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林业科学 ›› 2019, Vol. 55 ›› Issue (8): 176-183.doi: 10.11707/j.1001-7488.20190819

• 研究简报 • 上一篇    下一篇

天山云杉体细胞胚的成熟及萌发条件

依巴代提·木合旦尔1,2, 伊丽米努尔2, 张毓涛3, 刘康2   

  1. 1. 新疆师范大学生命科学学院 乌鲁木齐 830054;
    2. 新疆林业科学院造林治沙研究所 乌鲁木齐 830063;
    3. 新疆林业科学院森林生态研究所 乌鲁木齐 830063
  • 收稿日期:2018-12-02 修回日期:2019-03-17 发布日期:2019-09-05
  • 基金资助:
    新疆天山森林生态系统国家定位观测研究站资助项目;新疆维吾尔自治区自然科学基金项目(2013211A036);“十二五”农村领域国家科技计划课题(2015BAD07B03-03);2017年新疆林业科技项目。

Maturation and Germination Conditions of Somatic Embryos of Picea schrenkiana var. tianschanica

Yibadaiti Muhedaner1,2, Yiliminuer2, Zhang Yutao3, Liu Kang2   

  1. 1. College of Life Sciences, Xinjiang Normal University Urumqi 830054;
    2. Institute of Forestation and Desert ification Control, Xinjiang Academy of Forestry Science Urumqi 830063;
    3. Institute of Forest Ecology, Xinjiang Academy of Forestry Science Urumqi 830063
  • Received:2018-12-02 Revised:2019-03-17 Published:2019-09-05

摘要: [目的]建立天山云杉体细胞胚成熟及萌发阶段的适宜培养条件,为该树种的体细胞胚大规模繁殖和工厂化生产提供基础。[方法]以天山云杉成熟合子胚为外植体所诱导形成的胚性愈伤组织为材料,通过14个株系、4种成熟培养基、接种量(100、150、200 mg的新鲜胚性愈伤组织)、ABA浓度(1.9、3.8、7.6、19、38 μmol·L-1)等体细胞胚成熟培养条件,以及对不同起源(体细胞胚、合子胚和种子)的幼苗形态特征的比较,筛选出最优的天山云杉体细胞胚成熟与萌发条件。采用SPSS 18.0等统计软件对体细胞胚进行统计分析。[结果] 1)天山云杉14个株系诱导体细胞胚的结果表明,不同株系在同一成熟培养基上诱导体细胞胚的数量存在差异;其中4个株系及其接种量对体细胞胚形成数量影响的结果表明,不同株系及其接种量诱导体细胞胚的数量也存在差异,当接种量为100 mg时,T36株系的体细胞胚诱导数量最高,为3 470 ±546个,显著高于其他接种量和株系(P<0.05)。2)在4种不同成熟培养基上,均形成了成熟的体细胞胚,在1/2BLG+30 g·L-1麦芽糖培养基上形成的体细胞胚数量高于其他培养基,分别为2 044个(T6株系)和2 282个(T36株系),且与其他培养基差异显著(P<0.05)。3)不同ABA浓度对天山云杉体细胞胚诱导数量的影响无显著性差异(P>0.05),在ABA浓度为1.9~38 μmol·L-1时,其体细胞胚平均诱导数量为2 850~2 913个,其中以7.6 μmol·L-1 ABA的浓度处理效果较好,平均获得2 913个正常体细胞胚,高于其他处理。4)将不同起源(体细胞胚、合子胚和成熟种子)萌发的小植株进行对比,结果表明,体细胞胚和合子胚幼苗发育阶段基本一致,但合子胚的幼苗比体细胞胚的幼苗约大1倍,但二者间差异不显著(P>0.05);体细胞胚与合子胚子叶的平均数量一致,二者无显著差异(P>0.05),多数幼苗子叶数量为7~8个。[结论]天山云杉体细胞胚成熟阶段的适宜培养条件为胚性愈伤组织接种量100 mg,1/2BLG培养基添加7.6 μmol·L-1的ABA及75% PEG4000、30 g·L-1麦芽糖,并添加750 mg·L-1的L-谷氨酰胺或50 mg·L-1 L-天冬酰胺。不同株系在诱导体细胞胚的数量上存在差异。体细胞胚再生植株与合子胚植株在形态上相似。研究结果可为天山云杉体细胞胚的大规模繁殖和工厂化生产提供基础。

关键词: 天山云杉, 体细胞胚, 脱落酸

Abstract: [Objective] The suitable culture conditions for somatic embryos maturation and germination of Picea schrenkiana var. tianschanica were established to provide a basis for large-scale propagation and industrial production of somatic embryos of P. schrenkiana var. tianschanica.[Method] Embryogenic callus induced from mature zygotic embryos of P. schrenkiana var. tianschanica were cultured in 14 strains, 4 mature media and 100, 150, 200 mg fresh embryogenic callus. ABA concentrations (1.9, 3.8, 7.6, 19, 38 μmol·L-1) were used for mature culture of somatic embryos, and the morphological characteristics of seedlings from different origins (somatic embryos, zygotic embryos and seeds) were compared. The optimum conditions for somatic embryos maturation and germination of P. schrenkiana var. tianschanica were selected. The somatic embryos were statistically analyzed using SPSS 18.0 and other statistical software.[Result] 1) 14 strains of P. schrenkiana var. tianschanica were used to induce cell embryos. The results showed that there were differences in the number of induced cell embryos among different strains on the same mature medium. Different strains and different quantity of embryogenic callus used for culture resulted in different number of induced somatic embryos. When the quantity of embryogenic callus was 100 mg, the number of edsomatic embryos induced of strain T36 was the highest (3 470±546), significantly higher than those of the other strains. 2) Mature somatic embryos of P. schrenkiana var. tianschanica were formed on all the 4 different media for maturing. The media with 1/2BLG+30 g·L-1 maltose produced a higher number of somatic embryos than other media, at respectively 2 044(strain T6) and 2 282(strain T36), indicating a significant difference (P<0.05). 3) There was no significant difference in the number of imduced somatic embryos among different concentrations of ABA (P>0.05). When the concentration of ABA was 1.9-38 μmol·L-1, the average number of somatic embryogenesis was 2 850-2 913, among which 7.6 μmol·L-1 ABA was a better treatment. The average number of normal somatic embryos was 2 913, higher than that of the other treatments. 4) The germinating plantlets of different origins (somatic embryos, naked zygotic embryos and mature seed) were compared. The results showed that the seedling development stages of somatic embryos and naked zygotic embryos were basically the same. However, the plantlets of naked zygotic embryos were about twice as large as those of the somatic embryos, but there was no significant difference between them (P>0.05). There was no significant difference between somatic embryos and zygotic cotyledons (P>0.05). The number of cotyledons in most plantlets was 7-8.[Conclusion] The suitable condition for somatic embryos maturation of P. schrenkiana var. tianschanica was 100 mg embryogenic callus, 1/2BLG medium supplemented with 7.6 μmol·L-1 ABA and 75% PEG4000, 30 g·L-1 maltose, and added with 750 mg·L-1 L-glutamine or 50 mg·L-1 L-asparagine. In addition, there were differences in the number of induced cell embryos among different strains. Somatic embryos regeneration plant is similar in morphology to zygotic embryos plant, which can be used for large scale propagation and industrial production of somatic embryos of P. schrenkiana var. tianschanica.

Key words: Picea schrenkiana var. tianschanica, somatic embryos, abscisic acid

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