Knocking Out O-methyltransferase Targets on Chromosome 9
By researching maize, scientists have been able to manipulate plant genes to fight diseases and repel insects. In order to discover more about the corn leaf aphid resistance pathway involving benzoxazinoid biosynthesis, I tried to isolate mutant knockout alleles of the o-methyltransferase paralogs on Chromosome 9 using the transposable element Dissociation (Ds). This would help to determine the importance of each of these genes in insect resistance. Ac and Ds are transposable elements that move during DNA replication. If Ds inserts into a gene such as the o-methyltransferases, their expression can be disrupted and more can be learned about them. To carry out this project, I planted nearly one hundred plants from a genetically closely linked Ds testcross population and sampled tissue from each seedling in pools from which DNA was extracted. Then through multiple PCR reactions surveying the o-methyltranferase target genes and four others in the region, I was able to amplify the genes and search for Ds insertions. With the results of this experiment, more can be known about the genetic makeup of maize. We are one step closer to identifying the role that these genes play in benzoxazinoid biosynthesis.
Through this internship, I learned a lot about plant genetics and molecular biology and I acquired many new skills and techniques. I learned how to do technical lab work, such as DNA extractions and PCR reactions. My co-workers in the lab were absolutely amazing to work with and couldn’t have been more supportive. My mentor Kevin Ahern, gave me a significant amount of background information before I started my project, and helped me with any question I had along the way. I am now confident in successfully completing a DNA extraction as well as running PCR reactions. Because of this internship, I now know that I definitely want to major in biology, and pursue the subject as a future career.