摘要
研究了普通小麦(Triticum aestivum L.)、多花黑麦草(Lolium muiriflorum Lam.)和多年生黑麦草(Lolium perenne L.)原生质体电融合的适宜条件。结果表明:在同一电场条件下,不同物种原生质体的电融合反应有差异,在500kHz、100V/cm交流脉冲和1.5kV/cm、40μs直流脉冲下,原生质体排列成串,小麦、多花黑麦草和多年生黑麦草原生质体融合频率分别为80％,60％,30％;电场参数中的场强和直流脉冲幅宽对原生质体融合频率影响较大;融合液中CaCl_2浓度以0.5～1mmol/L为宜;渗透压较低的融合液有利于原生质体融合;在适宜的电脉冲处理后,原生质体存活率与对照相似,经融合处理的原生质体培养一个月后,其植板率有所提高。
The cell suspension lines of wheat (Triticum aestinon L.), annual ryegrass (Lolium multiflorum Lain.) and perennial ryegrass (L. perenne L.) were used to study the optimal condition of monocotyledonous protoplast electrofusion. The protoplasts were aligned in AC field of 500kHz and 100V/cm and fused in DC pulse of 1.5kV/cm and 40μs at fusion frequency of 80%, 60%, and 30% for wheat, annual ryegrass and perennial ryegrass, respectively. Field strength and DC pulse duration were more effective parameters for the protoplast fusion frequency. The presence of CaCl_2 at 0.5～1 mmol/L in fusion solut/on was optimal to increase the fusion frequency of the protoplasts. It was beneficial for increasing fusion yield to reduce the osmotic pressure of fusion solution. The protoplast viability was not decreased after being treated with shorter pulse duration, and the plate efficiency of the protoplasts with Fusion treatment was incrcased.
出处
《南京农业大学学报》
CAS
CSCD
北大核心
1991年第4期1-6,共6页
Journal of Nanjing Agricultural University
