Pesticide Side Effects in an Agricultural Soil Ecosystem as Measured by amoA Expression Quantification and Bacterial Diversity Changes.
METHODS
Assessing the effects of pesticide hazards on microbiological processes in the soil is currently based on analyses that provide limited insight into the ongoing processes. This study proposes a more comprehensive approach. The side effects of pesticides may appear as changes in the expression of specific microbial genes or as changes in diversity. To assess the impact of pesticides on gene expression, we focused on the amoA gene, which is involved in ammonia oxidation. We prepared soil microcosms and exposed them to dazomet, mancozeb or no pesticide. We hypothesized that the amount of amoA transcript decreases upon pesticide application, and to test this hypothesis, we used reverse-transcription qPCR. We also hypothesized that bacterial diversity is affected by pesticides. This hypothesis was investigated via 454 sequencing and diversity analysis of the 16S ribosomal RNA and RNA genes, representing the active and total soil bacterial communities, respectively.


CONCLUSIONS
Treatment with dazomet reduced both the bacterial and archaeal amoA transcript numbers by more than two log units and produced long-term effects for more than 28 days. Mancozeb also inhibited the numbers of amoA transcripts, but only transiently. The bacterial and archaeal amoA transcripts were both sensitive bioindicators of pesticide side effects. Additionally, the numbers of bacterial amoA transcripts correlated with nitrate production in N-amended microcosms. Dazomet reduced the total bacterial numbers by one log unit, but the population size was restored after twelve days. The diversity of the active soil bacteria also seemed to be re-established after twelve days. However, the total bacterial diversity as reflected in the 16S ribosomal RNA gene sequences was largely dominated by Firmicutes and Proteobacteria at day twelve, likely reflecting a halt in the growth of early opportunists and the re-establishment of a more diverse population. We observed no effects of mancozeb on diversity.
Geological Survey of Denmark and Greenland (GEUS), Department of Geochemistry, Copenhagen, Denmark.
Louise
L
Feld
Geological Survey of Denmark and Greenland (GEUS), Department of Geochemistry, Copenhagen, Denmark.
Mathis Hjort
MH
Hjelmsø
Geological Survey of Denmark and Greenland (GEUS), Department of Geochemistry, Copenhagen, Denmark.
Morten Schostag
MS
Nielsen
Geological Survey of Denmark and Greenland (GEUS), Department of Geochemistry, Copenhagen, Denmark.
Anne Dorthe
AD
Jacobsen
University of Copenhagen, Department of Biology, Section of Terrestrial Ecology, Copenhagen, Denmark.
Regin
R
Rønn
University of Copenhagen, Department of Biology, Section of Terrestrial Ecology, Copenhagen, Denmark.
Flemming
F
Ekelund
University of Aarhus, Department of Bioscience, Section of Soil Fauna Ecology and Ecotoxicology, Silkeborg, Denmark.
Paul Henning
PH
Krogh
University of Copenhagen, Department of Plant and Environmental Sciences, Section of Environmental Chemistry and Physics, Frederiksberg, Denmark.
Bjarne Westergaard
BW
Strobel
Geological Survey of Denmark and Greenland (GEUS), Department of Geochemistry, Copenhagen, Denmark.
Carsten Suhr
CS
Jacobsen
D000383:Agriculture; D000641:Ammonia / Q000378:metabolism; D001419:Bacteria / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D044822:Biodiversity; D017753:Ecosystem; D015870:Gene Expression; D009566:Nitrates / Q000378:metabolism; D010084:Oxidation-Reduction; D010575:Pesticides; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012987:Soil / Q000737:chemistry*; D012988:Soil Microbiology; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D009566:Nitrates; D010575:Pesticides; D012336:RNA, Ribosomal, 16S; D012987:Soil; D000641:Ammonia