Sun Jingshuang, Li Jiewen, Zhao Mengnan, Zheng Guangshun, Zhang Jing, Di Bao, Ma Wenjun, Wang Junhui, Hu Ruiyang
State Key Laboratory of Tree Genetics and Breeding, Experimental Center of Forestry in North China, Chinese Academy of Forestry, Beijing 100091, China.
National Forest Germplasm Resource Bank of Representative Plants in North China, Chinese Academy of Forestry, Beijing 102300, China.
Plants (Basel). 2025 Aug 28;14(17):2688. doi: 10.3390/plants14172688.
C.A.Mey is an economically significant deciduous tree valued for timber production and landscaping applications. An efficient regeneration system is crucial for clonal propagation and serves as a foundation for future molecular breeding in . This study established two in vitro regeneration pathways-indirect somatic embryogenesis and shoot organogenesis utilizing mature zygotic embryos as explants. Primary callus was induced from cotyledon, hypocotyl, and plumule explants. A high frequency (45.73%) of yellow-green compact callus was achieved on De-Klerk and Walton (DKW) medium supplemented with 2.0 mg/L 6-BA, 1.0 mg/L zeatin (ZT), and 0.1 mg/L NAA. Subsequent transfer to 1.5× Murashige and Skoog (MS) medium containing 1.5 mg/L 6-BA, 0.2 mg/L ZT, and 0.1 mg/L NAA yielded the highest embryogenic callus induction rate (16.67%). Embryogenic callus demonstrated bipotent potential, generating both adventitious shoots and somatic embryos under specific hormonal conditions. Histological analyses confirmed the typical developmental stages of somatic embryos, from globular to cotyledonary forms, validating the embryogenic origin of regenerated structures. Furthermore, hormone or osmotic additives such as abscisic acid (ABA), Phytagel, and polyethylene glycol 4000 (PEG4000) significantly enhanced somatic embryo induction, with Phytagel at 5.0 g/L achieving the highest rate (76.31%). For shoot organogenesis, the optimal hormonal combination of the 0.6 mg/L 6-BA, 0.4 mg/L KT, and 0.15 mg/L NAA achieved the highest bud induction rate (88.89%) and produced an average of 4.07 adventitious buds per explant. This study presents an efficient regeneration system for , providing a practical platform for large-scale propagation and basis for biotechnological applications in woody plants.
C.A.Mey是一种具有重要经济价值的落叶乔木，因其木材生产和景观美化用途而受到重视。高效的再生系统对于克隆繁殖至关重要，并且是未来分子育种的基础。本研究利用成熟合子胚作为外植体，建立了两条离体再生途径——间接体细胞胚胎发生和芽器官发生。从子叶、下胚轴和胚芽外植体诱导出初级愈伤组织。在添加2.0 mg/L 6-苄基腺嘌呤（6-BA）、1.0 mg/L玉米素（ZT）和0.1 mg/L萘乙酸（NAA）的德克勒克和沃尔顿（DKW）培养基上，获得了高频（45.73%）的黄绿色致密愈伤组织。随后转移到含有1.5 mg/L 6-BA、0.2 mg/L ZT和0.1 mg/L NAA的1.5倍Murashige和Skoog（MS）培养基上，产生了最高的胚性愈伤组织诱导率（16.67%）。胚性愈伤组织具有双能性，在特定激素条件下可产生不定芽和体细胞胚。组织学分析证实了体细胞胚从球形到子叶形的典型发育阶段，验证了再生结构的胚性起源。此外，脱落酸（ABA）、植物凝胶和聚乙二醇4000（PEG4000）等激素或渗透添加剂显著提高了体细胞胚的诱导率，其中5.0 g/L的植物凝胶诱导率最高（76.31%）。对于芽器官发生，0.6 mg/L 6-BA、0.4 mg/L激动素（KT）和0.15 mg/L NAA的最佳激素组合获得了最高的芽诱导率（88.89%），每个外植体平均产生4.07个不定芽。本研究提出了一种高效的再生系统，为大规模繁殖提供了实用平台，并为木本植物的生物技术应用奠定了基础。
