Effects of squirting cucumber (Ecballium elaterium) fruit juice onAgrobacterium tumefaciens-mediated transformation of plants

Authors: SANCAR FATİH ÖZCAN, MUSTAFA YILDIZ, HUSSEIN ABDULLAH AHMED AHMED, MUHAMMAD AASIM

Abstract: Abstract: Different concentrations of squirting cucumber (Ecballium elaterium (L.) A.Rich.) 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 (2.5-20 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 (0.2-1.6 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 perlite in growth cabinets for acclimatization. Transgenic plants grew normally and set seeds. The presence of T-DNA in these transformants was confirmed by PCR and GUS analysis.

Keywords: Genetic transformation, squirting cucumber, phenolics, potato, tobacco

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