响应面优化长白落叶松体胚形成早期培养条件的研究

以长白落叶松胚性愈伤组织为材料，通过Box-Behnken Design设计试验，建立以体胚发生量为响应值的响应面模型，优化长白落叶松体胚形成早期的培养条件。结果表明：相对离子浓度（P=0.007）、肌醇浓度（P=0.000）、培养周期（P=0.000）、肌醇浓度与培养周期的交互作用（P=0.007）对长白落叶松体细胞胚胎发生量的影响极为显著。长白落叶松体胚形成早期的最适培养条件为：相对离子浓度26.767 7% BM、肌醇浓度10.454 5 g·L^-1、培养周期12.656 6 d，预测体细胞胚胎的发生量为337.04 个·g^-1。为便于实际操作，验证试验将培养条件调整为：离子浓度25% BM、肌醇浓度10 g·L^-1、培养周期13 d，体胚平均发生量为336.46 个·g^-1，与预测值336.29 个·g^-1基本一致，体胚萌发率与生根率分别达100%和83.33±；5.78%，与前期试验相比，长白落叶松体胚发生的数量和质量均有显著提高。

Response Surface Optimization of Culture Conditions for the Formation of Early Embryos in Larix olgensis

With the larch embryogenic callus, through the Design Box-Behnken design test, we established a response surface model based on the somatic embryogenesis. The culture conditions of somatic embryo early maturation was optimized in Larix olgensis. The relative ion concentration(P=0.007), the inositol concentration(P=0.000), the culture period(P=0.000) and the interaction between inositol content and culture period(P=0.007) effect on the number of somatic embryos of L.olgensis were extremely significant. Cultivation of Larix somatic embryo maturation was 337.04 ind·g^-1 at the early stage of the most optimum conditions for relative ion content of 26.767 7% BM basic culture medium, inositol 10.454 5 g·L^-1, culture time 12.656 6 d, and the number of somatic embryos average. In order to facilitate the practical operation, the culture condition was changed to the relative ion concentration of 25% BM, inositol 10 g·L^-1, incubation period of 13 d, then the average number of somatic embryos was 336.46 ind·g^-1, that was consistent with the predicted value 336.29 ind·g^-1. Compared with previous test, the quantity and quality of L.olgensis somatic embryos were increased significantly.