二倍体草莓组培苗的培养和继代   

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摘要:二倍体草莓组培苗通常通过种子培养获得,基于植物的再生能力和植物细胞的全能性,可选择适龄的组培植株,在无菌环境中去除根部及叶片,将茎作为外植体置于适宜的继代培养基培养、分化即可获得无菌组培苗。继代培养可缩短草莓组培苗的培养周期,提高繁殖效率,是一种行之有效的草莓育种方式。

关键词: 草莓, 组织培养, 继代

材料与试剂

  1. 2 ml离心管
  2. 乙醇
  3. NaClO
  4. MS (Phytotechnology Laboratories, catalog number: M519)
  5. Glucose 
  6. Agar 
  7. IBA sigma
  8. NaOH (1 M)
  9. Sucrose
  10. 点种子培养基 (1 L) (见溶液配方)
  11. 继代培养基 (1 L) (见溶液配方)

仪器设备

  1. 剪刀
  2. 移液器

实验步骤

  1. 将草莓种子置于2 ml离心管内,用70%的乙醇清洗30 sec,吸出液体时注意不要吸掉种子。
  2. 10% NaClO 清洗8 min。
  3. 无菌水洗5次。
  4. 将种子均匀点在培养基上,避光7天左右移至光下培养,培养条件为相对湿度60%,光周期为16 h/8 h (日/夜),光密度为450 µmol m-2 s-1
  5. 草莓的继代培养
    将生长周期60天左右的无菌草莓组培苗的根及叶片叶柄剪去,将茎段插入继代培养基,每瓶约5个外植体。培养条件为相对湿度60%,光周期为16 h/8 h (日/夜),光密度为450 µmol m-2 s-1

溶液配方

  1. 点种子培养基 (1 L)
    MS
    2.2 g
    Glucose
    10 g
    Agar  7 g
    IBA
    0.01 mg/L
    用1 M NaOH调pH至5.8
  2. 继代培养基 (1 L)
    MS 4.4 g
    Sucrose
    30 g
    Agar
    7 g
    用1 M NaOH调pH至5.8
Copyright: © 2018 The Authors; exclusive licensee Bio-protocol LLC.
引用格式:魏灵芝 , 毛文文 , 李冰冰, 贾文锁. (2018). 二倍体草莓组培苗的培养和继代. Bio-101: e1010225. DOI: 10.21769/BioProtoc.1010225.
How to cite: Wei, L. Z., Mao, W. W., Li, B. B. and Jia, W. S. (2018). Production of Diploid Strawberry Seedlings by Tissue Culture and Subculture. Bio-101: e1010225. DOI: 10.21769/BioProtoc.1010225.
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