Medicinal Plant Research 2025, Vol.15, No.4, 151-160 http://hortherbpublisher.com/index.php/mpr 151 Research Perspective Open Access Physiological Study on Root Adaptation and Recovery of Tissue-Cultured Anoectochilus roxburghii Seedlings after Transplanting Huixian Li, Jianhui Li Institute of Life Science, Jiyang College of Zhejiang A&F University, Zhuji, 311800, Zhejiang, China Corresponding author: jianhui.li@jicat.org Medicinal Plant Research, 2025, Vol.15, No.4 doi: 10.5376/mpr.2025.15.0016 Received: 10 May, 2025 Accepted: 15 Jun., 2025 Published: 02 Jul., 2025 Copyright © 2025 Li and Li, This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Preferred citation for this article: Li H.X., and Li J.H., 2025, Physiological study on root adaptation and recovery of tissue-cultured Anoectochilus roxburghii seedlings after transplanting, Medicinal Plant Research, 15(4): 151-160 (doi: 10.5376/mpr.2025.15.0016) Abstract Anoectochilus roxburghii, a rare medicinal plant, has attracted widespread attention for its scarce resources and high cultivation difficulty. Tissue culture rapid propagation, has become the main seedling raising method, but the adaptation and recovery of the root system after transplantation remain a bottleneck. This study summarizes several regulatory mechanisms: the proportional relationship of hormones (IAA, CK, stellolactone), the accumulation of osmotic regulatory substances (proline, soluble sugar), and the activation of antioxidant systems (SOD, CAT, POD), which play an important role in alleviating transplant shock. Meanwhile, the upregulation of energy metabolism, the accumulation of secondary metabolites, and the absorption of mineral elements have been proven to be the key supports for the rejuvenation of root systems. Studies show that, a reasonable hormone ratio can shorten the recovery period of the root system. The accumulation of osmotic regulatory substances, and the enhancement of antioxidant enzyme activity, play a prominent role in the early recovery stage. In the later stage, light quality regulation and substrate improvement have shown obvious effects on the improvement of root function, and the overall adaptation of the plant. This study provides new ideas for the efficient cultivation and management of A. roxburghii, and also offers theoretical basis and guidance for its industrial promotion and resource protection. Keywords Anoectochilus roxburghii; Transplant shock; Root recovery; Osmotic regulation; Growth hormone 1 Introduction Anoectochilus roxburghii, often referred to as the "King Medicine" or "Golden Herb", is a highly valuable medicinal orchid. In traditional Chinese medicine, it has been widely applied, for its various pharmacological activities, including antioxidant, liver-protective, anti-diabetic and anti-inflammatory effects (Huang et al., 2022; Zhang et al., 2025a). It is rich in various bioactive components, like kinsenoside, flavonoids and polysaccharides, and is widely used in health foods, medicines and functional beverages (Chen et al., 2020; Zhang et al., 2020a; b; Huang et al., 2022). Because of the endangered status of wild A. roxburghii, and the need for stable quality, tissue culture has become the main propagation method. This method can achieve rapid reproduction, maintain genetic consistency, and produce disease-free plants (Wang et al., 2022), and plays a role in meeting industrialization demands, and protecting germplasm resources. But, when transplanting A. roxburghii tissue culture seedlings to a non-sterile environment, transplanting shock often occurs, containing root damage, water loss and impaired nutrient absorption, thereby significantly reducing the survival rate (Shao et al., 2014). The root systems of seedlings cultured in vitro, are fragile and are prone to be affected by environmental stress, at the time of the domestication process. Survival and growth after transplantation, are affected by various factors, like substrate composition, light intensity and quality, humidity, and the presence of beneficial or harmful microorganisms (Shao et al., 2014; Ye et al., 2020; Zhang et al., 2020a). If the conditions are not ideal, it will lead to poor adaptability of the root system, increased susceptibility to diseases, and a decrease in the accumulation of medicinal components. This study attempts to reveal the physiological, and molecular response mechanisms of the root system of A. roxburghii to transplanting stress, with a focus on the activity of antioxidant enzymes, the accumulation of
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