摘要
以贵州特有地方李资源——酥李为材料,探讨了超低温保存过程中海藻酸钠浓度、氯化钙浓度、渗透保护剂中蔗糖浓度和玻璃化液PVS2处理时间对茎尖存活的影响。结果表明:剥取长约2 mm的李茎尖悬浮于2%～3%海藻酸钠的溶液中,将该混合液滴于0.1 mol/L Ca2+溶液中,包埋液均含有2 mol/L甘油和0.4 mol/L蔗糖。包埋30 min后,在0℃下PVS2处理120 min后快速投入液氮(LN),24 h后取出。在37℃水浴中化冻3 min,用1.2 mol/L的蔗糖溶液洗涤2次,每次10 min,接种于恢复培养基中,暗培养1周后,转移至正常光下,茎尖存活率达21%。
Using local germplasm of crisp plum in Guizhou Province as materials, shoot tips was cryopreserved by encapsulation-vitrification, and effects of sodium alginate concentration, calcium chloride concentration, the sucrose concentration of osmotic protective agent, and exposure time to PVS2 on the survival rate of shoot tips were investigated in the present work. Precultured shoot tips were encapsulated and simultaneously osmoprotected with a loading solution of 2 moL/L glycerol and 0.4 mol/L sucrose for 30 min. Osmoprotected and encapsulated shoot tips were dehydrated with a highly concentrated vitrification solution for 120 min at 0℃ prior to direct immersion in liquid nitrogen for 24 h. After rapid thawing in water at 37℃ for 3 min, the shoot tips were rinsed in a 1.2 mol/L sucrose solution for twice at room temperature and transferred onto the recovery medium. The cultures were kept in dark for one week prior to exposure to the light. The best survival rate of encapsulation vitrified shoot tips amounted to 21% after recovery training.
出处
《山地农业生物学报》
2013年第3期276-279,共4页
Journal of Mountain Agriculture and Biology
基金
贵州省科学技术基金项目"贵州地方李资源遗传多样性研究"[黔科合J字(2010)2073]
贵州省工程技术研究中心专项"贵州喀斯特山区特异果树种质资源发掘与利用"[黔科合农G字(2009)4003]
关键词
酥李
茎尖
包埋玻璃化法
存活率
Crisp plum
shoot tips
Encapsulation-vitrification
survival rate
