油橄榄体细胞胚发生和植株再生  

Somatic Embryogenesis and Plant Regeneration in Olive

作　　者：樊育彤 崔祺禛 刘晴晴 李书媛 李雨霏 张建国[1,2,3] 饶国栋[1,2,3] FAN Yu-tong;CUI Qi-zhen;LIU Qing-qing;LI Shu-yuan;LI Yu-fei;ZHANG Jian-guo;RAO Guo-dong(State Key Laboratory of Tree Genetics and Breeding,Research Institute of Forestry,Chinese Academy of Forestry,Beijing 100091,China;Collaborative Innovation Center of Sustainable Forestry in Southern China,Nanjing Forestry University,Nanjing 210037,Jiangsu,China;Key Laboratory of Tree Breeding and Cultivation,National Forestry and Grassland Administration,Research Institute of Forestry,Chinese Academy of Forestry,Beijing 100091,China)

机构地区：[1]林木遗传育种全国重点实验室,中国林业科学研究院林业研究所,北京100091 [2]南京林业大学南方现代林业协同创新中心,江苏南京210037 [3]国家林业和草原局林木培育重点实验室,中国林业科学研究院林业研究所,北京100091

出　　处：《林业科学研究》2024年第6期104-111,共8页Forest Research

基　　金：中央级公益性科研院所专项资金项目(CAFYBB2023PA005-2);国家自然科学基金(32371837)。

摘　　要：[目的]以油橄榄的种胚为外植体,建立油橄榄‘Koroneiki’品种的体细胞胚发生及植株再生体系,旨在为油橄榄育种和遗传转化奠定基础。[方法]将油橄榄种胚及后续不同发育阶段的组织培养材料分别接种于愈伤组织诱导培养基、体细胞胚诱导培养基、成熟培养基、生芽培养基和植物生长培养基中,以诱导油橄榄体细胞胚和植株的再生,并对愈伤组织诱导率、体细胞胚诱导率和体细胞胚的成苗率进行统计。[结果]在添加0.5mg·L^(-1)2-iP和5 mg·L^(-1)IBA的OM培养基中,‘Koroneiki’的种胚外植体诱导出愈伤组织,愈伤组织诱导率为98.72%,随后将愈伤组织分离并转移到添加0.5 mg·L^(-1)IBA的OM培养基中,成功诱导出体细胞胚,诱导率为8.50%。体细胞胚在其发育过程中经历了球形胚、心形胚、鱼雷胚以及子叶胚的阶段。增殖培养基显著增加了胚的生成数量,增幅达到普通培养基诱导体细胞胚的2.06倍。将成熟的子叶胚移至生芽培养基和植物生长培养基中,成功培育出油橄榄的再生苗,成苗率达到80.73%。将再生苗移植至温室,经过2个月后能够稳定存活。[结论]初步建立了我国主栽油橄榄品种‘Koroneiki’的体细胞胚间接发生体系,研究结果为油橄榄种苗的商业化生产和分子育种提供了理论和技术基础。[Objective]A somatic embryogenesis and plant regeneration system for the'Koroneiki'variety of olive was established using olive seed embryos as explants.The objective of this study l was to establish a foundation for efficient breeding and genetic transformation of olive plants.[Methods]The embryogenic callus induction medium,somatic embryo induction medium,maturation medium,budding medium,and plant growth medium were inoculated with olive seed embryos and tissue culture materials at different developmental stages to induce somatic embryo and plant regeneration.The rates of callus induction,somatic embryo induction,and seedling formation from somatic embryos were then calculated.[Results]In the OM medium supplemented with 0.5 mg·L^(-1)2-iP and 5 mg·L^(-1)IBA,embryo explants of'Koroneiki'induced embryogenic callus with a callus induction rate of 98.72%.The callus was subsequently isolated and transferred to OM medium supplemented with 0.5 mg·L^(-1)IBA,resulting in the successful induction of somatic embryos with an induction rate of 8.50%.The somatic embryos progressed through stages including globular,heart-shaped,torpedo,and cotyledon embryos during their development.The use of proliferation medium significantly increased the number of embryos,with 2.06 times that of somatic embryos induced by the normal medium.After transferring mature cotyledon embryos to budding and plant growth media,regenerated olive seedlings were successfully cultivated,with a seedling formation rate of 80.73%.The regenerated seedlings were transplanted to the greenhouse and exhibited stable survival after 2 months.[Conclusion]In this study,an indirect somatic embryogenesis system for'Koroneiki',the main olive variety in China,has been successfully established.The results obtained provide a solid theoretical and technical foundation for the commercial production,as well as the biomolecular design and breeding,of olive seedlings.

关 键 词：油橄榄 愈伤组织 组织培养 体细胞胚 植株再生 

分 类 号：S718.46[农业科学—林学] S565.7