生物技术通报 ›› 2019, Vol. 35 ›› Issue (2): 16-22.doi: 10.13560/j.cnki.biotech.bull.1985.2018-0672

• 研究报告 • 上一篇    下一篇

银杏愈伤组织生长、褐化与黄酮积累研究

王俊燚1,3, 董金金1, 刘伟3, 曹福亮2, 汪贵斌2, 王义强1,3   

  1. 1. 中南林业科技大学林业生物技术湖南省重点实验室,长沙 410004;
    2. 南京林业大学林学院,南京 210037;
    3. 中南林业科技大学经济林培育与保护教育部重点实验室,长沙 410004
  • 收稿日期:2018-07-24 出版日期:2019-02-26 发布日期:2019-03-07
  • 作者简介:王俊燚,男,硕士研究生,研究方向:银杏资源综合利用;E-mail:wangjunyi22@163.com
  • 基金资助:
    国家重点研发计划专项(2017YFD0600701),国家自然科学基金项目(31570682)

Research on the Growth,Browning and Flavonoid Accumulation of Ginkgo biloba Callus

WANG Jun-yi1,3, DONG Jin-jin1, LIU Wei3, CAO Fu-liang2, WANG Gui-bin2, WANG Yi-qiang1,3   

  1. 1. Hunan Provincial Key Laboratory for Forestry Biotechnology,Central South University of Forestry and Technology,Changsha 410004;
    2. College of Forestry,Nanjing Forestry University,Nanjing 210037;
    3. Key Lab of Non-wood Forest Nurturing and Protection of National Ministry of Education,Central South University of Forestry and Technology,Changsha 410004
  • Received:2018-07-24 Published:2019-02-26 Online:2019-03-07

摘要: 旨在建立稳定的银杏愈伤组织继代体系,筛选细胞活性强且黄酮产量高的细胞培养条件,为工业化生产提供一定参考。以MS培养基为基本培养基,在40 d的继代周期内设计正交实验研究外植体月份、外源激素与抗褐化剂交互作用下的银杏愈伤组织生长、褐化与黄酮积累,并对比了4、5、6月中旬银杏叶片黄酮含量。结果表明,银杏愈伤组织继代中的最佳生长组合为:4月叶片诱导的愈伤组织+3.0 mg/L NAA+0.5 mg/L KT+5 mg/L VC;最佳抗褐化组合为:4月叶片诱导的愈伤组织+1.0 mg/L NAA+1.0 mg/L KT+10 mg/L VC;银杏叶黄酮含量从4-6月呈上升趋势,且不同月份叶片诱导的愈伤组织黄酮含量与之呈正相关,以6月叶片诱导的愈伤组织+2.0 mg/L NAA+1.0 mg/L KT+5 mg/L VC为最佳黄酮积累条件;结合愈伤组织干重,总黄酮实际产量最高组合为:5月叶片诱导的愈伤组织+2.0 mg/L NAA+0.5 mg/L KT+5 mg/L VC。银杏愈伤组织的适宜继代周期为24-32 d,生长量较大且褐化率低。

关键词: 银杏, 愈伤组织, 正交实验, 生长, 褐化, 黄酮

Abstract: The aim of this study is to establish a stable callus succession system of Ginkgo biloba,to screen the condition of culturing cells with high activity and flavonoids,and thus to provide references for industrial production. Using MS medium as the basic medium,we investigated the G. biloba callus growth,browning and flavonoid accumulation under the interactions among explant month,exogenous hormone and anti-browning agent by an orthogonal experiment of 40 d succession period,and compared with the contents of G. biloba leaves’ flavonoids in April,May,and June. The results showed that the condition for optimal growth of G. biloba callus was callus induced by April leaf + 3.0 mg/L NAA + 0.5 mg/L KT + 5 mg/L VC. The optimal anti-browning condition was callus induced by April leaf +1.0 mg/L NAA+1.0 mg/L KT+10 mg/L VC. The flavonoid content of G. biloba leaf tended to increase from April to June,and the flavonoid content of callus at different month was positively correlated with its explants,the condition of callus induced by June leaf + 2.0 mg/L NAA + 1.0 mg/L KT + 5 mg/L VC was the optimal for flavonoids accumulation. Combined with the dry weight of callus,the maximum flavonoid yield was from the combination of callus induced by May leaf + 2.0 mg/L NAA + 0.5 mg/L KT + 5 mg/L VC. The appropriate succession period for G. biloba callus was 24-32 d,with large growth and low browning rate.

Key words: Ginkgo biloba L., callus, orthogonal experiment, growth, browning, flavonoids