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本文以秦艽叶片和茎段作为外植体,通过离体培养对秦艽植株再生途径进行研究。愈伤组织在添加2mg/L 2,4-D和0.5mg/L BA的MS培养基上诱导,两周内可出现愈伤组织。愈伤组织在相同激素配比并附加500mg/L LH的MS培养基上继代。愈伤组织的分化在添加有0.1mg/L 2,4-D和0.5mg/L BA的MB培养基上进行。通过显微观测,疑似体细胞胚可以在叶片和茎段的愈伤组织上产生。形态学和组织学的分析进一步证实了秦艽离体再生过程中体细胞胚发生的现象。体细胞胚和合子胚一样,也经历球形、心形、鱼雷和子叶胚等发育时期。相对独立的结构说明秦艽的体细胞胚可能是单细胞来源。体细胞胚在愈伤组织的表面和内部都有出现。在本实验中,体细胞胚发生途径是在秦艽愈伤组织形成后观察到的唯一再生途径。
In this paper, the leaves and stem segments of Gentiana macrophylla were used as explants to study the regeneration pathway of Gentiana macrophylla by in vitro culture. Callus was induced on MS medium supplemented with 2 mg / L 2,4-D and 0.5 mg / L BA, and callus appeared within two weeks. Callus was subcultured on MS medium supplemented with 500 mg / L LH with the same hormone ratio. Differentiation of callus was performed on MB medium supplemented with 0.1 mg / L 2,4-D and 0.5 mg / L BA. By microscopic observation, suspected somatic embryos can be produced on callus of leaves and stem segments. Morphological and histological analysis further confirmed the phenomenon of somatic embryogenesis during the in vitro regeneration of Gentiana macrophylla. Somatic embryos, like zygotic embryos, also experience developmental stages such as globular, heart-shaped, torpedo and cotyledonary embryos. The relatively independent structure of Gentiana solanum cells may be a single cell source. Somatic embryos appear on the surface and inside the callus. In this experiment, the somatic embryogenesis pathway was the only regeneration pathway observed after the formation of callus of Gentiana macrophylla.