Agrobacterium-mediated transformation of cabbage (Brassica oleracea cv capitata L.) with soybean 13-1,3-endoglucanase cDNA
| Main Author: | Perpustakaan UGM, i-lib |
|---|---|
| Format: | Article NonPeerReviewed |
| Terbitan: |
[Yogyakarta] : Universitas Gadjah Mada
, 2003
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| Subjects: | |
| Online Access: |
https://repository.ugm.ac.id/23562/ http://i-lib.ugm.ac.id/jurnal/download.php?dataId=6513 |
Daftar Isi:
- Transgenic cabbage (Brassica oleracea cv. capitata L.) has been produced efficiently by inoculating different explants with Agrobacterium tumefaciens strain LBA4404 harbouring a binary vector pROKIa-EG containing a soybean 13- 1,3-endoglucanase cDNA and npfll (neomycin phosphotransferase) genes. Genetic transformation eficiency was found to be high on cotyledonary node explants followed by 7 days co-cultivation on MS medium supplemented with 2 mg/1 NAA and 4 mg/1 BAP and 2 days pre-cultured before co-cultivation with Agrobacterium. A concentration of 50 mg/1 kanamycin was added to regeneration medium in order to select the transformed shoot. The average percentage of kanamycin resistant of shoot regenerant after Agrobacterium infection and selection was 1.03% for hypocotyls explant and 10.13% for cotyledonary node explant. As a positive control the same transformation was observed on tobacco (Nicotiana tabacum cv. Besuki H283) leaf disc with 7 days co-cultivation on MS medium supplemented with 1 mg/1NAA and 3 mg/1 BAP and the selection medium containing 100 mg1/1 kanamycin. The average kanamycin resistant shoot regenerant was 27.36%. The presence of 0-1,3-endoglucanase gene in the putative transgenic lines was confirmed by PCR analysis, while the insertion of the 13-1,3-endogIucanase in the nuclear genome of transgenic plants was verified by dot blot hybridization analysis. Keywords: Agrobacterium � transformation �Brassica oleracea cv.'capitata L.-13-1,3-endoglucanase