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植物研究 ›› 2017, Vol. 37 ›› Issue (4): 613-620.doi: 10.7525/j.issn.1673-5102.2017.04.018

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响应面优化长白落叶松体胚形成早期培养条件的研究

宋跃, 李淑娟, 白晓明, 张含国   

  1. 东北林业大学林木遗传育种国家重点实验室, 哈尔滨 150040
  • 收稿日期:2017-01-06 出版日期:2017-07-15 发布日期:2017-07-22
  • 通讯作者: 李淑娟,E-mail:lishujuan@126.com E-mail:lishujuan@126.com
  • 作者简介:宋跃(1991-),男,硕士研究生,主要从事落叶松遗传育种方面的研究。
  • 基金资助:
    国家高技术研究发展计划("863"计划2013AA102704-04-03)

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

SONG Yue, LI Shu-Juan, BAI Xiao-Ming, ZHANG Han-Guo   

  1. State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin 150040
  • Received:2017-01-06 Online:2017-07-15 Published:2017-07-22
  • Supported by:
    National High-tech R&D Program("863" Program 2013AA102704-04-03)

摘要: 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。

关键词: 落叶松属, 长白落叶松, 响应面优化, 体胚发生, 胚胎形成早期

Abstract: 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.

Key words: Larix, Larix olgensis, response surface optimization, somatic embryogenesis, early embryo formation

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