Plant Cell, Tissue and Organ Culture

, Volume 78, Issue 3, pp 273-276

First online:

Micropropagation of Cypripedium macranthos var. rebunense through Protocorm-like Bodies Derived from Mature Seeds

  • Hanako ShimuraAffiliated withDepartment of Botany, Graduate School of Agriculture, Hokkaido University
  • , Yasunori KodaAffiliated withDepartment of Botany, Graduate School of Agriculture, Hokkaido University Email author 

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Cypripedium macranthos var. rebunense is the most famous terrestrial orchid in Japan, since the variety has large beautiful yellowish-white flowers and is threatened with extinction. Establishment of an efficient method for micropropagation is urgently needed. When imbibed mature seeds of the orchid, that had been pre-chilled at 4°C for 3 months, were sown onto Hyponex-peptone medium that contained both α-naphthaleneacetic acid (NAA) and cytokinin, protocorm-like bodies (PLBs) were formed from germinated seeds. Although the growth of PLBs was very slow, plantlets were easily regenerated from the PLBs on hormone-free medium. The PLBs were subcultured eight times along 2 years without loss of ability to regenerate plantlets, and one aggregate of PLBs (ca. 5 mm in diameter) produced ca. 10 plants within a year. A reduction of commercial value through a large-scale micropropagation by this method will be able to prevent illegal collection from the wild populations.

asymbiotic germination conservation of threatened orchid Cypripedium macranthos var. rebunense micropropagation protocorm-like bodies