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    吴夏雷, 韩超, 孙宇涵, 曹森, 胡瑞阳, 徐金良, 郑会全, 李云. 杉木体细胞胚胎发生胚性愈伤组织诱导条件的优化[J]. 北京林业大学学报, 2020, 42(2): 79-86. DOI: 10.12171/j.1000-1522.20190196
    引用本文: 吴夏雷, 韩超, 孙宇涵, 曹森, 胡瑞阳, 徐金良, 郑会全, 李云. 杉木体细胞胚胎发生胚性愈伤组织诱导条件的优化[J]. 北京林业大学学报, 2020, 42(2): 79-86. DOI: 10.12171/j.1000-1522.20190196
    Wu Xialei, Han Chao, Sun Yuhan, Cao Sen, Hu Ruiyang, Xu Jinliang, Zheng Huiquan, Li Yun. Optimization of induction conditions for embryogenic callus of somatic embryogenesis in Cunninghamia lanceolata[J]. Journal of Beijing Forestry University, 2020, 42(2): 79-86. DOI: 10.12171/j.1000-1522.20190196
    Citation: Wu Xialei, Han Chao, Sun Yuhan, Cao Sen, Hu Ruiyang, Xu Jinliang, Zheng Huiquan, Li Yun. Optimization of induction conditions for embryogenic callus of somatic embryogenesis in Cunninghamia lanceolata[J]. Journal of Beijing Forestry University, 2020, 42(2): 79-86. DOI: 10.12171/j.1000-1522.20190196

    杉木体细胞胚胎发生胚性愈伤组织诱导条件的优化

    Optimization of induction conditions for embryogenic callus of somatic embryogenesis in Cunninghamia lanceolata

    • 摘要:
      目的优化杉木体细胞胚胎发生过程中胚性愈伤组织的诱导条件,探究基本培养基、供体母株基因型和外源添加剂对杉木体胚诱导的影响。
      方法以3个供体母株基因型(Z1、Z2、Z3)的杉木雌配子体(其未成熟合子胚处于裂生多胚期)为外植体,采用6种基本培养基及不同浓度的外源添加剂塞苯隆(TDZ)和茉莉酸甲酯(MeJA)处理,并对诱导培养过程中的愈伤组织进行细胞学观察。
      结果以DCR为基本培养基,诱导培养效果最好,愈伤组织诱导率达70.74%,胚性愈伤组织的诱导率达17.36%;供体母株基因型对胚性愈伤组织诱导有较大影响,Z1基因型供体母株的雌配子体最适合用于诱导产生胚性愈伤组织,诱导率为14.73%;诱导培养时以DCR为基本培养基,添加蔗糖30 g/L、活性炭1 g/L、倍力凝5 g/L并附加植物生长调节剂2,4-D 1.5 mg/L、KT 0.4 mg/L、MeJA 1.2 μmol/L和TDZ 0.004 mg/L,杉木胚性愈伤组织的诱导率最高,达19.83%;仅需4 d就可诱导产生胚性愈伤组织,不同阶段原胚团的结构和极性特征有明显差异。
      结论基本培养基、供体母株基因型和外源添加剂都会明显影响杉木体胚诱导培养,以Z1供体母株基因型的杉木雌配子体为外植体、DCR为培养基并组合添加MeJA和TDZ可明显提高杉木体胚诱导率。

       

      Abstract:
      ObjectiveThis paper aims to optimize the induction culture of embryogenic callus in the somatic embryogenesis process of Cunninghamia lanceolata and explore the effects of basic medium, maternal genotype and exogenous additives on the induction of Cunninghamia lanceolata somatic embryo.
      MethodFemale gametophyte (the immature zygotic embryo is in the multi-embryo stage) of three parental genotypes (Z1, Z2, Z3) were used as explants, and used six basic media, different concentrations of exogenous additives of TDZ and MeJA, cytological observation during the induction culture.
      ResultWith DCR as the basic medium, the induction culture was best, the callus induction rate was 70.74%, and the induction rate of embryogenic callus was 17.36%, which was the best choice for the induction of immature embryo culture of Cunninghamia lanceolata; the maternal genotype had a greater impact on embryogenic callus induction, and the female gametophyte of the Z1 genotype tree was most suitable for inducing embryogenic callus; with DCR as the basic medium, adding sucrose 30 g/L, activated carbon 1 g/L, plant gel 5 g/L and adding plant growth regulator 2,4-D 1.5 mg/L, KT 0.4 mg/L, MeJA 1.2 μmol/L and TDZ 0.004 mg/L, for the embryogenic callus of Cunninghamia lanceolata, the induction rate was the highest, reaching 19.83%; embryogenic callus can be induced in only 4 days, and the structure and polarity characteristics of the proembryogenic masses in different stages were significantly different.
      ConclusionThe basic medium, maternal genotype and exogenous additives all significantly affected the induction culture of Cunninghamia lanceolata somatic embryos. Female gametophyte with Z1 donor parental genotype as explants, DCR as the basic medium and adding MeJA and TDZ in combination can significantly improve the somatic embryos induction rate of Cunninghamia lanceolata.

       

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