Anbazhakan Rengasamy, Zhu Xin-Meng, Li Neng-Qi, Poudel Brihaspati, Gao Jiang-Yun
Institute of Biodiversity, School of Ecology and Environmental Sciences, Yunnan University, Kunming 650500, China.
Plants (Basel). 2025 Jul 15;14(14):2186. doi: 10.3390/plants14142186.
Chun & S.K. Lee, an endemic monotypic species that belongs to the family Olacaceae, is under continuous pressure of decline owing to several ecological and physiological factors. The present study aimed to establish an efficient in vitro protocol for callus-mediated indirect somatic embryogenesis in by alleviating tissue browning. Internodes and leaves obtained from seedlings were used as explants. Antioxidant pre-treatment (ascorbic acid, AA) followed by different carbon sources (sucrose, maltose, glucose, and fructose) and plant growth regulators in various concentrations and combinations were employed in Woody Plant Medium (WPM) to alleviate explant browning and induce callus formation from the explants. AA pre-treatment and subsequent culture on maltose at a concentration of 116.8 mM were optimal for controlling phenolic exudation on >90% of both explants. The highest responses of 53.77% and 57.43% for embryogenic calli were induced from internode and leaf explants, respectively. The highest responses, 85.22% and 93.80%, were observed for somatic embryos that matured into the globular, heart-shaped and torpedo stages at different percentages on NAA 2.5 mg/L in combination with BA 1.0 mg/L for both explants. The matured somatic embryos were finally germinated at a maximum concentration of GA, 2.0 mg/L. All plantlets were successfully hardened and acclimatized under culture room conditions and then transferred to the greenhouse. The current study suggests an efficient protocol for indirect somatic embryogenesis by alleviating phenolic exudation from the explants of . This first successful report of in vitro culture establishment in may offer an effective alternative measure to conserve this species and provide a system for analyzing bioactive chemicals and for use in the oil industry.
春氏木犀榄（Chun & S.K. Lee）是木犀榄科的一个单型特有物种，由于多种生态和生理因素，其数量持续下降。本研究旨在通过减轻组织褐变，建立一种高效的愈伤组织介导的间接体细胞胚胎发生体外培养方案。以幼苗的节间和叶片为外植体。在木本植物培养基（WPM）中采用抗氧化剂预处理（抗坏血酸，AA），随后添加不同碳源（蔗糖、麦芽糖、葡萄糖和果糖）以及不同浓度和组合的植物生长调节剂，以减轻外植体褐变并诱导外植体形成愈伤组织。AA预处理并随后在浓度为116.8 mM的麦芽糖上培养，对于控制两种外植体>90%的酚类渗出最为理想。节间和叶片外植体诱导胚性愈伤组织的最高响应率分别为53.77%和57.43%。在两种外植体中，当萘乙酸（NAA）2.5 mg/L与苄氨基嘌呤（BA）1.0 mg/L组合时，体细胞胚胎成熟为球形、心形和鱼雷形阶段的最高响应率分别为85.22%和93.80%。成熟的体细胞胚胎最终在最高浓度为2.0 mg/L的赤霉素（GA）作用下萌发。所有小植株在培养室条件下成功炼苗并驯化，然后转移到温室。本研究提出了一种通过减轻外植体酚类渗出进行间接体细胞胚胎发生的高效方案。这是春氏木犀榄体外培养建立的首个成功报告，可能为保护该物种提供一种有效的替代措施，并为分析生物活性化学品和用于石油工业提供一个系统。
