Transgenic Canola Plants Expressing cry1Ia5 Gene Are Resistant to (Agrotis Ipsilon) Using Sonication Assisted Agrobacterium Mediated Transformation (SAAT) Technique

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Transgenic Canola Plants Expressing cry1Ia5 Gene Are Resistant to (Agrotis Ipsilon) Using Sonication Assisted Agrobacterium Mediated Transformation (SAAT) Technique

The aim of this work was to transfer the cry1Ia5 gene into the genome of two commercial canola cultivars using Sonication assisted Agrobacterium mediated transformation system and to test its toxicity against A. ipsilon. The integration and expression of the cry1Ia5 gene into the plants genome was confirmed using PCR, Southern blot and Northern blot analyses. The insecticidal activity of transgenic plants against A. ipsilon was studied. The 2nd and 4th A. ipsilon larval instars were fed on leaves from five transgenic lines (L1, L2, L3, L4 and L5) for one week of T1 plants. Transgenic L5 was found to be more toxic to the 2nd larval instar causing 60% mortality on the 3rd day followed by 100% on the 4th day post feeding. While the other transgenic lines L2, L3, L4 caused 60, 20, 80% mortality respectively on the 6th day, whereas L1 did not show any observed toxicity to A. ipsilon. Digestive and detoxification enzyme activity were examined in the body of 2nd instar of A. ipsilon, fed on bt canola, showing significantly lower trypsin, protease, lipase, esterase and acetylcholinesterase activities and significantly increase Glutathione-S-transferase and Chitinase activities than non-bt fed larvae. Whilst the 4th larval instar fed on bt canola showed cannibalism as change in the behavior. The results of the study imply that transgenic canola lines obtained varying in toxicity against A. ipsilon which could be used to construct new canola cultivars resistant to A. ipsilon.