“Exploring the Characteristics of Plant Nuclei”
While the structure of the nucleus is well-studied, we are interested in investigating the genomic characteristics that govern nuclear architecture and physical properties. This summer, we developed a protocol to prepare intact nuclei from tomato plants, and we also studied the changes in nuclear phenotypes caused by mutations in the tomato NMCP2 gene, which encodes a nuclear lamina protein.
The nucleus is an integral part of eukaryotic cells, as it contains, protects and organizes the genome. While the structure of the nucleus is well-studied, the genomic characteristics that govern the nuclear architecture and physical properties remain unclear. This summer, we developed a nuclear isolation method for future studies in nuclear biomechanics to probe how genome parameters affect nuclear properties. We developed a protocol to prepare intact nuclei from Solanum lycopersicum (tomato) plants on microscope slides and petri dishes with minimal debris. This procedure will facilitate further investigations on the mechanical properties of plant nuclei, and we can use this procedure to determine whether plant nuclei are pliable or rigid. We also studied the changes in nuclear phenotypes caused by mutations in the tomato NMCP2 gene, which encodes a nuclear lamina protein. In particular, we found that individuals homozygous for the nmcp2-3 hypomorphic allele had nuclei that were smaller and rounder than in wild-type individuals. Furthermore, we found some evidence for nuclear positional differences between wild-type individuals and nmcp2-3 mutants. In nmcp2-3 mutants, the nuclei in the two guard cells of each stomata tended to form a more drastic angle with the length of the stomata. This larger offset angle could be correlated to the smaller and rounder shape of the nuclei, and/or abnormal interactions between the nucleus and the cytoskeleton. Our discovery of a hypomorphic but viable nmcp2-3 allele (nmcp2 null alleles are lethal) will allow us to include study of the NMCP2 gene in our future investigation of nuclear biomechanics.
As an intern at BTI this summer, I gained experience in lab techniques including PCR and fluorescence microscopy. I also learned about general research practices, such as maintaining a good lab notebook and designing experiments. Going into the program, I had no previous research experience and was quite nervous about being in a lab environment, and I often worried that I was not cut out for biology research. However, with the support of my mentor and fellow interns, I quickly gained confidence and became comfortable with asking questions whenever I needed guidance. In addition, my graduate school mentors provided me with valuable advice on how to prepare for a career in scientific research. This program helped me hone the skills I have learned in previous biology coursework and apply my knowledge to real-life experiments, all while advancing my technical knowledge and allowing me to further explore different career options.