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组培水稻种子发芽   

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关键词: 水稻, 培养基, 发芽

材料与试剂

  1. 5 ml离心管
  2. 各种型号吸头 (20 μl, 200 μl, 1 ml)
  3. 培养皿
  4. 滤纸
  5. 水稻种子
  6. 75%乙醇
  7. HgCl2
  8. 蔗糖
  9. 生根培养基 (见溶液配方)
  10. MSmax(10x) (见溶液配方)
  11. MSmin母液(100x) (见溶液配方)
  12. Fe2+-EDTA(100x) (见溶液配方)
  13. 维生素(100x) (见溶液配方)

仪器设备

  1. 移液器
  2. 摇床
  3. 超净工作台

实验步骤

  1. 将种子去除颖壳,挑选质量好,无霉菌斑点的米粒。
  2. 75%乙醇漂洗1-2 min,漂洗过程中需要不断振荡。
  3. 1.5‰ HgCl2漂洗18-25 min,大约每隔3 min上下翻转振荡一次。
  4. 用灭过菌并放凉的ddH2O漂洗3~5次,去除表面残留的HgCl2
  5. 将处理好的种子放入生根培养基上,于光培养室培养,约半个月后可长成完整植株,再移入田间。

溶液配方

  1. 生根培养基
    MSmax母液(10x)
    50 ml
    MSmin母液(100x)
    5 ml
    Fe2-EDTA母液 (100x)
    10 ml
    Vitamin母液 (100x)
    10 ml
    蔗糖
    20 g
    Phytagel
    3 g
    加H2O 1,000 ml并用1 N KOH调节pH到5.8,煮开后分装到生根管中,封口膜封口灭菌。
  2. MSmax(10x)
    NH4NO3
    16.5 g
    KNO3
    19.0 g
    KH2PO4
    1.7 g
    MgSO4∙7H2O
    3.7 g
    CaCl2∙2H2O
    4.4 g 或 CaCl2 3.32g
    逐一溶解药品后,加dH2O定容到1,000 ml
  3. MSmin母液(100x)
    KI
    0.083 g
    H3BO3
    0.62 g
    MnSO4∙4H2O
    2.23 g或MnSO4∙H2O 1.69 g
    ZnSO4∙7H2O
    0.86 g
    Na2MoO4∙2H2O
    0.025 g
    CuSO4∙5H2O
    0.0025 g
    CoCl2∙6H2O
    0.0025 g
    注:Na2MoO4必须单独溶解后再与其它组分混合。
    加dH2O定容到1,000 ml室温保存
  4. Fe2+-EDTA (100x)
    取一个烧杯加300 ml dH2O以及FeSO4∙7H2O 2.78 g
    在另一个烧杯中也加300ml dH2O加热到70 °C后加入Na2 EDTA∙2H2O 3.73 g
  5. 维生素(100x)
    Nicotinic acid
    0.1 g
    Pyridoxine HCl (VB6)
    0.1 g
    Thiamine HCl (VB1)
    0.1 g
    Glycine
    0.2 g
    Inositol
    10 g
    加dH2O定容到1,000 ml 4 °C保存
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Copyright: © 2018 The Authors; exclusive licensee Bio-protocol LLC.
引用格式:李燕, 龙湍, 吴昌银. (2018). 组培水稻种子发芽. Bio-101: e1010183. DOI: 10.21769/BioProtoc.1010183.
How to cite: Li, Y., Long, T. and Wu, C. Y. (2018). Germination of Rice Seeds with Tissue Culture Method. Bio-101: e1010183. DOI: 10.21769/BioProtoc.1010183.
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