Throughout this experiment, specific sites and mutations on pol delta were observed to determine which would have the greatest effects on the final polymerase molecule. Ideally, these changes would positively impact DNA replication. For the physical plant science laboratory component, folic acid (a B vita min)’s presence was studied to determine its effect on tobacco seedling growth.

It was hypothesized that specific sites and mutations on the pol delta protein, for example G680V, would affect cassava resistance to CMD by altering the functionality of pol delta and therefore the reliable production of high-quality DNA. G680V in particular was researched for its potential to allow the “fin gers” of pol delta to move excessively and potentially interfere with DNA reproduction. Additionally, the hypothesis that folic acid would increase the growth of tobacco seedlings was tested.

Materials included computers with Visual Molecular Dynamics (VMD) software, Nicotiana tabacum seeds, Petri dishes, folic acid, potting mix, and a dissection microscope.

This project required the physical handling of Nicotiana tabacum, which has been known to cause nicotine poisoning in workers handling the crop. However, this only occurs when proper precautions are not taken and there is a lengthened exposure to the material. Thus, when performing this part of the procedure, all biological material was carefully disposed of in a manner comparable to if it were an invasive species—the plant matter will be bagged and thrown away, not burned. Biosafety Level 1 was observed for the duration of the laboratory.

References

• “Biosafety Level (BSL) 1: Hazard Control.” Blink, UC San Diego, 2 Mar. 2024, blink.ucsd.edu/safety/ research-lab/biosafety/containment/bsl-1.html. Accessed 12 Jan. 2025.
• “Cassava.” Florida Heritage Foods, floridaheritage-foods.com/latin-american-collection/cassava/.
• Chikoti, Patrick Chiza et al. “Cassava mosaic disease: a review of a threat to cassava production in Zambia.” Journal of plant-pathology : an international journal of the Italian
• Phytopathological Society vol. 101,3 (2019): 467 477. doi:10.1007/s42161-019-00255-0
• Crider, Krista S et al. “Folate and DNA methylation: a review of molecular mechanisms and the evidence for folate’s role.” Advances in nutrition (Bethesda, Md.) vol. 3,1 (2012): 21-38. doi:10.3945/ an.111.000992
• Diaz, Sylvia, et al. Uncovering the Mechanism of Cassava Mosaic Disease Resistance: A Computational Investigation of Cassava DNA Polymerase Delta and Mutant (V528L and G690V) Systems. 2024.
• Foley, M.C., Couto, L., Rauf, S. et al. Insights into DNA polymerase δ’s mechanism for accurate DNA replication. J Mol Model 25, 80 (2019). https://doi.org/10.1007/s00894-019-3957-z
• “Folic Acid Safety, Interactions, and Health Outcomes.” Folic Acid. U.S. Center for Disease Control, 15 May 2024, www.cdc.gov/folic-acid/about/safety.html.  Accessed 12 Jan. 2025.
• “Green Tobacco Sickness.” Occupational Safety and Health Administration: Safety and Health 10. Topics. U.S. Department of Labor, www.osha.gov/green-tobacco-sickness. Accessed 12 Jan. 2025.
• National Center for Biotechnology Information. “Pub Chem Compound Summary for CID135398658, Folic Acid” PubChem, https://pubchem.ncbi.nlm.nih.gov/compound/Folic-Acid. Accessed 27 Janu ary, 2025.
• Scholthof, Karen-Beth G. “Tobacco mosaic virus.” The American Phytopathological Society (APS), 1 Jan. 2005, www.apsnet.org/edcenter/disandpath/viral/pdlessons/Pages/TobaccoMosaic.aspx. Accessed 4 Oct. 2024.
• “Tobacco mosaic virus (TMV) Frequently Asked Questions.” 24 Feb. 2014. PDF