Efficient Production of Isoflavonoids by Astragalus membranaceus Hairy Root Cultures and Evaluation of Antioxidant Activities of Extracts

Abstract

In this study, Astragalus membranaceus hairy root cultures (AMHRCs) were established as an attractive alternative source for the efficient production of isoflavonoids (IF). A. membranaceus hairy root line II was screened as the most efficient line and was confirmed by PCR amplification of rolB, rolC and aux1 genes. Culture parameters of AMHRCs were systematically optimized, and five main IF constituents were quali-quantitatively determined by LC-MS/MS. Under optimal conditions, the total IF accumulation of 34 day old AMHRCs was 234.77 μg/g dry weight (DW). This yield was significantly higher compared to that of 3 year old field grown roots (187.38 μg/g DW). Additionally, in vitro antioxidant assays demonstrated that AMHRC extracts exhibited antioxidant activities with lower IC50 values (1.40 and 1.73 mg/mL) as compared to those of field grown roots (1.96 and 2.17 mg/mL). Overall, AMHRCs may offer a promising and continuous product platform for naturally derived, high quality and valuable nutraceuticals.

Cannabis sativa: The Plant of the Thousand and One Molecules

“…In this study hypocotyls were found to be the most responsive tissue for infection. The hairy root system is very interesting for the production of secondary metabolites in medicinal plants ( Jiao et al, 2014 ; Patra and Srivastava, 2014 ; Wawrosch et al, 2014 ; Gai et al, 2015 ; Tian, 2015 ) or to engineer model plants to secrete industrially valuable metabolites. For example, in tobacco transgenic hairy roots the production of THCA was successfully obtained by expressing hemp THCAS ( Sirikantaramas et al, 2007 ).…”

RolB gene-induced production of isoflavonoids in transformed Maackia amurensis cells

“…T-DNA carries the rol genes (rolA, rolB, rolC, and rolD), which together or separately cause the hairy root syndrome in host plants at sites of infection (White et al 1985). Transgenic hairy roots were shown to stably produce secondary metabolites at levels comparable to or greater than those of the intact plants (Swain et al 2012;Oksman-Caldentey and Hiltunen 1996;Giri and Narasu 2000;Grzegorczyk et al 2006;Tiwari et al 2008;Sheela 2012;Jiao et al 2014;Bansal et al 2014;Gai et al 2015).…”

Direct determination of astragalosides and isoflavonoids from fresh Astragalus membranaceus hairy root cultures by high speed homogenization coupled with cavitation-accelerated extraction followed by liquid chromatography-tandem mass spectrometry

“…8,9 Therefore, the mobile phase composition was simplied using an acetonitrile-water mixture for all analytes in this work. Optimization of LC conditions.…”

“…Optimization of LC conditions. 9 For the simultaneous determination of all target analytes, the ESI-MS/MS with negative-ion mode was eventually chosen to analyze the eleven target compounds in spite of sacricing the detection sensitivity of astragalosides. According to our previous report, the acetonitrile-water mobile phase is the best choice for the separation and ionization of astragalosides and isoavonoids.…”

“…[1][2][3][4] However, the limited supply (endemic plant species for East Asia) together with over-exploitation constitutes the most important hurdles for developing Radix astragali-based drugs or dietary supplements. [6][7][8][9] Astragalosides and isoavonoids, the principal active ingredients of Radix astragali, possess versatile biological activities as diverse as anti-tumor, cardiotonic, immunomodulatory, neuroprotective, antiviral, antioxidant, anti-inammatory, antiperspirant, hepatoprotective, antihypertensive, antifatigue, and anti-osteoarthritis effects. 4 In this context, A. membranaceus hairy root cultures (AMHRCs) generated from the genetic transformation of Agrobacterium rhizogenes, have emerged as attractive alternatives to produce active compounds effectively, economically and in an environmentally friendly way.…”