Enhanced micropropagation of Dendrobium huoshanense C.Z. Tang et S.J. Cheng through protocorm-like bodies: The effects of cytokinins, carbohydrate sources and cold pretreatment

The effects of cytokinins, carbohydrate sources and cold pretreatment on the conversion of protocorm-like bodies (PLBs) to shoots were investigated for the enhancement of micropropagation of Dendrobium huoshanense C.Z. Tang et S.J. Cheng, an endangered medicinal plant in China. The effects of cytokinins and carbohydrate sources on the conversion of PLBs to shoots depended on their types and concentrations. The best results in terms of shoot development from PLBs occurred on 1/2 MS medium supplemented with 20 μM kinetin and 10 g l−1 maltose. Cold pretreatment at 10 °C for 1–2 weeks significantly enhanced the conversion of PLBs to shoots, and over 1300 shoots were obtained from one gram of PLBs after 3 months of culture. The developed shoots were rooted on growth regulator-free MS medium supplemented with 8 g/l banana paste to give complete plantlets, which were successfully acclimatized with a survival rate of approximately 65%. The results indicate that a suitable cold pretreatment (10 °C for 1 week) followed by the use of 20 μM kinetin and 10 g/l maltose in 1/2 MS medium would produce a large number of shoots from PLBs for plantlet regeneration of D. huoshanense.