Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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A novel method for high-frequency transgenic shoot regeneration via Agrobacterium tumefaciens in flax (Linum usitatissimum L.)

  • Beyaz, Ramazan (Department of Soil Science and Plant Nutrition, Faculty of Agriculture, Ahi Evran University) ;
  • Darcin, E. Selcen (Department of Field Crops, Faculty of Agriculture and Natural Sciences, Bilecik Seyh Edebali University) ;
  • Aycan, Murat (Department of Field Crops, Graduate School of Natural and Applied Sciences, Ankara University) ;
  • Kayan, Mustafa (Department of Field Crops, Graduate School of Natural and Applied Sciences, Ankara University) ;
  • Yildiz, Mustafa (Department of Field Crops, Faculty of Agriculture, Ankara University)
  • Received : 2016.04.24
  • Accepted : 2016.04.25
  • Published : 2016.06.30
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Abstract

In this study, routinely used transformation method, which includes transferring explants onto co-cultivation medium after inoculating them with bacterial solution for a while, was compared with 3 different inoculation methods. In every 3 methods, hypocotyl explants excised from 7-day-old sterile flax seedlings having cotyledon leaves and no root system dried under air flow in sterile cabin for 35 min were inoculated with different volumes of bacterial solution at different inoculation periods. GV2260 line of Agrobacterium tumefaciens having 'pBIN 19' plasmid containing npt II (neomycin phosphotransferase II) gene and GUS reporter gene was used in transformation studies. After inoculation, hypocotyl segments of seedlings (0.5 cm in length) - were excised and left to co-cultivation for 2 days. Then, explants were transferred to regeneration medium supplemented with different antibiotics. The presence of npt-II and GUS genes in transformants was confirmed by PCR and GUS analysis. The highest results in all characters examined in all cultivars were obtained from the 2 inoculation method in which hypocotyls excised from seedlings inoculated with $500{\mu}l$ of bacterial solution after drying in sterile cabin for 35 min were used.

Keywords

References

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