Discipline: Biological Sciences
Subcategory: Plant Research
Javier Carrillo-Cortez - Humboldt State University
Co-Author(s): David A. Johnson, Samford University, Birmingham, AL
Members of the genus Cuscuta are heterotrophic parasitic plants that feed on a host plant, limiting host growth and lifespan. Individual species identification of Cuscuta is difficult during the short, species-specific flowering period, and nearly impossible when not in bloom. The aim of our project was to develop a new method to identify the species of Cuscuta found in North-Central Alabama through polymerase chain reaction (PCR) and restriction fragment length polymorphisms (RFLP). We propose that PCR amplification of three genome segments (using primer sets trnL/trnF, RBC-F/RBC-R, and ITS4/ITS5) followed by Mbol1 restriction enzyme digestion will create unique-sized fragments for each species. Results indicate a positive confirmation of species with trnL/trnF and ITS4/ITS5 primer sets using the identification protocol. Our protocol allows for a relatively simple, easily replicated identification method. Simplified identification of Cuscuta species will assist future research examining seed dispersal patterns to predict and control Cuscuta invasion of agricultural networks, as well as produce an identification method that could be adapted for other organisms.Not Submitted
Funder Acknowledgement(s): National Science Foundation (NSF) Research Experience for Undergraduates (REU) for funding this project. Samford University for offering me this research opportunity.
Faculty Advisor: David A. Johnson, firstname.lastname@example.org
Role: Research of primers using Geneious application, determination of usable restriction enzyme, PCR process (sample collection, DNA extraction and preparation) and subsequent enzyme digestion were all conducted by primary author, with secondary author/mentor assistance and advisement. Mentor contacted various sources for sample specimens, trained primary author in proper technique and protocol, offering invaluable guidance.