紫花苜蓿花药愈伤组织和幼苗诱导技术的研究

Study on Induction Technology of Anther Callus and Seedlings of Alfalfa.

在不同时期、不同花药低温预处理时间及热激处理、不同培养基、不同激素浓度组合、不同培养条件下对苜蓿花药进行了培养,结果表明:9月份较7月、8月份选取的花药愈伤组织诱导率高;花药低温预处理24～48h较72h处理对愈伤组织诱导的效果好,低温预处理与热激处理相结合比单用低温预处理其花药愈伤组织诱导率高;NB培养基对愈伤组织的诱导效果比B5培养基好,2,4-D浓度0.5～2.0mg/L、NAA浓度0.1～1.0mg/L、6-BA浓度0.5～1.0mg/L、KT浓度1.0～3.0mg/L都能诱导出愈伤组织,其最佳的浓度组合为NB+2,4-D 0.5mg/L+NAA 0.2mg/L+ 6-BA 0.5 mg/L+KT 3mg/L;暗培养结合光照培养较直接光照培养愈伤组织诱导率低,但绿苗率较高.

At different stages and in different low temperature preliminary treatment and heat stock treatment, the anther of alfalfa was cultured in various medium,different ratio of growth regulator and culture condition. The results indicated that anther selected in September was better than in July and August. Low temperature preliminary treatment for 24-48h was better than that of treatment for 72h for inducing callus. Frequency of inducing callus was higher by combining low temperature preliminary treatment with heat stock treatment. NB medium supplemented with 2,4 -D 0.5mg/L ,NAA 0. 2mg/L, 6-BA 0.5 mg/Land KT 3mg/L was optimal for induction of callus. Ratio of growth regulator such as 2,4--D 0.5-2.0mg/L,NAA0.1 1.0mg/L, 6 - BA0. 5 - 1.0mg/L and KT1.0 - 3.0mg/L can induce callus. Although the induction rate was low by combinating dark with light, percentage of plantlet was higher.