摘要
为获得高质量的原生质体,给光皮树转化系统的建立奠定基础,以光皮树花药愈伤组织为材料,研究了愈伤组织结构、酶液组合、酶解时间及渗透压对原生质体分离效果及活力的影响,从而确定适合光皮树花药愈伤组织原生质体最佳分离条件。结果表明:淡黄绿色、疏松的愈伤组织在CPW+2%纤维素酶R-10+0.3%蜗牛酶+0.25%果胶酶Y-23+13%甘露醇+0.6/L MES的酶解液中,黑暗条件下振荡酶解10 h,原生质体产量达到3.19×106个/g,具活力的原生质体为73.48%。
Anther callus of Swida wilsoniana were used as test materials to study the influencing factors on protoplast isolation,such as concentration and combination of enzyme,enzymolysis time and mannitol concentration,and ulteriorly,to determine the optimal separation condition that is appropriate for isolating protoplast of anther callus.The results show that the loose structure callus with light yellow and green color was mixed with enzyme liquid of CPW＋ 2% Cellulase R-10 ＋ 0.3% Snailase ＋0.25% Pectinase ＋13%mannitol ＋0.6/L MES,and was oscillated slowly for 10 hours in dark,resulting the highest yield of protoplast go up to 3.19×106 个 /g with viability of 73.48%.The research results would be a basis of establishing in vitro culture system of gene transform.
出处
《中南林业科技大学学报》
CAS
CSCD
北大核心
2012年第4期135-139,共5页
Journal of Central South University of Forestry & Technology
基金
国家林业公益项目(201004071)
国家科技支撑项目(2011BAD22B08)共同资助
关键词
光皮树
花药愈伤组织
原生质体
分离条件
Swida wilsoniana
anther callus
protoplast
Condition of isolation
