酶解时间对灯盏花原生质体产量及活力的影响  

Effect of Enzymatic Hydrolysis Time on Yield and Viability of Protoplast in Erigeron breviscapus

作　　者：李余润 王馨[1] 杨丽英[1] 杨斌[1] 宁玲 董志渊[1] LI Yurun;WANG Xin;YANG Liying;YANG Bin;NING Ling;DONG Zhiyuan(Institute of Medicinal Plants,Yunnan Academy of Agricultural Sciences,Kunming,Yunnan 650205,China;College of Tropic Crops,Yunnan Agriculture University,Puer,Yunnan 665099,China)

机构地区：[1]云南省农业科学院药用植物研究所,云南昆明650205 [2]云南农业大学热带作物学院,云南普洱665099

出　　处：《天津农业科学》2020年第6期12-14,20,共4页Tianjin Agricultural Sciences

基　　金：云南省科技计划项目重大科技专项(2017ZF002-3);云南省科技人才和平台计划(2017HB088);国家自然科学基金项目(31660422)。

摘　　要：原生质体是进行单细胞测序和基因编辑研究的理想材料。为了研究不同酶解时间对灯盏花原生质体分离的影响,采用纤维素酶1%+果胶酶0.3%+半纤维素酶0.5%酶液处理愈伤组织悬浮细胞2,3,4,5,6 h,纤维素酶0.5%+果胶酶0.2%+离析酶0.5%酶液处理幼叶4,6,8,10,12,14,16 h。结果表明,不同酶解时间,灯盏花悬浮细胞和幼叶的原生质体产量及活力均差异显著。悬浮细胞酶解5 h,原生质体的产量和活力达到最高,分别为3.73×105个·g-1、75.84%;幼叶酶解12 h,原生质体产量最高,达到1.17×106个·g-1,酶解10 h,原生质体活力最高,为80.98%。叶片的原生质体产量和活力明显高于悬浮细胞。Protoplast is an ideal material for single-cell sequencing and gene editing.In order to optimize enzymatic hydrolysis times for the protoplast isolation of Erigeron breviscapus,suspension cells were incubated in an enzyme solution containing 1%cellulase,0.3%pectinase and 0.5%hemicellulase for the 2,3,4,5,6 h and young leaves were incubated in an enzyme solution containing 0.5%cellulase,0.2%pectinase and 0.5%macerozyme for the 4,6,8,10,12,14,16 h.The results showed enzymatic hydrolysis times significantly affect protoplast yield and viability.Five-hour enzyme incubation produced the maximum yield(3.73×105 protoplasts·g-1)and viability(75.84%)of protoplasts from suspension cells.Using young leaves as materials,the maximum yield(1.17×106 protoplasts·g-1)of the protoplasts was obtained at 12 h and the maximum viability(80.98%)was obtained at 10 h.The yield and viability of protoplasts from young leaves were both higher than those from suspension cells.

关 键 词：灯盏花 原生质体 酶解时间 产量 活力 

分 类 号：S567.2[农业科学—中草药栽培]