ISHS Acta Horticulturae 706: IV International Symposium on Brassicas and XIV Crucifer Genetics Workshop
STUDY ON TRANSFORMATION OF CYSTEINE PROTEINASE INHIBITOR GENE INTO CABBAGE (BRASSICA OLERACEA VAR. CAPITATA L.) |
| Authors: | J.J. Lei, W.J. Yang, S.H. Yuan, F.Y. Ying, L.C. Qiong |
| Keywords: | Brassica oleracea var. capitata, cysteine proteinase inhibitor gene, transformation, Agrobacterium tumefaciens |
| Abstract:
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 3d preculture, the explants were inoculated with Agrobacterium and co-cultivated for 4d at 25°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•l-1 carbanicilin, 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•l-1 kanamycin 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. | |
Download Adobe Acrobat Reader (free software to read PDF files) | |
