Cry3A transgenic, bark beetle resistant spruce
description
Transcript of Cry3A transgenic, bark beetle resistant spruce
Cry3A transgenic, bark Cry3A transgenic, bark beetle resistant sprucebeetle resistant spruce
NAZV project NAZV project QH71290QH712902007-20112007-2011
Spruce bark beetle (Ips typographus)
Bark beetle galleries
Bacillus thuringiensiscrystal protein (Cry, δ-endoTOXin)
Agrobacterium interaction with the plant cell
• Spruce is difficult to transformSpruce is difficult to transform
• No known Bt toxin is toxic enough to bark No known Bt toxin is toxic enough to bark beetlebeetle
Major problems:Major problems:
Spruce callus cultures
Somatic embryogenesis
induction conversion cultivation
Modified Cry3A genes
Cry3A orig
Synt TOX-1 (602 aa)-codons Picea optimized
Ndel/sTOX-1
Cdel/sTOX-1 (602 aa)
Cdel/Ndel/sTOX-1 GUS
Refined modification of domain II
Expression vector
PAGE analysis of modified Cry3A expression
WESTERN detection of modified Cry3A proteins
Toxicity tests on live beetles
K o n e c p r e s e n t a c e