马铃薯叶肉原生质体电融合参数优化及杂种植株再生  

Optimization of Electrofusion Parameters from Mesophyll Protoplasts and Somatic Hybrids Regeneration in Potato

作　　者：蔡兴奎[1] 柳俊[2] 谢从华[1] 

机构地区：[1]华中农业大学园艺林学学院,武汉430070 [2]华中农业大学生命科学技术学院,武汉430070

出　　处：《华中农业大学学报》2003年第5期494-498,共5页Journal of Huazhong Agricultural University

基　　金：国家自然科学基金项目(39870516);"八六三"重大专项(2002AA207011)资助

摘　　要：以马铃薯栽培种“中薯2号”的无性系3#、8#和野生种Solanum chacoense叶肉原生质体为融合试材,优化马铃薯高效融合的电融合参数。研究表明,在交变电场(AC)100 V/cm、AC作用时间20 s、直流脉冲电压(DC)1 100 V/cm、DC脉冲时间60 μs、脉冲数1次时,双核融合率可达40％以上。融合产物经过4-5个月的培养获得再生植株。对首先再生的15个株系通过RAPD标记检测,证明均为体细胞杂种。所用引物C-01能够同时扩增出杂种植株中双单倍体亲本11 00 bp和670 bp的特征带以及野生种S.chacoense亲本510 bp的特征带。Somatic hybridization via protoplast fusion is of great advantages for use of potato wild species. This study was primarily attempted to optimize the electrofusion parameters, using mesophyll protoplasts isolated from dihaploid clones 3# and 8#, which were derived from potato cultivars 'Zhongshu 2' via anther culture, and diploid wild species Solanum chacoense. It was shown that binu-clear fusion frequency reached 40% with the following parameters: alternate current (AC) 100 V/cm, AC duration 20 s, direct current (DC) 1100 V/cm, 1 DC pulse with 60 s. The fusion products regenerated into plantlets within 4-5 months. 15 putative somatic hybrid lines were clarified by RAPD techniques with primer C-01, which amplified two specific bands (1 100 bp, 670 bp) of DH parent and 510 bp specific band of S. chacoense.

关 键 词：马铃薯 原生质体 电融合参数 RAPD 脉冲 

分 类 号：S532[农业科学—作物学]