Medicinal Plant Research 2025, Vol.15, No.2, 80-87 http://hortherbpublisher.com/index.php/mpr 82 substrates, for instance, pH and nutrient content, also plays a key role in seedling growth. A substrate pH of approximately 4.75 and suitable nutrient level is in favor of beneficial growth for A. roxburghii (Zhang et al., 2015). 3.2 Effects of water retention and aeration of substrates on survival Aeration and water storage are essential factors to be considered for the survival of A. roxburghii seedlings. Mediums that find equilibrium between water storage and aeration, i.e., mixtures of peat and vermiculite, have been documented to lead to maximum survival by preventing waterlogging and providing an efficient oxygenation of roots (Xu et al., 2016). Use of substrates like bark and peat, with a capacity to provide aeration in addition to controlling water-holding ability, has been associated with survivability levels of 98.1% and higher in certain research reports (Wang et al., 2022). It is this equilibrium that has resulted in environments which will induce root disease to be shunned and instead promote roots to grow favorably. 3.3 Correlation analysis between substrate types and survival rates Different substrates have been reported to be linked with varying survival rates of A. roxburghii seedlings. Experiments revealed that substrates that contain peat, river sand, and peanut shell in some ratio have been reported to affect survival rates and growth patterns. For instance, the 4:2:2 (peat:river sand:peanut shell) proportion of substrates has been associated with optimal survival and best-growth factors such as plant height and root elongation (Zhu et al., 2019; Wang et al., 2022). Similarly, peat and bark in the proportion of the 14:1:5 ratio have been highly successful, with the survival rate being as high as 98.1%3. These findings highlight the importance of careful substrate composition choice to maximize tissue culture seedling survival and growth (Figure 1). Figure 1 The effect of different matrix on the growth of Anoectochilus roxburghii (Adopted from Wang et al., 2022)
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