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- Title
Establishment of an efficient Agrobacterium tumefaciens -mediated leaf disc transformation of Thellungiella halophila.
- Authors
Hong-Qing Li; Jie Xu; Lei Chen; Mei-Ru Li
- Abstract
Abstract  Thellungiella halophila is a salt-tolerant close relative of Arabidopsis, which is adopted as a halophytic model for stress tolerance research. We established an Agrobacterium tumefaciens-mediated transformation procedure for T. halophila. Leaf explants of T. halophila were incubated with A. tumefaciens strain EHA105 containing a binary vector pCAMBIA1301 with the hpt gene as a selectable marker for hygromycin resistance and an intron-containing β-glucuronidase gene as a reporter gene. Following co-cultivation, leaf explants were cultured on selective medium containing 10 mg lâ1 hygromycin and 500 mg lâ1 cefotaxime. Hygromycin-resistant calluses were induced from the leaf explants after 3 weeks. Shoot regeneration was achieved after transferring the calluses onto fresh medium of the same composition. Finally, the shoots were rooted on half strength MS basal medium supplemented with 10 mg lâ1 hygromycin. Incorporation and expression of the transgenes were confirmed by PCR, Southern blot analysis and GUS histochemical assay. Using this protocol, transgenic T. halophila plants can be obtained in approximately 2 months with a high transformation frequency of 26%.
- Subjects
AGROBACTERIUM; ARABIDOPSIS; TRANSGENES; HALOPHILA
- Publication
Plant Cell Reports, 2007, Vol 26, Issue 10, p1785
- ISSN
0721-7714
- Publication type
Article
- DOI
10.1007/s00299-007-0391-y