biologia plantarum

International journal on Plant Life established by Bohumil Nìmec in 1959

Biologia plantarum 49:53-57, 2005 | DOI: 10.1007/s10535-005-3057-z

Influence of different co-cultivation temperatures, periods and media on Agrobacterium tumefaciens-mediated gene transfer

S. Uranbey1, C. S. Sevimay2, M. D. Kaya2, A. İpek2, C. Sancak2, D. Başalma2, C. Er2, S. Özcan2,*
1 Central Field Crops Research Institute, General Directorate of Agriculture Research, Ministry of Agriculture and Rural Affairs, Ankara, Turkey
2 Department of Field Crops, Faculty of Agriculture, University of Ankara, Ankara, Turkey

Tobacco leaf disc explants were inoculated with Agrobacterum tumefaciens strain GV2260 carrying p35S GUS-INT to determine the influence of different co-cultivation temperatures (18 - 26 °C), periods (24 - 96 h) and media (solid and liquid) on transformation efficiency. Kanamycin-resistant shoots developed on leaf discs inoculated with Agrobacterium after 4 weeks of culture initiation. Regenerated shoots were excised and rooted in the basal medium supplemented with 100 mg dm -3 kanamycin and 250 mg dm -3 augmentin. The rooted plantlets were finally transferred to compost and confirmed by GUS assay and PCR analysis. The highest transformation frequency was achieved from the explants co-cultivated with A. tumefaciens in liquid medium for 48 h at 22 or 24 °C.

Keywords: GUS activity; leaf disc; PCR; shoot regeneration; tobacco; transformation
Subjects: Agrobacterium tumefaciens; auxins; cytokinins; gene transfer; β-glucuronidase; Nicotiana tabacum; nutrient medium MS (Murashige and Skoog); polymerase chain reaction (PCR); tobacco, transgenic plants; transgenic plants

Received: October 6, 2003; Accepted: May 5, 2004; Published: March 1, 2005  Show citation

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Uranbey, S., Sevimay, C.S., Kaya, M.D., İpek, A., Sancak, C., Başalma, D., Er, C., & Özcan, S. (2005). Influence of different co-cultivation temperatures, periods and media on Agrobacterium tumefaciens-mediated gene transfer. Biologia plantarum, 49(1), 53-57. doi: 10.1007/s10535-005-3057-z
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