芝麻菜下胚轴原生质体的分离培养及再生植株  

Plant Regeneration and Hypocotyl Protoplasts Isolation of Eruca sativa

作　　者：张传利[1] 桂雪梅[1] 朱媛[1] 刘雅婷[1] 毛孝强[1] 林良斌[1] 

机构地区：[1]云南农业大学农学与生物技术学院

出　　处：《华南农业大学学报》2008年第2期63-68,共6页Journal of South China Agricultural University

基　　金：云南省自然科学基金(2004C0035M)

摘　　要：以芝麻菜Erucasativa Mill无菌苗下胚轴为材料，研究了光照条件、苗龄、酶液组合、酶解时间、酶液中甘露醇浓度及纯化条件对其原生质体分离制备的影响．以附加不同激素组合的改良KM8p培养基进行液体浅层培养．结果表明，无菌苗的下胚轴酶解10h，用“过滤-离心-漂浮法”进行纯化后，可高效分离出有活力的原生质体；在改良的KM8p培养基的进行原生质体培养，当密度为7×10^4mL^-1时，分裂频率最高，为24．8％．4周后形成大量的细胞团和肉眼可见的小愈伤组织，植板率为5．6％．然后转移到培养基上使其增殖，当愈伤组织长至3～5mm时，将其转到分化培养基上诱导芽的分化，芽分化率为33．6％．当芽长2～3cm时，将其切下插入生根培养基上诱导生根，可获得完整植株．The hypocotyls from aseptic seedlings of Eruca sativa Mill were used as materials for protoplast isolation. The effects of the light, concentration of mannitol, the seedling grown days, combinations of different enzyme, digesting time, the methods of protoplast purification, on protoplast isolation were studied. The protoplasts were cultured in modified KMSp medium supplemented with different phytohormone by ＂thin liquid layer＂ method. The results showed that the protoplasts with higher yield and quality were obtained by treating the hypocotyls for 10 h with the combination of enzyme. The hypocotyls were appropriate for protoplasts isolation. When the protoplasts were cultivated in the modified KMSp medium at the density of 7 × 10^4 mL^-1 , the division frequency was up to 24. 8%. The small calli could be obtained in four weeks, and then transferred to the proliferation medium, the plating frequency was about 5.6%. Calli of 3 - 5 mm in diameter were transferred on differentiation medium, the frequency of shoot regeneration was 33.6%. Planflets were obtained upon transferring 2 -3 cm shoots to 1/2 MS medium with 0. 5 mg/LIBA ＋ 0.2mg/L6-BA.

关 键 词：芝麻菜 下胚轴 分离 原生质体培养 再生植株 

分 类 号：Q942.5[生物学—植物学]