Fatty acid biomarker analysis to characterize soil microbial communities in soybean agroecosystems with Sclerotinia stem rot disease

Soybean (Glycine max (L.) Merr.) is one the major crops produced worldwide. However, soybean is susceptible to many diseases. Sclerotinia stem rot (SSR) disease caused by Sclerotinia sclerotiorum (Lib.) de Bary is considered one of the most important fungal diseases of soybean. It can be controlled by chemicals (e.g. fungicides), by breeding cultivars with disease resistance and by cultural control (e.g. increasing the width between rows, reducing plant populations). A promising and complementary method of controlling SSR disease in the field is the application of biological control agents. Biological control agents introduced in a soil environment will interact with other soil food web organisms, as do the pathogenic organism and infected plants, which may change the genetic and functional diversity in soil microbial communities. Profiling these changes may lead to an improved understanding of the interactions between these players (biological control agents, pathogens, soil biota and plants) in the biological control phenomenom, permiting us to exploit naturally-occurring ecological relationships and develop more sustainable approaches to control soybean diseases. Fatty acid biomarkers analysis was used to profile microbial communities in soils. Two laboratory studies were conducted to evaluate the methods used for extraction and profiling the fatty acid biomarkers from soil samples with a range of soil properties (clay content, organic matter content), The first study investigated the best solvent mixture for recovering fatty acid biomarkers from soil using an automated pressurized solvent extraction (PSE) system. Solvent mixtures containing chloroform and methanol were more efficient at extracting fatty acids from agricultural soils than hexane:2-propanol and acetone. The second study presented an exploratory pyrolysis-mass spectrometry technique to rapidly fingerprint soil lipids extracted from different agroecosystems. Pyrolysis-mass spectrometry discriminated among soils and crop production systems in the same way as the fatty acid profiling. I also report on the efficicacy of biological control agents to control Sclerotinia stem rot disease in soybean. A two-year study was conducted in soybean fields under conventional or no tillage to determine whether Trichoderma virens (SoilGard(TM)) and arbuscular mycorrhizal fungi (a mixture of Glomus intraradices and G. mosseae ), used alone or in combination, could reduce sclerotinia stem rot (SSR) disease incidence. Generally, SSR disease indicators, as well as the soybean yield, were not affected significantly by the biological control treatments. I then studied whether changes in microbial community composition were related to the inoculation of the biological control agents and the disease incidence in soybean fields. Inoculation of biological control agents changes the expression of many soil fatty acids during both years of the trial. Also, in the plots with severely diseased plants, fatty acids biomarkers of gram positive and actinomycetes bacteria were significantly greater than in plots with healthy plants. I conclude that further improvement in laboratory techniques and procedures will permit researchers to efficiently extract and characterize soil lipids, providing new insight into soil organic matter dynamics and soil microbial ecology. Further study will be needed to verify the efficacy and optimize the application method, dose and timing of biocontrol agents to provide protection against SSR disease in soybean fields.