CAJ | 학술논문

百合原生质体培养对百合远源杂交育种具有重要意义。本研究以东方百合‘Sorbonne’花托为外植体，诱导胚性愈伤组织，并对其原生质体提取和培养进行初步研究。结果表明：MS+Picloram 3 mg/L为胚性愈伤组织诱导最佳培养基；胚性愈伤组织在酶解液为2%纤维素酶R-10+0.5%离析酶R-10+0.05%果胶酶Y23+147 mg/L二水合氯化钙+976 mg/L 2-吗啉乙磺酸+0.6 mol/L甘露醇，酶解12 h，制备的原生质体产量最高达4×105个/mL，活性达52%；在胚性愈伤组织诱导培养基中进行原生质体的悬滴培养、液体培养、固体培养、固液结合培养和看护培养。结果表明，看护培养是最佳培养方法，并在改良MS+Picloram 2 mg/L的培养基中培养2~3 d观察到长细胞壁的原生质体，培养4~6 d，可见原生质体的第一次分裂，培养40~45 d，观察到原生质体分化成的细胞团。
백합원생질체배양대백합원원잡교육충구유중요의의。본연구이동방백합‘Sorbonne’화탁위외식체，유도배성유상조직，병대기원생질체제취화배양진행초보연구。결과표명：MS+Picloram 3 mg/L위배성유상조직유도최가배양기；배성유상조직재매해액위2%섬유소매R-10+0.5%리석매R-10+0.05%과효매Y23+147 mg/L이수합록화개+976 mg/L 2-마람을광산+0.6 mol/L감로순，매해12 h，제비적원생질체산량최고체4×105개/mL，활성체52%；재배성유상조직유도배양기중진행원생질체적현적배양、액체배양、고체배양、고액결합배양화간호배양。결과표명，간호배양시최가배양방법，병재개량MS+Picloram 2 mg/L적배양기중배양2~3 d관찰도장세포벽적원생질체，배양4~6 d，가견원생질체적제일차분렬，배양40~45 d，관찰도원생질체분화성적세포단。
It is sign ifican t to culture the protoplast for wide cross breeding of Lily. In this study, the embryonic callus of receptacle of lilium oriental hybrids‘Sorbonne’ were induced and protoplasts were isolated and cultiv-ated. The results showed that feasible culture medium of inductive embryonic callus was MS+Picloram 3 mg/L;The optimal enzyme solution used for protoplasts isolation with 12 hours enzymolysis was 2%Cellulase R-10+0.5%Macerozyme R-10+0.05%Pectolyase Y23+147 mg/L CaCl2·2H2O+976 mg/L MES+6 mol/L Sorbitol, and yield reached its highest, which was up to 4×105/mL, and the activity was up to 52%. Comparing with the hanging drop culture, liquid culture, solid culture, liquid and solid culture, the nurse culture was the best way to cultivate protoplast in feasible culture medium of inductive embryonic callus. Some changes can be observed in the process of protoplast culture with improved medium that is MS+Picloram 2 mg/L by the nurse culture, such as cell wall for 2~3 days, the first division of protoplast for 4~6 days, cell mass for 40~45 days.