花生原生质体分离方法探究  

Study on Protoplasts Isolation of Arachis hypogaea

作　　者：伍顺达 梁绮雯 孙敏 王思远 苏益[1] 肖浪涛[1] WU Shun-da;LIANG Qi-wen;SUN Min;WANG Si-yuan;SU Yi;XIAO Lang-tao(College of Bioscience and Biotechnology,Hunan Agricultural University,Changsha 410128,Hunan China)

机构地区：[1]湖南农业大学生物科学技术学院,湖南长沙410128

出　　处：《亚热带植物科学》2023年第3期220-227,共8页Subtropical Plant Science

基　　金：国家自然科学基金(32261143733);湖南省自然科学基金(2022JJ30294)。

摘　　要：以花生多粒型四粒红Arachis hypogaea var. fastigiate ‘Silihong’为材料,调节不同酶的浓度,优化原生质体分离条件。结果显示,混合酶解液的分离效果显著优于崩溃酶。制备花生60 d龄植株叶片原生质体的适宜条件为:2%纤维素酶+3%果胶酶+1%离析酶+0.02 mol·L^(-1) KCl+0.02 mol·L^(-1) MES+0.01 mol·L^(-1) CaCl_(2)+0.6 mol·L^(-1)甘露醇+0.1%BSA,28℃黑暗酶解3 h。在此条件下,花生叶片原生质体产量为2.965×10^(5)个·g^(-1),花瓣原生质体产量为2.55×10^(5)个·g^(-1)。此外,花瓣与黄化叶均能成功分离得到大量原生质体,而花生苗的下胚轴以及果针未分离出原生质体。Using leaves and petal of Arachis hypogaea var.fastigiate‘Silihong’as materials,protoplast isolation conditions were optimized through modifying the concentrations of different enzymes.The results showed that the protoplast outcome by using the mixed enzymes was significantly better than that by using driselase.The optimum conditions for the preparation of leaves of peanut at 60 days old protoplasts were 2%cellulase+3%pectinase+1%diastase+0.02 mol·L^(-1) KCl+0.02 mol·L^(-1) MES+0.01 mol·L^(-1) CaCl_(2)+0.6 mol·L^(-1) mannitol+0.1%BSA,28℃ for 3 hours in dark.Under this condition,the protoplast yield of peanut leaves was 2.965×10^(5) cells·g^(-1),and peanut petal protoplast yield was 2.55×10^(5) cells·g^(-1).In addition,A large number of protoplasts were successfully isolated from peanut petals and yellow leaves,but protoplasts were not isolated from hypocotyls of peanut seedlings and peanut pegs.

关 键 词：花生 原生质体 分离 混合酶 

分 类 号：S565.2[农业科学—作物学]