Title of Research Student Names

Bacteria Biodiversity from Soil of Different Land Managements at Furman UniversityNeely Wood, Clayton Bishop, Min-Ken LiaoDepartment of Biology, Furman UniversityThere are at least three different soil management practices on the Furman University campus: chemically managed (lawn on the Furman Mall), organically managed (Furman farm), and no management (untreated forested area). Based on the assumption that different land managements, or lack thereof, would create different ecosystems, we hypothesized that the bacterial communities in the soil of these land managements would be different. We isolated soil bacteria from different land managements and selected 6-10 isolates that are morphologically distinct to represent each site for further analysis. We amplified and sequenced the 16S rRNA gene of these representatives and taxonomically identified them to the genus level then categorized them at the family level. For each soil type, we concluded, then, that the family Bacillaceae was most prominent, followed by Paenibacilliceae, Flavobacteriaceae, and Pseudamonaceae for the forested area, the organically managed area, and the chemically managed area, respectively. Concurrently, we performed double restriction enzyme digests (HinP1I and MspI) on the 16S rRNA gene of the remaining bacterial isolates for a quick comparison of bacterial community diversity in different soil types. Our preliminary results based on double digest data of at least 15 individual isolates per site showed no significant difference in relative diversity. However, based on the sequencing analysis results and the Simpsons diversity tests, the soil bacteria community in the forested area was the most diverse, followed by the organically managed soil and, lastly, the chemically managed soil. More soil samples will be collected and analyzed in summer 2015. Soil Sampling. Soil samples were collected during June and July of 2014 from three differently managed soil sites at Furman University. These soil managements included organically managed (Figure 3a), chemically managed soil (Figure 2a), and unmanaged forested soil (Figure 1a).Community DNA Extraction. Community soil DNA was extracted using the PowerSoil DNA Isolation Kit and its protocol (MO BIO, Carlsbad, CA). Bacteria Culturing. Bacteria from each site was extracted from the soil, diluted 1:100, and was grown on both 0.1 TSA and R2A media. Representative isolates of each site were selected based on morphological differences and isolated. PCR Amplification for Identification. PCR amplification was used selecting for the 16S rDNA gene for each isolate. The isolates were then identified at the genus level.Restriction Enzyme Digest. Double restriction enzyme digests were performed on the 16S rDNA amplicons using HinP1I and MspI and subjected to gel electrophoresis.

AbstractMaterials and MethodsIntroductionResultsa)

SummaryAcknowledgementsThis project was funded by the Furman Advantage Research Fellowship.Figure 1b,c. Bacterial diversity in the nonmanaged, forested soil. Representative prevalent bacterial isolates were selected based on morphology, genetically identified at a genus level, and then grouped according to family (Figure 1b). Charts display percent total isolates identified. 15 bacterial isolates were double digested with restriction enzymes HinP1I and MspI at 37C for 3h, and sized by gel electrophoresis. Note: the first well did not produce a result (Figure 1c). According to Simpsons reciprocal index (1/D), the bacterial diversity at each of the three sites was determined to be in the following order from most to least diverse: unmanaged, forested soil (1/D = 3.42), organically managed soil (1/D = 2.15), and chemically managed soil (1/D = 1.57).The family Bacillaceae was most prominent throughout the three sites and the only family seen in all three land managements, followed by Paenibacilliceae, Flavobacteriaceae, and Pseudamonaceae for the forested area (Figure 1b), the organically managed area (Figure 3b), and the chemically managed area, respectively (Figure 2b).Based on the double restriction digest of the isolates, no plausible conclusion can be made about the relative diversity of the bacteria from the differently managed soil sites (Figures 1c, 2c, 3c, 3d). However, it can be tentatively concluded that between the differently managed soil sites, that there are varying and diverse bacterial populations.This preliminary study will be continued in the Summer of 2015.

Figure 1a. Figure 2a.Figure 3a.Figure 2b.Figure 2c.In soil ecosystems, bacteria play a crucial role in supporting all living organisms in the soil. However, the interactions between the below-ground and above-ground communities are not a one-way street. While the soil bacterial communities influence the biodiversity of the living communities above ground, the above ground biota reciprocally influence the below ground biota. To further complicate the interactions of these mutual drivers, soil ecosystems are often impacted by humans to various degrees, physically, chemically, and biologically. Different soil ecosystems thus create different environments that subsequently support different soil bacterial communities.At Furman University, at least three managements are present unmanaged (Figure 1a), chemically managed (Figure 2a), organically managed (Figure 3a). In this study, we examined the effects of different land management on soil bacterial community composition. We hypothesized that the differently managed soil on campus would entail variance between sites in the prevalence of different bacteria species. More specifically, we hypothesized that we would see the least bacterial diversity in the chemically managed soil, and the most in the organically managed soil. This diversity would be seen at both the taxonomic identification of bacterial family, as well as the diversity of to the 16S rDNA genes of the soil bacteriaFigure 2b,c. Bacterial diversity in the chemically managed soil. Representative prevalent bacterial isolates were selected based on morphology, genetically identified at a genus level, and then grouped according to family (Figure 2b). Charts display percent total isolates identified. 19 bacterial isolates were double digested with restriction enzymes HinP1I and MspI at 37C for 3h, and sized by gel electrophoresis (Figure 2c). Figure 3b,c,d. Bacterial diversity in the organically managed soil. Representative prevalent bacterial isolates were selected based on morphology, genetically identified at a genus level, and then grouped according to family (Figure 3b). Charts display percent total isolates identified. 54 total bacterial isolates, where farm beds supporting both maize and leaf vegetables were examined, were double digested with restriction enzymes HinP1I and MspI at 37C for 3h, and sized by gel electrophoresis (Figure 3c,d).

Figure 1b.Figure 1c.Figure 3b.Figure 3c.Figure 3d.