In vitro production of phenolic compounds and antioxidant activity in callus suspension cultures of Habenaria edgeworthii: A rare Himalayan medicinal orchid

作者：Giri Lalit; Dhyani Praveen; Rawat Sandeep; Bhatt Indra D^*; Nandi Shyamal K; Rawal Ranbeer S; Pande Veena

来源：Industrial Crops and Products, 2012, 39: 1-6.

DOI：10.1016/j.indcrop.2012.01.024

• Antioxidant activity
• Callus culture
• Habenaria edgeworthii
• Methyl jasmonate
• Phenol

An in vitro method was developed for the production of phenolic compounds from callus suspension cultures of Habenaria edgeworthii Hook. f. ex. Collett, a rare Himalayan medicinal orchid. Friable and light yellow coloured callus was induced within eight weeks when seeds were cultured on Murashige and Skoog (MS) or 1/2 MS medium supplemented with 1 mu M alpha-naphthaleneacetic acid (NAA). In order to increase the biomass, following another 2 weeks these calli were repeatedly subcultured (4 week interval) onto 1/2 MS liquid medium supplemented with 0.1 mu M 6-benzyladenine (BA).
The effect of BA (0.0-3.0 mu M) and methyl jasmonate (MeJA; 10-1000 mu M) treatment on callus growth, total phenolic content and corresponding antioxidant activity was also determined following 4 weeks of culture on 1/2 MS medium (liquid). Maximum callus biomass (2.61 g fresh weight) was obtained in medium containing 1.0 mu M BA. The total phenolic content varied significantly (p < 0.05) in callus grown on BA, and ranged between 10.33 and 14.30 mg gallic acid equivalent (GAE) per g dry weight (DW) (compared to control 10.17 mg GAE per g DW). Antioxidant activity, measured by three in vitro assays, i.e., 2,2'- azinobis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging, 1,1-dipheny1-2 picrylhydrazyl (DPPH) radical scavenging and ferric reducing antioxidant power (FRAP) assays showed maximum activity in callus grown on 3.0 mu M BA containing medium. High performance liquid chromatography (HPLC) analysis showed the presence of high gallic acid content (143.63 mg/100 g DW) when compared to wild tuber (5.5 mg/100 g DW). The result indicate that callus grown on medium supplemented with 3.0 mu M BA showed high phenolic content along with increased antioxidant activity, however, callus growth was higher with 1.0 mu M BA. The result of this investigation has commercial applications as increased biomass production will yield active compounds, much needed by the pharmaceutical and neutraceutical industries.

• 出版日期2012-9