1 1 azobis

Standards 2,3,7,8 TCDD and PCB-1 (2-monochlorobiphenyl), PCB-28 (2,4,4-tri-chlorobiphenyl), PCB-101 (2,2,4,5,5-pentachlorobiphenyl) and PCBs-77 (3,3′,4,5′-tetrachlorobiphenyl), and the organic solvents (AR grade) were purchased from Riedel-de-Haën (Seelze-Hannover, Germany). Pesticide quality solvents (n-hexane and chloroform) were purchased from Panreac (Barcelona, Spain). Chemicals for the synthesis of molecularly imprinted polymers (MIPs), which include, functional monomer methacrylic acids (100 g, 98% purity), porogen solvent chloroform anhydrous (1 L, GC grade 99%), Standard of dioxin (2,3,7,8 TCDD) (1 mL, 97.5% purity) which was used as a template, 1,1′^- Azobis (cylohexanecarbonitrile) (25 g, 98%) used as an initiator, cross linker ethylene glycol dimethacrylate (25 g, 98%), acetic acids (2.5 L, 100%), methanol (1 L, 99.8%), were all purchased from Sigma-Aldrich (St Louis, MO, USA). Nitrogen gas for purge was purchased from AFROX (Johannesburg, South Africa).

Seedlings of A. virginicus L. (2- to 3-week-old) were transferred to fresh 0.5 mM CaCl₂ solution (adjusted to pH 4.2) including 0 or 300 μM AlCl₃ for 8 or 24 h to investigate their gene-expressions. Young seedlings were also exposed to 35 μM CdCl₂, 50 μM CuSO₄, 300 μM PbCl₃, 200 μM ZnCl₂, 1.5 mM diamide (DM; 1,1′-Azobis (N,N-dimethyl formamide), Sigma-Aldrich, USA) or 1.5 mM H₂O₂ in 0.5 mM CaCl₂ solution at pH 5.7 for 8 h to determine gene-expression in each condition. Seedlings of A. thaliana grown in 1/6 MS medium (10-days to 2-weeks-old) were also exposed to 300 μM AlCl₃ in a fresh 1/6 MS medium (adjusted to pH 4.2) for 2 days or to 35 μM CdCl₂, 50 μM CuSO₄, 300 μM PbCl₃, 200 μM ZnCl₂, 1.5 mM DM or 1.5 mM H₂O₂ in a fresh 1/6 MS medium (adjusted to pH 5.7) for 2 days. Root length was randomly measured for more than 20 plants in each condition to calculate the relative root growth which was shown previously (Ezaki et al., 2000 (link)).

The monomer ethylene glycol dimethacrylate (EGDMA), the initiator 1,1′‐azobis (cyclohexanecarbonitrile) (ACHN), and the chain transfer agent 2‐Cyanopropan‐2‐yl benzodithioate (CPBD) were purchased from Sigma‐Aldrich (Sigma‐Aldrich, Munchen, Germany). The solvent toluene (HPLC grade, Sigma‐Aldrich) and precipitation agent n‐hexane (ACS reagent grade, Sigma‐Aldrich) were used as received. Anodized aluminium oxide porous membranes with an inner pore diameter of ~200 nm (Whatman, Maidstone, UK) were purchased from Sigma‐Aldrich. The photo‐initiator (2‐hydroy‐2‐methylpropiophenone) and dye (rhodamine 6G) were also purchased from Sigma‐Aldrich.