摘要
目的:以愈伤组织为材料,研究新疆杨原生质体的游离、纯化。方法:以新疆杨愈伤组织为材料,采用简单试验设计和方差分析方法,对新疆杨原生质体游离的影响因素进行研究,并利用二乙酸荧光素染色法观察原生质体活力。结果:适宜新疆杨愈伤组织原生质体游离的较适宜条件是:CPW+2.0%纤维素酶R-10+1.0%离析酶R-10+1.0%果胶酶Y-23+0.6 mol/L甘露醇,酶解温度27℃,酶解时间8 h。在此条件下,原生质体产量达8.5×106个/(g.FW),活力达83.6%。原生质体纯化可采用蔗糖等密度离心法,较适蔗糖浓度为30%。结论:研究筛选出的酶解因素组合与等密度离心条件较适宜新疆杨愈伤组织原生质体的游离和纯化。
Objective：Protoplasts of Populus alba L.var.pyramidalis were isolated from calli,isolation and purification of protoplasts were studied.Methods：Using simple experimental design and variance analysis method,according to the influence factors of protoplasts isolation,isolated time and mannitol concentration of protoplasts were studied,by using FDA staining method observing viability of calli protoplasts.Results：The more appropriate yield of protoplasts was produced by using CPW salts solution containing 2.0% cellulase R-10,1.0% macerozyme R-10,1.0% pectolase Y-23,0.6 mol/L mannitol which incubated at 27℃ for 8 hours.The protoplasts yield was 8.5×10^6/（g·FW） and the viability was 83.6% under the condition.Moreover,the more suitable protoplasts purification method was sucrose gradient centrifugation at concentration of 30%.Conclusion：The combination of different enzyme concentrations and factors is appropriate conducted on investigation into isolation and purification of calli-derived protoplasts.
出处
《生物技术通讯》
CAS
2011年第1期49-52,76,共5页
Letters in Biotechnology
基金
国家林业局专项(2002-66)
关键词
新疆杨
愈伤组织
原生质体
游离
纯化
Populus alba L. var. pyramidalis
calli
protoplast
isolation
purification
