Physiologia plantarum ·1 Jan 2026 ·Added 10 Nov 2026

Chromosomal Stability and Metabolomic Plasticity in a 65-Year-Old Panax ginseng Cell Culture: A Physiological Model of Long-Term In Vitro Adaptation.

E V Popova, A V Nosov, T M Tyurina, O N Prudnikova, M V Titova, A A Fomenkov, I E Kulichenko, E G Nikolaeva, N A Maslennikova, M S Borts, A M Nosov

DOI ↗

Abstract

The first plant cell cultures were established in the 1930-60s. Since then, they were widely used as research models and sustainable sources of bioactive phytochemicals. However, questions remain regarding the long-term stability of plant cell cultures during in vitro maintenance. This study explored the stability of the world's oldest (65-year) Panax ginseng cell culture, maintained by periodic subcultures since 1959 under stable conditions. Growth, cytology, chromosome number and the ability to produce ginsenosides of protopanaxadiol, protopanaxatriol, and oleanane groups were assessed periodically during the long-term maintenance, and in comparison with historical baseline data. The results revealed that chromosome range (30-90) and mean chromosome number (45 ± 14) of the 65-year-old cell culture remained stable and relative to those historically recorded (50-100 and 54.2 ± 4.5 respectively). By contrast, the ginsenoside profile shifted dramatically since culture establishment and the differences accumulated with the culture's age. The UPLC-ESI-TOF-MS analysis of 24 ginsenosides showed that the long-term cultured cells lost nearly all dammarane-type ginsenosides, with protopanaxadiol ginsenosides being the most affected. In contrast, oleanane-type ginsenosides such as Ro and chikusetsusaponin IVa were retained in substantial concentrations. As a result, the ginsenoside profiles of the 65-year-old cell culture differed significantly from those in rhizomes and roots of plantation-grown plants. Elicitation of cell culture with methyl jasmonate (25-100 μM) stimulated production of oleanane-type ginsenosides but failed to reactivate dammarane ginsenosides biosynthesis. These findings provide new insights into the age-related stability of cultured medicinal plant cells, in particular, revealing variable age-dependent performance among ginsenosides of different structural groups.