tailieunhanh - Effects of squirting cucumber (Ecballium elaterium) fruit juice on Agrobacterium tumefaciens-mediated transformation of plants

Different concentrations of squirting cucumber (Ecballium elaterium (L.) .) fruit juice were added to Agrobacterium tumefaciens growth, leaf disc inoculation, and cocultivation media, to investigate its effect on the transformation frequency of tobacco and potato. A. tumefaciens strain GV2260 harboring p35S GUS-INT and pAoPR1-GUS-INT plasmids were used separately in the transformation experiments. | Turkish Journal of Biology Turk J Biol (2015) 39: 790-799 © TÜBİTAK doi: Research Article Effects of squirting cucumber (Ecballium elaterium) fruit juice on Agrobacterium tumefaciens-mediated transformation of plants 1 2, 2 3 Sancar Fatih ÖZCAN , Mustafa YILDIZ *, Hussein Abdullah Ahmed AHMED , Muhammad AASIM 1 Central Research Institute for Field Crops, Ministry of Food, Agriculture, and Livestock, Ankara, Turkey 2 Department of Field Crops, Faculty of Agriculture, Ankara University, Ankara, Turkey 3 Department of Biology, Kamil Özdağ Faculty of Science, Karamanoğlu Mehmetbey University, Karaman, Turkey Received: Accepted/Published Online: Printed: Abstract: Different concentrations of squirting cucumber (Ecballium elaterium (L.) .) fruit juice were added to Agrobacterium tumefaciens growth, leaf disc inoculation, and cocultivation media, to investigate its effect on the transformation frequency of tobacco and potato. A. tumefaciens strain GV2260 harboring p35S GUS-INT and pAoPR1-GUS-INT plasmids were used separately in the transformation experiments. Neomycin phosphotransferase (NPT-II) gene was used as a plant selectable marker at a concentration of 100 mg L–1. The addition of 5–10 mg L–1 squirting cucumber fruit juice to bacterial nutrient medium increased A. tumefaciens growth significantly by 6 h. Moreover, the use of high concentrations (–20 mL L–1) of fruit juice resulted in excessive bacterial growth on cocultivation and selection media around the explants, which was difficult to eliminate by subculture or higher levels of antibiotics. On the other hand, lower concentrations (– mL L–1) of squirting cucumber fruit juice significantly increased the transformation frequency in both tobacco and potato. Kanamycin-resistant tobacco shoots, rooted in a medium containing 100 mg L–1 kanamycin, were transferred to pots containing organic soil and .