水稻愈伤组织超低温保存体系的建立  

Establishment of Rice Embryonic Callus Cryopreservation System

作　　者：李天菲[1] 林田[1] 滕小英 周丽[1] 韩静[1] 刘鸿艳[1] 罗利军[1] LI Tian-fei;LIN Tian;TENG Xiao-ying;ZHOU Li;HAN Jing;LIU Hong-yan;LUO Li-jun(Shanghai Agrobiological Gene Center,Shanghai 201106)

机构地区：[1]上海市农业生物基因中心,上海201106

出　　处：《植物遗传资源学报》2021年第5期1219-1227,共9页Journal of Plant Genetic Resources

基　　金：上海市农作物种质资源共享服务平台(18DZ2293700);国家农作物种质资源共享服务平台(上海)(NICGR2021-21)~~。

摘　　要：水稻种质资源的长期有效保存对于世界粮食安全至关重要,而超低温保存是实现这一目标的最佳方式。本研究以水稻的胚性愈伤组织为材料,利用冻存后恢复培养阶段的新生愈伤率和新生愈伤分化率来评价不同品种不同粒级大小的胚性愈伤组织、不同浓度蔗糖预培养,以及高糖预培养和高糖预培-玻璃化处理2种预处理方式对水稻愈伤组织超低温保存效率的影响,并选用自主开发的15对水稻SSR标记引物,对超低温保存前后的愈伤组织分化的幼苗进行遗传多样性鉴定。研究结果表明,粳稻日本晴和籼稻旱恢3号粒径为1.6~3.0 mm和>3.0 mm的愈伤组织超低温保存后的新生愈伤率都显著高于本品种粒径<1.6 mm的愈伤组织;旱恢3号愈伤组织经100 g/L和170 g/L蔗糖预培养后的新生愈伤率极显著高于240 g/L蔗糖预培养。粳稻WDR48、粳稻日本晴和籼稻旱恢3号仅采用高糖预培处理的新生愈伤率都极显著高于本品种玻璃化处理的新生愈伤率。新生愈伤分化率在处理间与品种间都无显著差异。简单序列长度多态性(SSLP,simple sequence length polymorphism)鉴定表明超低温保存前后的愈伤组织生成的幼苗遗传性稳定。本研究表明利用愈伤组织高糖预培养-直接超低温冻存是一种易操作、高通量、稳定可靠的中长期保存水稻种质资源的方法。Effective long-term conservation of rice germplasm resources is crucial to world food security,and cryopreservation is the best way to achieve this goal.In this study,a method with rice embryogenic callus high sugar preculture and direct liquid nitrogen cryopreservation was established,and the callus proliferation rate and the differentiation rate of proliferative calli were used to evaluate the cryopreservation efficiency directly.The results showed that the callus proliferation rates of japonica rice Nipponbare and indica rice Hanhui 3 with sizes of 1.6-3.0 mm and>3.0 mm was significantly higher than those of<1.6 mm of the same cultivar after cryopreservation.The callus proliferation rates of Hanhui 3 after preculture with 100 g/L sucrose and 170 g/L sucrose were significantly higher than those after preculture with 240 g/L sucrose.The callus proliferation rates of japonica rice WDR48,japonica rice Nipponbare and indica rice Hanhui 3 treated only with high sugar preculture were significantly higher than those treated with vitrification.There was no significant difference in callus differentiation rate among treatments and cultivars.Simple Sequence Length Polymorphism(SSLP)identification indicated that seedlings regenerated from calli either with or without cryopreservation were genetically stable.The study indicated that callus preculture with high sucrose and direct cryopreservation is an easy,high-throughput,stable and reliable method for long-term preservation of rice germplasm resources.

关 键 词：水稻 愈伤组织 超低温保存 

分 类 号：S511[农业科学—作物学]