Transcriptional repression plays a critical role in the regulation of various biological processes, including developmental pathways and disease progression. Corepressors are proteins recruited by partner proteins to negatively influence the transcription of genes. TPL is a corepressor from the model plant Arabidopsis thaliana, and is known to play a pivotal role in transcriptional repression. My project aims to identify other proteins that work with TPL to form a transcriptional repression complex at a single engineered locus. To further understand the function of corepressors, we built a synthetic repressor system, dCas9-TPL, designed to specifically repress the transcription of the RUBY reporter gene. When expressed, the RUBY reporter turns Arabidopsis thaliana pink. In previous phases of this research, we utilized the EMS (ethyl methanesulfonate) mutagenesis protocol to create a population of plants containing many random mutations. Screening these plants for increased RUBY expression, I successfully Identified promising homozygous lines where plants demonstrated bright pink flowers and unique phenotypes such as early flowering, light avoidance, and small stature. Last quarter, I sent five lines to be sequenced and each line displayed distinct mutations that I can further explore to pinpoint the exact TPL interactor responsible for its unique phenotype. I am also investigating known TPL interactors such as SPT4, SPT5, and MED21 by creating transgenic lines within my control dCas9-TPL + RUBY line. Through genetic screening, I have validated the phenotypes among these control experiments. By investigating the intricate network of interactions between these regulatory proteins, I aim to gain a deeper understanding of how gene expression is coordinated across different cell types and how this process controls complex developmental pathways.

The timing of flowering in Arabidopsis thaliana is tightly regulated by environmental cues that induce the expression of FLOWERING LOCUS T (FT), a mobile signal responsible for floral induction. Recent studies have identified FPF1-LIKE PROTEIN 1 (FLP1), a gene co-expressed with FT in phloem companion cells, as a potential regulator of flowering. Given FLP1’s genetic similarity to known flowering regulators, we hypothesize that it functions as a mobile flowering-promoting signal. Using predictive protein interaction modeling with AlphaFold 3, followed by Bimolecular Fluorescence Complementation (BiFC) and Yeast-Two Hybrid assays, we confirmed that FLP1 interacts with key proteins in the shoot apical meristem. Overexpression of FLP1 leads to early flowering in Arabidopsis, while loss-of-function mutants exhibit delayed flowering, supporting its role in floral induction. Interestingly, while FLP1 homologs generally promote flowering across species, a contrasting effect in Brachypodium distachyon has been observed, where overexpression of its homolog (BdFLP1/BdFLP7) delays flowering. This unexpected result raises critical questions about the molecular basis of functional divergence among FLP1 homologs. To investigate this, we are introducing BdFLP1 and BdFLP7 into Arabidopsis to assess their effects on flowering time and to determine whether structural differences or distinct protein interactions underlie their divergent functions. Through molecular cloning, transgenic expression analysis, and biochemical assays including western blotting, immunoprecipitation, and mass spectrometry, we aim to elucidate the role of FLP1 and its homologs in flowering regulation. Understanding these mechanisms will provide deeper insights into conserved and species-specific pathways controlling floral induction. This knowledge is essential for improving crop adaptation strategies in the face of climate change, highlighting the broader significance of our research in plant developmental biology.

Industrial presence has increased in the Ecuadorian Amazon over the past several decades, in the form of lucrative monoculture cultivation and petroleum extraction. Simultaneously, large quantities of land have been deforested. Using a combination of GIS and remote sensing tools such as the Normalized Difference Vegetation Index and supervised classification, this study assesses the changes in vegetation, land-use, and petroleum infrastructure in the surrounding area of the Kichwa community of Loma del Tigre, between the years 2011-2024. The interactions and connections between these three landscape factors are also considered. The study region encompasses growing urban areas, African palm plantations, pasture, petroleum infrastructure and forest, among other land-use types. Forest presence was found to decrease, while the other mentioned factors increased. These results suggest that ecosystem services have been reduced, while water and soil contamination has worsened. The findings of this study may be beneficial in the advocacy for the expansion of land protection programs, such as Socio Bosque. The resulting maps from this study may also be beneficial to the community of Loma del Tigre, in pinpointing the sources of contamination in their surroundings.

Plants' defense mechanism against herbivory is integral to both resistance in nature and the global food supply. Glycine max or soybean, is one of the most widely grown crops in the world, and suffers substantial losses from pests, including many Lepidopteran species. Related legumes, including cowpea, and common bean, can respond to Lepidopteran herbivory by detecting Inceptin-11 (In11), a short peptide found in larval oral secretions. The protein responsible for this ability, Inceptin Receptor (INR), is not found in soybean. The aims of this project are two fold, firstly introducing INR into soybean lines and testing for improved resistance to Lepidopteran herbivory, and secondly studying the effects of INR on defense gene expression in soybean, in order to better understand mechanisms of herbivory resistance. The first step in this project was to create soybean lines which consistently express INR. This was done by sending our INR construct to collaborators, who used it to inoculate multiple soybean lines, then breeding the corresponding lines until response to In11 was seen in all offspring. We will then test larval beet armyworms (Spodoptera exigua) on both INR- and INR+ lines. We expect the INR+ lines to have significantly lower S. exigua growth, indicating an improved immune response. We are also infiltrating INR- and INR+ lines with both In11 and flg22 (a well studied bacterial elicitor) for RNA sequencing of the early immune response. We expect genes involved specifically anti herbivory mechanisms being upregulated when compared to flg22. These two prongs allow us not just to demonstrate the viability of stable transgenic herbivory resistant lines, but to uncover the molecular mechanisms involved in that resistance, allowing for future scientists to better engineer the next wave of pest resistant crops.

The Manduca sexta hawkmoth, a proficient pollinator, employs its antennae to efficiently navigate its surroundings. The antennae have highly sensitive olfactory receptor cells, allowing the moths to have an acute olfactory sense and odor recognition ability, making the moth antennae an ideal candidate tissue for developing reliable biosensors. In contrast, commonly used portable artificial sensors are inefficient and inaccurate in chemical detection. To evaluate the moth antenna's effectiveness as a biosensor model, I assessed the longevity in neural activity of the antenna via an electroantennogram over 24-hour durations. To do this, I attached an excised antenna to a circuit board to measure voltage variations across the antennal nerves during odor stimulation to understand the baseline antennal lifespan. To increase the longevity of electrical activity, I formed a hydrogel solution to enclose the antenna to protect it from drying out, and Leibovitz's L-15 Medium so the antenna has access to Amino Acids to build and repair its tissues, serving as an energy source. Preliminary findings show a tradeoff between longevity and electrical activity, where the antenna-only trials had high voltage readings over 4 hours while the hydrogel antenna had less intense electrical readings over 8 to 12 hours. The hydrogel proved to be a quality medium for the diffusion of the L-15 media over long periods of time, preserving the antenna from dying prematurely. These results demonstrate that moth antennae are a suitable model for  highly accurate and efficient long-duration biosensors and support the feasibility of implementing them in devices that detect and identify substances of interest with a longer life span. Future work will apply machine learning methods for enhanced chemical discrimination in disease diagnosis.

One of the most deadly creatures, the Aedes aegypti mosquito, flies under the radar of the everyday person far too often. However, as known carriers of vector-borne diseases such as dengue fever, Zika and yellow fever, these insects pose a significant risk for disease transmission. Our study aims will investigate the role of human-associated odors and colors in mosquito learning. I plan to test hexanoic acid and acetophenone, both odors which are commonly found on human skin. Acetophenone in particular is found in increased levels in patients with dengue fever, so by studying that particular odor more can be learned about the spread of disease. Using aversive training, mosquitoes are exposed to these odors paired with a shaking stimulus that simulates defensive swatting. A human-like color (orange) will additionally be paired with the odor to assess whether visual and olfactory stimuli presented together will strengthen the learning. After training we will place them in a two-way maze to test whether the scent and/or color was learned or not. This research will contribute to understanding how mosquitoes associate human-related  visual and olfactory cues, which may help inform future research on mosquito behavior and control strategies.