Tawni Middleton

Tawni Middleton
Year: 2015
School: Colorado Mesa University

Factors that affect Agrobacterium tumefaciens-mediated transformation of Solanum prinophyllum

Project Summary

The human population is predicted to reach 9.6 billion and the food demand is predicted to increase 70 to 100 percent by 2050. Arable land is being lost to urbanization, water supply is slowly dissipating, and the climate is gradually changing. Genetic engineering and crop improvement will play a crucial role to meet the demands and alleviate crop damage from both biotic and abiotic stresses. One major plant family is Solanaceae, which contains approximately 2,700 species including many important food crops.  Solanum prinophyllum, a member of the Solanaceae, was the focus of this study.  The purpose of the experiment was to investigate factors that can affect the transformation of S. prinophyllum.  Factors included different Agrobacterium tumefaciens strains (AGL1 and LBA4404), two explant types (cotyledons and mature leaves) and selective plant regeneration medium that contained two different concentrations of the antibiotic kanamycin (75 mg/l and 100 mg/l). Tissue culture and biotechnology techniques were used throughout this study. To confirm the recovery of transgenic lines, a β-Glucuronidase (GUS) assay and Polymerase Chain Reaction (PCR) will be used. The ultimate goal of this study was to develop the first successful transformation methodology for S. prinophyllum. S. prinophyllum can then be used as a model plant system for a larger NSF funded project in the Van Eck lab to investigate the genes and networks involved in meristem maturation and transition from vegetative growth to reproductive growth in the Solanaceae.

My Experience

Being able to participate in the Plant Genome Research Project at the Boyce Thompson Institute for Plant Research has been the opportunity of a lifetime. Working with my mentors Cynthia Du and Sarika Gupta provided me with insight and hands on experience in the field I dream of working in. My advisor, Joyce Van Eck, was very welcoming and she taught me so much. I learned many new lab skills and techniques such as plant tissue culture. This internship has given me confidence for stepping into the graduate school world. I now know what I need to do to prepare for the next step in my career path. I had an amazing time this summer in Ithaca, meeting other summer interns and lab members. Thank you for this amazing opportunity BTI!