摘要
把茂木、尾早生、解放钟、长红等枇杷品种的心脏形至完全成熟的胚,离体培养于分别附加一定浓度植物生长调节剂的B_5,MS和改良MS(大量元素减半,简称mMS)的培养基中,诱导产生胚性愈伤组织.这种愈伤组织在添加低浓度的2,4-D的mMS培养基上产生体细胞胚.以初代和继代培养若干代的愈伤组织作为分离原生质体的材料来源.酶解液的主要成分为纤维素酶、离析酶和崩溃酶,附加0.6 mol/L的山梨醇作为渗透压稳定剂,原生质体的最高得率为1.65±0.6×10~7/g.原生质体经液体浅层培养或半固体培养4—5d后,观察到第1次细胞分裂,2周后形成小细胞团.培养2个月后,获得了愈伤组织.
Callus was induced from the young or mature embryos of Mogi, Moriowase Jiefangzhong and Changhong loquat in B_5 of MS (some treatments being half strength of macroelements, called mMS) media supplemented with 2, 4-D and BA 2-3 weeks after inoculation. Embryoids were induced in mMS media supplemented with lower level of 2,4-D. Protoplasts were isolated from the callus, which incubated in macerozyme, cellulase and driselase. The highest protoplast yield was 1.65±0.6×10~7/ g. The first mitosis was observed 3 days after culture in MS media added with 2,4-D (0.1 mg/ L) and BA (0.25 mg/ L), and sorbitol (0.06 mol / L). Callus was established from protoplasts cultured for 2 months.
基金
国家自然科学基金
关键词
楷杷
原生质体
培养
愈伤组织
loquat
embryo culture
embryo-forming callus
protoplast isolation
protoplast culture
callus
