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ที่มา:วิทยาสารเกษตรศาสตร์ สาขา วิทยาศาสตร์

หัวเรื่อง:ไม่มีชื่อไทย (ชื่ออังกฤษ : Protoplast Isolation and Culture of Dendrobium Sonia “Bom 17”)

ผู้เขียน:Imgยุพิน เคนตรี, ImgAmpaiwan Paradornuvat, Imgสุรียา ตันติวิวัฒน์, Imgดร.ศาลักษณ์ พรรณศิริ, Imgดร.นิพนธ์ ทวีชัย, ศาสตราจารย์



The optimization of Dendrobium Sonia “Bom 17” protoplast isolation and culture conditions were investigated. Protoplasts were successfully isolated from one-month-old leaves of in vitro-grown plantlets using an enzyme solution comprising 1% Cellulase Onozuka R-10, 0.2% Macerozyme, 0.3 M mannitol, 10 mM CaCl2.2H2O and 10 mM 2 (N-morpholino)-ethanesulfonic acid (MES) at pH 5.8. Approximately 3.97?0.63 ? 105 protoplasts per gram fresh weight were obtained when digested in enzyme solution for 4 h on a shaker under dark conditions, then washed with 0.3 M mannitol solution supplemented with 10 mM CaCl2.2H2O and 10 mM MES, and purified with 0.3 M sucrose solution. Freshly isolated protoplasts were then cultured at final density of 2 ? 105 protoplasts/ml in Kao & Michayluk medium (KM8P) by agarose-bead cultures under dark conditions at 25?C. First cell division was observed after culturing for 2 days and multicellular colonies (15-20 cells) were formed after 2 weeks.

Article Info
Agriculture and Natural Resources -- formerly Kasetsart Journal (Natural Science), Volume 040, Issue 2, Apr 06 - Jun 06, Page 361 - 369 |  PDF |  Page