# Nanoparticle Formulation Generated from DDGS and Its Anthraquinone Synthesis Elicitation in Rubia tinctorum Hairy Roots

## TL;DR

A nanoparticle formulation made from DDGS boosts anthraquinone production in Rubia tinctorum hairy roots without harming plant growth.

## Contribution

A novel nanoparticle formulation from DDGS is developed and shown to enhance anthraquinone synthesis in plant roots.

## Key findings

- DDGS-NPs increased anthraquinone production up to 9.3 µmol/gFW in hairy roots.
- The highest DDGS-NP concentration caused 18 µM extracellular AQ accumulation.
- Combining DDGS-NPs with methyl jasmonate maximized AQ accumulation.

## Abstract

A nanoparticle formulation was generated from distiller dried grains with solubles (DDGS), and its effect on the production of anthraquinones (AQs) was evaluated on Rubia tinctorum hairy roots. The DDGS material was washed with water and ethyl acetate to remove mainly the soluble organic/inorganic molecules and reduce the fat content, respectively, followed by an alkaline treatment to remove the polysaccharides. The resulting alkaline solutions were then lyophilized and redispersed in deionized water to generate a monodispersed nanoparticulate formulation (DDGS-NP) with a hydrodynamic diameter and zeta potential of 227 ± 42 nm and −53 ± 7 mV, respectively. The formulation demonstrated good colloidal stability over time, and sterilized DDGS-NPs maintained comparable physicochemical properties. The nanoparticles were enriched in protein fractions, unsaturated fatty acids, and orthophosphate anion components from DDGS, as determined by solid-state Nuclear Magnetic Resonance (NMR), X-ray photoelectron spectroscopy (XPS), organic elemental analysis (OEA), and inductively coupled plasma optical emission spectrometry (ICP-OES) techniques. The DDGS-NPs were tested at different concentrations on Rubia tinctorum hairy roots, in comparison to or in combination with methyl jasmonate (MeJ), for their capacity to induce the production of AQs. All DDGS-NP concentrations increased the production of specific AQs to 7.7 (100 mg L−1), 7.8 (200 mg L−1), and 9.3 µmol/gFW (500 mg L−1), with an extracellular AQ accumulation of 18 µM for the highest DDGS-NP concentration, in comparison with the control hairy roots (~2 µM AQ). The plant growth was not affected at any of the tested nanoparticle concentrations. Interestingly, the combination of DDGS-NPs and MeJ resulted in the highest extracellular AQ accumulation in R. tinctorum root cultures.