Ten polymorphic microsatellite loci were isolated and characterized from an enriched genomic library of Paphiopedilum concolor (Batem.) Pfitzer. The number of alleles per microsatellite locus ranged from three to 11 with an average of 6.4 in a sample of 30 individuals from three populations. The observed and expected heterozygosity ranged from 0.200 to 0.800 and from 0.544 to 0.827, respectively. These microsatellites can be used as tools to investigate the genetic structure of P. concolor populations and relationship patterns with closely related taxa.
Nothodoritis zhejiangensis Z. H. Tsi is a rare and endangered epiphytic orchid restricted to a narrow distribution in China. The species is threatened with extinction as a result of loss of suitable habitats. An efficient propagation system is part of this species’ conservation plan. Seed germination reached 64.7% on Knudson's C (KC) medium containing 1.0 mg·L−1 α-naphthaleneacetic acid (NAA), 10% coconut water, and 0.1% activated charcoal. After 50 days culture, most callus (71.3%) from seed-derived protocorms formed on KC medium with 1.0 mg·L−1 2,4-dichlorophenoxyacetic acid and 10% coconut water. Callus could be subcultured 12 times during ≈2 years with more than a 3.0-fold increase from the third to the twelfth subculture. Furthermore, 84% of callus from the tenth sub-culture on KC media supplemented with 1.0 mg·L−1 NAA, 5% coconut water, and 0.1% activated charcoal formed protocorm-like bodies (PLBs). Many (57%) protocorms on KC medium containing 1.0 mg·L−1 6-benzylaminopurine and 10% coconut water formed PLBs. Both callus and PLBs formed simultaneously from different protocorms on KC medium containing 0.01 to 1.0 mg·L−1 thidiazuron and 10% coconut water. PLBs were incubated 12 times during ≈2 years with an approximate 2.4-fold increase per sub-culture. Both callus and PLBs maintained their competence to regenerate plantlets. Hyponex N026 medium supplemented with 1.0 mg·L−1 NAA, 50 g·L−1 banana homogenate, and 0.1% activated charcoal was suitable for plantlet formation and growth of 95.5% of plantlets that developed from PLBs. The roots of plantlets 2 cm in height or taller were wrapped in Chilean sphagnum moss and fixed to a fir bark block; 69.3% of plantlets survived after 180 days in a greenhouse. This protocol is an efficient means for the large-scale propagation of this endangered orchid.