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| Content Provider | Springer Nature Link |
|---|---|
| Author | Celikkol Akcay, Ufuk Mahmoudian, M. Kamci, H. Yucel, M. Oktem, H. A. |
| Copyright Year | 2008 |
| Abstract | A simple and reproducible Agrobacterium-mediated transformation protocol for a recalcitrant legume plant, lentil (Lens culinaris M.) is reported. Application of wounding treatments and efficiencies of three Agrobacterium tumefaciens strains, EHA105, C58C1, and KYRT1 were compared for T-DNA delivery into lentil cotyledonary node tissues. KYRT1 was found to be on average 2.8-fold more efficient than both EHA105 and C58C1 for producing transient β-glucuronidase (GUS) gene (gus) expression on cotyledonary petioles. Wounding of the explants, use of an optimized transformation protocol with the application of acetosyringone and vacuum infiltration treatments in addition to the application of a gradually intensifying selection regime played significant roles in enhancing transformation frequency. Lentil explants were transformed by inoculation with Agrobacterium tumefaciens strain, KYRT1 harboring a binary vector pTJK136 that carried neomycin phosphotransferase gene (npt-II) and an intron containing gusA gene on its T-DNA region. GUS-positive shoots were micrografted on lentil rootstocks. Transgenic lentil plants were produced with an overall transformation frequency of 2.3%. The presence of the transgene in the lentil genome was confirmed by GUS assay, PCR, RT-PCR and Southern hybridization. The transgenic shoots grafted on rootstocks were successfully transferred to soil and grown to maturity in the greenhouse. GUS activity was detected in vegetative and reproductive organs of T0, T1, T2 and T3 plants. PCR assays of T1, T2 and T3 progenies confirmed the stable transmission of the transgene to the next generations. |
| Starting Page | 407 |
| Ending Page | 417 |
| Page Count | 11 |
| File Format | |
| ISSN | 07217714 |
| Journal | Plant Cell Reports |
| Volume Number | 28 |
| Issue Number | 3 |
| e-ISSN | 1432203X |
| Language | English |
| Publisher | Springer-Verlag |
| Publisher Date | 2008-12-16 |
| Publisher Place | Berlin, Heidelberg |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Agrobacterium tumefaciens Genetic transformation Micrograft Lentil Lens culinaris Transgenic plants Plant Biochemistry Biotechnology Plant Sciences Cell Biology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Plant Science Medicine Agronomy and Crop Science |
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