Medicinal Plant Research 2025, Vol.15, No.2, 80-87 http://hortherbpublisher.com/index.php/mpr 83 4 Effects of Transplanting Substrates on the Growth Quality of Tissue Culture Seedlings 4.1 Impact of different substrates on plant growth characteristics Anoectochilus roxburghii tissue culture seedling growth behavior is greatly reliant on transplanting media selection. Research has identified that peat and river sand media at a 2:1 ratio can result in maximum survival rates and improved parameters of growth such as plant height, stem diameter, and fresh weight (Ye et al., 2020). Additionally, peat, bark, and other materials substrate composition of 14:1:5 has been reported to be optimal for better growth conditions with 98.1% survival rate and mean plant height of 9.9 cm after 90 days (Li et al., 2020). These findings specify the role of substrate composition towards achieving the maximum growth quality of A. roxburghii seedlings. 4.2 Differences in root growth and biomass distribution The substrate also affects the root growth and biomass allocation of A. roxburghii seedlings. A 4:2:2 peat soil: river sand: peanut shell ratio has been found to enhance root growth, and the length of the longest root and the root diameter were significantly higher compared to other combinations of substrates. Such a substrate mix supports even-distributed biomass that results in better seedling vigor and overall seedling health. Moreover, with the use of a peat, muck, and vermiculite mix at a 1:1:1 proportion, there is indication that it can achieve a root rate of more than 95%, thus being effective in healthy root development (Zhang et al., 2015). 4.3 Relationship between substrates and accumulation of bioactive compounds The composition of bioactive compounds within A. roxburghii, such as polysaccharides, flavonoids, and polyphenols, varies according to substrate composition. Substrate ratio 4:2:2 had no major effect on polysaccharide content, while a 4:2:1 substrate ratio was observed to increase flavonoid content in A. formosanus. The 4:2:2 substrate ratio was also identified as being associated with increased content of polyphenols in A. formosanus and A. chapaensis, which suggests substrate can play a role in optimizing the drug value of the plant. The findings suggest that optimizing substrate blends could not just grow more in quality but optimize the pharmacology value of A. roxburghii (Gam et al., 2020). 5 Screening and Optimization of Transplanting Substrates 5.1 Optimization methods and experimental design for substrate formulation Substrate optimization of Anoectochilus roxburghii involves the testing of the various substrate mixtures to enhance the quality and survival rate of tissue culture seedlings. Trial experiments usually involve a change in the ratio of constituents such as peat, river sand, bark, and vermiculite. For instance, an experiment ascertained that a ratio of 2:1 of peat and river sand had optimum conditions for growth and high survival1. A single study indicated that a mix of 14:1:5 peat, bark, and other components was effective for 98.1% survival after 90 days (Gam et al., 2020). Orthogonal experiments in these cases are employed to determine the effects of different substrate components on growth parameters (Lei et al., 2017). 5.2 Comprehensive evaluation of growth quality across substrate combinations The quality of growth of Anoectochilus roxburghii in various substrate combinations is measured based on parameters including stem diameter, plant height, length of the roots, and fresh weight. With a combination of peat soil, river sand, and peanut shell in the ratio of 4:2:2, maximum plant height, stem diameter, and root length were observed. In addition, bark and peat substrates were also discovered as economically suitable substrates allowing for healthy growth, with significant variation in plant growth and survival rates. Such assessment is relevant in the determination of substrates that not only stimulate plant growth but also influence active compound concentrations such as flavonoids and polyphenols (Chen et al., 2021). 5.3 Economic and environmental benefits of selecting optimal substrates Optimum substrate choice for cultivation of Anoectochilus roxburghii has economic and ecological benefits. Economically, bark and peat substrates are low cost and readily available, therefore the best for large-scale production (Jin et al., 2018). Ecologically, high survival rate and plant quality are obtained through the use of substrates, hence doing away with chemical fertilizers and pesticides, therefore minimizing environmental
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