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甘农4号紫花苜蓿愈伤组织原生质体游离条件的研究

Study on protoplasts isolation conditions of callus of Medicago sativa cv. Gannong No.4
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摘要 以甘农4号紫花苜蓿下胚轴诱导所得愈伤组织为材料,研究酶液组合,渗透压调节物甘露醇浓度,酶解时间,愈伤组织继代时间及预处理措施对原生质体游离效果的影响。结果表明:当酶液组合为2%纤维素酶+0.5%果胶酶+0.3%半纤维素酶,渗透压为0.6mol/L甘露醇,愈伤组织继代培养时间为12d,预处理措施为在CPW-11溶液中预质壁分离1h,酶解时间10~12h时,游离出的原生质体产量最高,可达3.34×106个/g,原生质体活力高达82.8%。 The hypocotyl callus of Medicago sativa cv. Gannong No. 4 was used to study the effects of en- zyme combinations, mannitol concentration in the mixed enzyme solution, enzymolysis time, callus subculture time and pretreatment measures on hydrolysis isolation. Results showed that the optimal conditions for the high- est yield (3.34 ×106 g 1) and viability (82.8%) of protoplast were as follows:the optimum enzyme combinan- tiong was 2% cellulose onnzuka R-10+0.5% Pectinase Y-23+0.3% Henicellulase Onozuka,the optimal manni- tol concentration in enzyme solution was 0. 6 mol/L, the optimal callus subculture time was 12 days, the opti- mum enzymolysis time was 10 to 12 hours and the optimal pretreatment measure was 1 h in CPW-ll solution.
出处 《草原与草坪》 CAS 2013年第3期5-10,共6页 Grassland and Turf
基金 牧区优质高效饲草生产利用技术研究与示范(201003023)项目资助
关键词 甘农4号紫花苜蓿 愈伤组织 原生质体 酶解条件 Medicago sativa cv. Gannong No. 4 callus enzymolysis condition protoplast
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