RESUMEN

Rhizosphere pH determines nutrient bioavailability, but this pH is difficult to measure. Standard pH tests require adding water to growth media. This dilutes hydrogen ion activity and increases pH. We used a novel, in situ, pointed-tip electrode to estimate rhizosphere pH without dilution. Measurements from this electrode matched a research-grade pH meter in hydroponic nutrient solutions. We then compared measurements from this electrode to saturated paste and pour-through methods in peat moss, coconut coir, and pine bark. The pointed-tip electrode was unable to accurately measure pH in the highly-porous pine bark media. Adding deionized water to the other media at container capacity using the saturated paste method resulted in a pH that was 0.59 ± 0.30 units higher than the initial in situ measurement at the top of the container. This increase aligns with established solution chemistry principles. Measurements of pH using the pour-through method were 0.38 ± 0.24 pH units higher than in situ measurements at the bottom of the container. We conclude that in situ pH measurements are not subject to dilution and are thus more representative of the rhizosphere pH than the saturated paste and pour-through techniques.

RESUMEN

Silicon (Si) in plant tissues reduces abiotic and biotic stress, but it is incorporated as silica (SiO2), which is difficult to solubilize for analysis. We modified an oven-induced tissue-digestion and analysis method to improve Si solubilization and validated its accuracy by quantifying the mass-balance recovery of Si from the hydroponic solution and plant tissues of cucumber (Cucumis sativus). Leaf, stem, and root tissues were dried, finely-ground, and digested in 12.5 molar sodium hydroxide at 95°C for 4 hours. Solutions were then acidified with 6 molar hydrochloric acid to achieve a pH below 2 for measurement of Si using the molybdate blue colorimetric method. Interference of phosphorus in the analysis was minimized by increasing the addition of oxalic acid from 0.6 to 1.1 molar. We recovered 101% ± 13% of the expected Si, calculated using mass-balance recovery, in leaf, stem, and root tissues across 15 digestions. This Si recovery was fourteen-fold higher than the standard acid-extraction method and similar to a USDA-ARS alkaline-extraction method. Our procedure offers a low-cost, accurate method for extraction and analysis of Si in plant tissues.

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Germination and seedling establishment for transplanting into hydroponics often uses porous substrates, but fine roots grow into these substrates, and they cannot be removed without damaging these roots. Seedlings transplanted without removal of substrates can cause interactions with solution chemistry or addition of particulates to the nutrient solution. Germination of seeds on slant boards is clean, uniform, and reduces the time to transplanting. Slant boards facilitate development of long roots, which maximize exposure of the primary root to the nutrient solution after transplanting. The "boards" are made from thin acrylic or polycarbonate sheets with germination paper on top. Seeds are held in place by covering with thin paper before vertical placement of the boards in the container. Four to twelve days later, the seedlings with long roots can be removed from the paper without damage and transplanted into the hydroponic system. Here we describe slant board construction and procedures for rapid germination and transplanting in hydroponics.

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Urea is a byproduct of the urea cycle in metabolism and is excreted through urine and sweat. Ammonia, which is toxic at low levels, is converted to the safe storage form of urea, which represents the largest efflux of nitrogen from many organisms. Urea is an important nitrogen source in agriculture, is added to many industrial products, and is a large component in wastewater. The enzyme urease hydrolyzes urea to ammonia and bicarbonate. This reaction is microbially mediated in soils, hydroponic solutions, and wastewater recycling and is catalyzed in vivo in plants using native urease, making measurement of urea environmentally important. Both direct and indirect methods to measure urea exist. This protocol uses diacetyl monoxime to directly determine the concentration of urea in solution. The protocol provides repeatable results and stable reagents with good color stability and simple measurement techniques for use in any lab with a spectrophotometer. The reaction between diacetyl monoxime and urea in the presence of sulfuric acid, phosphoric acid, thiosemicarbazide, and ferric chloride produces a chromophore with a peak absorbance at 520 nm and a linear relationship between concentration and absorbance from 0.4 to 5.0 mM urea in this protocol. The lack of detectable interferences makes this protocol suitable for the determination of millimolar levels of urea in wastewater streams and hydroponic solutions.

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