Medicinal Plant Research 2025, Vol.15, No.3, 119-128 http://hortherbpublisher.com/index.php/mpr 119 Feature Review Open Access Effects of Growth Regulators on Root Development and Antioxidant Capacity of Anoectochilus roxburghii Tissue Culture Seedlings Chuchu Liu 1, Yuhong Huang 2 1 Institute of Life Science, Jiyang College of Zhejiang A&F University, Zhuji, 311800, Zhejiang, China 2 Traditional Chinese Medicine Research Center, Cuixi Academy of Biotechnology, Zhuji, 311800, Zhejiang, China Corresponding author: chuchu.liu@jicat.org Medicinal Plant Research, 2025, Vol.15, No.3 doi: 10.5376/mpr.2025.15.0013 Received: 10 Apr., 2025 Accepted: 28 May, 2025 Published: 07 Jun., 2025 Copyright © 2025 Liu and Huang, 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: Liu C.C., and Huang Y.H., 2025, Effects of growth regulators on root development and antioxidant capacity of Anoectochilus roxburghii tissue culture seedlings, Medicinal Plant Research, 15(3): 119-128 (doi: 10.5376/mpr.2025.15.0013) Abstract The wild resources of Anoectochilus roxburghii are becoming increasingly scarce, and breakthroughs are still needed in tissue culture rapid propagation in terms of root growth, and stress resistance. This study sorted out the effects of auxin (IBA, NAA), cytokinin (6-BA), novel regulatory factors, and light quality on root development and physiological functions. The results showed that, the rooting rate of MS+1.0 mg/L IBA+1.0 mg/L NAA was as high as 92%, and the average number of roots was 2.62. 1.0 mg/L 6-BA+0.05 mg/L NAA, can promote bud cluster formation. In proliferation culture, B5 medium and red light are more beneficial. The 1:1 combination of red and blue light can simultaneously optimize growth and polyamine metabolism, while blue light enhances the activities of SOD, POD and CAT. When the transplanting substrate is peat to vermiculite in a ratio of 3:1, the survival rate reaches 80%. Regulators can also promote the accumulation of flavonoids and polysaccharides, and form a synergistic effect with exogenous polyamines and strigolactone, enhancing antioxidant capacity and drought resistance. This study provides a technical path for the sustainable cultivation and improvement of medicinal quality of A. roxburghii, and also offers a reference for the rapid propagation and resource conservation of related medicinal plants. Keywords Anoectochilus roxburghii; Growth regulator; Tissue culture; Growth hormone; Antioxidant capacity; Medicinal quality 1 Introduction Anoectochilus roxburghii, also known as "Jewel Orchid", is a highly regarded medicinal and edible plant in traditional Chinese medicine, renowned for its efficacy in treating hypertension, diabetes, liver diseases, and as a tonic ingredient (Gam et al., 2020; Ye et al., 2020). Its pharmacological benefits are mainly attributed to its rich bioactive components, including flavonoids, kinsenoside and polysaccharides (Ye et al., 2017; 2020). However, excessive digging and slow natural reproduction rate have led to its endangered status, and conservation and sustainable cultivation efforts are urgently needed (Gam et al., 2020). Tissue culture has become an important technical means for the large-scale propagation and resource conservation of A. roxburghii. It can not only obtain high-quality and disease-free seedlings, but effectively preserve valuable germplasm resources (Gam et al., 2020; Zhang et al., 2025a). By optimizing specific growth regulators and medium formulations, researchers enhanced bud cluster proliferation, root induction efficiency, and transplanting survival rate, thereby simultaneously supporting commercial production and germplasm conservation (Zhang et al., 2025a). Root development is the foundation of plant health, and directly affects the absorption of nutrients and water, biomass accumulation and overall growth performance (Ye et al., 2020; Zhang et al., 2025a; Zhong et al., 2025). A well-developed root system, manifested as more lateral roots and root hairs, has been proven to contribute to enhancing plant vitality and the accumulation of secondary metabolites (Ye et al., 2020). Antioxidant capacity plays a role in the process of plants adapting to adverse conditions, mainly relying on the activities of antioxidant enzymes, like superoxide dismutase, catalase and peroxidase. These enzymes can alleviate oxidative stress, and enhance the stress resistance of plants in adverse environments, such as drought, salt stress and extreme temperature (Sun et al., 2023; Zhong et al., 2025). In A. roxburghii, the enhancement of
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