Rocío Medina

Remediation of a soil chronically contaminated with hydrocarbons through persulfate oxidation and bioremediation

The impact of remediation combining chemical oxidation followed by biological treatment on soil matrix and microbial community was studied, of a chronically hydrocarbon contaminated soil sourced from a landfarming treatment. Oxidation by ammonium persulfate produced a significant elimination of polycyclic aromatic hydrocarbons (PAHs) and an increase in PAH bioavailability. Organic-matter oxidation mobilized nutrients from the soil matrix. The bacterial populations were affected negatively, with a marked diminution in the diversity indices. In this combined treatment with oxidation and bioremediation working in tandem, the aliphatic-hydrocarbon fractions were largely eliminated along with additional PAHs. The chemical and spectroscopic analyses indicated a change in soil nutrients. In spite of the high residual-sulfate concentration, a rapid recovery of the cultivable bacterial population and the establishment of a diverse and equitable microbial community were obtained. Pyrosequencing analysis demonstrated a marked succession throughout this twofold intervention in accordance with the chemical and biologic shifts observed. These remediation steps produced different effects on the soil physiology. Spectroscopic analysis became a useful tool for following and comparing those treatments, which involved acute changes in a matrix of such chronically hydrocarbon-contaminated soil. The combined treatment increased the elimination efficiency of both the aliphatic hydrocarbons and the PAHs at the expense of the mobilized organic matter, thus sustaining the recovery of the resilient populations throughout the treatment. The high-throughput-DNA-sequencing techniques enabled the identification of the predominant populations that were associated with the changes observed during the treatments.

Assessment of the Responsiveness to Different Stresses of the Microbial Community from Long-Term Hydrocarbon-Contaminated Soils

Soils exposed to long-term contamination with hydrocarbons may present extreme challenges to maintain the biological resilience to the stress. To elucidate the relationships between the initial event of contamination and the responsiveness to the stress, we investigated the extent of the microbial resilience of biological functions from two contaminated soils sampled from a petrochemical area (S1, underwent diffuse hydrocarbon contamination, and S2, from a land farming unit where an alkaline petrochemical sludge was treated) after the Cd, saline, and acid stresses. Both contaminated soils were characterized by low organic matter content compared with a pristine soil. Although similar Shannon diversity index and heterotrophic bacterial count were observed, different bacterial community structures (PCR-DGGE) and less enzymatic activities characterized the contaminated soils. Particularly, functional diversity determined by Biolog EcoPlates™ was not detected in S2 soil. Only the S1 soil showed resilience of the enzymatic activities and functional diversity, suggesting the presence of a well-adapted microbial community able to face with the stresses. The S2 was the most disturbed and less responsive soil. However, an increase in the functional diversity was evidenced after acidification, and it is possible to correlate this responsiveness with the sludge properties treated in the land farming unit. In addition, if the selected stress can reverse the soil condition provoked for the first disturbance, responsiveness could be expected.

Draft Genome Sequence and Gene Annotation of Stemphylium lycopersici Strain CIDEFI-216

ABSTRACT Stemphylium lycopersici is a plant-pathogenic fungus that is widely distributed throughout the world. In tomatoes, it is one of the etiological agents of gray leaf spot disease. Here, we report the first draft genome sequence of S. lycopersici, including its gene structure and functional annotation.

The mitochondrial genome of the plantpathogenic fungus Stemphylium lycopersici uncovers a dynamic structure due to repetitive and mobile elements

Stemphylium lycopersici (Pleosporales) is a plant-pathogenic fungus that has been associated with a broad range of plant-hosts worldwide. It is one of the causative agents of gray leaf spot disease in tomato and pepper. The aim of this work was to characterize the mitochondrial genome of S. lycopersici CIDEFI-216, to use it to trace taxonomic relationships with other fungal taxa and to get insights into the evolutionary history of this phytopathogen. The complete mitochondrial genome was assembled into a circular double-stranded DNA molecule of 75,911 bp that harbors a set of 37 protein-coding genes, 2 rRNA genes (rns and rnl) and 28 tRNA genes, which are transcribed from both sense and antisense strands. Remarkably, its gene repertoire lacks both atp8 and atp9, contains a free-standing gene for the ribosomal protein S3 (rps3) and includes 13 genes with homing endonuclease domains that are mostly located within its 15 group I introns. Strikingly, subunits 1 and 2 of cytochrome oxidase are encoded by a single continuous open reading frame (ORF). A comparative mitogenomic analysis revealed the large extent of structural rearrangements among representatives of Pleosporales, showing the plasticity of their mitochondrial genomes. Finally, an exhaustive phylogenetic analysis of the subphylum Pezizomycotina based on mitochondrial data reconstructed their relationships in concordance with several studies based on nuclear data. This is the first report of a mitochondrial genome belonging to a representative of the family Pleosporaceae.

A survey on occurrence of Cladosporium fulvum identifies race 0 and race 2 in tomato-growing areas of Argentina

The presence of Cladosporium fulvum (syn. Passalora fulva), causal agent of tomato leaf mold, was confirmed in the two main greenhouse-production areas for tomato in Argentina. Using both morphological characters and internal transcribed spacer sequencing, we confirmed the presence of physiological races of this pathogen. A diagnostic multiplex polymerase chain reaction (PCR) was also developed, using primers derived from C. fulvum avirulence (Avr) genes. In all, 20 isolates of Cladosporium spp. were obtained as monospore cultures and 12 were identified as C. fulvum. By this method, we showed that, of these 12 isolates, 5 were race 0 (carrying functional Avr2, Avr4, Avr4E, and Avr9 genes) and 7 were race 2 (lacking the Avr2 gene). Race identity was confirmed by testing their virulence on a set of tomato differentials carrying different Cf resistance genes. All Avr genes could be amplified in single or multiplex PCR using DNA isolated from in vitro grown monospore cultures but only three Avr could be amplified when genomic DNA was isolated from C. fulvum-infected necrotic leaf tissue.