13 to 33.10% of total fatty acids when the incubation temperature was raised from 15 to 40 °C, respectively. The increase of temperature from 15 to 40 °C decreased the poly-unsaturated fatty acids from 75.15% to 43.10% of total fatty acids, respectively. Conclusions. The present study concluded that incubation temperature was a critical parameter for quantitative and qualitative fatty acid compositions of S. acutus. In addition, the type and proportion of individual fatty acids, which interfere with biodiesel quality, can be modified using different incubation temperatures in order to meet the biodiesel international standards

Data from: Genome-wide association analyses in the model rhizobium Ensifer meliloti

Genome-wide association studies (GWAS) can identify genetic variants responsible for naturally occurring and quantitative phenotypic variation and therefore provide a powerful complement to approaches that rely on de novo mutations for characterizing gene function. Although bacteria should be amenable to GWAS, few GWAS have been conducted on bacteria, and the extent to which non-independence among genomic variants (e.g. linkage disequilibrium, LD) and the genetic architecture of phenotypic traits will affect GWAS performance is unclear. We apply association analyses to identify candidate genes underlying variation in 20 biochemical, growth, and symbiotic phenotypes among 153 stains of Ensifer meliloti. For 10 traits we find genotype-phenotype associations that are stronger than expected by chance, with the candidates in relatively small linkage groups, indicating that LD does not preclude resolving association candidates to relatively small genomic regions. The significant candidates show an enrichment for nucleotide polymorphisms (SNPs) over gene presence-absence variation (PAV), and for five traits, candidates are enriched in large linkage groups, a possible signature of epistasis. Many of the variants most strongly associated with symbiosis phenotypes were in genes previously known to be involved in nitrogen-fixation or nodulation. For other traits, apparently strong associations were not stronger than the range of associations detected in permuted data. In sum, our data show that GWAS in bacteria may be a powerful tool for characterizing genetic architecture and identifying genes responsible for phenotypic variation, however, careful evaluation of candidates is necessary to avoid false signals of association

Reference Genome

Copy of reference genome sequence and annotation