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Geosciences 2018, 8, 56 5 of 18 2. Materials and Methods 2.1. Reagents Na salt based chelators were used in this work and were of analytical grade. Disodium ethylenediaminetetraacetate dihydrate, EDTA disodium salt (C10H14N2Na2O8 · 2H2O) (E6635 Sigma Aldrich Ireland Ltd., Vale Road, Arklow, Wicklow, Ireland) was used to prepare all EDTA solutions at 0.05 M. This product is partially soluble in water at room temperature, the gradual addition of a concentrated NaOH solution was used to solubilise the salt. All EDDS solutions were made from a [SS]-EDDS trisodium salt solution in at 0.05M (C10H13N2Na3O8) (92698 Sigma Aldrich Ireland Ltd., Vale Road, Arklow, Wicklow, Ireland). Li solutions for soil spiking were prepared from LiCl salt (203637 Aldrich), ≥99.99% trace metals basis. Li (59916 Sigma Aldrich Ireland Ltd., Vale Road, Arklow, Wicklow, Ireland), K (96665 Sigma-Aldrich) and Na (02397 Sigma-Aldrich), standards (certified reference material), TraceCERT®, 1000 mg/L in HNO3 were used to prepare all working standards for FES determinations. Plants were fertilised using the commercially available fertiliser Miracle-Gro® (Scotts Miracle-grow, Marysville, Ohio, USA)) water soluble all-purpose plant food, Total N—24%, 3.5% ammoniacal nitrogen, 20.5% urea nitrogen, available phosphate—8%, soluble potash—16%, B—0.02%, Cu—0.07%, Fe—0.15%, Mn—0.05%, Mo—0.0005%, Zn—0.06%. Plant and soil digestions were carried out using, nitric acid (225711 Aldrich) (HNO3), 70%, purified by redistillation, ≥99.999% trace metals basis, hydrogen peroxide solution (95321 Sigma-Aldrich) (H2O2), ≥30%, for trace analysis and hydrochloric acid (435570 Sigma-Aldrich) (HCl) reagent grade, 37%. Glassware was thoroughly soaked in dilute HNO3 and rinsed several times with deionized water before use. HNO3 and H2O2 were used as matrix modifiers in all plant working standards. All reagents used were commercially available from Sigma Aldrich Ireland Ltd., Vale Road, Arklow, Wicklow, Ireland apart from Miracle-Gro® which is widely available. Soil used in these experiments was obtained from Westland Horticulture Ltd., 14 Granville Industrial Estate Granville Road Dungannon County Tyrone BT70 1NJ. 2.2. Germination Trials Seed germination trials were carried out to assess the total germination percentage of 34 plant species in the presence of Li and selected chelating agents. Most plants show tolerances to Li because of the similarity of the element to K and Na which are ubiquitous in all plants. Li also has comparable effects on the metabolic processes of plants to K and Na. Taking this into account the information obtained from these germination trials can only be viewed as general. The trials were conducted this way to narrow down the number of initial plant species to carry on to plant trials. The trials were carried out over 21 days at a temperature of 19 ± 1 ◦C. Test groups contained 10 replicates of 4 seeds, for each concentration of Li tested (n = 400 seeds per plant tested). Seeds were placed on cotton wool in a petri dish along with 4 mL of a specific concentration of Li solution. The Li solutions ranged from 20 mg/L to 1000 mg/L with controls. Thirty-four species of plants were selected for germination trials, firstly in the presence of Li and then in the presence of Li and EDTA and Li and EDDS. The plants used were: Brassica napus, Brassica oleracea var. capitata, Helianthus annuus, Solanum lycopersicum, Nicotiana tabacum, Lolium perenne, Pisum sativum, Vicia faba, Phaseolus coccineus, Hordeum vulgare, Avena sativa, Beta vulgaris, Daucus carota, Allium ampeloprasum, Mentha spicate, Cucumis melo, Spinacia oleracea, Brassica oleracea var. italica, Brassica oleracea var. gemmifera, Brassica oleracea gongylodes group, Brassica hirta, Brassica oleracea var. botrytis, Brassica rapa subsp. rapa, Brassica rapa subsp. chinensis, Salvia splendens, Brassica hirta, Raphanus raphanistrum, Brassica oleracea var. sabellica, Brassica oleracea borecole, Cardamine hirsuta, Nasturtium officinale, Eruca sativa, Lepidium sativum and Cardamine hirsuta. 2.3. Soil Preparation Li occurs in most soils in small quantities and was present in the soils used in these experiments. All soil used in these experiments was a commercially obtained top soil, consisting of a sieved dark brown/black rich clay loam soil with a high humus content. Ten 1 g soil samples from our bulk soilPDF Image | Induced Plant Accumulation of Lithium
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