An in vitro slow-growth callus conservation strategy for several medicinal plants using response surface methodology and machine learning Ye-Won Kim, Han-Sol Lee, Hosakatte Niranjana Murthy, Hyoshin Lee, So-Young Park BMC Biotechnology, 2026 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 5 °C 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 (3–9%), MeJA (0-200 µM), and CTR (0–20 mg/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. 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 µM MeJA, and 20 mg/L CTR, while the highest TSSC was achieved using the species-specific proliferation medium supplemented with 9% sucrose and 200 µM MeJA. When the storage temperature was set to 15 °C 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. This study demonstrates that long-term callus conservation is closely associated with AC and TPC. Medium supplemented with sucrose 3%, 135 µM MeJA, and 20 mg/L CTR, followed by storage at 15 °C, 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.
Comprehensive transcriptomic and functional characterization of protoplast regeneration in Angelica gigas Nakai Han-Sol Lee, Jong-Eun Han, Donghwan Shim, Bae Young Choi, Hyunwoo Cho, Hosakatte Niranjana Murthy, So-Young Park BMC Plant Biology, 2026 Protoplasts that are isolated from various plant sources, including leaf mesophyll tissue, callus, will continue to have the ability to take part in cell wall regeneration, division, and expression of totipotency. In model systems like Arabidopsis thaliana, it has been thoroughly established. Meanwhile, the fate of protoplasts isolated from both embryogenic (EC) and non-embryogenic callus (NEC) in other plants is unknown. Thus, we conducted transcriptome analyses of protoplasts produced from both EC and NEC in Angelica gigas in the present investigation. To achieve this, three stages of RNA sequencing were carried out: (1) EC, (2) freshly isolated protoplasts (Pt), and (3) cells undergoing cell division (CD) during A. gigas in vitro protoplast regeneration. Different gene expression programs were identified across stages through transcriptome profiling, which highlighted early stress responses, transcriptional changes, and the acquisition of stem cell identity following protoplast isolation. A pre-existing stem cell-like condition was shown by the strong expression of WUSCHEL-RELATED HOMEOBOX5 (WOX5) and CUP-SHAPED COTYLEDON2 (CUC2) genes at the EC stage. Important genes linked to stress reactions and cellular transcriptional changes, as WOX13 and WOUND INDUCED DEDIFFERENTIATION1 (WIND1), were significantly elevated at the Pt stage. Additionally, during this phase, genes associated with the cell cycle, auxin and cytokinin signaling, and cell wall regeneration were also active, indicating a dynamic shift toward regaining stem cell identity. Key regulators of stem cell maintenance and proliferation, such as LATERAL ORGAN BOUNDARIES DOMAIN16 (LBD16) and ENHANCER OF SHOOT REGENERATION2 (ESR2), were then substantially expressed at the CD stage, encouraging the start of cell division. The dynamic regulation of embryogenesis-related genes, including SOMACTIC EMBRYOGENESIS RECEPTOR-LIKE KINASE 2 (SERK2), LBD29, ESR2, LBD16, D-TYPE CYCLINS (CYCD3-2), and WOX1, during protoplast culture was validated by real-time quantitative polymerase chain reaction (RT-qPCR). WOX5 and CUC2 genes were found to be significantly expressed at the EC stage, suggesting a pre-existing stem cell-like state. Important genes linked to stress reactions and cellular transcriptional changes, as WOX13 and WIND1, were clearly elevated at the Pt stage. The activation of genes linked to cell wall renewal, auxin and cytokinin signaling, and the cell cycle during this phase also suggested a dynamic transition toward regaining stem cell identity. Additionally, the findings previously reported show that the EC-derived protoplasts successfully underwent cell division and contributed to the development of somatic embryos. Although the NEC-generated protoplasts did not divide upon culture, these results show that embryogenic cells maintain their embryogenic potential even after protoplast isolation and culture.
Micropropagation of Salix species – a review Hosakatte Niranjana Murthy, Praveen Nagella, Kadanthottu Sebastien Joseph, Kee Yoeup Paek, So-Young Park, Yaser Hassan Dewir, Maria V. Titova, Elena V. Popova Plant Cell Tissue and Organ Culture, 2026
Preface Bioactive Compounds in Edible Flowers, 2026
Phytochemicals and Biological Activities of Flowers of Clitoria ternatea (Butterfly Pea) Shrinivas Lamani, Hosakatte Niranjana Murthy Bioactive Compounds in Edible Flowers, 2026 In recent years, many countries have witnessed a significant rise in the adoption of plant-based remedies, leading to a substantial increase in the global demand for herbal resources. This chapter explores Clitoria ternatea, a species with remarkable potential due to its bioactive constituents and diverse biological activities. Most of the unique secondary metabolites and their analogues identified in this plant are anthocyanins, including ternatins, preternatins, cyanidin, and delphinidin, along with phenolic acids, terpenoids, and phytosterols derived from the flowers. Altogether, 32 anthocyanins and 19 additional metabolites have been documented across various studies. Different extracts of C. ternatea have demonstrated a wide range of biological properties, notably antioxidant, antimicrobial, anti-inflammatory, antidiabetic, cytotoxic, and anticancer activities. To support young researchers investigating potential chemotherapeutic agents for the treatment and prevention of cancer and other diseases, this chapter focuses on the biological functions of C. ternatea, with particular emphasis on its anticancer and antidiabetic benefits.
INTEGRATION OF SALICYLIC ACID IN IN VITRO ROOTING MEDIUM IMPROVES PHOTOSYNTHESIS AND GROWTH DURING ACCLIMATIZATION OF PHILODENDRON BIPINNATIFIDUM SCHOTT EX ENDL. PLANTLETS Propagation of Ornamental Plants, 2020
In Vitro Propagation of Artemisia japonica Smita Shinde, Pooja R. Katewal, Divya D. Shanbhag, Kadanthottu Sebastian Joseph, H. N. Murthy Journal of Herbs Spices and Medicinal Plants, 2017
Safety issues of food ingredients from plant cell, tissue, and organ cultures: An explication Food Toxicology, 2016
Atomic force microscopy studies for antibacterial activity of Garcinia xanthochymus seed extracts International Journal of Pharma and Bio Sciences, 2016
Characterization of nutritional constituents of Garcinia morella seeds and seed oil International Food Research Journal, 2016
Atomic force microscopic study for the antibacterial study of Garcinia Xanthochymus hook. F. leaf extract International Journal of Pharmacy and Pharmaceutical Sciences, 2015
Effect of mutagens on the in vitro adventitious shoot growth and bacoside a accumulation in Bacopa monnieri (L.) International Journal of Pharma and Bio Sciences, 2012
The effects of gamma and ethylmethanesulphonate treatments on agronomical traits of niger (Guizotia abyssinica Cass.) African Journal of Biotechnology, 2009
Direct somatic embryogenesis and plantlet regeneration from leaf explants of niger, Guizotia abyssinica (L.f.) Cass. Indian Journal of Experimental Biology, 2000
Plant regeneration from leaf cultures of Guizotia abyssinica (Niger) and Guizotia scabra Phytomorphology an International Journal of Plant Morphology, 1998
