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Dendrobium is a large genus in the family Orchidaceae that exhibits vast diversity in floral characteristics, which is of considerable importance to orchid breeders, biotechnologists and collectors. Native species have high value as a result of their medicinal properties, while their hybrids are important as ornamental commodities, either as cut flowers or potted plants and are thus veritable industrial crops. Thus, preservation of Dendrobium germplasm is valuable for species conservation, breeding programs and the floriculture industry. Cryopreservation represents the only safe, efficient and cost-effective long-term storage option to facilitate the conservation of genetic resources of plant species. This review highlights 16 years of literature related to the preservation of Dendrobium germplasm and comprises the most comprehensive assessment of thorough studies performed to date, which shows reliable and reproducible results. Air-drying, encapsulation-dehydration, encapsulation-vitrification, vitrification and droplet-vitrification are the current cryopreservation methodologies that have been used to cryopreserve Dendrobium germplasm. Mature seeds, pollen, protoplasts, shoot primordia, protocorms and somatic embryos or protocorm-like bodies (PLBs) have been cryopreserved with different levels of success. Encapsulation-vitrification and encapsulation-dehydration are the most used protocol, while PLBs represent the main explant explored.

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Objetivou-se neste trabalho estudar a influência de diferentes concentrações de sacarose no crescimento in vitro de Caularthron bicornutum (Hook.) Raf. O experimento foi conduzido no Departamento de Tecnologia da Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista - Unesp/FCAV, em Jaboticabal, São Paulo (SP), no período de janeiro a outubro de 2009. O delineamento experimental foi inteiramente casualizado. Foram seis tratamentos (cinco concentrações de sacarose: 10; 20; 30; 40 e 50g L-1 e ausência de sacarose no meio de cultura) e cinco repetições, totalizando 30 parcelas e 12 plântulas/parcela. A semeadura foi feita em meio de cultura MS e, após 90 dias, estas foram selecionadas e transferidas para frascos contendo meio de cultura com metade da concentração de macronutrientes (½ MS), com as diferentes concentrações de sacarose. Aos 150 dias após a semeadura, foi realizado o segundo subcultivo, cujos procedimentos foram semelhantes aos do primeiro, sendo as plântulas reinoculadas para renovação do meio. Após 210 dias da semeadura, foi realizada avaliação. Verificou-se que a sacarose não influenciou a porcentagem de enraizamento, porém foi efetiva no número e crescimento de raízes e da parte aérea de plântulas de Caularthron bicornutum, em concentrações que variaram de 13 a 29g L-1 de sacarose.

Vitrification, a simple, fast, and recommended cryopreservation method for orchid germplasm conservation, was evaluated for Dendrobium hybrid "Dong Yai" mature seeds. The genetic stability of regenerated seedlings was also evaluated using flow cytometry. Mature seeds from this hybrid were submitted to plant vitrification solution (PVS2) for 0, 0.5, 1, 2, 3, 4, 5, or 6 h at 0 °C. Subsequently, they were plunged into liquid nitrogen (LN) at -196 °C for 1 h and recovered in half-strength Murashige and Skoog culture medium (1/2 MS), and seed germination was evaluated after 30 days. Seeds directly submitted to LN did not germinate after cryopreservation. Seeds treated with PVS2 between 1 and 3 h presented the best germination (between 51 and 58%), although longer exposure to PVS2 returned moderated germination (39%). Germinated seeds were further subcultured in P-723 culture medium and developed whole seedlings in vitro after 180 days, with no abnormal characteristics, diseases, or nutritional deficiencies. Seedlings were successfully acclimatized under greenhouse conditions with over 80% survival. Flow cytometry analysis revealed no chromosomal changes on vitrified seedlings, as well as seedlings germinated from the control treatment (direct exposure to LN). These findings indicate that vitrification is a feasible and safe germplasm cryopreservation method for commercial Dendrobium orchid hybrid conservation.