A novel method for high-frequency transgenic shoot regeneration via Agrobacterium tumefaciens in flax (Linum usitatissimum L.)


Creative Commons License

Beyaz R., Selcen Darcin E., Aycan M., Kayan M., YILDIZ M.

Journal of Plant Biotechnology, cilt.43, sa.2, ss.240-247, 2016 (Scopus) identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 43 Sayı: 2
  • Basım Tarihi: 2016
  • Doi Numarası: 10.5010/jpb.2016.43.2.240
  • Dergi Adı: Journal of Plant Biotechnology
  • Derginin Tarandığı İndeksler: Scopus
  • Sayfa Sayıları: ss.240-247
  • Anahtar Kelimeler: Agrobacterium tumefaciens, Flax, Gene transfer, Inoculation method, Transgenic
  • Ankara Üniversitesi Adresli: Evet

Özet

© Korean Society for Plant Biotechnology.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 μl of bacterial solution after drying in sterile cabin for 35 min were used.