Hafeez, Zaki et al. (2021) International Worm Meeting "Investigating the Diversity and Distribution of Caenorhabditis elegans in Georgia"

Shaver, Amanda, Hafeez, Zaki, Edison, Arthur, Anderson, Lindsay, Mohamed, Youssef, Spencer, Deontis, Shah, Man, Asif, Muhammad Zaka, Muzio, Cole

[International Worm Meeting, 2021]

The Caenorhabditis elegans Natural Diversity Resource (CeNDR) maintains a library of strains of C. elegans collected from every continent except Antarctica (Caenorhabditis elegans Natural Diversity Resource [CeNDR], 2020). However, currently, there is only one C. elegans strain from the state of Georgia cataloged in CeNDR (CeNDR, 2020). In an attempt to expand upon the diversity of these collected strains and in an effort to study the distribution of C. elegans in Georgia, we set forth to collect a number of wild nematode isolates. Samples of rotting and decaying vegetation were collected from a variety of locations across Georgia. Data such as temperature, location, and sample type were recorded along with images of each collection site using the Nematode Field Sampling app within the data collection app Fulcrum. Nematodes were isolated from these samples and screened by visual inspection for morphological similarity to C. elegans. Potential C. elegans strains were then cultivated. To confirm that an isolate is indeed C. elegans, we will perform PCR and gel electrophoresis. For worms with an appropriately sized PCR band, we intend to submit samples of the PCR product for Sanger sequencing. We will then use NCBI BLAST to compare the sequencing results of the wild isolates with known species. Finally, we will submit frozen isolates of C. elegans to CeNDR for cataloging and whole-genome sequencing. Caenorhabditis elegans Natural Diversity Resource. (2020, August 30). Global Strain Map [Interactive Map]. Retrieved March 24, 2021 from https://www.elegansvariation.org/strain/global-strain-map

Asif, Muhammad Zaka et al. (2021) International Worm Meeting "Determining the Effects of 1-Hydroxyphenazine Exposure on UGT Mutants in Caenorhabditis elegans"

Shah, Man, Hafeez, Zaki, Nocilla, Kelsey, Asif, Muhammad Zaka, Edison, Arthur

[International Worm Meeting, 2021]

Caenorhabditis elegans is an ideal model organism for studying the xenobiotic detoxification pathways of various natural and synthetic toxins. One such toxin that has been shown to cause death in C. elegans is 1-hydroxyphenazine (1-HP), a molecule produced by the bacterium Pseudomonas aeruginosa. Prior research in our lab has shown the median lethal dose (LD50) for 1-HP in C elegans is 179 muM in PD1074 and between 150-200muM in N2.(Asif et al., 2021; Stupp et al., 2013). Prior research has also shown that C. elegans detoxifies 1-HP by glycosylating it with one, two, or three glucose molecules in N2 worms.(Stupp et al., 2013) We hypothesize that UDP-Glucuronosyltransferase (UGT) enzymes are responsible for glycosylating 1-HP in C. elegans. To identify UGT enzymes implicated in the glycosylation of 1-HP in PD1074, we have implemented our plate-based toxicity assay developed in our prior work on available UGT strains. We began by testing eight UGT mutants, UGT-1, UGT-6, UGT-9, UGT-23, UGT-49, UGT-60, UGT-62, and UGT-66, at the LD50 concentration of 1-HP in PD1074. We screened for mutants with a different mortality rate to N2 and PD1074 worms. Additionally, we will perform HPLC/UV analysis and NMR analysis in order to describe the differences in glycosylation patterns and the ratios of glycosylated and unglycosylated products in mutant strains with differential susceptibility to 1-HP than N2 and PD1074 worms. This could help explain the variation in mortality rates between the different strains and help us understand the complexity of UGTs in C. elegans.