摘要
为保持甜樱桃矮化砧木吉塞拉的遗传稳定性和避免种质遗传资源丢失,以玻璃化法超低温冷冻保存过程中涉及的主要因子设计正交试验,摸索适合吉塞拉的最佳保存体系;设计单因子试验,观察不同单因子对吉塞拉超低温冷冻保存的影响。结果表明,取生长90 d的组培苗于4℃低温锻炼3周或4周,剥取茎尖放于含0.3 mol/L蔗糖的预培养液中预培养1 d,在装载液中渗透30 min,用PVS3玻璃化试剂0℃处理60min后投入到液氮中保存24 h,取出后经40℃水浴快速化冻1 min,卸载液洗涤两次,每次10 min,后置于恢复培养基上,茎尖的存活率最高且遗传稳定性好。本试验成功建立了甜樱桃矮化砧木吉塞拉的玻璃化法超低温保存技术,为甜樱桃种质资源的长期保存提供了一条有效途径。
To maintain the genetic stability and avoid the loss of germplasm resources of sweet cherry rootstock Gisela,based on the main factors involved in the process of cryopreservation by vitrification,the orthogonal experiment was designed to obtain the optimal preservation system for Gisela. The effect of single factor on cryopreservation of Gisela was observed by single factor test. The results showed that after the 90-day- old tissue culture seedlings were acclimatized at 4 ℃ for 3 ~ 4 weeks,the shoot tips were precultured in preculture solution containing 0. 3 mol / L sucrose for 1 day,then permeated in loading solution for 30 minutes,vitrified by PVS3 at 0 ℃ for 60 minutes,stored in liquid nitrogen for 24 hours,rapidly thawed in a water bath at 40 ℃ for 1 minute,washed by unloading solution for twice and each time for 10 minutes,cultured in recovering medium and then the shoot tips got the best survival rate and genetic stability. The vitrification cryopreservation technique for sweet cherry dwarf rootstock Gisela was built in this study. It provided an efficient way to conserve the germplasm resources of sweet cherry for a long term.
出处
《山东农业科学》
2016年第10期134-139,144,共7页
Shandong Agricultural Sciences
基金
"十二五"国家科技支撑计划"主要果树新品种选育"项目"樱桃新品种选育"子课题(2013BAD02B03-3-2)
中俄国际科技合作专项"野果树优异种质资源及种质库技术的引进"(2012DFR30700)
山东省农业良种工程项目"果树种质资源挖掘与种质创新利用研究"[鲁科字(2014)96]
山东省农业科学院重大科技成果培育计划项目"甜樱桃矮化砧木新品种选育及推广利用"(2015CGPY01)
