Light-induced production of antidepressant compounds in etiolated shoot cultures of Hypericum hookerianum Wight & Arn. (Hypericaceae)

Abstract

Hypericum hookerianum is a lesser known ethnomedicinal plant having wound healing, antitumor and anti-HSV-1 properties. Isolated nodes of in vitro shoots sub-cultured in the dark for 4 weeks on half strength Murashige and Skoog medium solidified with Gelzan (1.5 g l−1), and supplemented with 2.325 μM kinetin produced 8.0 ± 0.40 etiolated shoots of 5.0 ± 0.62 cm length at 74 % efficiency versus 9.2 ± 0.6 healthy shoots of 4.4 ± 0.5 cm obtained from nodes in light at 96 % efficiency. Low concentrations of hypericin were found in wild plant [0.35 ± 0.09 mg g−1 dry weight (DW)] and control green shoot cultures (0.91 ± 0.03 mg g−1 DW). Etiolated shoots exposed to a 12 h photoperiod (50 μmol m−2 s−1) through 1–25 days turned red incrementally due to synthesis and accumulation of 0.1–3.83 mg g−1 DW hypericin in sub-epidermal cortical cells of the stem and varied shaped cells of the distorted mesophyll. Flavonoid and anthocyanin concentrations of the etiolated shoots subjected to the 12 h photoperiod were 3–5 fold higher than the control shoot cultures while total chlorophylls [1.97 ± 0.05 mg g−1 fresh weight (FW)] of the light exposed shoots were significantly less compared to the control (2.86 ± 0.18 mg g−1 FW) and natural plant (6.82 ± 0.29 mg g−1 FW). HPLC analysis of shoot extracts revealed the presence of 0.14 ± 0.03, 0.16 ± 0.02 and 1.45 ± 0.16 mg g−1 DW hyperforin in wild plant, control shoot cultures and etiolated shoot cultures illuminated for 25 days, respectively. Despite a reasonable presence in etiolated shoots (0.61 ± 0.15 g−1 FW), total phenols did not increase significantly during illumination. The results indicate light induced synthesis of anti-depressant phenolic derivatives (hypericin, hyperforin and flavonoids) in etiolated shoot cultures of H. hookerianum.