Elite chemotype selection, antipathogenic activities, secondary metabolite fingerprinting of in vitro regenerated Tinospora cordifolia (Willd.) Hook.f. & Thomson – a plant with multipurpose therapeutic significance

Tinospora cordifolia (Willd.) Hook.f. & Thomson is an ethnomedicinal plant belonging to the family Menispermaceae, known for its multi-disciplinary use. The present study is focused on selecting an elite plant chemotype through phytocompound quantification and antimicrobial activity assay against multi-drug-resistant (MDR) urinary-tract-infecting (UTI) pathogens, being a growing concern for mortality and antimicrobial resistance nowadays. Twelve accessions of this plant were collected from different agro-climatic zones of West Bengal, and crude extraction was done by solvent optimization. Sample TC 07 resulted in maximum crude extract yield in methanol. All twelve sample extracts showed antimicrobial efficacy against eight pathogens producing significant inhibition zones, while TC 07 resulting maximum inhibition zone of 16.37 ± 0.12 mm against Staphylococcus aureus. Extracts were evaluated through high-performance thin-layer chromatography, which resulted in optimum amount of berberine (5.05 ± 0.05 mg/g) and palmatine (3.00 ± 0.11 mg/g) in TC 07, significantly higher compared to other accessions. Therefore, comparing the data, TC 07 was considered as the elite chemotype and introduced in in vitro culture for conservation. The maximum number of shoot (16.39 ± 0.09) production was obtained in MS medium containing 2.0 mg/l 6-Benzylaminopurine and 1.0 mg/l meta-Topolin. The plant was acclimatized in ex-vitro condition and confirmed for optimum bioactive compound production through high-performance liquid chromatography. The cytogenetic stability of the regenerated plantlets was ensured using chromosome number determination, and through start codon targeted (SCoT) polymorphism. Therefore, this study provides a simple, validated and reproducible method for optimal berberine and palmatine production and controlling MDR-UTI pathogens.