Environmental impact on the insulin signaling pathway during overwintering: Part two ;ate season diapausers

The production of alfalfa seed is significantly enhanced by bee pollination; within the United States and Canada the primary alfalfa pollinator is the alfalfa leafcutting bee (ALCB), Megachile rotundata. However, ALCB populations are not sustainable in the United States, indicating that the current management practices are not conducive for maintaining a stable commercial population of these bees. Since the ALCB spends 9 to 10 months a year in storage during management practices, optimizing storage parameters for diapausing bees may help to improve overall ALCB survival. To ensure high quality bees, storage conditions may need to be adjusted as the physiology of bees changes over the course of diapause development during winter storage. Therefore, to better understand how diapause physiology progresses during storage we screened key diapause-regulatory genes in early and late season diapausing ALCB stored either under current management conditions or field conditions. This second part focuses on only late season diapausing ALCB. Classic techniques such as a qPCR and RNA-seq have limitations for studying gene expression in non-model organisms such as ALCB. We used a novel approach for studying insect genetics by using nCounter technology, a massive parallel hybridization technique, to conduct this screening. Expression of members of the cyclin family, and the insulin pathway regulator FOXO We found that nCounter technique was easier to carry out and analyze than either qPCR or RNA-seq, and gave results equivalent to our previous work.