长白落叶松体胚发生再生体系优化  

Optimization of the Regeneration System from Somatic Embryogenesis in Larix olgensis

作　　者：刘建飞 刘炎 刘克俭 池阳 霍志发 霍永洪 由香玲[1] Jianfei Liu;Yan Liu;Kejian Liu;Yang Chi;Zhifa Huo;Yonghong Huo;Xiangling You(College of Life Sciences,Northeast Forestry University,Harbin 150040,China;State-owned Forest General Factory of Yongji County,Jilin 132000,China;Siping State-owned Farm Shilingzi Larch Nation Improved Seed Base,Siping 136000,China)

机构地区：[1]东北林业大学生命科学学院,哈尔滨150040 [2]永吉县国有林总厂,吉林132000 [3]四平市国有林总场石岭子落叶松国家良种基地,四平136000

出　　处：《植物学报》2020年第5期605-612,共8页Chinese Bulletin of Botany

基　　金：中央高校基金(No.2572018CL02)。

摘　　要：以长白落叶松(Larix olgensis)未成熟合子胚为外植体诱导胚性愈伤组织,通过调节影响体胚发生的营养物质和植物生长调节剂配比,进行愈伤组织的胚性恢复与保持以及体胚发生再生体系的优化。结果表明:不同无性系之间胚性愈伤组织诱导率差异显著,胚性愈伤组织在S+0.2 mg·L^-1 NAA+0.5mg·L^-1 BA+0.5mg·L^-1 KT+0.5g·L^-1谷氨酰胺+0.5 g·L^-1水解酪蛋白+30g·L^-1蔗糖及3.0g·L^-1植物凝胶培养条件下,可以恢复胚性并长久保持。在S+20mg·L^-1 ABA+60g·L^-1PEG4000+60g·L^-1蔗糖及3.0g·L^-1植物凝胶条件下分化培养6周,体胚发生率可达100%。将正常发育的体胚先在WPM+6 mg·L^-1间苯三酚+1.0g·L^-1活性炭+3.0mg·L^-1 VB1+20g·L^-1蔗糖及3.0g·L^-1植物凝胶条件下培养2周,再转接至B5+0.4 mg·L^-1 NAA+1.0 mg·L^-1 IBA+0.5 mg·L^-1 GA3+2.0 mg·L^-1 VB1+1.0 g·L^-1活性炭+20 g·L^-1蔗糖及3.0 g·L^-1植物凝胶条件下培养2周,可见子叶舒展、下胚轴伸长且根系正常的体胚苗。该研究建立了长白落叶松胚性愈伤组织胚性恢复与保持方法,并进一步优化了体胚发生的植株再生体系,为林木资源快速繁育和遗传改良奠定了基础。In this study,immature zygotic embryos of Larix olgensis were used as explants to induce embryogenic callus and optimize the regeneration system from somatic embryogenesis.The rejuvenation and preservation of the embryogenic potential of embryogenic callus,the somatic embryogenesis and plant regeneration were investigated through adjusting the nutrition and plant growth regulator.The results showed that the generation rates of embryogenic callus were significantly different among different lines.Under the conditions of S+0.2 mg·L^-1 NAA+0.5 mg·L^-1 BA+0.5 mg·L^-1 KT+0.5 g·L^-1 glutamine+0.5 g·L^-1 hydrolyzed casein+30 g·L^-1 sucrose and 3.0 g·L^-1 vegetable gel,the embryogenic potential of embryonic callus could be recovered and maintained for a long time.Somatic embryogenesis were induced from embryonic callus cultured in S+20 mg·L^-1 ABA+60 g·L^-1 PEG4000+60 g·L^-1 sucrose and 3.0 g·L^-1 vegetable gel for 6 weeks,and the generation rate of somatic embryo reached 100%.The normal somatic embryos were first cultured for 2 weeks under the conditions of WPM+6 mg·L^-1 phloglucinol+1.0 g·L^-1 active carbon+3.0 mg·L^-1 VB1+20 g·L^-1 sucrose and 3.0 g·L^-1 vegetable gel,and then transferred to B5+0.4 mg·L^-1 NAA+1.0 mg·L^-1 IBA+0.5 mg·L^-1 GA3+2.0 mg·L^-1 VB1+1.0 g·L^-1 active carbon+20 g·L^-1 sucrose and 3.0 g·L^-1 vegetable gel.After 2 weeks,somatic embryo plantlets with cotyledon stretch,hypocotyl elongation and normal root system were observed.This study established a method for the recovery and maintenance of embryogenic callus from larch,and further optimized the somatic embryogenic pathway,which will lay a foundation for the rapid breeding and genetic improvement of L.olgensis.

关 键 词：长白落叶松 胚性愈伤组织 胚性恢复 体胚发生优化 

分 类 号：S791.22[农业科学—林木遗传育种]