摘要
为改良培育苜蓿(Medicago)新品种,拟建立苜蓿原生质体融合体系。通过电融合方法,使俄罗斯杂花苜蓿(M.varia)原生质体和甘农4号紫花苜蓿(M.sativa‘Gannong No.4’)原生质体进行非对称融合,获得了杂交细胞,研究不同失活处理条件、电场条件、原生质体密度对苜蓿原生质体融合的影响。结果表明:经过紫外灯辐射5min的俄罗斯杂花苜蓿原生质体和6mmol.L-1 IOA处理的甘农4号紫花苜蓿原生质体,密度调至3×105～5×105个.mL-1,以交流电场强度15～20V.cm-1、交流频率2000～2500kHz,直流脉冲场强200～250V.cm-1、脉冲宽幅40μs、脉冲个数为3的条件为最适电融合条件,此时一对一有效融合率最高可达13.8%。
The somatic hybridization of asymmetric fusion between protoplasts isolated from callus of Rus- sian variegated alfalfa and ＇Gannong No. 4＇ alfalfa was obtained through electrofusion method. The effects of different inactivation pretreatments, electric field conditions and protoplast densities on alfalfa protoplast fusion were studied. Test results showed that the optimum inactivation pretreatments were that Russian variegated alfalfa protoplast was treated with ultraviolet radiation by 5 min and ＇Gannong No. 4 ~ alfalfa protoplast was treated with 6 mmol ~ L-1 iodoacetamide. The optimum electrofusion parameters were AC electric field intensity for 15~20 V ~ cm 1, frequency for 2000~2500 kHz, DC electric field intensity for 200~250 V cm-1 , pulse width for 40 ~s and pulse 3 times. The optimum protoplast density is 3 X 105 -5 X 105 mL-1. The highest protoplast binary fusion frequency reached 13.8% with above conditions.
出处
《草地学报》
CAS
CSCD
北大核心
2013年第4期769-775,共7页
Acta Agrestia Sinica
基金
牧区优质高效饲草生产利用技术研究与示范项目(201003023)
国家牧草产业体系项目(CARS-35)资助
关键词
苜蓿
原生质体
细胞融合
电融合
适宜条件
Alfalfa
Protoplast
Cell fusion
Electrofusion
Optimum condition
作者简介
张凌云（1987-），女，甘肃敦煌人，硕士研究生，研究方向为牧草种植资源与育种，E-mail：zlingyun0408@163．com
通信作者Authorforcorrespondence，E-mail：shisl@gsau．edu．cn
