植物学报 ›› 2022, Vol. 57 ›› Issue (2): 227-235.DOI: 10.11983/CBB21069

• 技术方法 • 上一篇    下一篇

樟叶越桔细胞悬浮培养条件的优化

李楚然1,2, 付羚3, 刘云1,3, 杨晓琴1, 朱国磊1, 解思达1, 马焕成2, 赵平1,3,*()   

  1. 1西南林业大学西南地区林业生物质资源高效利用国家林业和草原局重点实验室, 昆明 650224
    2西南林业大学西南地区生物多样性保育国家林业和草原局重点实验室, 昆明 650224
    3云南森林资源培育与利用协同创新中心, 昆明 650224
  • 收稿日期:2021-04-27 接受日期:2021-11-24 出版日期:2022-03-01 发布日期:2022-03-24
  • 通讯作者: 赵平
  • 作者简介:*E-mail: hypzhao2022@163.com
  • 基金资助:
    国家自然科学基金(32060107);国家自然科学基金(32060327);云南省农业基础研究联合专项重点项目(2017FG001-016);西南林业大学木棉纤维人工林产业化培育云南省创新团队(云科人发[2017] 7号)

Optimization of Cell Suspension Culture Conditions of Vaccinium dunalianum

Churan Li1,2, Ling Fu3, Yun Liu1,3, Xiaoqin Yang1, Guolei Zhu1, Sida Xie1, Huancheng Ma2, Ping Zhao1,3,*()   

  1. 1Key Laboratory of State Forestry and Grassland Administration on Highly-Efficient Utilization of Forestry Biomass Resources in Southwest China, Southwest Forestry University, Kunming 650224, China
    2Key Laboratory of State Forestry and Grassland Administration on Biodiversity Conservation in Southwest China, Southwest Forestry University, Kunming 650224, China
    3Collaborative Innovation Center of Forest Resources Breeding and Utilization in Yunnan, Kunming 650224, China
  • Received:2021-04-27 Accepted:2021-11-24 Online:2022-03-01 Published:2022-03-24
  • Contact: Ping Zhao

摘要: 为了提高樟叶越桔(Vaccinium dunalianum)悬浮培养细胞的生物量, 以樟叶越桔叶片愈伤组织为试材, 通过单因素试验探究不同蔗糖浓度、培养基pH值、培养基体积、初始接种量和摇床转速对悬浮培养细胞生长的影响, 并根据响应面法Box-Behnken试验设计原理进行组合试验以优化培养条件。结果显示, 以改良WPM培养基为基础培养基, 樟叶越桔细胞悬浮培养的最优条件为40 g·L-1蔗糖、培养基pH5.2、培养基体积45 mL、初始接种量2.64 g和摇床转速为149 r·min-1, 其细胞生物量干重为0.184 4 g, 与理论预测值0.184 5 g较为接近, 且细胞的生长曲线呈S型。研究结果为樟叶越桔悬浮培养细胞次生代谢产物的生产调控奠定了技术基础。

关键词: 樟叶越桔, 愈伤组织, 细胞悬浮培养, 培养条件, 响应面法

Abstract: To increase the biomass of suspension culture of Vaccinium dunalianum, we used callus induced from leaves as start material, and investigated the effects of sucrose concentration, pH value, culture medium volume, initial inoculation size, and shaking speed using single factor experiment method. We optimized the suspension culture conditions using Box-Behnken response surface methodology. Our results showed that the optimal conditions for cell suspension culture were: 40 g·L-1 sucrose, pH5.2, culture medium volume 45 mL, callus inoculation size of 2.64 g, and shaking speed of 149 r·min-1 based on the improved WPM medium. Under these optimal conditions, the dried cell biomass was 0.184 4 g which was close to the theoretical prediction of 0.184 5 g, and the growth curve of cells showed ‘S’ type. These results provide a basis for further study on the regulation of the secondary metabolites of cell suspension culture in V. dunalianum.

Key words: Vaccinium dunalianum, callus, cell suspension culture, culture condition, response surface methodology