Optimal inductive and cultural conditions of Polygonum multiflorum transgenic hairy roots mediated with Agrobacterium rhizogenes R1601 and an analysis of their anthraquinone constituents

Abstract

Background:Polygonum multiflorum is an important medicinal plant. Hairy roots systems obtained by transforming plant tissues with the natural genetic engineer Agrobacterium rhizogenes can produce valuable biological active substances, which have immense potential in the pharmaceutical industry.Objective:To optimize the inductive and cultural conditions of P. multiflorum hairy roots and to identify the major active secondary metabolites in hairy roots.Materials and Methods:P. multiflorum hairy root were mediated with A. rhizogenes R1601 to induce hairy roots. Four combinations, including Murashige–Skoog (MS), 1/2 MS, B5, and White, were investigated to optimize the culture medium. MS medium was selected for the growth measurement. The qualitative and quantitative determinations of free anthraquinone in hairy roots were compared with the calli and aseptic plantlets using high-performance liquid chromatography.Results:The inductive rates of hairy roots by leaves were higher than for any other explants. The presence of agropine in the P. multiflorum hairy roots confirmed that they were indeed transgenic. MS medium was the most suitable of the four media for hairy root growth. Meanwhile, the growth kinetics and nutrient consumption results showed that the hairy roots displayed a sigmoidal growth curve and that their optimal inoculation time was 18-21 days. The determination of the anthraquinone constituents indicated that the rhein content of the hairy roots reached 2.495 μg g−1 and was 2.55-fold higher than that of natural plants.Conclusion:Transgenic hairy roots of P. multiflorum could be one of the most potent materials for industrial-scale production of bioactive anthraquinone constituents.

Jasmonates promote enhanced production of bioactive caffeoylquinic acid derivative in Eclipta prostrata (L.) L. hairy roots

“…Obtained results indicated the 21st day of culture growth as the most effective day for starting elicitation. The growth profile of E. prostrata (L.) L. hairy roots resemble Polygonum multiflorum hairy roots where it was found that the ideal time for the induction of anthraquinones was between the 18th and 21st days of elicitation (Huang et al 2014 ). Similarly, studies have reported the 21st day of elicitation as the end of the exponential growth phase of Artemisia annua hairy roots (Sivakumar et al 2010 ).…”

“…Eclipta prostrata (L.) L. produces phenolic acids such as chlorogenic acid and its derivative 3,5-di- O -caffeoylquinic acid (3,5-diCQA) (Lee et al 2010 ). Chlorogenic acid and its derivatives have demonstrated several biological activities, such as antioxidant, antibiosis, anti-inflammatory, antivirus, and antitumor activities (Huang et al 2014 ; Ali et al 2017 ; Liu et al 2018 ; Devrnja et al 2020 ). More recently, studies by molecular docking using a serine-type protease (Mpro), a SARS-CoV-2 virus polyprotein, showed that the obtained binding energies of 3,5-diCQA were closed to remdesivir (antiviral compound), indicating stronger binding to Mpro protein and promising antiviral compounds that could treat Covid-19 (Shah et al 2021 ).…”

Discovery and resupply of pharmacologically active plant-derived natural products: A review

“…As with cell cultures, secondary metabolite production can often be enhanced through optimized growth conditions or by using elicitors, and through metabolic engineering by transferring heterologous genes with engineered A. rhizogenes strains (Georgiev et al, 2012). Recent research on the production through hairy roots of lignans (Wawrosch et al, 2014), steroids (Doma et al, 2012), anthraquinones (Huang et al, 2014), or alkaloids (Pandey et al, 2014) clearly indicates that optimized hairy root culture systems can be used for the sustainable biotechnological production of biologically active compounds. Regarding the commercial application, the Swiss company ROOTec Bioactives AG (http://www.rootec.com) has developed an optimized bioreactor system for large-scale cultivation of hairy roots with potential to produce a number of pharmacologically active compounds (Table 6).…”

In vitro plant tissue culture: means for production of biological active compounds

“…Some of the advantages provided by hairy root cultures include high growth rates without the need for plant growth regulators, genetically and biochemically stable cultures, and a similar capacity for production of secondary metabolites than cell suspension cultures (Guillon et al 2006;Georgiev et al 2012;Mora-Pale et al 2014). A wide range of natural products has been produced using this system, including lignans, steroids, anthraquinones and alkaloids (Doma et al 2012;Huang et al 2014;Pandey et al 2014;Wawrosch et al 2014). However, metabolite 1 3 production requires that the compound of interest be one that is normally synthesized within the roots of the source plant, so this limits the versatility of the hairy root system (Ochoa-Villarreal et al 2016).…”