Workshop: Fundamentals of Agrobacterium and its applications in plant biotechnology research
July 27th, 2023
9:00 AM CDT/10:00 AM EDT/3:00 PM BST
ABOUT THE WORKSHOP
Agrobacterium is a genus of soilborne Gram-negative bacteria that have the natural ability to transfer some of its own DNA to host genomes. Despite over 40 years of research, only a limited number of Agrobacterium strains are utilized for the generation of transgenic organisms. In the past, major problems associated with Agrobacterium- mediated transformation include improving the efficiency and quality of the transformation event, broadening the host range, and preventing recombination and rearrangement of plasmids introduced into Agrobacterium. The aim of this workshop is to understand the fundamentals of Agrobacterium handling and manipulation for plant transformation, share best practices for handling Agrobacterium, and create a list of potential webinar series topics to learn more about various aspects of Agrobacterium.
The live workshop will be 2 hours followed by a 30+ minute roundtable discussion, which is an opportunity to meet fellow PlantGENE members and discuss the workshop topics. The workshop will be recorded and posted here after the event.
Certificates of participation can be obtained here.
WATCH THE VIDEOS
Download Stan Gelvin's presentation slides:
Download Erh-Min Lai’s presentation slides: Related work:
DISCUSSION FROM THE WORKSHOP
These articles and questions/answers are from the Zoom chat. Thank you to the many participants who contributed!
Articles and helpful links |
|---|
Co–Transformation of Unlinked Foreign Genes into Plants by Direct Gene Transfer |
ASPB: Teaching Tools in Plant Biology Free Plantae account required for login. Teaching Tools in Plant Biology, published by the American Society of Plant Biologists, combines up-to-date peer-reviewed research-based content with flexible presentation components that can be used alone or integrated into your lesson plans so that you can confidently present these exciting topics in your classroom. |
“Agrolistic” transformation of plant cells: Integration of T-strands generated in planta |
Funding Opportunities |
DOE Proposal: DELIVERY TECHNOLOGIES FOR GENETIC ENGINEERING BIOENERGY CROPS (see page 52) |
Questions and answers from the workshop |
|---|
Q: Are there strains that are better suited to avoid recombination of plasmids when transformed into Agrobacterium? A: Yes, strain AGL1 is a recA- mutant, which is meant to improve construct stability |
Q: Are there any patents for strains C58C1(pGV2260)H and EHA105 (for commercial use)? A: EHA105 is not patented and is free to use by anyone. |
Q: If most of the disarmed strains have C58 chromosomal background, do you think the disarmed strains we have are enough to solve the bottlenecks of Agrobacterium-mediated plant transformation? If not, what direction should we follow? A: Great question, I guess the answer is more research is required and probably a webinar on diversity in Agro will help understand current knowledge around different Agro strains. Only a handful of Agro are used across transformation labs: is it due to the convenience, availability of strains? A: I suspect is it the difficulty of disarming Ti oncogenes and further "domesticating" wild-type strains. And then convenience of using these few original lab strains for transformation of new plant genotypes led to their use everywhere and where we stand today not utilizing the full genetic diversity of the Agrobacterium genus. |
Q: Does anyone have experience in comparing C58-based strains with LBA4404? I guess there are quite some differences e.g. I recall the presence of a cytokinin synthase in the chromosome of LBA. Callus induction is usually depending on Auxin/Cytokinins, so there should be difference, I guess. A: It depends on the application how the differences manifest, but they are for sure present. My work with LBA4404 and AGL1 in teff callus infection showed that AGL1 was too aggressively virulent and damaged the fragile tissue, whereas LBA4404 was less aggressive but also was less capable of inducing transformation. I think it varies a lot between different plant systems at the end of the day though. A: There is a lot of evidence in the literature that the transformation efficiency of different strains varies for different target species and genotypes. |
ABOUT OUR WORKSHOP SPEAKERS
WORKSHOP ORGANIZERS
Keunsub Lee
Iowa State University
PlantGENE Steering Committee Member