Seed storage method for in vitro germination and propagation from leaves of Brasiliorchis picta was developed. Seeds were harvested and stored at -20 and -80°C for 1, 3, 6, and 12 months and were germinated on Knudson C (KC), Murashige and Skoog (MS), half-strength MS (½ MS macro-and micronutrients), and woody plant medium (WPM). Seeds stored at -20°C, the recommended temperature for seed banks, had a high germination rate (76.0%) when cultivated in WPM after 12 months of storage. WPM is the

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... t medium for seed germination and seedling development for both harvested and stored seeds, regardless of storage time and storage temperature. Whole leaf and leaf transversal thin cell layers (tTCL) from 3-month-old in vitro grown protocorms were cultured in ½ MS supplemented with 6benzyladenine (BA; 2.5, 5.0 and 10.0 µM) and thidiazuron (TDZ; 3.0, 6.0 and 9.0 µM) for 12 weeks. The highest frequency of regenerated protocorm-like bodies (PLBs) from explants (70.0%) occurred when whole leaves were cultured in medium containing 5.0 µM BA, whereas the best response for leaf TCL was with the basal section in medium containing 9 µM TDZ, in which PLBs developed in all regions of leaves. Plantlets were successfully acclimatized (with a survival rate of 97%) when vermiculite was used as a substrate.