ItemArticle

An in vitro slow-growth callus conservation strategy for several medicinal plants using response surface methodology and machine learning

Title
An in vitro slow-growth callus conservation strategy for several medicinal plants using response surface methodology and machine learning
Creator
Kim, Ye-Won; Lee, Han-Sol; Murthy, Hosakatte Niranjana; Lee, Hyoshin; Park, So-Young
Description
Background: In vitro culture of callus is an effective method for conserving the genetic resources of economically important crops. However, continuous subculturing is a costly and labor-intensive process. Therefore, establishing an efficient in vitro long-term conservation system applicable to various plant species is required. In this study, calli derived from five medicinal plant species, Camellia japonica (Cj), Centella asiatica (Ca), Ligusticum afficinale (Lo), Panax ginseng (Pg), and Sageratia thea (St) were used to optimize storage conditions and establish a suitable in vitro conservation strategy. Calli cultures were maintained on the appropriate culture medium at 5C for 120 days. Cell viability and regrowth rate were assessed during the storage period, and correlations between growth and antioxidant traits were examined. Subsequently, pretreatment optimization using sucrose (39%), MeJA (0-200 M), and CTR (020mg/L) was performed using RSM, and the effects of pretreatment and storage temperature on callus conservation were evaluated. In addition, machine learning models such as GRNN, MLP, RF, SVR, and XGBoost were applied to the experimental data. Results: The findings demonstrated that, in comparison to Ca and St, Lo, Pg, and Cj exhibited noticeably higher antioxidant capacity while maintaining high cell viability and regrowth rates. Interestingly, Ca and St drastically decreased viability and regrowth after 60 days, whereas Lo, Pg, and Cj maintained viability and regeneration for up to 90 days. Both TPC and AC (measured by FRAP assay) showed a high positive correlation with cell viability and growth rate, according to correlation analysis. RSM predicted that the optimal pretreatment medium for enhancing antioxidant capacity was the species-specific proliferation medium supplemented with 3% sucrose, 135 M MeJA, and 20mg/L CTR, while the highest TSSC was achieved using the species-specific proliferation medium supplemented with 9% sucrose and 200 M MeJA. When the storage temperature was set to 15C following the antioxidant-enhancing pretreatment derived from the RSM optimization, all five species showed improved cell viability and regrowth rates, among the storage methods. Among the ML models tested, XGBoost demonstrated the most stable and accurate predictive performance for both viability and regrowth during in vitro conservation. SHAP-based analysis of the XGBoost model, focusing on regrowth rate, revealed that storage duration was the most influential factor affecting regrowth prediction, followed by storage temperature, while pretreatment conditions showed secondary but meaningful contributions. Conclusions: This study demonstrates that long-term callus conservation is closely associated with AC and TPC. Medium supplemented with sucrose 3%, 135 M MeJA, and 20mg/L CTR, followed by storage at 15C, significantly improved viability and regrowth, and calli could be maintained up to 120 days without subculturing. This approach provides an efficient and broadly applicable in vitro strategy for the conservation of diverse plant genetic resources. The Author(s) 2026.
Source
BMC Biotechnology;Volume;26;Issue;1;Article No.;49;
Date
01-01-2026
Publisher
BioMed Central Ltd
Subject
Antioxidant capacity; Citric acid; In vitro conservation; Methyl jasmonate; Plant cell culture; Response surface methodology
Coverage
Kim Y.-W., Department of Horticultural Science, Division of Animal, Horticultural and Food Sciences, Chungbuk National University, Cheongju, 28644, South Korea; Lee H.-S., Department of Horticultural Science, Division of Animal, Horticultural and Food Sciences, Chungbuk National University, Cheongju, 28644, South Korea; Murthy H.N., Department of Horticultural Science, Division of Animal, Horticultural and Food Sciences, Chungbuk National University, Cheongju, 28644, South Korea, Department of Life Sciences, Christ University, Bengaluru, 560029, India; Lee H., Department of Horticultural Science, Division of Animal, Horticultural and Food Sciences, Chungbuk National University, Cheongju, 28644, South Korea; Park S.-Y., Department of Horticultural Science, Division of Animal, Horticultural and Food Sciences, Chungbuk National University, Cheongju, 28644, South Korea
Rights
All Open Access; Gold Open Access; Green Open Access
Relation
ISSN: 14726750; CODEN: BBMIE
Format
online
Language
English
Type
Article

Citation

Kim, Ye-Won; Lee, Han-Sol; Murthy, Hosakatte Niranjana; Lee, Hyoshin; Park, So-Young, “An in vitro slow-growth callus conservation strategy for several medicinal plants using response surface methodology and machine learning,” CHRIST (Deemed To Be University) Institutional Repository, accessed June 18, 2026, https://archives.christuniversity.in/items/show/23199.

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