Wax On: Amplification of Waxy Cuticle Layer Genes CER1 and SHINE3 in \u3cem\u3eArceuthobium americanum\u3c/em\u3e

Arceuthobium americanum (lodgepole pine dwarf mistletoe) is a parasitic flowering plant that infects coniferous trees in Canadian forests along the Pacific Northwest Coast. Through the invasion of the host’s vascular tissue, dwarf mistletoes significantly reduce timber value by stunting the trees growth. A unique feature of dwarf mistletoe is its seed dispersal mechanism, explosive discharge. It is believed that the explosive discharge is made possible using thermogenesis and hydrostatic pressure to fire the seed onto a nearby host. This study was designed to examine genes involved in the production of a waxy cuticle layer on the outside of the fruit (CER1 and SHINE3). Primers were developed, using results from an Affymetrix microarray and gene libraries of other plants. Polymerase chain reaction (PCR) was used to amplify the genes of interest, but due to A. americanum being a particularly recalcitrant organism, successful amplification has been difficult. As such, preliminary results and troubleshooting methods will be discussed at length. Once successfully amplified, the genes will be cloned into a bacterial plasmid for sequencing. By sequencing these two genes we can better understand the fruit’s development and what roles these genes play in the explosive seed discharge. The productions of these waxes may play a critical role in the discharge of the seed by developing a waxy cuticle preventing water loss during fruit maturation