| 摘要: |
| 为了提高白及组培过程中愈伤的增殖系数，以白及种子为外植体，利用响应面法对诱导愈伤的增殖培养基中的激素水平进行优化。在添加不同浓度的6-BA（6-苄氨基嘌呤）和2,4-D（2,4-二氯苯氧乙酸）的正交试验诱导愈伤基础上，根据Box-Benhnken（BBD）试验设计原理，采用三因素三水平的方法对3个激素的不同浓度水平进行组合实验。结果显示，通过回归模型分析获得增殖诱导的最佳培养基为MS+0.801 mg/L 6-BA（6-苄氨基嘌呤）+1.192 mg/L 2,4-D（2,4-二氯苯氧乙酸）+0.724 mg/L NAA（萘乙酸）。以优化后培养基进行愈伤组织继代发现其增殖系数为6.154 2，为理论预测值的99.74%。利用响应面法对愈伤组织增殖培养基进行优化，为以愈伤组织为研究材料的细胞悬浮培养、遗传转化、种苗快繁等研究奠定了基础。 |
| 关键词: 白及 愈伤组织 增殖 培养基配方 Box-Benhnken（BBD）响应面法 |
| DOI：10.14188/j.ajsh.2020.01.018 |
| 分类号:S567.23+9 |
| 基金项目:国家自然科学基金项目（31560079、31560087），贵州省科学技术基金项目（黔科合LH字[2014]7549号），贵州省中药现代化科技产业研究开发专项项目（黔科合ZY字【2013】3002号），遵义市2014年度“15851”人才工程项目（201424），遵义医科大学2018年研究生社会实践项目（zy-yjs2018001） |
|
| Optimization of Bletilla striata proliferation medium formula by Box-Benhnken response surface methodology |
|
SHANGGUAN Yanni1,LI Lin1,PAN Yinchi1,Zhao Ziqi1,Liu Houbo1,XU Delin1
|
|
Department of Cell Biology, Zunyi Medical University, Zunyi 563099, Guizhou, China
|
| Abstract: |
| To improve the proliferation effeciency of callus during tissue culturing in Bletilla striata, the seeds were used as explants, and the hormone level in the proliferation medium for inducing callus was optimized by the response surface methodology. On the basis of callus induction by orthogonal experiments with 6-BA and 2,4-D at different concentrations, combined experiments were carried out with three factors and three levels according to the design principle of Box-Benhnken (BBD) experiment. The optimal medium for proliferation induction was MS+0.169 mg/L 6-BA+1.635 mg/L 2,4-D+0.631 mg/L NAA through regression analysis and the multiplication factor was 6.1542, which was 99.74% of the theoretical predicted value. The response surface methodology was used to optimize the callus proliferation medium, which laid a foundation for researches of cell suspension culture, genetic transformation and seedling rapid propagation based on callus. |
| Key words: Bletilla striata callus proliferation medium formula Box-Benhnken response surface methodology |
