森林工程2026，Vol.42Issue(3)：452-462,11.DOI:10.7525/j.issn.1006-8023.2026.03.002
偃松体细胞胚胎发生与遗传转化技术体系的建立
Establishment of a Technical System for Somatic Embryogenesis and Genetic Transformation in Pinus pumila
摘要
Abstract
Immature zygotic embryos from immature seeds of Pinus pumila were used as explants,with DCR as the basal medium.Somatic embryogenesis was induced by adjusting the concentrations of plant growth regulators(6-BA,NAA,and ABA),leading to the acquisition of 3 embryogenic cell lines.Based on this somatic embryogenesis system,an Agro-bacterium-mediated genetic transformation system suitable for P.pumila was finally established through the determina-tion of kanamycin selection concentration,infection time,and co-cultivation duration,and resistant calli of P.pumila were successfully obtained using this system.The results indicated that the somatic embryogenesis system of P.pumila included 6 stages:disinfection of immature seeds,induction of embryogenic callus,proliferation of embryogenic callus,induction of somatic embryos,post-germination culture,and rooting culture.The optimal medium for embryogenic cal-lus induction was DCR+1 mg/L 6-BA+2 mg/L NAA,with an induction rate of 1.11%;the optimal medium for embryo-genic callus proliferation was DCR+0.35 mg/L 6-BA+0.5 mg/L NAA,achieving a proliferation multiple of 3.73;the op-timal medium for somatic embryo induction was DCR+20 mg/L ABA,with an average of 86.67 somatic embryos per gram of embryogenic callus;after somatic enbryos post-germination,DCR+activated carbon was used for root induction,resulting in a rooting rate of 58%;and the optimal medium for rooting culture was WPM+0.1 mg/L IBA.For genetic transformation,the optimal conditions were a kanamycin concentration of 60 mg/L,an infection time of 15 minutes,and an Agrobacterium suspension concentration of OD₆₀₀ is 0.4;after 7 days of recovery culture,the tissues were transferred to a medium containing 60 mg/L kanamycin for selection and callus proliferation induction.Through verification by DNA-level identification and GUS histochemical staining,transgenic calli were successfully obtained,and a genetic transfor-mation system for P.pumila was initially established.关键词
偃松/体细胞胚胎发生/遗传转化/农杆菌介导转化/胚性愈伤组织/离体再生/卡那霉素筛选/GUS组织化学染色Key words
Pinus pumila/somatic embryogenesis/agrobacterium-mediated transformation/embryogenic callus/plant regeneration in vitro/kanamycin screening/GUS histochemical staining/genetic transformation分类
农业科技引用本文复制引用
陈鑫,申丽诗,李伟,周晨光,李爽..偃松体细胞胚胎发生与遗传转化技术体系的建立[J].森林工程,2026,42(3):452-462,11.基金项目
农业生物育种国家科技重大专项(2023ZD0405904). （2023ZD0405904）
