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<p><i>Hedychium spicatum</i> has gained prominence in recent years for its terpenoid-rich essential oil, which is widely used as a flavoring and coloring agent in agricultural applications, cosmeceuticals, and pharmaceutical formulations, thereby significantly enhancing its market value. However, the availability of the species in the wild is decreasing. Therefore, the present study aimed to optimize terpenoid-rich essential oil production through in vitro callus culture followed by the establishment of a cell suspension culture. Callus was induced from early-stage germinated seeds (2 days old) on solid MS medium and subsequently used to develop a cell suspension culture in liquid medium. Five macro and micronutrient combinations with plant growth regulators (PGRs, 2.5 µM thidiazuron, and 3 µM 1-naphthaleneacetic acid) were used to optimize the Murashige and Skoog (MS) medium concentration. Phytochemical analysis was performed on the 30<sup>th</sup> day during the stationary growth phase of the cell suspension culture. The highest biomass (15.74 g FW) was achieved, along with maximum yields of phenolic (8.71 mg GAE/g DW), tannin (19.1 mg TAE/g DW), flavonoid (9.91 mg QE/g DW), antioxidant (ABTS activity - 25.25 mg AAE/g DW, and DPPH - 39.4 mg AAE/g DW) in optimized MS medium concentration. Comparative analysis indicated a higher relative abundance of major monoterpenes in the optimized medium, including 1,8-cineole (13.28%), sabinene (9.96%), terpinen-4-ol (7.79%), and linalool (5.04%). Sesquiterpenes such as caryophyllene (10.2%) and β-farnesene (9.24%) also exhibited higher relative proportions in the optimized medium, whereas α-cadinol (9.53%) and α-eudesmol (3.72%) were relatively more abundant in the basal medium (control). This method ensures superior and consistent yields with enhanced quality throughout the year while minimizing dependence on wild-harvested rhizomes.</p>

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Optimization of MS medium concentration for callus-derived cell suspension culture of Hedychium spicatum for the production of terpenoid, phenolic, flavonoid, and antioxidant activity using response surface methodology

  • 期刊论文

  • 发表日期：

    2026年1月31日

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  • Vibhash Dhyani,
  • Indra D. Bhatt,
  • Veena Pande

摘要

Hedychium spicatum has gained prominence in recent years for its terpenoid-rich essential oil, which is widely used as a flavoring and coloring agent in agricultural applications, cosmeceuticals, and pharmaceutical formulations, thereby significantly enhancing its market value. However, the availability of the species in the wild is decreasing. Therefore, the present study aimed to optimize terpenoid-rich essential oil production through in vitro callus culture followed by the establishment of a cell suspension culture. Callus was induced from early-stage germinated seeds (2 days old) on solid MS medium and subsequently used to develop a cell suspension culture in liquid medium. Five macro and micronutrient combinations with plant growth regulators (PGRs, 2.5 µM thidiazuron, and 3 µM 1-naphthaleneacetic acid) were used to optimize the Murashige and Skoog (MS) medium concentration. Phytochemical analysis was performed on the 30th day during the stationary growth phase of the cell suspension culture. The highest biomass (15.74 g FW) was achieved, along with maximum yields of phenolic (8.71 mg GAE/g DW), tannin (19.1 mg TAE/g DW), flavonoid (9.91 mg QE/g DW), antioxidant (ABTS activity - 25.25 mg AAE/g DW, and DPPH - 39.4 mg AAE/g DW) in optimized MS medium concentration. Comparative analysis indicated a higher relative abundance of major monoterpenes in the optimized medium, including 1,8-cineole (13.28%), sabinene (9.96%), terpinen-4-ol (7.79%), and linalool (5.04%). Sesquiterpenes such as caryophyllene (10.2%) and β-farnesene (9.24%) also exhibited higher relative proportions in the optimized medium, whereas α-cadinol (9.53%) and α-eudesmol (3.72%) were relatively more abundant in the basal medium (control). This method ensures superior and consistent yields with enhanced quality throughout the year while minimizing dependence on wild-harvested rhizomes.