Gene Gap Investigation of Chalcone Synthase In California Tarweeds and the Hawaiian Silverswords

Complex organisms such as plants have many different metabolic pathways that influence substrate specific reactions. Changes in these lead to phenotypic differences related to speciation events. The anthocyanin biosynthetic pathway (ABP), which is a part of the Phenylpropanoid pathway, controls many internal and external mechanisms that regulate temperature, UV protection, microbial invasion, fruit, seed and flower coloring. Anthocyanin production is important to attract pollinators for the proliferation and evolution of the species. In the ABP, Chalcone synthase (CHS) is an enzyme that is responsible for catalyzing the first condensation reaction and essential for other genes further down the pathway such as, dihydroflavonol-4-reductase (DFR), which is a middle gene, and anthocyanidin synthase (ANS), which is a late gene in the pathway. The Hawaiian Silverswords are of great interest due to the fact that the species under went adaptive radiation and have large phenotypic differences but little genotypic differences from their ancestor, the California Tarweeds. In previous studies copies of CHS in Dubautia scabra, Dubautia linearis, and Wilkesia gymnoxiphium in the Hawaiian Silverswords as well as Madia elegans, Madia gracilis, and Holocarpha macradenia, which are apart of the California tarweeds, have been partially sequenced and analyzed. The gaps that remain are the focus of this study looking at sequence included in exon 1 and 2 of the CHS gene for Wilkasea gymnoxiphium, and Dubautia linearis. The current strategy will be to use polymerase chain reaction, cloning, sequencing, and further analysis of genotypic differences will be done using bioinformatics.