Horizontal genetic transfer in asexual fungi

Four aspects of horizontal genetic transfer during heterokaryon formation were examined in the asexual pathogen Fusarium oxysporum f.sp. cubense (Foc): (1) variability based on method of heterokaryon formation; (2) differences in nuclear and mitochondrial inheritance; (3) the occurrence of recombination without nuclear fusion; (4) the occurrence of horizontal genetic transfer between distantly related isolates. The use of non-pathogenic strains of Fusarium oxysporum as biocontrol agents warrants a closer examination at the reproductive life cycle of this fungus, particularly if drug resistance or pathogenicity genes can be transmitted horizontally. Experiments were divided into three phases. Phase I looked at heterokaryon formation by hyphal anastomosis and protoplast fusion. Phase II was a time course of heterokaryon formation to look at patterns of nuclear and mitochondrial inheritance. Phase III examined the genetic relatedness of the different vegetative compatibility groups using a multilocus analysis approach. Heterokaryon formation was evident within and between vegetative compatibility groups. Observation of non-parental genotypes after heterokaryon formation confirmed that, although a rare event, horizontal genetic transfer occurred during heterokaryon formation. Uniparental mitochondria inheritance was observed in heterokaryons formed either by hyphal anastomosis or protoplast fusion. Drug resistance was expressed during heterokaryon formation, even across greater genetic distances than those distances imposed by vegetative compatibility. Phylogenies inferred from different molecular markers were incongruent at a significant level, challenging the clonal origins of Foc. Mating type genes were identified in this asexual pathogen Polymorphisms were detected within a Vegetative Compatibility Group (VCG) suggesting non-clonal inheritance and/or sexual recombination in Foc. This research was funded in part by a NIH-NIGMS (National Institutes of Health-National Institute of General Medical Sciences) Grant through the MBRS (Minority Biomedical Research Support), the Department of Biological Sciences and the Tropical Biology Program at FIU. ^

Determination of vertical and horizontal pathways of injected fresh wastewater into a deep saline aquifer (Florida, USA) using natural chemical tracers

Two deep-well injection sites in south Florida, USA, inject an average of 430 million liters per day (MLD) of treated domestic fresh wastewater into a deep saline aquifer 900 m below land surface. Elevated levels of NH3 (highest concentration 939 µmol) in the overlying aquifer above ambient concentrations (concentration less than 30 µmol) were evidence of the upward migration of injected fluids. Three pathways were distinguished based on ammonium, chloride and bromide ratios, and temperature. At the South District Wastewater Treatment Plant, the tracer ratios showed that the injectate remained chemically distinct as it migrated upwards through rapid vertical pathways via density-driven buoyancy. The warmer injectate (mean 28°C) retained the temperature signal as it vertically migrated upwards; however, the temperature signal did not persist as the injectate moved horizontally into the overlying aquifers. Once introduced, the injectate moved slowly horizontally through the aquifer and mixed with ambient water. At the North District Wastewater Treatment Plant, data provide strong evidence of a one-time pulse of injectate into the overlying aquifers due to improper well construction. No evidence of rapid vertical pathways was observed at the North District Wastewater Treatment Plant.