1. Cry3A transgenic, bark beetle resistant spruce NAZV project QH71290 2007-2011

2. Spruce bark beetle (Ips typographus)

3. Bark beetle galleries

6. Bacillus thuringiensis crystal protein (Cry, δ-endoTOXin)

7. Agrobacterium interaction with the plant cell

8. Spruce is difficult to transformSpruce is difficult to transform No known Bt toxin is toxic enough to bark beetleNo known Bt toxin is toxic enough to bark beetle Major problems:

9. Spruce callus cultures

10. Somatic embryogenesis induction conversion cultivation

11. Modified Cry3A genes Cry3A orig Synt TOX-1 (602 aa)-codons Picea optimized Ndel/sTOX-1 Cdel/sTOX-1 (602 aa) Cdel/Ndel/sTOX-1GUS

12. Refined modification of domain II

13. Expression vector

14. PAGE analysis of modified Cry3A expression

15. WESTERN detection of modified Cry3A proteins

16. Toxicity tests on live beetles

17. K o n e c p r e s e n t a c e