Organic solvents

2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), Ellman’s reagent—5,5′-dithiobis(2-nitrobenzoic acid) (DTNB), sodium dodecyl sulfate (SDS), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), 2-thiobarbituric acid (TBA), 1,1,3,3-tetramethoxypropane, phenolic compounds and organic solvents of HPLC grade were purchased from Sigma-Aldrich (Steinheim, Germany). All other chemical reagents and solvents used in this study were analytical grade and obtained from POCh (Gliwice, Poland). All agars for microbiological analyses were purchased from Yongxin Biological Technology Co., Ltd. (Yixing, Jiangsu, China).

Certified analytical standards of mycotoxins (fumonisins B_1-3 and beauvericin), other reagents, water and organic solvents of high purify for the chromatographic analysis were purchased from Sigma-Aldrich (Steinheim, Germany). A small amount (0.20 g) of each plant root was extracted using 1.0 mL of methanol:water (3:1, v/v) by shaking on an orbital shaker (24 h) and by sonication for 20 min. After centrifugation (Eppendorf, Hamburg, Germany) at 15,000× g for 10 min, the extracts were filtered through a 0.22-μm membrane (Chromafil PET 20/15/MS, Macherey-Nagel, Germany) and transferred to chromatography vials. The mycotoxin concentration was analyzed for each variant of the plant material using an UPLC™ system (Acquity, Waters, Milford, MA, USA) connected with a photodiode array detector (PDA) and a triple-quadrupole mass spectrometer (TQD; Waters Micromass, Manchester, UK) according to the methods described in detail earlier [56 (link),57 (link)]. The samples were collected from biological triplicates, and qualitative and quantitative analyses of the mycotoxins were performed in three analytical replicates.

Organic solvents were from Sigma-Aldrich (St Quentin Fallavier France) and of 99% purity. Cholesterol, POPC and POPS were from Avanti Polar Lipids (Birmingham, AL).

Organic solvents used for extraction were purchased from Sigma Aldrich (Castle Hill, NSW, Australia). Other chemicals and standards of analytical grade or higher were also sourced from Sigma Aldrich, such as 2,2′-diphenyl-1-picrylhydrazyl (DPPH), 2′-azino-di-(3-ethylbenzthiazoline sulfonic acid) (ABTS), 3-(2-Pyridyl)-5,4,4-dimethoxybenzaldehyde,4-triazine-p,5,6-diphenyl-1,6-hydroxy-2,6-Tris(2-pyridyl)-s-triazine (TPTZ), 7,8-tetramethylchroman-2-carboxylic acid (Trolox), aluminium trichloride, anhydrous sodium carbonate, catechin hydrate, disodium ethylenediaminetetraacetic acid (EDTA-Na₂), ferrous chloride, Folin–Ciocalteu reagent, gallic acid monohydrate, phloroglucinol, potassium persulfate, p’-disulfonic acid monosodium salt hydrate (Ferrozine), sodium acetate, trisodium phosphate, and quercetin. HPLC grade standards including phloroglucinol, gallic acid, chlorogenic acid, syringic acid, synaptic acid, catechin, epicatechin, and epigallocatechin were purchased from Sigma Aldrich (Castle Hill, NSW, Australia). Vanillin was obtained from Chem-Supply Pty Let., Adelaide, SA, Australia. The Milli-Q water used was obtained from Millipore Milli-Q Gradient Water Purification System (Darmstadt, Germany).

All chemicals, reagents, and organic solvents of the highest grade available were purchased from Sigma-Aldrich (St. Louis, MO, USA) unless otherwise stated. Trifluoroacetic acid (TFA) was supplied by Romil Ltd. (Cambridge, UK). Ultrapure water was prepared by an Arium 611 VF system from Sartorius (Göttingen, Germany). Mass grade solvents used for medium-sized peptides were purchased from VWR International (Milan, Italy). Optima LC-MS grade water and acetonitrile (ACN), used for short peptide analysis, were purchased from Thermo Fisher Scientific (Waltham, MA, USA).

Leaf tissue from plants grown in 2019, 2020, and 2021 field experiments was harvested at market maturity, weighted, lyophilized, and shipped to Rutgers University (New Brunswick, NJ, USA) for flavonoid quantification. Laboratory analyses were performed using analytical standards >98% purity quercetin 3-glucoside (16654), cyanidin chloride (94099), and ≥97% purity kaempferol (60010) purchased from Sigma (St. Louis, MO, USA), PTFE filters (0.45 μm, 89041-306) and tert-butylhydroquinone purchased from VWR (Radnor, PA, USA), and organic solvents purchased from Sigma Aldrich (St Louis, MO, USA).