As a child born and raised in the agricultural areas of the Terai region in Nepal, I began to connect with plants in my early childhood. Since then, I’ve considered agriculture as a meaningful and versatile vocation. When I picked a biology-focused high school curriculum and learned the fundamentals of gene function, hereditary traits, gene control, and expression, I started to address how plants feed billions of people throughout the globe.
I began to ponder how to increase agricultural output in order to feed more people in the future without damaging the environment. If improvement in humans and plants is solely genetic, then what genes are responsible for the changes? How are genetic pathways regulated? At what stage of development are these genes turned ON or OFF? To better comprehend the central dogma of molecular biology and how it differs in different organisms, I pursued biotechnology at university to better understand these questions.
At the beginning of my funded undergraduate education, I took rigorous courses in cell and molecular biology, as well as plant biotechnology, bioinformatics, chemistry, physics, biochemistry, and genetics. I took part in scientific tutorials as a freshman, where I learned presenting skills, scientific writing, and applying mathematics in research. In the later years of my degree, I began to comprehend how cells function at the molecular level. I participated in 4 weeks of labwork to study the effect of IAA on reactive oxygen species in tomato seedlings exposed to cadmium stress, which reinforced my desire and conviction to pursue research in plant sciences.
It was the first time I used PCR to clone a stress-related gene (AtFC1) and fluorescence microscopy to see root stains. Despite the fact that the experiments were not a huge success, I realized that failure in research is an opportunity to learn and improve. I also got the opportunity to work with [redacted], who was working as a bioinformatician at Cambridge University on a project to study the structural and functional role of conserved water molecules in bovine Lactoperoxidase when he started his position back home in All India Institute of Medical Science. From that moment, I realized how bioinformatics research is heavily reliant on, and linked to, molecular genetics. I learned how a heme group can operate as an enzyme’s fundamental structural unit and how amino acids and water molecules combine to build a stable structure for an enzyme. This opportunity taught me to be patience, research motivated, and an eagerness to learn about the broad branches of science.
Later, I presented the study at a national conference, where I was able to hone my presenting abilities and network with a variety of scholars. The undergraduate courses and several labs enabled me to master a plethora of techniques such as transformation, PCR, cloning, western blot, HPLC, growth media, and buffer preparations, protein purification, etc. In addition, a project on phytochemical analysis in the medicinally valuable plant Trachyspermum ammi, as well as lessons in scientific writing, increased my enthusiasm for plant science research and review article writing. Following graduation, I wanted to pursue more plant-related molecular research, particularly with wet-lab techniques in well-equipped labs around the world. Thus, my motivation led me to pursue a Master’s degree in plant science at the University of Bonn, Germany.
From the very first semester, I started getting involved in both coursework and lab work. My experimental journey began with lab work on plant signal transduction, where my fellow students and I attempted to find the calmodulin-binding protein Tic32 and its calcium-induced shift. I learned techniques such as expression and purification of protein from E. coli, affinity chromatography, and SDS-PAGE. There was one particular moment in one of the lab courses that really stood out to me, and that was the study related to the change in lipid content in Arabidopsis thaliana when treated with plant nematodes. I was astounded to observe how different the plants treated with nematodes looked from the wild type. Since then, I have started to develop my passion in the field of plant-microbiome interactions.
My motivation to pursue a project in this area for master’s thesis strengthened by open conversations with colleagues over lunch breaks about plant-fungal research in partnering labs. I chose to pursue a career in arbuscular mycorrhizal symbiosis after that. I worked in the plant genetics lab of Prof. Dr. [redacted] at the Technical University of Munich, where our goal was to find out how lipids are transferred from plants to mycorrhiza. We found that two half ABCG transporters, STR/STR2- localized in the peri-arbuscular membrane and expressed in the roots during symbiosis, are essential for maintaining mycorrhizal symbiosis in Lotus japonicus. Because the closest ABCG homologs were examined to transport lipid monomers across the plant cell plasma membrane, which polymerizes to create the thick cell wall, we speculated that STR/STR2 might be involved in lipid transport. My 8 months of study will be recognized as co-authorship after the project is completed and submitted to the journal.
During the project, I employed AM phenotyping, genotyping of mutant plants, golden gate cloning, hairy-root transformation, BiFC, complementation assay, promoter-GUS assay, transactivation assay, phylogenetics, confocal and fluorescence microscopy. If AM symbiosis is to be successfully employed to improve agriculture, this research into the transport and regulatory processes underlying plant control over fungal colonization would undoubtedly be beneficial. This opportunity marked a watershed moment in my professional life. I was eager to learn, eager to put in long hours and weekends, and I developed a strong interest in a new study area. From that point onward, I started following Prof. Dr. [redacted]’s work on molecular aspects of AM symbiosis research.
Following the completion of my Master’s degree, I began working as a research trainee in Prof. [redacted]’s lab at the University of Cologne in January 2021 to investigate how a fungal effector expressed in planta can cause the death of an important plant, such as a tomato. Recently, I found that the effector secreted in planta is localized in the nucleus of a plant cell. I and colleagues discovered that the putative interactors of the fungal effector protein are transcription factors involved in auxin signaling utilizing techniques such as pull-down assay and LC-MS. I discovered that the hypothesized fungal effector interacts with Auxin Response Factors using the CO-IP and Y2H assay. We predicted that the binding would alter auxin-responsive genes regulation downstream.
Therefore, future research will focus on determining which genes are affected and how gene up- or down-regulation leads to fungal susceptibility. Once the project is completed, I’m aiming to get a co-authorship. I am taking advantage of this opportunity to learn new skills in the field of plant-fungal molecular investigations, as well as to prepare for a PhD program, work independently, and broaden my scientific network. In terms of my career, I intend to remain in academia and contribute to hunger alleviation for years, if not decades, by enhancing crop output and inspiring new generations to pursue plant molecular research through teaching. I hope to eventually manage my own lab. A Ph.D. at one of the high research-based university would ideally be the ideal next career step for me to accomplish my dream, demonstrate my intellectual capacity, and live out my enthusiasm for plant-fungal studies.
The University of Miami biology program looms largely in my mind because of its outstanding faculty and interdisciplinary projects in plant microbiome research. In my own quest of finding a suitable graduate faculty, Prof. Dr. [redacted]’s research on understanding the molecular processes underpinning plant control over AM symbiosis has piqued my attention. I’ve worked on AM symbioses before and will add the majority of the needed expertise and knowledge to the project. The lab investigates the molecular function of symbiotic peptide signals using targeted mutagenesis, transcriptomics, genetics, genomics, and microscopy methods, which is related to my research interests.
Prof. Dr. and I keep in touch on a regular basis, and we discuss research ideas that I might be able to pursue during my PhD if I am accepted. CLE53 signaling governs AM symbiosis, according to her and her colleagues, although the molecular mechanisms of CLE53 signal induction, transcriptional regulators of CLE53, and downstream signals are yet unknown. I would be extremely interested in putting my research experience, commitment, and enthusiasm to understand AM symbiotic autoregulation. Aside from the chance to learn from experts at the University of Miami, I feel that my PhD teaching experience will be an asset to my academic future.
After speaking with graduate students, I discovered that this university’s program is tailored to fulfill the demands of life science research. The exchange of ideas across the seven groups focusing on symbiosis will undoubtedly aid me in answering scientific issues and forging significant scientific linkages. I’ve learned from my prior experiences that every profession may be difficult and demands sacrifice. If given the chance, I am certain that I have the will and aptitude to thrive in this lucrative and intellectually stimulating profession. I appreciate your time and hope to receive favourable attention. I am excited to collaborate with scientific professionals at the University of Miami.
