Surface Response Optimization of Culture Conditions for Dendrobium nobile Lindl Protoplasts
Dendrobium nobile Lindl is a well-known endangered Chinese medicinal herb. It is therefore important to develop a germplasm resource for this species, using either cell engineering or other approaches. Our previous study found that concentrations of 2,4-dichlorophenoxyacetic
acid (S
1), 6-benzylaminopurine (S
2), and 1-naphthaleneacetic (S
3), and alsoincubation time (S
4) were four essential parameters for liquid culturing of D. nobile protoplasts. To identify the optimal values for these
four parameters, we in this study conducted experiments using three experimental designs. The experiment based on univariate analysis defined the value ranges for each parameter. The orthogonal experiment then identified 80.08% vitality rate for the cultured protoplasts was achieved
at the following optimal conditions: 1.50 mg/L for S
1, 1.00 mg/L for S
2, 1.00 mg/L for, and 24 h for S
4. Finally, response surface model (RSM) experiment showed that the vitality rate was significantly increased to 85.69% and death
rate was lowered below 13.37% when the values for S
1–4were 1.42 mg/L, 1.02 mg/L, 3.18 mg/L, and 19.44 h, respectively. The RSM experiment also showed that the vitality rate was significantly affected by synergistic effects between S
1 and S
2,
S
1 and S
3, and S
2 and S
3. These results thus allowed us to establish a reliable method for culturing the high-quality D. nobile protoplasts. The results also showed that the RSM was the best method for analyzing results
from the three experimental designs in identifying optimum values for four key parameters.
Keywords: DENDROBIUM NOBILE LINDL; ORTHOGONAL TEST DESIGN; PROTOPLAST CULTURE; RESPONSE SURFACE DESIGN; VITALITY RATE
Document Type: Research Article
Publication date: 01 May 2016
- Nanoscience and Nanotechnology Letters (NNL) is a multidisciplinary peer-reviewed journal consolidating nanoscale research activities in all disciplines of science, engineering and medicine into a single and unique reference source. NNL provides the means for scientists, engineers, medical experts and technocrats to publish original short research articles as communications/letters of important new scientific and technological findings, encompassing the fundamental and applied research in all disciplines of the physical sciences, engineering and medicine.
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