Medicinal Plant Research 2025, Vol.15, No.2, 80-87 http://hortherbpublisher.com/index.php/mpr 85 7 Case Studies and Strategy Analysis 7.1 Successful cases of substrate selection for Anoectochilus roxburghii cultivation domestically and internationally Optimal substrate selection has been reported in most studies for Anoectochilus roxburghii culture. Locally, under experiments, one study showed that the use of peat and river sand substrate mixture in a 2:1 ratio significantly improved plantlet percentage survival and quality of A. roxburghii plantlets with the best performance at 3 cm x 3 cm plant density and 70% shading rate (Karimi et al., 2021). Internationally, in another study, peat with bark and other material at 14:1:5 ratio was effective with an excellent survival rate of 98.1% and excellent growing conditions. Perlite blended with peat soil in the ratio of 4:1 was also found to be effective for growth of A. roxburghii with survival up to 97.6%. 7.2 Strategies for integrating substrate selection and cultivation management Along with substrate choice, cultivation control includes optimizing the environment and substrate mix for improved plant development. An article recommended a peat, muck, and vermiculite mixture substrate in a 1:1:1 ratio for transplanting tissue culture plantlets in controlled environments for survival at 87.6% (Jiang., 2024). It also involves the alteration of the shade and nutrient solution concentration, as confirmed using the application of a 1/4 MS nutrient solution and 70% shading rate to result in an excellent improvement of plantlet growth and survival. In addition, there is a need for the provision of adequate environmental conditions such as temperature, pH, and humidity, as is the case with the successful culture of A. roxburghii under a temperature of 23°C to 28°C and relative humidity of about 90% (Ahmed et al., 2024). 7.3 Recommendations for applying substrate optimization based on this study According to the result of this study, a substrate mixture ratio of 2 parts river sand to 1 part peat is suggested to be used in Anoectochilus roxburghii tissue culture seedling growth. It has been established to possess a high survival rate and improved quality growth. Furthermore, using a density of plant of 3 cm x 3 cm and a 70% shading level, additional enhancement of plantlet development can be achieved. For large-scale cultivation, a sterile blend of peat soil and sand in a ratio of 2:1 with a cover layer of live moss is suggested to achieve a high survival rate of 90.2% (Xu et al., 2017). Optimization techniques of substrate, along with proper management of the provided environment, can boost the yields of A. roxburghii cultivation considerably. 8 Concluding Remarks The research on Anoectochilus roxburghii has substantiated that transplanting substrate selection significantly affects the quality of growth and seedling survival rate. Studies have verified that a peat and river sand mixture at a 2:1 ratio, combined with certain planting density and shading, yields high levels of survival and optimal conditions for growth. Besides, substrate mix comprising peat, bark, and plant material in the ratio 14:1:5 has been used with high success and a survival rate of 98.1% after 90 days. Incorporation of endophytic fungi into peat and vermiculite medium has also been proved to induce survival percentages as well as fresh and dry weights of the seedlings. Even with the promising results, there are restrictions to the study. The study inclines to investigate a limited series of substrate compositions without extensive cross-experimentation of potential materials and their reactions. Additional study should also consider long-term effects of these substrates on the active compounds of Anoectochilus roxburghii, such as the polysaccharides and flavonoids. Future research should seek to test broader substrate compositions and long-term action of these compounds on plant growth as well as biochemical composition. To ensure effective cultivation and industrialization of Anoectochilus roxburghii, optimizing the use of effective substrate combinations such as peat and river sand or peat and bark that have been previously proven to ensure high survival and healthy growth is recommended. In addition, inclusion of endophytic fungi in cultivation will further enhance plant vigor and yield. It is also essential that thorough research on the scalability and cost-effectiveness of such methods is undertaken so that they are effectively scaled up.
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