In vitro propagation of Liparis nervosa (Thunb.) Lindl., an endangered medicinal orchid

Abstract

In vitro regeneration was studied to protect the rare Chinese medicinal orchid Liparis nervosa (Thunb.) Lindl. The mixtures of protocorm and seeding and the stem tip were used as explants. The results revealed that the best essential medium for L. nervosa growth was 1/3 MS medium with 25 g · L–1 sucrose, 50 g · L–1 banana puree, 40 g · L–1 mashed potato, and 1.0 g · L–1 AC (MS1); MS1 medium with 0.5 mg · L–1 BA, 0.05 mg · L–1 2,4-D, and 1.5 mg · L–1 NAA was optimal for proliferation. When stem tips were cultured in a proliferation medium, four types of proliferation occurred: basal stem cluster bud (occurring at the basal node), tiller bud (occurring at the root), protocorm-like body (occurring at the plant’s base incision), and high-position bud (occurring on plant stem nodes other than the basal nodes). Four methods produced 10.12 proliferation coefficients. In the MS1 medium with 0.5 mg · L−1 NAA, the plantlets rooted 100%, and the rooted plantlets survived 100% after domestication and transplantation.