Medicinal Plant Research 2024, Vol.14, No.6, 334-344 http://hortherbpublisher.com/index.php/mpr 340 evidenced by increased root number, elevated biomass, and increased number of new shoots (Figure 2). A strong root system is essential for transplanting of tissue culture seedlings, as a good root structure improves water and nutrient uptake, thereby increasing the plant's ability to adapt to changes in the external environment. In addition, the changes in leaf color indicated that Ceratobasidium sp. AR2 induced the accumulation of flavonoids in A. roxburghii. These secondary metabolites may play an important role in improving antioxidant capacity and enhancing stress tolerance, which may help the plants to better adapt to environmental stresses after transplanting. Figure 2 A. roxburghii inoculated with AR2 and uninoculation. (A) The semi-thin section of root of A. roxburghii after 1 week of symbiotic cultivation; arrows represent peloton, scale bar = 50 mm. (B) A. roxburghii grown in tissue culture bottle for 4 months inoculated with AR2 (Section II) and uninoculation (Section I). Section I represents the upper plantlets of the medium for the control; Section II represents the upper plantlets of the medium for the treatment; scale bar = 1 cm. (C) Local morphology from the plantlet of the treatment group; arrows represent new buds, scale bar = 1 cm. (D) Contrast of leave color between the control group (the left) and treatment group (the right) after 4 months of culture, scale bar = 1 cm (Adopted from Zhang et al., 2020) This study provides a new strategy for the artificial cultivation of A. roxburghii, i.e., by adjusting the symbiotic microorganisms and transplanting environmental conditions, such as suitable soil substrate and water and fertiliser management, to improve the health status of the tissue culture seedlings and the survival rate of transplanting. In the future, the mutualistic mechanism between mycorrhizal fungi and A. roxburghii can be further explored and combined with environmental optimisation techniques to enhance the benefits of large-scale cultivation of A. roxburghii. 6.2 Commercial-scale transplantation and survival enhancement strategies On a commercial scale, the transplant success rate of Anoectochilus roxburghii has been significantly improved through various strategies. For example, Wang et al. (2022) enhanced the tissue culture efficiency of A. roxburghii by optimizing the induction, proliferation, and regeneration (IPR-PLB) system of protocorm-like bodies (PLBs). The study found that using a culture medium containing MS + 3 mg/L 6-BA + 0.5 mg/L NAA + 0.8 mg/L ZT + 0.2 mg/L 2,4-D increased the PLB induction rate to 89%, with a secondary PLB induction rate of 120% and proliferation rates (in terms of both quantity and biomass) reaching 400% and 350%, respectively (Figure 3). The experiments demonstrated that PLBs proliferate optimally under dark or low-light conditions, while light exposure facilitates their differentiation into complete plantlets. This study established an efficient IPR-PLB system, improving the micropropagation efficiency of A. roxburghii, contributing to the resolution of wild resource shortages, and promoting its industrial-scale cultivation.
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