Research Assistant University of Idaho Benton City, Washington
Body of Abstract: Globadera pallida(pale cyst nematode) is an international threat to the potato industry, leading to losses up to 80% in solanaceous crops. Trap crops, such as Solanum sisymbriifolium, offer an effective control for G. pallida. Being a member of the family Solanaceae, S. sisymbriifolium produces hatching stimulants recognized by G. pallida, but resists infestation, unlike potato and tomato. To combat G. pallida, it is important to understand how and with which genes the S. sisymbriifolium resistance pathways function. The goal of this research is to identify, isolate, and characterize the regulatory regions for eight S. sisymbriifoliumgenes activated in the presence of G. pallida. These genes included an oxidative burst gene, a cell wall-associated kinase, a putative protease inhibitor, four receptor-like protein kinases, and a homologue to a tomato verticillium wilt disease resistance protein.Using the Amplicon Express Matrix Technology Seven Plate Superpool system, these genes were located within an S. sisymbriifoliumbacterial artificial chromosome library containing 96,768 clones. One to three bacterial artificial chromosomes were sequenced per gene. Vector DNA was removed and the open reading frame was aligned within the genomic sequence. Next, the 3kb region upstream was selected. This region is expected to contain the 5’ untranslated region of each gene, the promoter, and regulatory elements that can be used to study how these genes are induced by the presence of G. pallida. Characterization will be performed by transferring these 3 kb fragments into pCAMBIA 1391z, a GUS-encoding binary vector. When transformed into potato, GUS expression will indicate when regulatory regions are activated in the presence or absence of G. pallida.