Abstract

Cypripedium guttatum is one of the most endangered orchids in Japan. In vitro propagation is an important approach for conservation of the plant species. Generally, Cypripedium in vitro propagation is unstable, especially using immature seeds. Therefore, an in vitro propagation system that protocorm produces stable germination is required.In this study, one of the weather factors was tested to obtain stable immmature seeds for seedling production. From statistical analysis of the weather data, it was suggested that solar radiation affected the ripening of Cypripedium seed. From simulations of cumulative solar radiation, a seed 46 days after pollination is suitable for in vitro propagation in Akita.Using a seed 47 days after pollination of Cypripedium guttatum, seedlings were grown in hormone free 1/3 MS agar medium. In the case of adding 0.2 mg/L BAP, multiple shoots were observed. For root elongation of the seedling, hormone free medium was preferable to adding 0.2mg/L NAA.

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