Nicola Lorenz

Glyphosate effects on soil rhizosphere-associated bacterial communities.

Glyphosate is one of the most widely used herbicides in agriculture with predictions that 1.35 million metric tons will be used annually by 2017. With the advent of glyphosate tolerant (GT) cropping more than 10 years ago, there is now concern for non-target effects on soil microbial communities that has potential to negatively affect soil functions, plant health, and crop productivity. Although extensive research has been done on short-term response to glyphosate, relatively little information is available on long-term effects. Therefore, the overall objective was to investigate shifts in the rhizosphere bacterial community following long-term glyphosate application on GT corn and soybean in the greenhouse. In this study, rhizosphere soil was sampled from rhizoboxes following 4 growth periods, and bacterial community composition was compared between glyphosate treated and untreated rhizospheres using next-generation barcoded sequencing. In the presence or absence of glyphosate, corn and soybean rhizospheres were dominated by members of the phyla Proteobacteria, Acidobacteria, and Actinobacteria. Proteobacteria (particularly gammaproteobacteria) increased in relative abundance for both crops following glyphosate exposure, and the relative abundance of Acidobacteria decreased in response to glyphosate exposure. Given that some members of the Acidobacteria are involved in biogeochemical processes, a decrease in their abundance could lead to significant changes in nutrient status of the rhizosphere. Our results also highlight the need for applying culture-independent approaches in studying the effects of pesticides on the soil and rhizosphere microbial community.

Changes in rhizosphere bacterial gene expression following glyphosate treatment.

In commercial agriculture, populations and interactions of rhizosphere microflora are potentially affected by the use of specific agrichemicals, possibly by affecting gene expression in these organisms. To investigate this, we examined changes in bacterial gene expression within the rhizosphere of glyphosate-tolerant corn (Zea mays) and soybean (Glycine max) in response to long-term glyphosate (PowerMAX™, Monsanto Company, MO, USA) treatment. A long-term glyphosate application study was carried out using rhizoboxes under greenhouse conditions with soil previously having no history of glyphosate exposure. Rhizosphere soil was collected from the rhizoboxes after four growing periods. Soil microbial community composition was analyzed using microbial phospholipid fatty acid (PLFA) analysis. Total RNA was extracted from rhizosphere soil, and samples were analyzed using RNA-Seq analysis. A total of 20-28 million bacterial sequences were obtained for each sample. Transcript abundance was compared between control and glyphosate-treated samples using edgeR. Overall rhizosphere bacterial metatranscriptomes were dominated by transcripts related to RNA and carbohydrate metabolism. We identified 67 differentially expressed bacterial transcripts from the rhizosphere. Transcripts downregulated following glyphosate treatment involved carbohydrate and amino acid metabolism, and upregulated transcripts involved protein metabolism and respiration. Additionally, bacterial transcripts involving nutrients, including iron, nitrogen, phosphorus, and potassium, were also affected by long-term glyphosate application. Overall, most bacterial and all fungal PLFA biomarkers decreased after glyphosate treatment compared to the control. These results demonstrate that long-term glyphosate use can affect rhizosphere bacterial activities and potentially shift bacterial community composition favoring more glyphosate-tolerant bacteria.

Microbial Profiles of Rhizosphere and Bulk Soil Microbial Communities of Biofuel Crops Switchgrass (Panicum virgatum L.) and Jatropha (Jatropha curcas L.)

The production of biofuels from the low-input energy crops, switchgrass (Panicum virgatum L.) and jatropha (Jatropha curcas L.), is a sustainable approach that can provide more usable energy and environmental benefits than food-based biofuels. Plant rhizosphere affects the microbial community structure due to variations in root exudation rates and residue chemistry. The objective of this investigation was to determine the profiles of microbial communities associated with rhizosphere and bulk soils of switchgrass or jatropha using phospholipid fatty acid (PLFA) analysis and length heterogeneity PCR (LH-PCR). Switchgrass soil contained a significantly (Pl0.05) higher abundance of Gram-positive (i14:0, i15:0, a15:0), Gram-negative (16:1ω5c, 16:1ω7c, 18:1ω5c), and saturated (14:0, 15:0) PLFAs compared to jatropha soil, whereas jatropha had a higher abundance of fungal (18:2ω6, 9c), 18:1ω9c, 20:1ω9c, and 18:0 PLFAs compared to switchgrass soil. Irrespective of plant type, rhizosphere soil contained a significantly (Pl0.05) higher abundance of saturated PLFAs (16:0, 18:0, 20:0), actinomycetes (10Me17:0), and fungal (18:2ω6, 9c) PLFAs compared to bulk soil; whereas bulk soil had higher abundance of saturated (14:0), Gram-negative (16:1ω9c, 16:1ω5c, 16:1ω7c), and 18:1ω9c PLFAs compared to rhizosphere soil. Multivariate principle component analysis of PLFAs and LH-PCR percent relative peak areas successfully differentiated the microbial communities of rhizosphere and bulk soils of switchgrass and jatropha.

Fame Profiling and Activity of Microbial Communities During jatropha curcas L. Residue Decomposition in Semiarid Soils

There is widespread interest in the renewable biofuel potential of Jatropha curcas L., a shrub that exists under indigenous conditions in India. Following oil extraction from seeds, the remaining shell and cake residue can be used as a soil amendment. Microbial decomposition of organic residues can improve the quality of soils, but little is known about Jatropha as a soil amendment. Therefore, the objective of this study was to determine the influence of Jatropha residue on soil microbial community structure during decomposition. Soil microcosms amended with Jatropha leaves, cake, or fruit shell or an unamended control were incubated in the laboratory and destructively sampled at 3, 10, 20, 40, and 60 days. At each sampling, soil microbial communities were analyzed using fatty acid methyl ester (FAME) profiles, and enzyme assays performed for &bgr;-glucosidase, cellulase, and urease. Microbial biomass (FAMEtot) and bacterial and fungal FAME concentrations were significantly higher in cake-amended soil compared with other treatments. Averaged across all treatments, FAMEtot and fungal and bacterial FAMEs were highest on Day 3 and subsequently decreased significantly. Higher fungal-to-bacterial FAME ratios were found in residue-amended soils over control, with the highest ratio occurring in fruit shell-amended soil (except Day 3). A significantly lower ratio of the stress indicator (saturated to monounsaturated FAMEs) was found in cake-amended soil compared with other amendments. Fungal, bacterial, and actinomycetes biomarkers were significantly correlated with &bgr;-glucosidase and cellulase activities. The study shows that decomposition of Jatropha affected the soil microbial community, differently depending on residue type, but overall had positive effects in promoting the microbial diversity and activity, thus making Jatropha residue a favorable amendment for soils.