生物技术通报 ›› 2015, Vol. 31 ›› Issue (9): 111-118.doi: 10.13560/j.cnki.biotech.bull.1985.2015.09.015

• 研究报告 • 上一篇    下一篇

高温胁迫诱导对白桦悬浮细胞三萜积累及防御酶活性的影响

常志凯1, 白祎杰1, 董恒1, 尹静1, 詹亚光1,2   

  1. (1.东北林业大学生命科学学院,哈尔滨 150040;2.东北林业大学 林木遗传育种与生物技术国家重点实验室,哈尔滨 150040)
  • 收稿日期:2014-12-24 出版日期:2015-09-15 发布日期:2015-09-16
  • 作者简介:常志凯,男,研究方向:植物细胞工程;E-mail:1014381994@qq.com
  • 基金资助:
    国家基础科学人才培养基金项目(J1210053),国家自然科学基金项目(31070531)

The Effects of High Temperature Stress on the Accumulation of Triterpenoid and the Activity of Defense Enzyme in the Suspension Cells of Birch

Chang Zhikai1, Bai Yijie1, Dong Heng1, Yin Jing1, Zhan Yaguang1,2   

  1. (1. College of Life Sciences,Northeast Forestry University,Harbin 150040;2. State Key Laboratory of Forest Tree Genetic Improvement and Biotechnology,Northeast Forestry University,Harbin 150040)
  • Received:2014-12-24 Published:2015-09-15 Online:2015-09-16

摘要: 通过研究不同高温胁迫下白桦细胞的细胞活力,防御酶活性和总三萜含量的变化,确定诱导白桦细胞合成三萜的最适高温胁迫条件。35℃、40℃、45℃、50℃和55℃胁迫白桦细胞1、2和4 h,测定其细胞活力,防御酶(SOD、CAT和PAL)活性和总三萜含量。结果显示,45-55℃高温胁迫下,细胞活力显著下降,以50℃ 2 h处理下降最显著,是对照细胞活力的6.8%;SOD和PAL活性显著增加,以55℃ 4 h处理后24 h时SOD和PAL活性最高,分别较对照提高164.9%和159.6%;CAT活性受到抑制,以55℃ 2 h和4 h处理在12 h后取样最低。50℃处理2 h后24 h取样,细胞总三萜含量最高,浓度为76.66 mg/g,比对照提高105.6%。最终确定50℃胁迫2 h后24 h取样为利用高温胁迫诱导白桦悬浮细胞合成三萜类物质的最适温度胁迫条件。

关键词: 白桦, 防御酶, 三萜, 温度胁迫

Abstract: This work aims to study how the high temperature stress affects the cell vitality, the activity of defense enzyme and the contents of total triterpenoid in the suspension culture cells of birch, and determine the optimal heat stress treatment by which birch cells can be induced to synthesize triterpenoid. We set temperature of 35℃, 40℃, 45℃, 50℃ and 55℃ treating birch cells for 1, 2 and 4 h respectively, then measured the cells’ vitalities, the activities of defense enzymes’(SOD, CAT and PAL), and the contents of total triterpenoid. Under the treatments of 45-55℃, cell's vitality reduced significantly, and at 50℃ for 2 h was only 6.8% of the control group;meanwhile, the activities of SOD and PAL increased significantly and reached the highest while treated at 55℃ for 4 h and measured at 24 h, i.e., they increased 164.9% and 159.6% compared to the control group. However, the CAT activity was strongly inhibited, in lowest while treated at 55℃ for 2 h and 4 h and measured at 12 h. When treated at 50℃ for 2 h and sampled at 24 h, the contents of total triterpenoid were the highest with 76.66 mg/g, increased 105.6% compared to the control. Finally, we determined that the condition of treatment at 50℃ for 2 h and measured after 24 h was the optimal to induce the birch suspension cells to produce the triterpenoid.

Key words: birch, defense enzyme, triterpenoid, temperature stress birch, defense