<?xml version="1.0" encoding="UTF-8"?><!DOCTYPE article  PUBLIC "-//NLM//DTD Journal Publishing DTD v3.0 20080202//EN" "http://dtd.nlm.nih.gov/publishing/3.0/journalpublishing3.dtd"><article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" dtd-version="3.0" xml:lang="en" article-type="research article"><front><journal-meta><journal-id journal-id-type="publisher-id">JACEN</journal-id><journal-title-group><journal-title>Journal of Agricultural Chemistry and Environment</journal-title></journal-title-group><issn pub-type="epub">2325-7458</issn><publisher><publisher-name>Scientific Research Publishing</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="doi">10.4236/jacen.2022.113014</article-id><article-id pub-id-type="publisher-id">JACEN-119184</article-id><article-categories><subj-group subj-group-type="heading"><subject>Articles</subject></subj-group><subj-group subj-group-type="Discipline-v2"><subject>Chemistry&amp;Materials Science</subject><subject> Earth&amp;Environmental Sciences</subject></subj-group></article-categories><title-group><article-title>
 
 
  Effect of Steel Slag on Soil Fertility and Plant Growth
 
</article-title></title-group><contrib-group><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Zakirul</surname><given-names>Islam</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref><xref ref-type="corresp" rid="cor1"><sup>*</sup></xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Quoc</surname><given-names>Thinh Tran</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Shohei</surname><given-names>Koizumi</given-names></name><xref ref-type="aff" rid="aff2"><sup>2</sup></xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Fumitaka</surname><given-names>Kato</given-names></name><xref ref-type="aff" rid="aff2"><sup>2</sup></xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Kimio</surname><given-names>Ito</given-names></name><xref ref-type="aff" rid="aff3"><sup>3</sup></xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Kiwako</surname><given-names>S. Araki</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Motoki</surname><given-names>Kubo</given-names></name><xref ref-type="aff" rid="aff1"><sup>1</sup></xref></contrib></contrib-group><aff id="aff2"><addr-line>Advanced Technology Research Laboratories, Research and Development, Nippon Steel Corporation, 20-1 Shintomi, Futtsu, Chiba, Japan</addr-line></aff><aff id="aff3"><addr-line>Resource and Process Solution Division, Mineral Resources Department, Nippon Steel Corporation, 20-1 Shintomi, Futtsu, Chiba, Japan</addr-line></aff><aff id="aff1"><addr-line>Department of Biotechnology, Faculty of Life Sciences, Ritsumeikan University, Noji Higashi 1-1-1 Kusatsu, Shiga, Japan</addr-line></aff><pub-date pub-type="epub"><day>05</day><month>07</month><year>2022</year></pub-date><volume>11</volume><issue>03</issue><fpage>209</fpage><lpage>221</lpage><history><date date-type="received"><day>5,</day>	<month>July</month>	<year>2022</year></date><date date-type="rev-recd"><day>13,</day>	<month>August</month>	<year>2022</year>	</date><date date-type="accepted"><day>16,</day>	<month>August</month>	<year>2022</year></date></history><permissions><copyright-statement>&#169; Copyright  2014 by authors and Scientific Research Publishing Inc. </copyright-statement><copyright-year>2014</copyright-year><license><license-p>This work is licensed under the Creative Commons Attribution International License (CC BY). http://creativecommons.org/licenses/by/4.0/</license-p></license></permissions><abstract><p>
 
 
  The effective utilization of steel slag, a byproduct produced in large quantities from the steel refining process, is an important issue. Because steel slag contains abundant mineral components, the effects of steel slag on soil bacterial biomass and plant mineral uptake were analyzed in this study. The soil pH increased in proportion to the amount of steel slag added. A lower concentration (0.2% to 1%) of steel slag addition did not change the bacterial biomass. However, a higher concentration of steel slag (above 1%) had a negative effect on bacterial biomass. A lower amount of steel slag (0.2% to 1%) addition in soil leads to increased mineral (Ca, Mg, and Fe) uptake and plant growth in 
  <em>Brassica rapa </em>var.
  <em> periviridis </em>and 
  <em>Spinacia oleracea</em> L. However, mineral uptake by the plants decreased when a large amount of steel slag (above 1%) was added to the soil. Low concentrations of steel slag (0.2% to 1%) in soil had positive effects on plant growth, mineral uptake of plants, and bacterial biomass.
 
</p></abstract><kwd-group><kwd>Bacterial Biomass</kwd><kwd> Mineral Uptake</kwd><kwd> Nitrogen Circulation</kwd><kwd> Phosphorus Circulation</kwd><kwd> Plant Growth</kwd><kwd> Steel Slag</kwd></kwd-group></article-meta></front><body><sec id="s1"><title>1. Introduction</title><p>Over the last century, chemical fertilizers have been developed to enhance agricultural activities, and crop and vegetable yields have been substantially enhanced [<xref ref-type="bibr" rid="scirp.119184-ref1">1</xref>]. Chemical fertilizers contain mineralized compounds, such as ammonium sulfate, phosphoric acid, and potassium, therefore, plants easily take up these elements [<xref ref-type="bibr" rid="scirp.119184-ref2">2</xref>]. Since chemical fertilizers contain fewer minerals than organic fertilizer, micronutrients in soil have been reduced gradually in this 100-year period [<xref ref-type="bibr" rid="scirp.119184-ref3">3</xref>].</p><p>In recent years, organic agriculture has been promoted from the viewpoint of environmental issues and health consciousness [<xref ref-type="bibr" rid="scirp.119184-ref4">4</xref>] [<xref ref-type="bibr" rid="scirp.119184-ref5">5</xref>]. Manure is mainly used as an organic fertilizer, and unfermented materials such as fish meal and bone meal are added as supplements of mineral components [<xref ref-type="bibr" rid="scirp.119184-ref6">6</xref>]. However, the mineral components contained in these materials are unstable and expensive, therefore, the search for new alternative materials is important.</p><p>The steel industry discharges a large volume of waste materials, and steel slag is a byproduct of the steel refining process [<xref ref-type="bibr" rid="scirp.119184-ref7">7</xref>] [<xref ref-type="bibr" rid="scirp.119184-ref8">8</xref>] [<xref ref-type="bibr" rid="scirp.119184-ref9">9</xref>]. Since steel slag contains high levels of minerals (Ca, Mg, Fe, Mn, Zn, and Si) and acceptable low ranges of heavy metals (Pb and Cd), steel slag is used for agriculture [<xref ref-type="bibr" rid="scirp.119184-ref10">10</xref>]. Besga et al., [<xref ref-type="bibr" rid="scirp.119184-ref11">11</xref>] reported that the addition of steel slag to soil promotes exchangeable Ca and Mg in the soil. In addition, steel slag addition to soil can be used to increase pH of acidic soils [<xref ref-type="bibr" rid="scirp.119184-ref12">12</xref>] [<xref ref-type="bibr" rid="scirp.119184-ref13">13</xref>]. These studies reported that addition of steel slag enhances mineral uptake by plants. However, the effects of steel slag in organic cultivation system on bacterial biomass and nutrient circulation need to be investigated.</p><p>In this study, nutrient concentration and application of steel slag were analyzed for organic agriculture. Plants growth and mineral uptake by plant from steel slag were also evaluated. Furthermore, the effects of steel slag on the soil bacterial biomass and nutrient circulation in the organic soil environment were also investigated.</p></sec><sec id="s2"><title>2. Materials and Methods</title><sec id="s2_1"><title>2.1. Soil and Slag Use</title><p>Previously constructed woodchip-based organic soil was used for this experiment [<xref ref-type="bibr" rid="scirp.119184-ref14">14</xref>] Base soil was constructed using wood chips, mountain soil, black soil, and peat moss at a ratio of 5:3:1:1 (v/v). Organic fertilizers (cow manure 5%, oil cake 0.25%, soybean 0.25%, and bone meal 0.05%, w/w) were mixed with the base soil. <xref ref-type="table" rid="table1">Table 1</xref> shows the properties of the soil. A specimen of steel slag was provided by a steel company (Nippon Steel Corporation, Japan). Total carbon (TC), total nitrogen (TN), total phosphorus (TP), and total potassium (TK) and minerals e.g., calcium (Ca), magnesium (Mg), Iron (Fe), and manganese (Mn) of slag were analyzed.</p></sec><sec id="s2_2"><title>2.2. Plant Cultivation</title><p>Brassica rapa var. periviridis and Spinacia oleracea L. seeds were purchased from Takii &amp; Co., Ltd. (Kyoto, Japan). The pot experiments were carried out in a plant incubation room located at Ritsumeikan University, Shiga, Japan (34˚58'58.0''N 135˚57'49.2''E). The temperature was constant at 23˚C, and the interval of</p><table-wrap id="table1" ><label><xref ref-type="table" rid="table1">Table 1</xref></label><caption><title> Properties of soil at 30% water content after 1 week incubation</title></caption><table><tbody><thead><tr><th align="center" valign="middle" >Parameter</th><th align="center" valign="middle" >Value</th></tr></thead><tr><td align="center" valign="middle" >TC (mg/kg)</td><td align="center" valign="middle" >38,000</td></tr><tr><td align="center" valign="middle" >TN (mg/kg)</td><td align="center" valign="middle" >1380</td></tr><tr><td align="center" valign="middle" >TP (mg/kg)</td><td align="center" valign="middle" >850</td></tr><tr><td align="center" valign="middle" >TK (mg/kg)</td><td align="center" valign="middle" >5100</td></tr><tr><td align="center" valign="middle" >Bacterial biomass (&#215;10<sup>8</sup> cells/g-soil)</td><td align="center" valign="middle" >12.6</td></tr><tr><td align="center" valign="middle" >Nitrogen circulation activity (point)</td><td align="center" valign="middle" >32</td></tr><tr><td align="center" valign="middle" >Phosphorus circulation activity (point)</td><td align="center" valign="middle" >67</td></tr><tr><td align="center" valign="middle" >pH</td><td align="center" valign="middle" >6</td></tr><tr><td align="center" valign="middle" >EC (ds/cm)</td><td align="center" valign="middle" >0.7</td></tr><tr><td align="center" valign="middle" >Water holding capacity (ml/kg)</td><td align="center" valign="middle" >1500</td></tr><tr><td align="center" valign="middle" >Bulk density (g/cm<sup>3</sup>)</td><td align="center" valign="middle" >0.55</td></tr></tbody></table></table-wrap><p>light and dark periods was 12 h/12 h. Two experiments were conducted in this study.</p><p>The first experiment was conducted to cultivate B. rapa at different application rates (0%, 0.2%, 0.4%, 0.6%, 0.8%, 1%, 1.2%, and 1.4%) of steel slag. One seedling of B. rapa was transplanted into each small pot that contained 200 g of the soil. The water content of the soil was maintained at 30% during the cultivation period. This experiment was carried out in one replicate for each treatment. The bacterial biomass and pH value of soil with each steel slag concentration were analyzed at the start point (0 week) and after plant cultivation (4 weeks). Fresh shoot weight as plant growth and mineral concentrations (Ca, Mg, Fe, and Mn) of dried shoots were measured.</p><p>In the second experiment, two different plant species (B. rapa and S. oleracea) were cultivated in the soil with 0% (T<sub>0</sub>), 0.5% (T<sub>0.5</sub>), and 1% (T<sub>1</sub>) steel slag in triplicate. A total of 2.5 kg of soil was put in a Wagner pot (1/5000a) and maintained at 30% water content. Three seedlings of B. rapa and S. oleracea were transplanted into each pot. Bacterial biomass, nitrogen circulation activity, and phosphorus circulation activity in the soil were measured at the initial experiment and after plant harvest. After 4 weeks for B. rapa and 6 weeks for S. oleracea fresh shoot weight, SPAD value, nitrate, and mineral concentrations (Ca, Mg, Fe, and Mn) of dried shoots were analyzed.</p></sec><sec id="s2_3"><title>2.3. Analytical Methods</title><p>TC was analyzed by using a Total Organic Carbon Analyser (TOC-VCPH, Shimadzu, Kyoto, Japan). TN, TP, and TK were extracted with CuSO<sub>4.</sub> 5H<sub>2</sub>O, H<sub>2</sub>SO<sub>4,</sub> and H<sub>2</sub>O<sub>2</sub> at 420˚C [<xref ref-type="bibr" rid="scirp.119184-ref15">15</xref>]. Ca, Mg, Fe, and Mn were extracted with HNO<sub>3</sub> and HClO<sub>4</sub> at 215˚C. After extraction, TN and TP were determined by the indophenol blue method [<xref ref-type="bibr" rid="scirp.119184-ref16">16</xref>] and molybdenum blue method [<xref ref-type="bibr" rid="scirp.119184-ref17">17</xref>] respectively. The TK, Ca, Mg, Fe, and Mn concentrations in the extracts were measured by usinga an atomic absorption spectrophotometer (Hitachi Z2300, Tokyo, Japan). Total bacterial biomass was analyzed by quantification of environmental DNA (eDNA) extracted by the slow-stirring method [<xref ref-type="bibr" rid="scirp.119184-ref18">18</xref>].</p><p>The evaluation of P circulation activities was carried out by following our previous procedure [<xref ref-type="bibr" rid="scirp.119184-ref19">19</xref>] [<xref ref-type="bibr" rid="scirp.119184-ref20">20</xref>]. A soil sample of 1 g was added to a phytic acid solution (pH 7) containing 3.9 mg of P and incubated for 3 days at 25˚C. Control treatment with only distilled water was also prepared simultaneously. Water-soluble phosphorous (SP) was extracted with 20 mL of distilled water and analyzed by the molybdenum blue method. The increase in SP was defined as the P circulation activity. The following formula was used to calculate P circulation and expressed in points (0 to 100).</p><p>Pcirculationactivity ( point ) = ( S P inP 3 − S P inP 0 ) ( S P inW 3 − S P in   W 0 ) TotaladdedP &#215; 100</p><p>where, P<sub>0</sub> and W<sub>0</sub> denote phytic acid and distilled water added tube at day 0, and P<sub>3</sub> and W<sub>3</sub> denote phytic acid and distilled water added tube at day 3.</p><p>Nitrogen (N) circulation activity was analyzed based on the bacterial biomass, ammonium oxidation rate and nitrite oxidation rate and expressed in points [<xref ref-type="bibr" rid="scirp.119184-ref21">21</xref>]. The bacterial biomass of 6.0 &#215; 10<sup>8</sup> cells g<sup>−1</sup> was defined as 100 points. Soil mixed with ammonium sulfate or sodium nitrite (60 &#181;g N g<sup>−1</sup> dry soil) was incubated at 25˚C and after 3 days of incubation, the percentage of reduction in the added N was defined as the ammonium or nitrite oxidation rate. Using the scores of bacterial biomasses, ammonium oxidation rate and nitrite oxidation rate, a radar chart was constructed, and the relative area of the inner triangle is expressed as the N circulation activity.</p><p>The area of the inner triangle in the radar chart is calculated as follows</p><p>Area = ( a &#215; b ) + ( b &#215; c ) + ( c &#215; a ) 4 &#215; 3 100</p><p>where a, b, and c denote scores of bacterial numbers, ammonium oxidation rate and nitrite oxidation rate, respectively. Nitrogen circulation activity was analyzed by calculating the relative area of the inner triangle as follows</p><p>Ncirculationactivity   ( point ) = Areaoftheinnertriangle Areaoftheoutertriangle &#215; 100</p><p>Soil pH was analyzed at a 1:2.5 ratio of soil and distilled water using a pH metre (LAQUA F-72, Horiba, Kyoto, Japan). The chlorophyll content in the leaves was measured by a Soil Plant Analysis Development (SPAD) meter (SPAD-502, Konica Minolta Sensing, Osaka, Japan). The SPAD value was an average value of 10 measurements. For nitrate analysis, fresh plants were mixed at a ratio of 1:5 (w/v) of plant and distilled water by using waring blender for 1 min. Subsequently, the supernatant was collected for nitrate measurement after centrifuging at 14,000 rpm for 5 min. Nitrate was analyzed following the brucine method with slight modification [<xref ref-type="bibr" rid="scirp.119184-ref22">22</xref>] by using spectrophotometer (Hitachi U-1900, Tokyo, Japan).</p></sec><sec id="s2_4"><title>2.4. Statistical Analysis</title><p>Mean and standard deviation was performed using SPSS 25.0 software (Armond, NY, USA). Significant differences were determined by one-way analysis of variance (ANOVA, p &lt; 0.05), followed by Tukey’s post-hoc test.</p></sec></sec><sec id="s3"><title>3. Results</title><sec id="s3_1"><title>3.1. Mineral Concentrations of Steel Slag</title><p>To understand the features of steel slag, the chemical composition was analyzed. The steel slag mainly contained Ca (228,000 mg/kg) and Fe (210,000 mg/kg) followed by Mn (40,400 mg/kg) and Mg (35,100 mg/kg) (<xref ref-type="table" rid="table2">Table 2</xref>). This result indicates that steel slag contains minerals and has the possibility of increasing micronutrients in agricultural soil.</p></sec><sec id="s3_2"><title>3.2. Effect of Steel Slag Application on Soil Bacterial Biomass and</title>pH<p><xref ref-type="table" rid="table3">Table 3</xref> shows the effect of steel slag on the soil bacterial biomass and pH value at 0 and 4 weeks. The steel slag additions from 0.2% to 1% did not change bacterial biomass compared to soil without steel slag. However, higher steel slag concentrations (1.2% and 1.4%) decreased bacterial biomass in the soil at weeks 0 and 4. The pH values of the soil gradually increased with the increase in steel slag addition.</p></sec><sec id="s3_3"><title>3.3. Effect Steel Slag on Plant (B. rapa)</title><p>The growth and mineral uptake of B. rapa were analyzed to determine the suitable concentration of steel slag addition to soil. When 0.2% to 1.4% steel slag were added to the soil, the shoot weight of the plants increased. Steel slag concentrations of 0.4% to 1.4% seem to be suitable for plant growth (<xref ref-type="fig" rid="fig1">Figure 1</xref>).</p><table-wrap id="table2" ><label><xref ref-type="table" rid="table2">Table 2</xref></label><caption><title> Properties of slag</title></caption><table><tbody><thead><tr><th align="center" valign="middle" >Parameter</th><th align="center" valign="middle" >Value</th></tr></thead><tr><td align="center" valign="middle" >TC (mg/kg)</td><td align="center" valign="middle" >12,130</td></tr><tr><td align="center" valign="middle" >TN (mg/kg)</td><td align="center" valign="middle" >80</td></tr><tr><td align="center" valign="middle" >TP (mg/kg)</td><td align="center" valign="middle" >20,500</td></tr><tr><td align="center" valign="middle" >TK (mg/kg)</td><td align="center" valign="middle" >15,220</td></tr><tr><td align="center" valign="middle" >Ca (mg/kg)</td><td align="center" valign="middle" >228,000</td></tr><tr><td align="center" valign="middle" >Mg (mg/kg)</td><td align="center" valign="middle" >35,100</td></tr><tr><td align="center" valign="middle" >Fe (mg/kg)</td><td align="center" valign="middle" >210,000</td></tr><tr><td align="center" valign="middle" >Mn (mg/kg)</td><td align="center" valign="middle" >40,400</td></tr></tbody></table></table-wrap><table-wrap id="table3" ><label><xref ref-type="table" rid="table3">Table 3</xref></label><caption><title> Bacterial biomass, and pH values after the addition of steel slag to the soil at weeks 0 and 4</title></caption><table><tbody><thead><tr><th align="center" valign="middle"  rowspan="2"  >Steel slag concentration (%)</th><th align="center" valign="middle"  colspan="2"  >Bacterial biomass (&#215;10<sup>8</sup> cells/g)</th><th align="center" valign="middle"  colspan="2"  >pH</th></tr></thead><tr><td align="center" valign="middle" >Week 0</td><td align="center" valign="middle" >Week 4</td><td align="center" valign="middle" >Week 0</td><td align="center" valign="middle" >Week 4</td></tr><tr><td align="center" valign="middle" >0</td><td align="center" valign="middle" >9.0</td><td align="center" valign="middle" >10.2</td><td align="center" valign="middle" >5.9</td><td align="center" valign="middle" >7.1</td></tr><tr><td align="center" valign="middle" >0.2</td><td align="center" valign="middle" >10.0</td><td align="center" valign="middle" >9.8</td><td align="center" valign="middle" >6.4</td><td align="center" valign="middle" >7.6</td></tr><tr><td align="center" valign="middle" >0.4</td><td align="center" valign="middle" >10.5</td><td align="center" valign="middle" >9.9</td><td align="center" valign="middle" >6.7</td><td align="center" valign="middle" >7.7</td></tr><tr><td align="center" valign="middle" >0.6</td><td align="center" valign="middle" >10.5</td><td align="center" valign="middle" >10.3</td><td align="center" valign="middle" >6.6</td><td align="center" valign="middle" >8.1</td></tr><tr><td align="center" valign="middle" >0.8</td><td align="center" valign="middle" >10.0</td><td align="center" valign="middle" >10.2</td><td align="center" valign="middle" >7.1</td><td align="center" valign="middle" >8.2</td></tr><tr><td align="center" valign="middle" >1.0</td><td align="center" valign="middle" >10.4</td><td align="center" valign="middle" >9.6</td><td align="center" valign="middle" >7.2</td><td align="center" valign="middle" >8.3</td></tr><tr><td align="center" valign="middle" >1.2</td><td align="center" valign="middle" >7.9</td><td align="center" valign="middle" >7.3</td><td align="center" valign="middle" >7.4</td><td align="center" valign="middle" >8.5</td></tr><tr><td align="center" valign="middle" >1.4</td><td align="center" valign="middle" >7.6</td><td align="center" valign="middle" >7.4</td><td align="center" valign="middle" >7.0</td><td align="center" valign="middle" >8.3</td></tr></tbody></table></table-wrap><p>Calcium uptake by B. rapa proportionally increased when the steel slag was added at 0.2% to 1%, and the highest calcium uptake was 24,980 mg/kg at 1% steel slag (<xref ref-type="table" rid="table4">Table 4</xref>). However, the addition of a high concentration of steel slag (above 1%) did not increase the uptake of Ca. A similar tendency was observed in the case of Mg, Fe, and Mn uptake by B. rapa.</p></sec><sec id="s3_4"><title>3.4. Cultivation of Two Different Species of Plants (B. rapa and</title>S. oleracea) with a Suitable Concentration of Steel Slag<p>Three different treatments (T<sub>0</sub>, T<sub>0.5</sub>, and T<sub>1</sub>) were carried out for the evaluation bacterial biomass, N-circulation activity, P-circulation activity, and growth of B. rapa and S. oleracea. The effects of steel slag on soil fertility are shown on <xref ref-type="table" rid="table5">Table 5</xref> and <xref ref-type="table" rid="table6">Table 6</xref>. The bacterial biomass in the soil was almost similar in all treatments. The addition of steel slag did not change N-circulation and P-circulation significantly at week 0 (p &gt; 0.05). Meanwhile, a significant increase in N-circulation activities was observed at weeks 4 and 6 after harvest B. rapa and S. oleracea respectively. On the other hand, P-circulation activities decreased at weeks 4 and 6 (p &gt; 0.05).</p><table-wrap id="table4" ><label><xref ref-type="table" rid="table4">Table 4</xref></label><caption><title> Mineral concentrations in the dried plant shoot (Brassica rapa)</title></caption><table><tbody><thead><tr><th align="center" valign="middle" >Steel slag concentration (%)</th><th align="center" valign="middle" >Ca (mg/kg)</th><th align="center" valign="middle" >Mg (mg/kg)</th><th align="center" valign="middle" >Fe (mg/kg)</th><th align="center" valign="middle" >Mn (mg/kg)</th></tr></thead><tr><td align="center" valign="middle" >0</td><td align="center" valign="middle" >10,780</td><td align="center" valign="middle" >2580</td><td align="center" valign="middle" >1040</td><td align="center" valign="middle" >380</td></tr><tr><td align="center" valign="middle" >0.2</td><td align="center" valign="middle" >13,500</td><td align="center" valign="middle" >2880</td><td align="center" valign="middle" >1040</td><td align="center" valign="middle" >430</td></tr><tr><td align="center" valign="middle" >0.4</td><td align="center" valign="middle" >16,740</td><td align="center" valign="middle" >2860</td><td align="center" valign="middle" >2020</td><td align="center" valign="middle" >440</td></tr><tr><td align="center" valign="middle" >0.6</td><td align="center" valign="middle" >15,000</td><td align="center" valign="middle" >2860</td><td align="center" valign="middle" >2300</td><td align="center" valign="middle" >480</td></tr><tr><td align="center" valign="middle" >0.8</td><td align="center" valign="middle" >20,260</td><td align="center" valign="middle" >3040</td><td align="center" valign="middle" >2500</td><td align="center" valign="middle" >480</td></tr><tr><td align="center" valign="middle" >1.0</td><td align="center" valign="middle" >24,980</td><td align="center" valign="middle" >3140</td><td align="center" valign="middle" >2800</td><td align="center" valign="middle" >460</td></tr><tr><td align="center" valign="middle" >1.2</td><td align="center" valign="middle" >22,800</td><td align="center" valign="middle" >2900</td><td align="center" valign="middle" >2700</td><td align="center" valign="middle" >480</td></tr><tr><td align="center" valign="middle" >1.4</td><td align="center" valign="middle" >22,820</td><td align="center" valign="middle" >2880</td><td align="center" valign="middle" >2420</td><td align="center" valign="middle" >540</td></tr></tbody></table></table-wrap><table-wrap id="table5" ><label><xref ref-type="table" rid="table5">Table 5</xref></label><caption><title> Soil bacterial biomass, N circulation activity, and P circulation activity in three treatments of Brassica rapa cultivated soil at 0 and 4 weeks</title></caption><table><tbody><thead><tr><th align="center" valign="middle"  rowspan="2"  >Treatment</th><th align="center" valign="middle"  colspan="2"  >Bacterial biomass (&#215;10<sup>8</sup> cells/g)</th><th align="center" valign="middle"  colspan="2"  >N-circulation activity (point)</th><th align="center" valign="middle"  colspan="2"  >P-circulation activity (point)</th></tr></thead><tr><td align="center" valign="middle" >Week 0</td><td align="center" valign="middle" >Week 4</td><td align="center" valign="middle" >Week 0</td><td align="center" valign="middle" >Week 4</td><td align="center" valign="middle" >Week 0</td><td align="center" valign="middle" >Week 4</td></tr><tr><td align="center" valign="middle" >T<sub>0</sub></td><td align="center" valign="middle" >9.6 &#177; 0.2 <sup>a</sup></td><td align="center" valign="middle" >8.7 &#177; 0.2 <sup>a</sup></td><td align="center" valign="middle" >37.3 &#177; 2.5 <sup>a</sup></td><td align="center" valign="middle" >49.6 &#177; 5.3 <sup>c</sup></td><td align="center" valign="middle" >53.3 &#177; 1.7 <sup>a</sup></td><td align="center" valign="middle" >19.3 &#177; 3.3 <sup>a</sup></td></tr><tr><td align="center" valign="middle" >T<sub>0.5</sub></td><td align="center" valign="middle" >9.3 &#177; 0.2 <sup>a</sup></td><td align="center" valign="middle" >8.4 &#177; 0.3 <sup>a</sup></td><td align="center" valign="middle" >38.3 &#177; 7.1 <sup>a</sup></td><td align="center" valign="middle" >92 &#177; 3.5 <sup>b</sup></td><td align="center" valign="middle" >50.6 &#177; 6.3 <sup>a</sup></td><td align="center" valign="middle" >8.0 &#177; 2.1 <sup>c</sup></td></tr><tr><td align="center" valign="middle" >T<sub>1</sub></td><td align="center" valign="middle" >9.4 &#177; 0.1 <sup>a</sup></td><td align="center" valign="middle" >8.5 &#177; 0.5 <sup>a</sup></td><td align="center" valign="middle" >37.6 &#177; 2.6 <sup>a</sup></td><td align="center" valign="middle" >95.7 &#177; 2.3 <sup>a</sup></td><td align="center" valign="middle" >50 &#177; 6.4 <sup>a</sup></td><td align="center" valign="middle" >12.6 &#177; 4.1 <sup>b</sup></td></tr></tbody></table></table-wrap><p>Means followed by different letters within a column are significantly different at p &lt; 0.05.</p><table-wrap id="table6" ><label><xref ref-type="table" rid="table6">Table 6</xref></label><caption><title> Soil bacterial biomass, N circulation activity, and P circulation activity in three treatments of Spinacia oleracea cultivated soil at weeks 0 and 6</title></caption><table><tbody><thead><tr><th align="center" valign="middle"  rowspan="2"  >Treatment</th><th align="center" valign="middle"  colspan="2"  >Bacterial biomass (&#215;10<sup>8</sup> cells/g)</th><th align="center" valign="middle"  colspan="2"  >N-circulation activity (point)</th><th align="center" valign="middle"  colspan="2"  >P-circulation activity (point)</th></tr></thead><tr><td align="center" valign="middle" >Week 0</td><td align="center" valign="middle" >Week 6</td><td align="center" valign="middle" >Week 0</td><td align="center" valign="middle" >Week 6</td><td align="center" valign="middle" >Week 0</td><td align="center" valign="middle" >Week 6</td></tr><tr><td align="center" valign="middle" >T<sub>0</sub></td><td align="center" valign="middle" >9.3 &#177; 0.21 <sup>a</sup></td><td align="center" valign="middle" >8.4 &#177; 0.04 <sup>a</sup></td><td align="center" valign="middle" >37.3 &#177; 2.4 <sup>a</sup></td><td align="center" valign="middle" >50.0 &#177; 5.3 <sup>c</sup></td><td align="center" valign="middle" >53.3 &#177; 1.7 <sup>a</sup></td><td align="center" valign="middle" >19.3 &#177; 3.3 <sup>a</sup></td></tr><tr><td align="center" valign="middle" >T<sub>0.5</sub></td><td align="center" valign="middle" >9.6 &#177; 0.34 <sup>a</sup></td><td align="center" valign="middle" >8.1 &#177; 0.41 <sup>a</sup></td><td align="center" valign="middle" >38.3 &#177; 7.1 <sup>a</sup></td><td align="center" valign="middle" >93.7 &#177; 4.0 <sup>b</sup></td><td align="center" valign="middle" >50.6 &#177; 6.3 <sup>a</sup></td><td align="center" valign="middle" >8.0 &#177; 2.2 <sup>c</sup></td></tr><tr><td align="center" valign="middle" >T<sub>1</sub></td><td align="center" valign="middle" >9.5 &#177; 0.16 <sup>a</sup></td><td align="center" valign="middle" >8.2 &#177; 0.26 <sup>a</sup></td><td align="center" valign="middle" >39.6 &#177; 6.8 <sup>a</sup></td><td align="center" valign="middle" >95.7 &#177; 2.4 <sup>a</sup></td><td align="center" valign="middle" >49.6 &#177; 6.8 <sup>a</sup></td><td align="center" valign="middle" >12.7 &#177; 4.1 <sup>b</sup></td></tr></tbody></table></table-wrap><p>Means followed by different letters within a column are significantly different at p &lt; 0.05.</p><p><xref ref-type="fig" rid="fig2">Figure 2</xref> and <xref ref-type="fig" rid="fig3">Figure 3</xref> show the growth of B. rapa and S. oleracea under the three different treatments. The fresh shoot weight in the T<sub>1</sub> treatment was significantly higher than those in the T<sub>0</sub> and T<sub>0.5</sub> treatments (p &lt; 0.05) (<xref ref-type="fig" rid="fig4">Figure 4</xref>(a)). The SPAD values of both plant species were the highest in the T<sub>1</sub> treatment (<xref ref-type="fig" rid="fig4">Figure 4</xref>(b)). The addition of the steel slag to the soil did not show any significant differences with nitrate uptake in the two plant species (p &gt; 0.05) (<xref ref-type="fig" rid="fig4">Figure 4</xref>(c)).</p><p>The uptake of Ca, Mg, and Fe increased in B. rapa and S. oleracea with increasing steel slag concentration in the soil. The highest contents of Ca (16,333 mg/kg), Mg (4166 mg/kg), and Fe (1460 mg/kg) in B. rapa were observed in the T<sub>1</sub> treatment (Figures 5(a)-(c)). Similarly, the same tendency was found in the case of S. oleracea. These results suggest that 0.5% to 1.0% steel slag addition seems to be a suitable condition not only plant growth but also for nutrient circulation in soil.</p></sec></sec><sec id="s4"><title>4. Discussion</title><p>Organic agriculture has been promoted, however the mineral components contained in the main organic fertilizers such as manure are limited. In this research, the fertilization possibility of steel slag, which is byproduct from the steel industry, for organic agriculture was investigated.</p><p>Low concentrations of steel slag addition did not have a negative effect on soil bacterial biomass. Previous studies reported that slag addition in soil increases total bacterial biomass, but decreases the relative abundance of Genus Acidobacteria, Bacteriodates, Nitrospirae, and Chloroflexi [<xref ref-type="bibr" rid="scirp.119184-ref23">23</xref>] [<xref ref-type="bibr" rid="scirp.119184-ref24">24</xref>]. The interaction of steel slag and microorganisms in soil is still unclear, and more investigation is needed to determine the relationship between soil microbes and slag addition.</p><p>The addition of steel slag leads to an increase in soil pH, and the pH values of the soil gradually increased with increasing steel slag addition. When slag is added to soil, CaO and MgO dissolve with water and release OH<sup>–</sup> which increases the pH values of the soil [<xref ref-type="bibr" rid="scirp.119184-ref25">25</xref>]. The minerals in steel slag seem to elute slowly in the soil environment and the effect upon 1% addition continues for approximately 1 year.</p><p>On the other hand, addition of steel slag into soil negatively affects P-circulation activity because increased calcium ions precipitate with phosphate in soil [<xref ref-type="bibr" rid="scirp.119184-ref26">26</xref>]. However, a positive effect on N-circulation activity was observed. This effect may be caused by the activation of ammonia and nitrite oxidizing bacteria in soil [<xref ref-type="bibr" rid="scirp.119184-ref24">24</xref>]. The effective addition of steel slag should consider the balance between P-circulation and N-circulation activities.</p><p>Plant growth and mineral uptake significantly increased when steel slag was added to the soil. This result supports previous studies [<xref ref-type="bibr" rid="scirp.119184-ref27">27</xref>] [<xref ref-type="bibr" rid="scirp.119184-ref28">28</xref>] [<xref ref-type="bibr" rid="scirp.119184-ref29">29</xref>]. Enhanced N circulation activity among treatments facilitated N uptake by plant which could be a reason for plant growth. The increase in mineral uptake by plants suggests that steel slag contains exchangeable mineral or could be mineralized by microorganisms.</p><p>Ionization of steel slag is promoted by a large number of soil microorganisms, therefore, the effect of steel slag may be more effective in organic agricultural fields [<xref ref-type="bibr" rid="scirp.119184-ref30">30</xref>]. Since, the bacterial biomass is lower in the agricultural field cultivated with chemical fertilizers, so the effect of slag may be lower compared to the organic cultivation system. Further investigation is progressing to determine the effect of slag in conventional cutivation system.</p></sec><sec id="s5"><title>5. Conclusion</title><p>Mineral uptake by the plants decreased when a large amount of steel slag (above 1%) was added to the soil, however, a lower amount of steel slag (0.2% to 1%) addition to the soil led to increased mineral (Ca, Mg, and Fe) uptake and plant growth. Low concentrations of steel slag (0.5% to 1%) in soil had positive effects on plant growth, mineral uptake of plants, and bacterial biomass during short-term cultivation practices. Further studies are suggested on the long-term cultivation and repetitive application of slag.</p></sec><sec id="s6"><title>Funding</title><p>This work was supported by Nippon Steel Corporation, Japan.</p></sec><sec id="s7"><title>Conflicts of Interest</title><p>The authors declare that they have no known competing interests.</p></sec><sec id="s8"><title>Cite this paper</title><p>Islam, Z., Tran, Q.T., Koizumi, S., Kato, F., Ito, K., Araki, K.S. and Kubo, M. (2022) Effect of Steel Slag on Soil Fertility and Plant Growth. 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