Conservation of Anoectochilus formosanus Hayata by artificial cross-pollination and in vitro culture of seeds

Anoectochilus formosanus is an important ethnomedicinal plant of Taiwan. We have optimized a method for mass propagation of A. formosanus by artificial cross-pollination and asymbiotic germination of seeds. The success of pollination and fruit set was found to be dependent on the developmental stage of male and female gametophytes. Fruit set of hand-pollinated flowers was 86.7%. The seeds from 7-week-old capsules were germinated by culturing on half-strength Murashige and Skoog’s (MS) medium supplemented with 0.2% activated charcoal and 8% banana homogenate for four months. Germinated seedlings were cultured in half-strength liquid MS medium containing 2 mg/l N-benzyladenine (BA) in 125-ml Erlenmeyer flasks for two months. Before ex vitro establishment of seedlings, seedlings with well-developed rhizomes and shoots were cultured on half-strength MS medium with 0.2% activated charcoal, 8% banana homogenate, 2 mg/l BA and 0.5 mg/l a-naphthaleneacetic acid (NAA) for further growth. Around 90% seed-derived plants survived two months after transfer to peat moss:vermiculite potting mixture and incubation in the growth chamber.