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章志宏, 胡中立. 水稻单倍体不定芽超低温保存和植株再生及其遗传稳定性研究[J]. 植物科学学报, 2000, 18(3): 169-173.
引用本文: 章志宏, 胡中立. 水稻单倍体不定芽超低温保存和植株再生及其遗传稳定性研究[J]. 植物科学学报, 2000, 18(3): 169-173.
Zhang Zhihong, Hu Zhongli. REGENERATING PLANTS FROM CRYOPRESERVATED ADVENTITIOUS BUDS OF HAPLOIDS AND THEIR GENETIC STABILITY IN RICE[J]. Plant Science Journal, 2000, 18(3): 169-173.
Citation: Zhang Zhihong, Hu Zhongli. REGENERATING PLANTS FROM CRYOPRESERVATED ADVENTITIOUS BUDS OF HAPLOIDS AND THEIR GENETIC STABILITY IN RICE[J]. Plant Science Journal, 2000, 18(3): 169-173.

水稻单倍体不定芽超低温保存和植株再生及其遗传稳定性研究

REGENERATING PLANTS FROM CRYOPRESERVATED ADVENTITIOUS BUDS OF HAPLOIDS AND THEIR GENETIC STABILITY IN RICE

  • 摘要: 通过水稻单倍体幼穗离体培养, 诱导形成了大量的不定芽。将3mm左右的不定芽转入半固体继代培养基继代培养7d, 而后转入1.8mL的安瓿瓶中, 冰浴条件下加入预冻了的冰冻保护剂淹没材料, 在冰上平衡45~60min后, 转入程序降温仪, 以1.0℃/min的速率从4℃降至-40℃, 在-40℃停留1h后, 投入液氮保存。将液氮保存30d左右的不定芽于38~40℃的水浴中快速解冻, 随即转入半固体的再生培养基, 以待恢复生长并再生植株。结果显示: 经继代培养的不定芽超低温保存后的成活率为21%~29%, 再生出苗率为14%~18%, 而未经继代培养的不定芽超低温保存后的成活率仅为8%, 且均不能再生成苗。继代培养基MS+蔗糖3%+山梨醇4%或马铃薯提取液20%均具有良好的继代培养效果, 有效的冰冻保护剂为10%DMSO+0.5mol/L山梨醇。对不定芽经液氮超低温保存的再生植株进行染色体镜检及RAPD分析, 结果表明再生植株在染色体倍性及DNA水平上仍具有较好的遗传稳定性。

     

    Abstract: Alarge number of adventitious buds were induced from in vitro cultured young panicles of haploids of rice.The adventitious buds subcultured on solidified subculture media at 26℃ for 7 days were loaded into 18 mLplastic cryotubes with enough cryoprotectant and kept on ice for 45~60 min.After cooled at a rate of 1.0℃/min down to -40℃, the samples were plunged into liquid nitrogen.The adventitious buds cryopreserved for about 30 days were thawed rapidly in 38~40℃ water and then plated on solidified MSmedium containing 3% sucrose, 0.5 mg/L NAAand 2.0 mg/Lkinetin.After plated, 21%~29% of adventitious buds resumed growth and 14%~18% regenerated plantlets.The results of this work indicated that selecting adventitious buds derived from in vitro cultured young panicles is a critical factor for the success and subculturing adventitious buds on MSmedium containing 3% sucrose and 4% sorbitol or 20% potato extract is essential to the procedure.The effective cryoprotectant is 10% DMSO (dimethyl sulfoximide)+0.5 mol/Lsorbitol.Chromosome identifying of root tip cell and RAPDanalysis of regenerating plants indicated that the regenerating plants have good genetic stability.

     

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