青扦胚性细胞悬浮培养中影响体细胞胚发生因素的研究

Studies on the Factors Affecting of Somatic Embryogenesis in Embryonic Cell Suspension Culture of Picea wilsonii

试验以青扦（Ｐｉｃｅａｗｉｌｓｏｎｉ）的胚性愈伤组织为材料，以改良５９基本成分附加２４－Ｄ１ｍｇ／Ｌ及ＫＴ１ｍｇ／Ｌ为培养介质，比较了液体悬浮与半固体二种培养方式对胚性愈伤组织增殖和体细胞发生的影响，研究了液体悬浮培养过程中影响体细胞胚发生的因素。结果表明：液体悬浮培养好于半固体培养，它的胚性愈伤组织的生长率为２６８％，是半固体培养的１２４倍；体细胞胚的分化率为９３％，是半固体培养的２２倍；悬浮培养较佳的培养条件为：初始细胞密度为２％（鲜重），蔗糖浓度为２０ｇ／Ｌ，摇床转速为１００ｒ／ｍｉｎ，ｐＨ为５８。经过两个月悬浮培养，将培养物转至１／２改良５９附加ＡＢＡ１ｍｇ／Ｌ的分化培养基上，３个月后每ｇ培养物上可获得２８５个正常的子叶期体细胞胚。

The embryonic callus induced from zygotic embryos of Picea wilsonii were used as initial culture and cultured on the modified 59 medium supplemented with 2 4-D 1 mg/L and KT 1 mg/L. The effect of the semi-solid and the suspension culture on embryonic callus proliferation and their embryogenesis have been compared. The result showed that the method of suspension culture is better than that of semi solid culture, the growth rate of embryonic callus in suspension culture is 268 5%; as 12 4 times as that of the semi-solid medium; its frequency of embryogenesis is 93%, as 2 2 times as that of semi-solid medium. In order to get suitable condition for large scale propagation of somatic embryos, the factors affecting somatic embryogenesis in embryonic cell suspension culture were investigated. The results are: the optimum values of the initial cell density and the initial pH and concentration of sucrose of medium, the shaking rate were 2 0%(fresh weight), 5 8, 20 g/L and 100 r/min respectively. After 2 month suspension culture, the cultures were translated into 1/2 strength modified 59 medium with 1 mg/L ABA, 286 normal somatic embryos with cotyledon per gram embryonic tissue were obtained in 3 months.