Medicinal Plant Research 2024, Vol.14, No.6, 334-344 http://hortherbpublisher.com/index.php/mpr 335 type, lighting conditions, and air humidity on seedling survival from multiple perspectives (Zhu, 2015; Chen et al., 2021). It has found that using a combination of red and blue LED light sources can promote plant growth, and increase the content of beneficial active ingredients in their bodies (Gam et al., 2020). And the selection of soil type is also crucial. If the soil properties used are similar to those in its natural habitat, it can better promote root development and enhance the water retention capacity of the substrate (Wang et al., 2022; Al Alwani et al., 2023). In the transplanting process, maintaining appropriate air humidity and taking reasonable shading measures can also help reduce the stress of external environment on plants, enable them to adapt to the new environment more smoothly as well (Zhang et al., 2020). However, although these improvement measures have shown positive effects in some studies, there are still studies indicating that they have not significantly improved the survival rate of seedlings. Thereby, it is currently unclear which method is most effective, and further research and verification are needed on response strategies under different environmental conditions. In this study, we want to figure that out. We will test different conditions during transplanting to see how they affect the growth and survival of A. roxburghii seedlings from tissue culture. Our goal is to find simple and useful ways to help more of them live. Even though we focus on A. roxburghii, our results may also help with transplanting other types of medicinal plants grown in a similar way. 2 Tissue Culture Propagation of Anoectochilus roxburghii 2.1 Tissue culture techniques for A. roxburghii Since A. roxburghii is a precious medicinal plant and its wild resources are becoming increasingly scarce, more and more people are beginning to propagate it artificially through tissue culture. This technology can not only quickly and safely reproduce plants in large quantities, which is beneficial to the protection of the species, but also promotes its industrial development. At present, most tissue culture experiments use Murashige and Skoog (MS) medium, a special medium containing a variety of nutrients. In order to improve its effect, researchers usually add plant growth regulators to the culture medium. Some studies have found that adding rare earth elements such as La(NO3)3 andCe(NO3)3 can promote the growth of buds and make seedlings stronger. But the amount of addition is crucial - the right amount is beneficial, but excessive amounts can harm the plants (Xu et al., 2016). The basic steps of tissue culture are relatively simple. The seeds are sterilized and then cultured in a sterile experimental environment. When the seeds germinate, healthy seedlings are selected and replicated in large numbers through asexual reproduction to obtain enough plants for large-scale cultivation (Chen and Liu, 2015). In order to further improve the development of buds and roots, researchers are also constantly optimizing culture conditions. Some studies have shown that using low-concentration MS culture medium and rationally regulating the content of plant hormones such as cytokinins and auxins can help promote the growth of buds and roots (Zhang et al., 2015). However, due to different experimental conditions, research results often vary, so a method that works in one laboratory may not work well in another laboratory. 2.2 Common issues affecting tissue-cultured seedlings' survival rate Although there have been some advances in tissue culture techniques for A. roxburghii, successfully transplanting seedlings from the laboratory into soil still faces many challenges. The main problem occurs when the seedlings are exposed to drastic changes in outdoor humidity and temperature after leaving the stable laboratory environment. If these environmental differences cannot be properly addressed, many seedlings may wither and die (Chen et al., 2017). Zou (2015) pointed out that selecting appropriate soil ratios, such as mixing fine sand with peat, can promote better root growth. This substrate is closer to the natural growth environment of A. roxburghii, which helps to improve the survival rate after transplantation. In addition to environmental stress, diseases are also a key issue. Seedlings cultured in tissue culture are particularly sensitive to external pathogens and are particularly susceptible to bacterial infections from unclean tools or materials (Zhang et al., 2021). Therefore, in the early stages of tissue culture and domestication, it is necessary to strictly ensure aseptic operation and finely control the culture environment.
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