Professor Northeastern University Boston, Massachusetts
Body of Abstract: Despite successes in genome editing of model organisms and major crops, tools to study gene function in non-model plant species lags, limiting our understanding of the wider biodiversity of gene function. Using the medicinal plant Catharanthus roseus as a case study, I will share what we have learned during our extensive optimization and validation of the EASI (Efficient Agro-mediated Seedling Infiltration) method. The EASI method utilizes vacuum infiltration to introduce Agrobacterium tumefaciens into seedling tissue. A. tumefaciens can introduce a variety of plasmids designed to overexpress or silence genes of interest in vivo. Following transient transformation, many downstream analyses are possible, such as transcriptomics, proteomics, or metabolomics. This allows researchers to probe gene function in a native environment without the need for constructing a transgenic plant. The success of our EASI method in C. roseus highlights the possibility and utility of in vivo transformations of non-model plant species.
In this talk, I will walk through the EASI method in detail, showing data at each step to highlight parameters that are critical for success, such as the developmental state of the seedling, or a pre-infiltration dark incubation, and areas that are less important and thus allow flexibility, like the A. tumefaciens growth stage. I will then give a brief example of how we apply this method to studying regulation of specialized metabolism in C. roseus. This plant is the sole source of the chemotherapy medicines, vinblastine and vincristine. The EASI method has allowed us to study the regulation of this unique biosynthetic pathway in vivo. I hope that these lessons will help you transform your favorite non-model plant species, and thus expand the capabilities of what plant biologists can study.