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Dean W. Gabriel

GM Citrus with Potential to Control Greening. Dean W. Gabriel. Plant Pathology Department University of Florida, Gainesville. and. Integrated Plant Genetics, Inc. Alachua. Strategies for greening disease control. No practical resistance breeding. Eradication impossible.

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Dean W. Gabriel

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  1. GM Citrus with Potential to Control Greening Dean W. Gabriel Plant Pathology Department University of Florida, Gainesville and Integrated Plant Genetics, Inc. Alachua

  2. Strategies for greening disease control • No practical resistance breeding. • Eradication impossible. • Chemical control of insects may require 25 sprays per year. • Genetically modified (GM) citrus (add a resistance gene).

  3. Citrus greening, likely trapped in dooryard citrus, nationwide. 6 year lag from psyllid establishment to greening diagnosis. APHIS/PPQ intercepted the vector 40 times in US ports between 1985-98.

  4. Why don't chemicals work on greening? Canker bacteria Greening bacteria

  5. How to make a "GMO" Nucleus 1 2 1. Start with DNA cloning vector. 2. Add gene of interest + plant selection gene. 3. Add cloned genes on vector to specialized bacterial pathogen. 4. Contact pathogen with target plant cell. 5. The pathogen injects the cloned genes into the plant cell, and the genes are guided to the nucleus by DNA vector.

  6.  Transformation process for GM Citrus • 1. In a test tube, construct a gene encoded on vector DNA. • 2. Add the vector DNA to a specialized bacterial pathogen. • 3. Contact wounded citrus with the pathogen; the vector • DNA is injected by the pathogen into the plant cell. --Essenberg & Richardson

  7.  Transformation process for GM Citrus 1 2 Greenhouse testing Bacteria with cloned genes applied to cut end of citrus stem section. Antibiotic selection applied. Grafting onto rootstock

  8. 4 weeks --- 26% juvenile stem pieces survive; 1/2 of these are transformed

  9. Mature Valencia & Hamlin regenerated

  10. Juvenile grapefruit after grafting

  11. May, 2008: Growth of transgenic citrus trees, each with 1 of 3 IPG DiseaseBlock genes.

  12. Current status: GM citrus trees currently being evaluated by IPG. Technology (IP) is co-owned by UF.

  13. Citrus has natural defenses Transgenes that interfere with defense are more susceptible...

  14. Citrus has natural defenses Harder to prove are transgenes that enhance defense... a negative result (determined by PCR). To date, negative. IPG DiseaseBlock® added

  15. Transgenic tobacco as proxy... to date negative IPG DiseaseBlock® added

  16. GMOs can be accepted in the U.S. Transgenic and nontransgenic papaya in Hawaii. Work by: D. Gonsalves, Cornell U.; S. Ferreira and R. Manshardt, U.Hawaii; M. Fitch, USDA; J. Slightom, Pharmacia

  17. Transgenic 'UH Rainbow' to be graded in a commercial packinghouse.

  18. The Regulatory Process---Who's minding the Store? • NIH mandated IBC review • USDA-APHIS facility review • USDA-APHIS field-release approval • USDA-APHIS shipping authority approval • USDA-APHIS permission to commercialize or non-regulatory status (public comment, 12-24 mo)

  19. The Regulatory Process • EPA experimental use permit (if > 10 acres) • EPA determines if limits needed on % gene product (public comment, 24-30 mo.) • EPA product registration (public comment) • FDA review (public comment (12-18 mo.) Earliest time to market: 2015

  20. Integrated Plant Genetics, Inc. IPG Research Sponsored by: Southern Gardens Citrus USDA-APHIS NSF Fischer USA Goldsmith Plants 12085 Research Drive Alachua, Florida 32615 USA http://www.ipgenetics.com gabriel@ipgenetics.com

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