Advancing Mitochondrial DNA Visualization and Dynamics in Plants: Application of the MT-HI-NESS Fluorescent Marker

Authors

Madeleine H. Faber, University of Nebraska - LincolnFollow

First Advisor

Alan C. Christensen

Second Advisor

Wayne Riekhof

Date of this Version

5-2025

Document Type

Thesis

Citation

Faber, M.H. (2025) Advancing Mitochondrial DNA Visualization and Dynamics: Application of the MT-HI-NESS Fluorescent Marker. Undergraduate Honors Thesis. University of Nebraska-Lincoln.

Comments

Copyright Madeleine Faber 2025.

Abstract

Mitochondrial DNA (mtDNA) plays a crucial role in cellular energy production and inheritance, yet its dynamics and exchange remain poorly understood in plant systems due to limitations in visualization techniques. Traditional visualization methods such as exogenous DNA stains and fluorescent protein markers present challenges in specificity and live-cell imaging. To address this challenge, we adapted the MT-HI-NESS fluorescent marker, originally developed for mammalian cells, for use in plant mitochondrial studies. Using Arabidopsis thaliana as a model system, we employed mitochondrial targeting sequences (IVD and AOX) to ensure accurate localization and to confirm mtDNA binding specificity. The validation process included fluorescence-based localization analysis and physiological assessments to determine the impact of MT-HI-NESS on plant growth and mitochondrial function. The results showed successful marker integration without adverse physiological effects, paving the way for future studies on mitochondrial dynamics, mtDNA exchange, and plant bioenergetics. This advancement provides a powerful tool for exploring mitochondrial biology in plants and expands the potential for more studies at the cellular level.