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›› 1994, Vol. 30 ›› Issue (5): 412-418.

• 论文及研究报告 • 上一篇    下一篇

群众杨悬浮细胞系的建立和耐盐体细胞变异体的初步筛选

张望东 张绮纹   

  1. 中国林业科学研究院林业研究所,北京100091
  • 收稿日期:1994-01-20 修回日期:1900-01-01 出版日期:1994-09-25 发布日期:1994-09-25

ESTABLISHMENT OF CELL SUSPENSION CULTURE AND SCREENING OF SALINE-TOLERANT SOMATIC VARIANTSOF POPULUS POPULARIS“39”

Zhang Wangdong,Zhang Qiwen   

  1. The Research Institute of Forestry, CAF Beijing 100091
  • Received:1994-01-20 Revised:1900-01-01 Online:1994-09-25 Published:1994-09-25

摘要: 对群众杨首次进行了悬浮细胞培养,对悬浮细胞系的生长进行测定,同时对耐盐体细胞变异体进行了初步筛选,获得了耐3.0‰-4.0‰不同NaCl水平的不定芽。获得适宜悬浮、松脆、易分散的愈伤组织是悬浮的关键,对不同供试脱分化激素配比的试验结果表明,诱导愈伤组织的MS+2,4-D(0.45mg/L)+NAA(0.3mg/L)+Kinetin(0.1mg/L)为最好.合适的液体培养基成份和培养技术也是悬浮细胞培养成败的主要因素。液体培养基中附加NAA或Kinetin,会使游离细胞中管状和液泡化成热细胞增加,同时,悬液中畸形细胞和细胞碎片也大量增加。液体培养基以Ms+2,4-D(0.5mg/L)+蔗糖(30g/L),另附加适量谷氨酸、天冬氨酸、精氨酸为最好。悬浮细胞在一个培养周期(14天)中,平均每个细胞分裂3—4次;细胞增长同pH变化呈明显相关。

关键词: 群众杨, 细胞悬浮培养, 耐盐细胞系, 不定芽

Abstract: Cell suspension cultures are successfully established from the friable calli which are produced mainly on a medium(M4:Ms + 0.45mg/L 2,4-D +0.3mg/L NAA + 0.1mg/L Kinetin). The best suspension medium is:Ms + 2,4D(0.5mg/L) + sucrose (30g/L). Addition of glutamic acid, aspartic acid and arginine into the liquid medium was beneficial for the growth of the suspension cell. In a culture cycle, there is a direct relationship between the growth rate of suspension cell and the change of the medium pH.Several NaCl-tolerant (3.0-4.5‰ NaCl) cell lines were obtained by direct selection and the adventitious buds had grown out from these NaCl-tolerant cell lines.

Key words: Populus popularis “39”, Cell suspension culture, NaCl-tolerant cell lines, Adventitious buds