Study on transformation of cysteine proteinase inhibitor gene into cabbage (Brassica oleracea var. capitata L.).

摘要

The plant expression vector pBI121-OCI was constructed, with OC-I gene under the control of CaMV35S promoter and Nos terminator. LBA4404 strain containing this plasmid was used in genetic transformation of cabbage. After 3 days preculture, the explants were inoculated with Agrobacterium and co-cultivated for 4 days at 25 degrees C. When a slight development of the bacteria was visible on the wound of the explants, they were then transferred to regeneration medium supplied with 500 mg carbanicilin/litre, and cultured for 7 days to kill the Agrobacterium. To select for transferred cells, explants were transferred to the same medium supplemented with 5 mg kanamycin/litre for 6-8 weeks. During the selection, the explants were transferred to fresh medium every 2 weeks. The putative transformants were assayed by PCR and Southern blot analysis. The result showed that OC-I gene was transferred into cabbage successfully. The resistance of transgenic plants to insects was stronger compared to the control in the field..