红皮云杉胚性愈伤组织诱导技术研究

Induction Technique of Picea koraiensis Nakai Embryogenic Callus

以红皮云杉未成熟胚为外植体进行胚性愈伤组织诱导实验,利用L16(42×2)混合水平正交设计研究基础培养基、光照条件、未成熟胚采集时期对胚性愈伤组织诱导的影响,以此为基础对不同的培养温度梯度进行了筛选。结果表明:改良RJW基本培养基为最适宜的基础培养基,光照条件以暗培养为宜,未成熟胚的最适宜的采集时间7月20日,适宜培养温度为22℃。当未成熟胚在添加1.0mg·L-1BA,5.0mg·L-1NAA,20g.L-1蔗糖,450mg·L-1L-谷氨酰胺、750mg·L-1水解酪蛋白的改良RJW培养基,22℃下暗培养时,胚性愈伤组织诱导率最高,达到81.3%。

Culture conditions for embryogenic callus induction from immature zygotic embryos of Picea koraiensis were studied. The effect of basal medium, light conditions, collection period of immature seeds on induction rate of embryogenic callus were compared through L16 （4^2 × 2） mixed level orthogonal experiment, subsequently culture temperature optimization experiment were also performed. The results were as follows： the optimal basal medium and culture conditions were improved RJW medium, dark culture, zygotic embryo collection date of July 20, culture temperature of 22 ℃.The highest embryogenic callus induction rate（81.3% ） was achieved on the improved RJW medium supplemented with 1.0 mg · L^- 1 BA, 5.0 mg · L^- 1 NAA, 20 g· L ^- 1 sucrose, 450 mg · L ^- 1 L-glutamine, 750 mg · L^-1 casein hydrolysate, 2.0 g · L^-1 gelrite, dark culture under 22℃, the average induction frequency was 81.3%.