In order to establish a high frequency and synchronous somatic embryo culture system, immature embryos of Black Locust(Robinia pseudoacacia L.)were used as explants to study the synchronization of somatic embryogenesis induction. Somatic embryogenesis synchronization system was established by suspension culture, osmotic pressure choice and exogenous hormone adjustment and low temperature processing method. The results showed that 1)callus stayed in low temperature(4 ℃)for 7 days showed- DOI:10.3969/j.issn.1000-1565.2020.03.009刺槐体细胞胚发生的同步化调控习洋1,孙宇涵2,李云2(1.国家知识产权局专利局专利审查协作北京中心 医药生物部,北京 100160;2.北京林业大学 生物科学与技术学院,北京 100083)摘 要:为了建立一个高频率、同步化发生的体细胞胚培养系统,以二倍体刺槐为材料,对影响刺槐体细胞胚同步化发生的培养基渗透压、培养基中添加的植物生长调节剂、低温等培养环境条件以及悬浮培养结合看护培养方法进行了研究,建立了刺槐体细胞胚同步发生体系.结果表明:1)4 ℃低温条件下处理7 d的胚性愈伤组织,最有利于刺槐体细胞胚的同步化诱导,同步化率可达37.3%;2)培养基中蔗糖质量浓度为50 g/L最适宜调控刺槐的体细胞胚同步化发生,同步化率可达35.0%;3)培养基中的2,4-二氯苯氧乙酸(2,4-dichlorophenoxyacetic acid,2,4-D)质量浓度为0.1 mg/L时最适宜体细胞胚同步化的调控,体细胞胚发生的同步化率可达43.6%,同时对畸形胚的发生有较好的控制作用;4)将悬浮培养的胚性细胞用50目筛(孔径0.28 mm)过滤,将留在筛上的细胞团接种到铺有滤纸的半固体体细胞胚诱导培养基上(培养基为MS(Murashige & Skoog)+NAA(naphthylacetic acid)0.5 mg/L+ BA(benzyladenine)0.5 mg/L+2-吗啉乙磺酸(MES)500 mg/L+谷氨酰胺 500 mg/L+水解酪蛋白500 mg/L+蔗糖 30 g/L+琼脂3 g/L),培养21 d后,球形胚大量发生,占到总胚状体数目的67.3%.本结果可为其他植物种类的体细胞胚同步化调控研究提供参考.关键词:刺槐;体细胞胚;同步化中图分类号:S792.27 文献标志码:A 文章编号:1000-1565(2020)03-0283-08Synchronization regulation of somatic embryogenesis in Black LocustXI Yang1, SUN Yuhan2, LI Yun2(1.The Pharmaceutical and Biological Invention Examination Department, Patent Examination Cooperation(Beijing)Center of the Patent Office, CNIPA, Beijing 100160, China; 2.College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China)Abstract: In order to establish a high frequency and synchronous somatic embryo culture system, immature embryos of Black Locust(Robinia pseudoacacia L.)were used as explants to study the synchronization of somatic embryogenesis induction. Somatic embryogenesis synchronization system was established by suspension culture, osmotic pressure choice and exogenous hormone adjustment and low temperature processing method. The results showed that 1)callus stayed in low temperature(4 ℃)for 7 days showed- 收稿日期:2019-11-09 基金项目:国家自然科学基金资助项目( 31170629) 第一作者:习洋(1986—),女,河北石家庄人,国家知识产权局助理研究员,博士,主要从事与生物技术相关的林木遗传研究及医药生物发明专利审查.E-mail: judyxi2011@163.com 通信作者:李云(1963—),男,河北蔚县人,北京林业大学教授,主要从事与生物技术相关的林木遗传研究.E-mail:yunli63@163.com第3期习洋等:刺槐体细胞胚发生的同步化调控the highest synchronization rate, which reached 37.3%;2)somatic embryogenesis medium which had sucrose 50 g/L showed the highest synchronization rate, which reached 35.0%;3)somatic embryogenesis medium which had 2,4-D 0.1 mg/L effectivelly controlled the occurrence of abnormal embryo and showed the highest synchronization rate, which reached 43.6%;4)cells from embryogenic cell suspension were cultured on the semi-solid medium, which was MS+NAA 0.5 mg/L+BA 0.5 mg/L+MES 500 mg/L+glutamine 500 mg/L+hydrolyzed casein 500 mg/L+sucrose 30 g/L+agar 3 g/L, for 21 days and formed a lot of globular-stage embryo at frequency of 67.3%.This study can provide theoretical support for somatic embryo synchronization regulation in other plant species.
XI Yang
,
SUN Yuhan
,
LI Yun
. Synchronization regulation of somatic embryogenesis in Black Locust[J]. Journal of Hebei University(Natural Science Edition), 2020
, 40(3)
: 283
-290
.
DOI: 10.3969/j.issn.1000-1565.2020.03.009
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