快速建立胚性细胞悬浮系的培养程序初探  

Elementary Exploration about the Cultural Procedure for Rapidly Establishing Cell Embryogenic Suspension Clones

作　　者：余舜武[1] 朱永生[1] 余毓君[1] 章荣德[1] 张端品[1] 

机构地区：[1]华中农业大学作物遗传改良国家重点实验室,武汉430070

出　　处：《华中农业大学学报》2001年第4期325-328,共4页Journal of Huazhong Agricultural University

基　　金："九五"国家科技攻关农业项目 ( 96 0 0 2 0 2 0 2 2 )资助

摘　　要：为了缩短建立胚性细胞悬浮系的进程 ,快速获得松脆胚性愈伤组织 ,以普通小麦、大麦和多花黑麦草为材料 ,采用能促进植株再生的高浓度铜离子作为毒害筛选因子 ,以MSD培养基为基本配方 ,2 .5mg L的 2 ,4 D浓度 ,不同梯度浓度的铜离子处理愈伤组织。结果表明 ,0 .1mmol L的Cu2 + 对愈伤组织具有较好的筛选作用 ,能使松脆胚性愈伤组织保存下来 ,而其它类型愈伤组织褐化死亡。产生的松脆性愈伤组织能很快适应悬浮培养 ,从而快速建立胚性细胞悬浮系。用密实细胞体积及其斜面高度评价细胞悬浮系的特点 ,发现悬浮细胞不仅具有很强的生长势 ,提高了愈伤对营养缺乏的忍受度 。In order to rapidly acquire crisp calli and reduce the span for establishing embryogenic cell suspension clones, a method of a reverse train was applied to common wheat, barley and ryegrass. These calli were treated with a series of media based on MSD medium and supplemented with 2.5 mg/L 2,4?D and various concentration of copper. The results revealed that high concentration (0.1～1 mmol/L) copper , which enhanced plant regeneration in callus culture, was appended in the medium as a toxic filtrating factor to conserve the crisp embryogenic calli and make other types brown and dead. The preferentially conserved calli would be suitable for suspension culture. Packed cell volume and slant height of cell suspension showed that the suspension culture clones not only had very strong potential of growth and tolerance to the deficiency of nutrition, but also kept the embryogenic nature with high differentiational potential.

关 键 词：细胞悬浮培养 胚性愈伤组织 铜离子 MSD培养基 斜面高度 培养程序 筛选 原生质体再生植株 禾谷类作物 

分 类 号：S512.103.53[农业科学—作物学]