|CAMBRON, LIZZETTE - North Dakota State University|
|GREENLEE, KENDRA - North Dakota State University|
Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 5/24/2019
Publication Date: 11/17/2019
Citation: Cambron, L.D., Yocum, G.D., Greenlee, K.J. 2019. New techniques for answering old questions: Measuring gene expression in diapausing Megachile rotundata [abstract]. Entomological Society of America Annual Meeting. Nov. 17-20, 2019. St. Louis, MO. Poster No. D3072.
Technical Abstract: The alfalfa leafcutting bee, Megachile rotundata, undergoes diapause to survive the winter. Diapause, a non-feeding state, limits energetic resources for metabolism. Insulin signaling is involved with regulation of cell growth and overall metabolism, making it an obvious target for diapause regulation. We hypothesize that the insulin signaling pathway regulates metabolism and development during overwintering. To test this, we need to measure gene expression. The gold standard for measuring gene expression has been quantitative real-time polymerase chain reaction (qRT-PCR). However, qRT-PCR has disadvantages. One drawback is the requirement for having known control genes which can be difficult for non-model organisms. It also limits how many genes can be tested at once. An emerging multiplex technique, NanoString™, works by hybridization and does not require amplification or large quantities of RNA. To test our hypothesis and compare these three techniques, we measured expression of genes in the insulin pathway in either lab- or field-reared M. rotundata during overwintering. First we conducted an Illumina study to get a global view of overall gene expression. Then we performed qRT-PCR on genes in the insulin pathway. Lastly, for NanoString™, we created a custom probe set for genes in the insulin pathway and used the same samples as in the other techniques. If NanoString™ confirms the results of both the Illumina and qRT-PCR studies, the benefits of this emerging technology will allow researchers to more quickly answer questions about how whole pathways change during overwintering and across development, to move the field of insect physiology forward.