Medicinal Plant Research 2025, Vol.15, No.2, 62-70 http://hortherbpublisher.com/index.php/mpr 65 45 °C is the optimal for suppressing most viral infections in plants. Longer exposure for extended durations at higher temperatures, for instance, 45 °C for 15-30 minutes, is able to enhance detoxification more effectively but may cause reduced survival rates of sensitive plant tissues (Torres et al., 2000). The duration of heat treatment of Lindera aggregata is also crucial. One must find a balance between the time for inactivation of the virus and avoid thermal stress. For example, in the thermotherapy of garlic, optimum detoxification was at 37 °C for 35 days (Lizárraga et al., 2017). 3.3 Effects of heat treatment onlindera aggregata growth Although heat treatment has been proven to reduce viral loads greatly, it may have some effect on Lindera aggregata development and vigor. Excessive exposure to high temperature may lead to reduced survival rates of seedlings and delayed growth. Studies in other plant species showed that the intensity and duration of heat treatment can be calibrated precisely to reduce these side effects. For instance, water treatments under heat and follow-up tissue culture have been used for vigour restoration and viability maintenance in treated plants (Langens-Gerrits et al., 2004). In Lindera aggregata, combining mild heat treatment with employing fast propagation techniques, such as tissue culture, enables proper recovery of the plant. The two-approach method minimizes the adverse impacts of heat stress while effectively eliminating viruses, making way for disease-free healthy seedlings with an enhanced capability for growth. 4 Application of Tissue Culture Technology in Healthy Seedling Propagation 4.1 Principles and processes of tissue culture Tissue culture is an aseptic method where cells, tissues, or organs are propagated in regulated environmental conditions (Grout, 2017). For Lindera aggregata, meristem culture and shoot tip culture are the two prominent detoxification methods. These methods are based on the fact that meristematic tissues will have lesser opportunities of virus harbouring, and virus-free plants will be produced. The tissue culture process entails explant harvesting and sterilization, culture medium preparation, and differentiation induction and proliferation. Microelements, PGRs such as cytokinins (e.g., 6-benzylaminopurine [6-BA]), and auxins (e.g., naphthaleneacetic acid [NAA]) are all significant constituents to be incorporated in inducing shoot development and initiation (Espinosa-Leal et al., 2018). 4.2 Optimization of culture media Optimization of the culture medium composition is to be made for enhanced efficiency in Lindera aggregata tissue culture. The concentration of sucrose as a source of energy and agar as structural support for the plantlet are the most significant. Concentrations of these have been found to play a vital role in inducing growth, differentiation, and proliferation of the tissues in culture. Besides, some PGR concentrations, such as 6-BA for shoot and NAA for rooting, also enhance the efficiency of regeneration and quality of detoxified seedlings (Hussain et al., 2012). 4.3 Detoxification efficiency of tissue culture Tissue culture improves detoxification effectiveness significantly by regeneration of virus-free plants from meristematic tissues. The efficiency of removal of viruses by detection through reverse transcription PCR (RT-PCR) in pre- and post-tissue culture has established high levels of removal of viruses from treated crops. Tissue culture also facilitates the promotion of quick development in seedlings to allow for the production of healthy plants on a regular basis. In Lindera aggregata, higher rates of proliferation and survival of detoxified seedlings have been established through maximized tissue culture protocols (Thorpe, 2012). 5 Application of Combined Heat Treatment and Tissue Culture Technology 5.1 Implementation process of combined technology The combined use of heat treatment and tissue culture represents a promising detoxification process for Lindera aggregata. Heat-treated shoot tips or stem segments are utilized as beginning materials, where virus is inhibited or
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