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Agrobacterium tumefaciens-mediated transformation of drumstick (Moringa oleifera Lam.)
oleh: Junjie Zhang, Mengfei Lin, Hanbin Chen, Xiaoyang Chen
| Format: | Article |
|---|---|
| Diterbitkan: | Taylor & Francis Group 2017-11-01 |
Deskripsi
Drumstick (Moringa oleifera Lam.) is an industrially important tree species. Due to the availability of an efficient regeneration system, the present study was undertaken to develop a reliable and stable Agrobacterium tumefaciens-mediated transformation system for the genetic improvement of drumstick. A. tumefaciens strain EHA105, harbouring the plasmid pCAMBIA1301 containing the hygromycin phosphotransferase II and ß-glucuronidase genes driven by the CaMV 35S promoter, was used to establish a transformation system in drumstick. The use of vacuum infiltration-assisted Agrobacterium infection (excluding a pre-culture step) and co-cultivation for two days significantly increased the transformation frequency. Genomic integration and transgene expression were confirmed by β-glucuronidase (GUS) assays, polymerase chain reaction (PCR), Southern blot analysis and real-time quantitative PCR (RT-qPCR). This transformation protocol provides a basis for the future development of genetic engineering techniques to improve the performance of drumstick.