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<p>The developmental requirements of <i>Platanthera bifolia</i> (L.) Rich. in asymbiotic culture are poorly understood. In this study, the factors determining post-germination development to the seedling stage in vitro were investigated. We hypothesized that seasonally changing growth cycles necessitate the use of variable conditions when growing this boreal orchid species in vitro. In the Central Siberian Botanical Garden, the effects of germination environment on achieving high germination rates and seedling growth of <i>P. bifolia</i> were tested. Temperature and photoperiod were considered as a complex factor with two gradations corresponding to the species’ habitat: (1) cold stratification in the dark at 5&#xa0;°C, based on winter temperatures in situ under snow cover; (2) warm incubation at 21&#xa0;°C under a photoperiod of 16/8 to obtain functional seedlings. The total germination percentages assessed at 6 months after inoculation increased significantly to 70.5% if the microenvironmental conditions were changed together with the growth media applied. The combined effects of in vitro microenvironmental conditions (temperature and light), close to those experienced by the species in situ, and optimal composition of growth media affected the development of functional seedlings: shoot height, number and length of roots. Thus, the efficient protocol of <i>P. bifolia</i> seed germination was developed by sequentially replacing the media and under changing growing conditions during 16 months of cultivation, in comparison with long-term maintenance of seeds at constant values ​​of these parameters. This new technique offers the chance to improve survival rate of other Holarctic orchid species in in vitro culture.</p>

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Comparative study of asymbiotic seed germination and seedling development of Platanthera bifolia (Orchidaceae) under different factors

  • 期刊论文

  • 发表日期：

    2025年7月21日

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  • Aleksandra Yu. Nabieva,
  • Elena V. Zhmud,
  • Irina N. Kuban,
  • Olga V. Dorogina

摘要

The developmental requirements of Platanthera bifolia (L.) Rich. in asymbiotic culture are poorly understood. In this study, the factors determining post-germination development to the seedling stage in vitro were investigated. We hypothesized that seasonally changing growth cycles necessitate the use of variable conditions when growing this boreal orchid species in vitro. In the Central Siberian Botanical Garden, the effects of germination environment on achieving high germination rates and seedling growth of P. bifolia were tested. Temperature and photoperiod were considered as a complex factor with two gradations corresponding to the species’ habitat: (1) cold stratification in the dark at 5 °C, based on winter temperatures in situ under snow cover; (2) warm incubation at 21 °C under a photoperiod of 16/8 to obtain functional seedlings. The total germination percentages assessed at 6 months after inoculation increased significantly to 70.5% if the microenvironmental conditions were changed together with the growth media applied. The combined effects of in vitro microenvironmental conditions (temperature and light), close to those experienced by the species in situ, and optimal composition of growth media affected the development of functional seedlings: shoot height, number and length of roots. Thus, the efficient protocol of P. bifolia seed germination was developed by sequentially replacing the media and under changing growing conditions during 16 months of cultivation, in comparison with long-term maintenance of seeds at constant values ​​of these parameters. This new technique offers the chance to improve survival rate of other Holarctic orchid species in in vitro culture.