Exploring the effect of soil amelioration on wheat growth, nutrition, and soil microbiomes in acidic sandy soil

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Asaduzzaman Prodhan, Gaus Azam, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7136714/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background and aims Soil acidity poses a significant global challenge to soil health and the sustainability of agricultural production. In Western Australia’s grain belt, soil acidification—exacerbated by crop removal and the use of acidifying fertilisers—reduces land productivity. A combination of organic and inorganic soil amendments including lime, gypsum, clay, compost, and synthetic fertilisers could improve soil health and microbial function. Methods A glasshouse experiment was conducted using undisturbed soil cores amended three years earlier in a “re-engineering soils” field trial, which comprised four soil treatments: Control, CLG (clay, lime, gypsum), CLG + NPK (CLG + nitrogen, phosphorus, and potassium fertilisers), and CLG + Compost. The study followed a completely randomised design with three replications. Plant growth, nutrient uptake, and soil chemical and microbial properties were assessed at three key wheat growth stages, according to Zadoks’ scale: Z23, Z61, and Z92. Results These amendments improved plant growth, nutrient uptake, grain yield, and nutrient status, while also enhancing soil chemical properties, microbial biomass carbon (MBC), and microbial composition. CLG was the main amendment used to improve soil conditions, and its application with compost showed greater effectiveness compared to its use with NPK fertilisers. Conclusion This study offers insights into the benefits of organic and inorganic amendments for managing acidic sandy soil. The findings support the development of sustainable soil management strategies. Future work should explore the long-term impacts of these amendments across different soil types and cropping systems. Soil amendments Wheat yield Nutrient content Soil health Soil microbiomes Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Introduction Wheat ( Triticum aestivum L.) is a major crop in Australia, significantly contributing to the nation’s economy. However, soil acidity significantly constrains productivity, particularly in the Western Australian grain belt region, where over 70% of topsoils and 50% of subsoils fall below recommended pH level 5.5 and 4.8, respectively (Gazey et al., 2019 ). Acidic subsoils significantly impact wheat production, causing annual economic losses AU $ 1.6 billion (Herbert, 2009 ; Petersen, 2015 ). This decline in soil quality has broader implications for food security, biodiversity, and ecosystem function. Both organic and inorganic soil amendments are used to improve soil fertility. Common organic materials include manures, composts, plant residues, household waste, sewage sludge and biochar, while inorganic amendments include lime, gypsum, clay minerals. These materials can improve nutrient and water availability, stimulate microbial activity, and enhance plant nutrient absorption (Tejada et al., 2009 ; Xu et al., 2015 ). Lime, compost, and inorganic fertilisers have been widely studies for their capacity to enhance soil fertility and crop productivity (Marschner, 2011 ), improve soil aggregation and bulk density, and optimise air–water relations (Manirakiza & Şeker, 2020 ; Tejada et al., 2009 ). Liming is a proven strategy for neutralising soil acidity, increasing pH and improving fertility (Orton et al., 2018 ). It enhances the availability of key nutrients such as calcium (Ca) and magnesium (Mg) (Bossolani et al., 2020 ), both essential for plant and microbial development (Guo et al., 2019 ; Jha et al., 2020 ; Liu et al., 2020 ). Liming reduces the toxicity of aluminium ions (Al 3+ ) by replacing them with calcium ions (Ca 2+ ), thereby raising soil pH to enhance nutrient availability (Paradelo et al., 2015 ). In non-tillage systems, surface lime application is effective for ameliorating surface acidity (Carmeis Filho et al., 2017 ; Joris et al., 2016 ; Tiritan et al., 2016 ), but less for acidic subsoil (Azam & Gazey, 2020 ). Combining deep tillage with lime incorporation has been shown to improve rooting depth, water use efficiency, and wheat grain yield by promoting root development (Scanlan et al., 2014 ) and increasing tolerance to moisture stress during grain filling (Azam & Gazey, 2019 ). Gypsum (CaSO 4 .2H 2 O), often applied with lime, helps displace excess sodium ions (Na + ) in saline soils and enhances water infiltration while also supplying Ca (Oster, 1982 ). In Finland, gypsum application reduced phosphorus (P) runoff to surface waters by up to 50% (Ollikainen et al., 2020 ). Applying lime together with gypsum effectively reduces subsoil acidity particularly by facilitating the movement of basic cations into the subsoil (Tang et al., 2013 ). However, lime alone may not fully restore degraded sandy soil prone to nutrient leaching (Hamza & Anderson, 2003 ). Organic amendments like compost and manure can improve soil structure, increase organic matter, and support microbial activity—key factors for long-term soil health (Lal, 2009 ). These amendments stabilise soil aggregates, enhance soil water-holding capacity, and slowly release nutrients, complementing the benefits of lime (Whalen et al., 2000 ). Compost increases soil organic matter, improves physical properties such as infiltration, aeration and porosity, and supports microbial growth by increasing biomass, and biodiversity (Irshad et al., 2013 ; Sharif et al., 2014 ). It also supports essential nutrients (N, P, K, Ca, and Mg) for plants to absorb through their roots (Irshad et al., 2013 ) and reduce soil erosion. In a US study, composted sandy clay loam retained 80% of simulated rainfall and reduced runoff by 60% (Faucette et al., 2007 ). Organic matter can also raise soil pH and reduced Al 3+ toxicity by enhancing buffering capacity and forming Al-chelates (Wang et al., 2014 ). Clay amendment offers another option for improving sandy soils by increasing water and nutrient retention, crop growth and soil carbon content (Davenport et al., 2011 ; Schapel et al., 2017 ; Ye et al., 2020 ). Fine particles such as clay and silt contribute to better aggregate formation, higher nutrient availability, and improved soil organic carbon (SOC) stability (Bronick & Lal, 2005 ; Feng et al., 2014 ; Regelink et al., 2015 ), and they also enhance cation exchange capacity (Chorom et al., 1994 ). Soil microbial communities are crucial in nutrient cycling and soil function. Soil harbours a rich diversity of microbes (Azim, 2019 ), including bacteria, fungi, and algae, making up about 2% of SOC (Lakshmipathi et al., 2019 ). These microorganisms help decompose organic matter, recycle nutrients, and support soil structure and function (Heikkinen et al., 2021 ; Lazcano et al., 2013 ). However, soil acidity can reduce microbial activity and diversity (Kuramae et al., 2012 ; Navarrete et al., 2013 ) by lowering carbon availability, restricting nutrient, and increasing toxic elements like aluminium (Al) and manganese (Mn) (Bowman et al., 2008 ; Tian et al., 2019 ). Amendments such as lime and compost can counter these effects by enhancing microbial biomass and diversity (Sun et al., 2015 ; Zhao et al., 2016 ) and, in turn, improving nutrient availability and plant uptake (Zhang et al., 2018 ). While many studies have examined individual amendments like lime, compost, or biochar, relatively few have explored the combined effects of multiple amendments on plant growth, soil health and microbial communities in sandy soil. Addressing this gap is essential for developing sustainable management practices for acidic sandy soil. The field experiment of “re-engineering soils” project aimed to address multiple soil constraints including acidity, compaction, poor water-holding capacity, water repellence, and low fertility—through soil amelioration with lime, clay, gypsum, fertilisers, and organic matter to enhance water use efficiency of grain crops in Western Australia. This study investigates the effects of soil amelioration on wheat growth, nutrition and soil microbiomes through a glasshouse experiment. The specific objective of the study is to assess the effects of soil amendments on soil health, plant performances, and soil microbiomes. We hypothesise that combinations of different soil amendments will enhance wheat productivity by improving soil physiochemical properties and supporting diverse microbial interactions. These findings will inform sustainable management strategies to improve crop performance and soil health in acidic sandy soils. Materials and methods Site description and soil collection Undisturbed soil cores were collected from a field trial conducted by the Department of Primary Industries and Regional Development (DPIRD) as part of the re-engineering soil project in Bolgart, Western Australia (31°18’59.9”S, 116°34’37.5”E). The site had previously received following four soil treatments (Table 1 ) to address multiple soil constraints. In total, 36 intact soil cores (4 treatments × 3 replications × 3 destructive harvests) were collected using cylindrical PVC pots (10 cm diameter, and 20 cm height) with sharpened bases, hydraulically press into the soil. Cores were transported to The University of Western Australia (UWA) for glasshouse experiment. Baseline soil chemical properties are detailed in Table S1 . The Bolgart region has a Mediterranean-type climate, characterised by hot, dry summer and cool, wet winters, with an average annual rainfall of 612 mm. Temperatures range from 10.5–26.7°C. The soil was moderately acidic (topsoil pH 5.2, subsurface pH 4.6) with a loamy sand texture, and low to medium Al (~ 6 mg kg –1 ). The field was heavily compacted, with penetrometer resistance readings between 3 and 4 MPa (Azam et al., 2024 ) before the application of soil amendments. Table 1 Experimental treatments applied to the top 10 cm of field soil Treatments Description T 1 (Control) No amendment. T 2 (CLG) Clay @ 110 t ha − 1 , Lime @ 1.5 t ha − 1 , Gypsum @ 1 t ha − 1 . T 3 (CLG + NPK) Clay @110 t ha − 1 , Lime @ 1.5 t ha − 1 , Gypsum @ 1 t ha − 1 , Nitrogen (N) @ 40kg ha − 1 , Phosphorous (P) @12.5 kg ha − 1 and Potassium (K) @15 kg ha − 1 . T 4 (CLG + Compost) Clay@110 t ha − 1 , [email protected] t ha − 1 , Gypsum@1 t ha − 1 , Compost@42t ha − 1 . Experimental design and glasshouse set up The 36 PVC soil cores were arranged in a completely randomised design into the UWA glasshouse incorporating the same four treatments and three harvest stages, each replicated three times. Five wheat seeds (cv. Scepter) were sown per pot and thinned to two plants eight days after sowing. Moisture content was maintained at 75% of field capacity during germination and increased to 100% during active growth. Glasshouse temperatures were maintained at 20–26°C, with relative humidity ranging from 34.9–84.7%. Plants were grown from 1 May to 10 September 2024. Crop development stages were recorded following the Zadoks’ decimal growth scale (Zadoks et al., 1974 ), with sampling focused on three key stages Z23 (main stem and three tillers), Z61 (beginning of anthesis), and Z92 (Grain hard, not dented by thumbnail). Plant growth and nutrient analysis Plants were harvested at Z23, Z61 and Z92 (12 pots per stage). Measurements included plant height, tiller number, leaf area, chlorophyll content, and shoot and root dry weights. Chlorophyll content was assessed using a SPAD meter (SPAD-502, Spectrum® Tech, Inc., Ramsey, NJ, USA) on the youngest fully expanded flag leaves. Leaf area was determined using ImageJ software (Schneider et al., 2012 ). Roots were washed under running tap water, and both roots and shoots were oven-dried at 70°C for four days to constant weight. At maturity, grain yield per plant and 1000-grain weight were also recorded. The 1000-grain weight was estimated by weighing 100 grains and multiplying by 10. Harvest index (HI) was calculated as: HI = [grain yield/ (grain yield + shoot dry weight)] × 100 (Metho & Hammes, 2000 ). Dried shoots and grains were ground using a mini grinder for nutrient analysis. Macronutrients such as Nitrogen (N), Phosphorous (P), Potassium (K), Calcium (Ca), Magnesium (Mg), Sulphur (S), and micronutrients like Zinc (Zn), Manganese (Mn), Copper (Cu), Iron (Fe) were measured. A 0.15 g subsample was digested using a 3:1 (v/v) mixture of HNO 3 and HClO 4 , following the method of (Simmons & Loneragan, 1975 ). The digests were analysed via inductively coupled plasma optical emission spectroscopy (ICP-OES,Optima 5300 DV, USA). Each batch included blanks and two standard references materials for quality control. Total nitrogen (N) was analysed in shoot and grain using the Dumas method (Elementar vario MACRO CNS; Elementar, Germany) (Rayment & Lyons, 2011 ), with 35 mg of ground material. Soil chemical analysis Rhizosphere soil samples (three replicates per treatment) were collected at each harvest and stored at 4°C for later analysis. Analysed parameters included pH (in H 2 O and 0.01M CaCl 2 ), electrical conductivity (EC), ammonium nitrogen (NH 4 + -N), nitrate nitrogen (NO 3 − -N), dissolved organic carbon (DOC), and microbial biomass carbon (MBC). Soil pH and EC were measured with a 1:5 soil water ratio (Rayment & Lyons, 2011 ), using a calibrated pH meter (Thermo Scientific Orion Star A111). NH 4 + -N and NO 3 − -N were measured using standard colorimetric methods (Srivastava et al., 2023 ). Soil MBC was assessed using the chloroform fumigation-extraction method (Vance et al., 1987 ). Soil samples were pre-incubated at 25°C for 7 days at 45% water-holding capacity. Moist sub-samples (10 g dry weight equivalent) were fumigated with alcohol-free chloroform (CHCl₃) for 24 hours, then extracted with 0.5 M potassium sulphate (pH 8.5). Non-fumigated sub-samples were extracted using the method of (Hargreaves et al., 2003 ) followed by shaking for one hour. Extracts were syringe filtered (0.45 µm, Filtropur S) and analysed using an automated total organic carbon analyser (Shimadzu, TOC-500, Tokyo, Japan). Data are expressed on an oven-dry basis (105°C, 24 h) and represent the mean of three replicates per treatment (n = 3). Soil microbiome analysis Rhizosheath soil—soil tightly adhering to the roots (Pang et al., 2018 ), was collected by gently shaking the roots for about 30 seconds. Soil samples were stored in sterile 20 mL tubes at − 20°C prior to DNA extraction and metagenomic sequencing. DNA extraction and quantification DNA was extracted from 0.25 g of rhizosphere soil using the DNeasy PowerSoil Pro® DNA Isolation Kit (Qiagen, Germantown, MD, USA), following the manufacturer's protocol with a modified lysis step. Cell lysis was performed using a FastPrep bead-beating system (Bio-101, Vista, CA, USA) at 5.5 m/s for 30 seconds. Extracted DNA was stored at − 20°C until further analysis. DNA quality and concentration were assessed using Qubit Fluorometer (Thermo Fisher Scientific Inc., Waltham, MA) and NanoDrop spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA). DNA libraries were prepared using the Oxford Nanopore Technologies’ Ligation Sequencing gDNA - Native Barcoding Kit (SQKNBD112.24) and sequenced using a DNA sequencer (MinION™ Mk1B, Oxford Nanopore Technologies). Sequencing data analysis Nanopore electrical signals were basecalled (in ‘sup’ mode) and demultiplexed using Dorado (v0.7.2, https://github.com/nanoporetech/dorado ). Reads were trimmed for adapter sequences, demultiplexed and filtered for quality reads (Q > 10, length > 200 bp) using NanoFilt (v2.8.0). High-quality reads were taxonomically classifid using Kraken2 1 (v2.1.2) with the “core nt” reference database https://benlangmead.github.io/aws-indexes/k2 , created on 12/28/2024). Kraken2 outputs were converted to BIOM format using kraken-biom ( https://github.com/smdabdoub/kraken-biom ). Microbial diversity analyses and visualisations were conducted using the R packages phyloseq 2 , ggplot2 3 and VennDiagram 4 . Statistical analysis Statistical analyses were conducted separately for each harvest using Genstat and R (R Core Team, 2020 ). Normality of data checked prior to analysis. One-way analysis of variance (ANOVA) was used to compare plant growth and nutrient data across treatments. Tukey’s Honest Significant Difference test was used for post-hoc comparisons. Statistical significance was accepted at p ≤ 0.05. Alpha diversity of microbial communities was analysed using permutational multivariate analysis of variance (PERMANOVA). Results Plant growth Figure 1 shows the growth performance of wheat (cv. Scepter) under four soil treatments at three key growth stages: (a) Z23, (b) Z61, and (c) Z92. Visual differences in plant height, leaf area, chlorophyll content, and shoot development were apparent among treatments (Fig. 1 ). Plants in amended soils appeared taller, more vigorous and greener than those in the control treatments, which exhibited poor growth, pale leaves, and reduced stature. At Z23 growth stage, significant differences in growth parameters were observed (Fig. 2 ). Although tiller number per plant did not differ significantly between treatments, all amended soils produced significantly taller plants with greater leaf area and significantly higher shoot and root dry weights than the control. Among treatments, The CLG + Compost had the highest chlorophyll content (SPAD value), followed by CLG and CLG + NPK. Table S2 presents detailed physiological parameters at growth stage Z23. Soil amendments significantly enhanced all growth metrics—except tiller number—relative to the control. Plants in the CLG + Compost treatment reached an average height of 36.33 cm with maximum leaf area (118.7 cm²), chlorophyll content (40.69), shoot dry weight (1.92 g plant − 1 ) and root dry weight (0.807 g plant − 1 ). ANOVA confirmed significant treatments effects on plant height (p = 0.0007), leaf area (p = 0.002), chlorophyll content (p = 0.023), shoot dry weight (p = 0.016), and root dry weight (p = 0.032). Tiller number did not vary significantly among treatments (p = 0.142). At Z61growth stage, similar trends were observed (Fig. 3 ). While plant height and tiller number did not significantly differ from the control (Table S2), all amended treatments resulted in significantly larger leaf area (p = 0.00015), higher chlorophyll content (p = 0.024), and greater shoot and root dry weight (p = 0.0134 and p = 0.007 respectively). At maturity (i.e., Z92 growth stage), the amended treatments significantly increased tiller numbers, shoot and root dry weights, 1000-grain weight, and grain yield compared to the control (Fig. 4 ). The CLG + Compost treatment resulted in the highest yield metrics. Table 2 summarises plant physiological and yield parameters at Z92 growth stage. Although plant height tended to increase across treatments—from 68.33 cm in the control to 73.33 cm in CLG + Compost—these differences were not statistically significant (p = 0.079). However, all other parameters showed marked improvements with soil amendments. The CLG + Compost treatment achieved the highest tiller number (6.33; p = 0.0025), shoot dry weight (9.40 g plant − 1 ; p = 0.013), and root dry weight (2.96 g plant − 1 ; p = 0.0041), all significantly greater than the control. Treatment also significantly influenced grain yield (p = 0.00016), with CLG + Compost producing the highest yield (7.70 g plant − 1 ), followed by CLG (6.99 g plant − 1 ) and CLG + NPK (6.96 g plant − 1 ). All amended treatments significantly outperformed the control (5.99 g plant − 1 ). The CLG + Compost treatment also had the highest 1000-grain weight (48.96 g), again outperforming the control (42.49 g) (p = 0.0082). Similarly, HI, a measure of grain production efficiency, was significantly higher in the amended soils (p = 0.0025), with CLG + Compost recording the highest value (45.04%), followed by CLG (44.24%), CLG + NPK (44.07%), and the control (41.31%). Table 2 Plant physiological and yield parameters of wheat (cv. Scepter) at growth stage Z92 (grain hard, not dented by thumbnail) under four soil treatments. Stage Treatments Plant physiological and yield parameters Plant height (cm) Tiller number (plant –1 ) Grain yield (g plant –1 ) 1000-grain weight (g) Shoot dry weight (g plant –1 ) Root dry weight (g plant –1 ) Harvest index (%) Z92 Control 68.33 ± 2.31a 4.67 ± 0.58a 5.9 ± 0.021a 42.49 ± 1.12a 8.52 ± 0.31a 2.13 ± 0.05a 41.31 ± 0.77a CLG 71 ± 1.0a 5.67 ± 0.58b 6.99 ± 0.18b 46.75 ± 1.23b 8.82 ± 0.07ab 2.73 ± 0.27b 44.24 ± 0.78b CLG + NPK 69.67 ± 2.08a 5.67 ± 0.58b 6.96 ± 0.10b 46.73 ± 2.15b 8.83 ± 0.12ab 2.83 ± 0.14b 44.07 ± 0.06b CLG + Compost 73.33 ± 0.58a 6.33 ± 0.58b 7.70 ± 0.13c 48.96 ± 0.42b 9.40 ± 0.20b 2.96 ± 0.12b 45.04 ± 0.64b P value 0.079 NS 0.0025** 0.00016*** 0.0082** 0.013* 0.0041** 0.0025** Control no amendment; CLG clay + lime + gypsum; CLG + NPK clay + lime + gypsum + N, P, K fertilisers; and CLG + Compost clay + lime + gypsum + compost. Data are means ± standard error (n = 3), *** p < 0.001, ** p < 0.01, * p < 0.05, NS - not significant. Different letters within a column indicate significant differences (p < 0.05) among treatments. Plant shoot and grain nutrient Table 3 summarises the shoot nutrient concentrations at the vegetative (Z23) and anthesis (Z61) stages under the four soil treatments. Significant treatment effects were observed for several nutrients at Z23 growth stage. All amended treatments had significantly higher N levels than the control. The CLG + Compost treatment had the highest P (4.89 mg plant − 1 ) and K (30.04 mg plant − 1 ), followed by CLG and CLG + NPK. The control had the lowest P (3.15 mg plant − 1 ) and K (8.19 mg plant − 1 ). The CLG + Compost treatment also had the highest Ca and S levels, while Mg did not significantly differ across treatments (p = 0.311). Among the micronutrients, CLG + Compost had significantly higher Zn levels (0.08 mg plant − 1 ) than the control (0.05 mg plant − 1 ; p = 0.036), while Mn, Cu, and Fe did not significantly vary among treatments. Nutrient trends in plant shoot at Z61 growth stage, largely mirrored those at Z23 growth stage (Table 3 ). Nitrogen concentration remained highest in the CLG + Compost treatment (126.39 mg plant − 1 ), followed by CLG (121.26 mg plant − 1 ) and CLG + NPK (117.26 mg plant − 1 ), all significantly higher than the control (107.10 mg plant − 1 ). The CLG + Compost treatment also had the highest P and K levels, followed by CLG and CLG + NPK, all significantly greater than the control. The amended treatments produced significantly higher Mg, Zn and Cu levels than the control (p = 0.017), with no significant differences for S, Mn, and Fe. Statistical analysis of the nutrient content in wheat grain revealed that all the treatments increased nutrient concentrations compared to the control, although the extent of enhancement varied between the nutrients and treatments (Table 4 ). Most of the nutrients showed highly significant differences due to the treatment effects. The CLG + Compost treatment consistently yielded the highest nutrient accumulation across all elements measured, followed by the CLG + NPK and CLG treatment. Macronutrients such as P, Ca, Zn exhibited very highly significant responses (p = 0.00013, 0.00035, and 0.00012, respectively), indicating strong treatment impacts on their availability or uptake. N, K, S, and Mg also showed significant differences (p = 0.0036, 0.0048, 0.0012, and 0.024, respectively). On the other hand, Mn and Cu showed no significant differences (p = 0.06 and 0.455, respectively), suggesting that the treatment had little to no effect on these two micronutrients. Table 3 Nutrient content in wheat (cv. Scepter) shoots at growth stage Z23 (main stem and three tillers) and Z61 (beginning of anthesis) under four soil treatments. Macronutrients (N, P, K, Ca, Mg, S) and micronutrients (Zn, Mn, Cu, Fe) were measured. Stage Treatments Plant shoot nutrient (mg plant –1 ) N P K Ca Mg S Zn Mn Cu Fe Z23 Control 22.62 ± 2.47a 3.15 ± 0.55a 8.19 ± 1.74a 9.89 ± 0.53a 2.76 ± 0.34a 1.33 ± 0.24a 0.05 ± 0.01a 0.09 ± 0.04a 0.01 ± 0.00a 0.34 ± 0.16a CLG 31.64 ± 2.32b 3.39 ± 0.27a 22.06 ± 2.83b 11.93 ± 0.89b 3.23 ± 0.32a 2.65 ± 0.31b 0.07a ± 0.01b 0.12 ± 0.03a 0.01 ± 0.00a 0.29 ± 0.12a CLG + NPK 32.43 ± 3.53b 3.67 ± 0.70ab 19.47 ± 3.83b 11.97 ± 0.95b 3.56 ± 1.26a 2.67 ± 0.28b 0.06a ± 0.01b 0.10 ± 0.02a 0.01 ± 0.00a 0.36 ± 0.23a CLG + Compost 35.26 ± 2.73b 4.89 ± 0.41b 30.04 ± 4.49c 14.80 ± 0.30c 3.85 ± 0.38a 3.09 ± 0.31c 0.08 ± 0.02b 0.11 ± 0.01a 0.01 ± 0.00a 0.46 ± 0.14a P value 0.009** 0.021* 0.0003*** 0.0007*** 0.311NS 1.47e-06*** 0.036* 0.650NS 0.455NS 0.709NS Z61 Control 107.10 ± 7.43a 18.27 ± 1.30a 119.68 ± 5.97a 12.22 ± 0.95a 5.04 ± 1.08a 11.63 ± 2.07a 0.27 ± 0.06a 0.37 ± 0.09a 0.02 ± 0.00a 2.55 ± 1.13a CLG 121.26 ± 2.94ab 22.62 ± 1.94ab 151.25b ± 3.57c 20.72b ± 2.07c 7.49 ± 0.20ab 14.52 ± 1.46a 0.38 ± 0.06ab 0.48 ± 0.09a 0.03 ± 0.0b 1.74 ± 0.55a CLG + NPK 117.26 ± 5.56ab 21.04 ± 1.33ab 142.79 ± 4.84b 18.29 ± 2.20ab 6.35 ± 1.15ab 12.85 ± 0.95a 0.30 ± 0.02ab 0.39 ± 0.05a 0.02 ± 0.0a 1.4 ± 0.21a CLG + Compost 126.39 ± 1.45b 25.95 ± 1.48b 155.79 ± 3.82c 26.11 ± 3.13c 8.75 ± 0.89b 15.25 ± 1.81a 0.45 ± 0.07c 0.49 ± 0.06a 0.023 ± 0.0ab 0.60 ± 0.12a P value 0.023* 0.009** 0.0003*** 0.002** 0.017* 0.136 NS 0.038* 0.291 NS 0.016* 0.073 NS Control no amendment; CLG clay + lime + gypsum; CLG + NPK clay + lime + gypsum + N, P, K fertilisers; and CLG + Compost clay + lime + gypsum + compost. Data are means ± standard errors (n = 3); *** p < 0.001, ** p < 0.01, * p < 0.05, NS - not significant. Different letters within a column and stages indicate significant differences (p < 0.05) among treatments. Table 4 Nutrient content in wheat (cv. Scepter) grain at growth stage Z92 (grain hard, not dented by thumbnail) under four soil treatments. Macronutrients (N, P, K, Ca, Mg, S) and micronutrients (Zn, Mn, Cu) were measured. Stage Treatments Grain nutrient (mg plant –1 ) N P K Ca Mg S Zn Mn Cu Z92 Control 169.29 ± 6.34a 22.26 ± 0.55a 34.03 ± 0.82a 1.11 ± 0.04a 6.60 ± 0.60a 8.10 ± 0.35a 0.15 ± 0.01a 0.17 ± 0.05b 0.03 ± 0.01a CLG 208.7 ± 8.41b 26.80 ± 0.41b 38.20 ± 3.35ab 1.38 ± 0.05b 7.41 ± 0.45ab 9.43 ± 0.21b 0.18 ± 0.01a 0.09 ± 0.02a 0.04 ± 0.006a CLG + NPK 203.50 ± 11.0b 25.99 ± 0.93b 36.97 ± 1.94a 1.53 ± 0.05b 7.09 ± 0.33ab 9.71 ± 0.51bc 0.19 ± 0.02a 0.11 ± 0.03ab 0.04 ± 0.006a CLG + Compost 229.85 ± 14.47b 30.04 ± 1.18c 42.56 ± 1.66b 1.80 ± 0.15c 8.25 ± 0.37b 10.89 ± 0.54c 0.32 ± 0.03b 0.12 ± 0.02ab 0.04 ± 0.00a P value 0.0036** 0.00013*** 0.0048** 0.00035*** 0.024* 0.0012** 0.00012*** 0.06 NS 0.455 NS Control no amendment; CLG clay + lime + gypsum; CLG + NPK clay + lime + gypsum + N, P, K fertilisers; and CLG + Compost clay + lime + gypsum + compost. Data are means ± standard errors (n = 3); *** p < 0.001, ** p < 0.01, * p < 0.05, NS - not significant. Different letters within a column and stages indicate significant differences (p < 0.05) among treatments. Soil chemical properties The organic and inorganic amendments significantly influenced properties at Z61 growth stage, including pH, EC, ammonium nitrogen (NH₄⁺-N), nitrate nitrogen (NO₃⁻-N), dissolved organic carbon (DOC), and MBC (Fig. 5 ). Soil pH and electrical conductivity (EC) Statistical analysis of soil chemical properties (Table S1 ) showed that the control had the lowest pH (7.1 in H 2 O), while the CLG + Compost treatment recorded the highest pH (7.91). Intermediate values were observed in the CLG (7.50) and CLG + NPK (7.62) treatments. Both CLG + NPK and CLG + Compost significantly increased soil EC (0.075 and 0.074 dS m⁻¹, respectively), whereas CLG (0.048 dS m⁻¹ and 0.055 dS m⁻¹) was statistically similar to the control (0.055 dS m⁻¹). Soil NH 4 + -N and NO 3 − -N concentrations Soil treatments significantly affected NH₄⁺-N and NO₃⁻-N concentrations. NH₄⁺-N levels remained unchanged in the control and CLG treatments (0.02 mg kg⁻¹), while CLG + Compost significantly increased NH₄⁺-N (0.047 mg kg⁻¹) compared to CLG + NPK (0.03 mg kg⁻¹). NO₃⁻-N levels also varied significantly (p = 1.71× 10 –7 ), with values increasing across treatments: control (0.61 mg kg⁻¹), CLG (0.86 mg kg⁻¹), CLG + NPK (0.82 mg kg⁻¹), and CLG + Compost (0.93 mg kg⁻¹). CO 2 respiration and soil microbial biomass carbon The CLG + Compost treatment recorded the highest CO 2 respiration rates (31.24 mg kg⁻¹ C), followed by CLG + NPK (18.76 mg kg⁻¹ C), both significantly greater than CLG (14.50 mg kg⁻¹ C), and the control (14.69 mg kg⁻¹ C). A similar trend was observed for MBC, where MBC was lowest in the control (9.56 mg C kg⁻¹ soil) and significantly increased in CLG (14.15 mg C kg⁻¹), CLG + NPK (16.7 mg C kg⁻¹), and CLG + Compost (21.71 mg C kg⁻¹) (p = 0.0003). Soil microbial diversity The Venn diagrams in Fig. 6 illustrate the number of unique and shared operational taxonomic units (OTUs) of bacteria, archaea, and fungi across treatments, revealing distinct microbial community structures at Z61. Shared OTUs represent the core microbiome, while unique OTUs reflect treatment-specific effects. This study identified 6382 bacterial, 112 archaeal, and 314 fungal OTUs across the four treatments. Bacterial OTUs (Fig. 6 a) showed the greatest diversity and highest proportion of shared taxa compared to archaeal (Fig. 6 b), and fungal communities (Fig. 6 c). The CLG + Compost treatment harboured the greatest number of unique OTUs, suggesting enhanced microbial diversity driven by organic matter and nutrient availability. Microbial beat-diversity analysis (Fig. 7 ) showed clear compositional shifts in microbial communities among treatments. Non-metric multi-dimensional scaling revealed that different soil amendments significantly altered bacterial (p = 0.001) and archaeal (p = 0.009) community structures. However, alpha-diversity metrics (Fig. S2) indicated that microbial richness and evenness were not significantly affected by the treatments (Tukey’s test, p > 0.05). Discussion Soil acidification constrains plant growth and microbial activity, reducing crop productivity. Understanding how plants respond to the remediation of acidic soils—particularly the role of rhizosphere microbial communities—is vital, given the central role of microbial functioning in maintaining soil health (Zhao et al., 2020 ). This study evaluated the effect of various soil amendments on wheat growth, nutrient uptake, and rhizosphere microbial communities in acidic soil. The results indicate that applying soil amendments enhances wheat performance and soil microbial characteristics by improving microbial activity in the rhizosphere and modifying soil pH and other physiochemical properties. The CLG, CLG + NPK and CLG + Compost treatment significantly improved plant growth, nutrient concentrations, soil chemical attributes, and microbial activity, underscoring their potential for ameliorating acidic soils. These outcomes support previous findings on the benefits of organic and inorganic soil amendments for enhancing crop productivity and soil health (Chen et al., 2024 ; Liu et al., 2017 ). Among the treatments, CLG + Compost had the strongest positive effect—likely due to the labile organic carbon in compost, which supports microbial taxa associated with soil remediation (Benbi et al., 2015 ; Li et al., 2018 ). Improved plant growth and biomass accumulation All soil amendments enhanced wheat growth, with the CLG + Compost treatment yielding the most pronounced improvements. Across all growth stages, this treatment produced significantly greater shoot and root biomass, leaf area, and chlorophyll content than the control (Table S2). These results align with previous studies highlighting how organic amendments promote plant growth by improving soil structure, nutrient availability, and water retention (Agegnehu et al., 2016 ). The increased plant biomass observed in the CLG and CLG + NPK treatments illustrates the contribution of inorganic amendments in supplying essential nutrients in readily available forms (Goyal et al., 1999 ). However, the superior performance of CLG + Compost suggests that organic inputs offer additional benefits, such as enhanced soil organic matter, greater microbial activity, and improved nutrient uptake. These combined effects, including more dynamic rhizosphere microbial interactions likely explain the enhanced biomass accumulation (O'Connor et al., 2024 ). Enhanced shoot nutrient uptake Nutrient analyses revealed significantly higher N, P, and K uptake in the soil amendment treatments—particularly CLG + Compost—at Z23 and Z61 growth stages. These macronutrients are essential for processes like photosynthesis, protein synthesis, and ATP production, contributing directly to increased growth and biomass (Marschner, 2011 ). The elevated N content aligns with prior research indicating that organic amendments stimulate N mineralisation and improve N availability (Azeez & Van Averbeke, 2010 ). Increased P and K levels (Table S1 ) further support evidence that organic matter enhances nutrient retention and reduces nutrient losses (Adesemoye et al., 2009 ). The increase in Zn content, especially in the CLG + Compost treatment, suggests improved micronutrient availability, which is essential for enzyme function and metabolic regulation. The lack of significant changes in Mg levels (Table S1 ) could be due to competitive cation uptake, where Mg absorption is reduced in the presence of higher levels of other cations like Ca and K (Anderson et al., 2017 ), warranting further investigation. Enhanced grain nutrient content All soil treatments significantly increased nutrient concentrations in wheat grain grown on acidified soils (Table 4 ). Notably, N, P, and K levels were significantly higher in grain from amended soils, suggesting improved nutrient supply and uptake. While CLG and CLG + NPK offered substantial improvements, CLG + Compost produced the greatest benefits, indicating that although inorganic amendments address immediate nutrient deficiencies, compost provides a more balanced nutrient profile and supports long-term soil health. Compost improves soil structure and moisture retention while stimulating microbial activity and nutrient cycling, which are crucial for maintaining soil fertility (Ollikainen et al., 2020 ). These properties likely contributed to the superior grain nutrient levels in CLG + Compost, consistent with findings that organic amendments enhance the rhizosphere environment, promote root development and enable more efficient nutrient absorption (Scanlan et al., 2014 ). Additionally, the significant increases in grain K, Ca, and Zn levels under CLG + Compost underscore its value in addressing micronutrient deficiencies common in acidic soils (Whitten, 2002 ). These findings also emphasise the importance of addressing surface and subsurface acidity for sustainable production, as surface-level treatments alone may be insufficient (Azam & Gazey, 2023 ). Increased grain yield and harvest index All soil treatments increased grain yield and HI, with the CLG + Compost treatment achieving the highest yield, supporting prior research showing that combining organic and inorganic amendments improves yield by enhancing soil fertility, nutrient availability, and water retention (O'Connor et al., 2024 ). The increased HI, particularly in the CLG + NPK and CLG + Compost treatments, suggests a more efficient partitioning of biomass towards reproductive growth—a key factor in crop productivity (Farooqi et al., 2023 ). (Gupta et al., 2020 ) also reported increased HI with integrated nutrient management strategies. The synergy between organic and inorganic inputs likely ensures vegetative and reproductive development towards reproductive growth—a key factor in crop productivity (Farooqi et al., 2023 ). (Gupta et al., 2020 ) also reported increased HI with integrated nutrient management strategies. The synergy between organic and inorganic inputs likely ensures vegetative and reproductive development (Iqbal et al., 2021 ). Enhanced soil chemical properties and microbial biomass carbon The soil amendments significantly improved soil chemical properties, including pH and EC. The slight increase in pH in the CLG + NPK and CLG + Compost treatments indicates their capacity to buffer acidity, enhancing nutrient availability and supporting plant growth in acidic soils (Rengel, 2003 ). Higher EC values suggest increased ion concentrations and nutrient availability, particularly for P (Chen et al., 2021 ). Elevated NH 4 + -N levels and NO 3 − -N in the CLG + Compost and CLG + NPK treatments indicate enhanced N mineralisation and nitrification, consistent with previous findings showing that compost improves microbial activity and provides a slow-release N source (Azeez & Van Averbeke, 2010 ). The significant increases in CO 2 respiration and MBC in CLG + Compost indicate enhanced organic matter turnover and a more biologically active soil ecosystem (Lal, 2015 ). These results are consistent with findings by (Owen et al., 2015 ), who reported that organic amendments increase microbial abundance and activity by supplying labile organic carbon. The highest MBC observed in CLG + Compost confirms the positive influence on microbial communities and their vital role in nutrient cycling and organic matter decomposition (Lal et al., 2004 ). Shifted microbiome composition Beta-diversity analysis revealed shifts in microbial community composition in response to soil amendments, as evidenced by treatment-specific taxa in Venn diagrams (Fig. 6 ). Although alpha-diversity remained largely unchanged—consistent with previous findings (Ali et al., 2022 ; Siedt et al., 2024 )—these results suggest that the amendments altered community composition without affecting overall microbial richness. However, the functional implications of these community shifts, warrant further investigation. Conclusion This study demonstrates the effectiveness of combining organic and inorganic amendments such as clay, lime, gypsum, NPK fertilisers, and compost—in improving wheat growth, nutrient uptake, soil chemical properties, and rhizosphere microbiomes. While the CLG amendment effectively alleviated soil constraints, its overall benefits for soil health and agronomic value were significantly enhanced when combined with compost compared to NPK fertilizers. These findings enhance our understanding of how soil management practices influence soil health and microbial diversity, supporting of more sustainable farming systems in acidic sandy agroecosystems. The results suggest that optimising rhizosphere microbial communities through targeted soil amendments could offer a regenerative strategy to boost grain production and promote food security in the wheatbelt region of Western Australia. However, the findings are based on a single soil type, which may limit their applicability to other soil types and environmental conditions. Moreover, socioeconomic aspects—such as the cost-effectiveness and labour demands of implementing these amendments, were not addressed. Future research should investigate the long-term impacts of these soil amendments, assess their effectiveness across diverse soil types and cropping systems, and evaluate their role in mitigating soil degradation and enhancing resilience under the increasing environmental stresses associated with climate change. Declarations Authors’ contribution: Shompa Akter : Conducted the glasshouse experiment, collected, analysed and interpreted the plant and soil data, collected soil samples for microbiome study, extracted DNA, prepared DNA library, and performed Oxford Nanopore Sequencing using MinION, drafted original manuscript. Md Sultan Mia: Conceived the original idea, developed the research concept, and designed the experimental framework, supervised glasshouse experiment, data collection and curation, DNA extraction, library preparation, sequencing, and sequencing data acquisition from the MinION. Provided sequencing facilities, reviewed the drafts and final manuscript. M. Asaduzzaman Prodhan: Supervised library preparation, MinION sequencing, and carried out the bioinformatics workflow for soil microbiome analysis, provided editorial comments. Gaus Azam: Conception and design, supervised the experiment, provided with the experimental soils, and funding for the sequencing facilities, provided editorial comments. Zakaria M. Solaiman: Supervised the glasshouse experiment, plant & soil data collection and analysis, critically discussed the results and substantially revised the manuscript. Kadambot H.M. Siddique: Supervision, coordination, provided funding for plant and soil analysis, discussed the results and discussion sections and substantially reviewed and edited the final manuscript for important intellectual content. Funding information DPIRD/GRDC funded project DAW1902_003RTX UWA Institute of Agriculture UWA School of Agriculture and Environment Acknowledgements Authors of this study are thankful to The UWA Institute of Agriculture and the UWA School of Agriculture and Environment for research support and facilities. We acknowledge “Soil Re-engineering Project” funded by the Department of Primary Industries and Regional Development (DPIRD), and the Grains Research and Development Corporation (GRDC) for research funding.Shompa Akther would like to express her sincere gratitude to “Australia Award Scholarship” funded by the Department of Foreign Affairs and Trade (DFAT) of the Australian Government for sponsor her to study at UWA. We thank Professor Nanthi Bolan for critical comments and suggestions on the manuscript. Data availability The datasets that support the findings are available from the corresponding authors on reasonable request. 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J Environ Manage 344:118392. https://doi.org/10.1016/j.jenvman.2023.118392 Sun R, Zhang X-X, Guo X, Wang D, Chu H (2015) Bacterial diversity in soils subjected to long-term chemical fertilization can be more stably maintained with the addition of livestock manure than wheat straw. Soil Biol Biochem 88:9–18. https://doi.org/10.1016/j.soilbio.2015.05.007 Tang C, Weligama C, Sale P (2013) Subsurface soil acidification in farming systems: its possible causes and management options. Mol Environ soil Sci 389–412. https://doi.org/10.1007/978-94-007-4177-5_13 Tejada M, Hernandez M, Garcia C (2009) Soil restoration using composted plant residues: Effects on soil properties. Soil Tillage Res 102(1):109–117. https://doi.org/10.1016/j.still.2008.08.004 Tian J, Dungait JA, Lu X, Yang Y, Hartley IP, Zhang W, Mo J, Yu G, Zhou J, Kuzyakov Y (2019) Long-term nitrogen addition modifies microbial composition and functions for slow carbon cycling and increased sequestration in tropical forest soil. Glob Change Biol 25(10):3267–3281. https://doi.org/10.1111/gcb.14750 Tiritan CS, Büll LT, Crusciol CA, Filho C, Fernandes AC, D. M., Nascente AS (2016) Tillage system and lime application in a tropical region: Soil chemical fertility and corn yield in succession to degraded pastures. Soil Tillage Res 155:437–447. https://doi.org/10.1016/j.still.2015.06.012 Vance ED, Brookes PC, Jenkinson DS (1987) An extraction method for measuring soil microbial biomass C. Soil Biol Biochem 19(6):703–707. https://doi.org/10.1016/0038-0717(87)90052-6 Wang L, Butterly C, Wang Y, Herath H, Xi Y, Xiao X (2014) Effect of crop residue biochar on soil acidity amelioration in strongly acidic tea garden soils. Soil Use Manag 30(1):119–128. https://doi.org/10.1111/sum.12096 Whalen JK, Chang C, Clayton GW, Carefoot JP (2000) Cattle manure amendments can increase the pH of acid soils. Soil Sci Soc Am J 64(3):962–966. https://doi.org/10.2136/sssaj2000.643962x Whitten M (2002) Amelioration and prevention of agriculturally generated subsurface acidity in sandy soils in Western Australia Xu S, Zhang L, McLaughlin NB, Mi J, Chen Q, Liu J (2015) Effect of synthetic and natural water absorbing soil amendment soil physical properties under potato production in a semi-arid region. Soil Tillage Res 148:31–39. https://doi.org/10.1016/j.still.2014.10.002 Ye L, Camps-Arbestain M, Shen Q, Lehmann J, Singh B, Sabir M (2020) Biochar effects on crop yields with and without fertilizer: A meta‐analysis of field studies using separate controls. Soil Use Manag 36(1):2–18. https://doi.org/10.1111/sum.12546 Zadoks JC, Chang TT, Konzak CF (1974) A decimal code for the growth stages of cereals. Weed Res 14(6):415–421. https://doi.org/10.1111/j.1365-3180.1974.tb01084.x Zhang D, Wang C, Li X, Yang X, Zhao L, Liu L, Zhu C, Li R (2018) Linking plant ecological stoichiometry with soil nutrient and bacterial communities in apple orchards. Appl Soil Ecol 126:1–10. https://doi.org/10.1016/j.apsoil.2017.12.017 Zhao J, Ni T, Li J, Lu Q, Fang Z, Huang Q, Zhang R, Li R, Shen B, Shen Q (2016) Effects of organic–inorganic compound fertilizer with reduced chemical fertilizer application on crop yields, soil biological activity and bacterial community structure in a rice–wheat cropping system. Appl Soil Ecol 99:1–12. https://doi.org/10.1016/j.apsoil.2015.11.006 Zhao Z-B, He J-Z, Quan Z, Wu C-F, Sheng R, Zhang L-M, Geisen S (2020) Fertilization changes soil microbiome functioning, especially phagotrophic protists. Soil Biol Biochem 148:107863. https://doi.org/10.1016/j.soilbio.2020.107863 Supplementary Files Supplementaryfiguresandtables.docx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7136714","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":490664501,"identity":"a5336657-4095-4a60-877d-af327fc17fcf","order_by":0,"name":"Shompa Akter","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Shompa","middleName":"","lastName":"Akter","suffix":""},{"id":490664502,"identity":"5bd65463-4248-4d62-a384-b956b9b8966b","order_by":1,"name":"Md Sultan Mia","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA6ElEQVRIiWNgGAWjYFAC5gYwxcbAfAAicICgFsbGBogWtgQStTAw8BgQp0V+RmL7g587GKL52M98k+apYZDju5HA+JkHjxaDG4mNjb1nGHLbeHK3SfMcYzCWvJHALI1Xi0RiYwNvG1ALA1BLDhtD4oYbCQx4tQAd1tj4F6SF/80z6Zx/DPVALcy/8WlhADqsGWyLRA6bNNCuBIMbCWz4HXbmYeNs2TYJoJZnxtZ/+yQMZ5552GY5B5/D2pMPfHzbZpM7vz/54c0Z32zk+Y4nH77xBp/DIEACmcHYQFjDKBgFo2AUjAK8AAB3W0wsmIIMewAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0003-0762-5727","institution":"University of Western Australia","correspondingAuthor":true,"prefix":"","firstName":"Md","middleName":"Sultan","lastName":"Mia","suffix":""},{"id":490664503,"identity":"bec20246-7a30-44c0-a1e6-fa1b76571e9c","order_by":2,"name":"M. Asaduzzaman Prodhan","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"M.","middleName":"Asaduzzaman","lastName":"Prodhan","suffix":""},{"id":490664504,"identity":"09a2413e-e8da-42b7-a7d8-266b5d120d4d","order_by":3,"name":"Gaus Azam","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Gaus","middleName":"","lastName":"Azam","suffix":""},{"id":490664505,"identity":"6567e7af-3510-4918-83de-551b030cd6d7","order_by":4,"name":"Zakaria M. Solaiman","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Zakaria","middleName":"M.","lastName":"Solaiman","suffix":""},{"id":490664507,"identity":"187079de-20e6-4ca6-9baf-457e135f2dcb","order_by":5,"name":"Kadambot H.M. Siddique","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Kadambot","middleName":"H.M.","lastName":"Siddique","suffix":""}],"badges":[],"createdAt":"2025-07-16 06:55:07","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7136714/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7136714/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":87742943,"identity":"347b474d-d4e5-4b41-a88f-4b90ba790047","added_by":"auto","created_at":"2025-07-28 13:51:18","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":2254625,"visible":true,"origin":"","legend":"\u003cp\u003eVisual assessment of wheat (cv. Scepter) growth under four different soil treatments at three key wheat growth stages, according to Zadoks’ scale: (a) Z23 (main stem and three tillers), (b) Z61 (beginning of anthesis), and (c) Z92 (grain hard, not dented by thumbnail). Four soil treatments: Control (no amendment), CLG (clay + lime + gypsum), CLG + NPK (clay + lime + gypsum + N, P, K fertilisers), and CLG + Compost (clay + lime + gypsum + compost).\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-7136714/v1/0c4164eaef02b583085e244d.png"},{"id":87742944,"identity":"bbeda9e8-858b-44ca-b7e0-704d062375e8","added_by":"auto","created_at":"2025-07-28 13:51:18","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":213791,"visible":true,"origin":"","legend":"\u003cp\u003eGrowth parameters of wheat (cv. Scepter) at Z23 (main stem and three tillers), under four soil treatments. Measured parameters include (a) plant height (b) tiller number, (c) leaf area, (d) chlorophyll content (e) shoot dry weight and (f) root dry weight. Different letters above bars indicate significant differences (p \u0026lt; 0.05) among treatments. Error bars are standard deviations (n=3). Four soil treatments: Control (no amendment), CLG (clay + lime + gypsum), CLG + NPK (clay + lime + gypsum + N, P, K fertilisers), and CLG + Compost (clay + lime + gypsum + compost).\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-7136714/v1/4cb7e0736668d53ec8e10bba.png"},{"id":87744403,"identity":"ddad9cb7-e65e-4ed8-b4a1-4b811c60fadd","added_by":"auto","created_at":"2025-07-28 13:59:18","extension":"jpeg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":507330,"visible":true,"origin":"","legend":"\u003cp\u003eGrowth performance of wheat (cv. Scepter) at Z61 stage (beginning of anthesis), under four soil treatments. Measured parameters include (a) leaf area, (b) chlorophyll content (c) shoot dry weight and (d) root dry weight. Different letters above bars indicate significant differences (p \u0026lt; 0.05) among treatments. Error bars are standard deviations (n=3). Four soil treatments: Control (no amendment), CLG (clay + lime + gypsum), CLG + NPK (clay + lime + gypsum + N, P, K fertilisers), and CLG + Compost (clay + lime + gypsum + compost).\u003c/p\u003e","description":"","filename":"floatimage3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7136714/v1/1eb9e8824f59078c3ae809c0.jpeg"},{"id":87742965,"identity":"33cebfb0-6717-478c-b0e6-06d0fa4c87d8","added_by":"auto","created_at":"2025-07-28 13:51:19","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":219218,"visible":true,"origin":"","legend":"\u003cp\u003eGrowth performance of wheat (cv. Scepter) at Z92 (grain hard, not dented by thumbnail) under four soil treatments. Measured parameters include (a) tiller number, (b) 1000-grain weight, (c) grain yield, (d) harvest index, (e) shoot dry weight and (f) root dry weight. Different letters above bars indicate significant differences (p \u0026lt; 0.05) among treatments. Error bars are standard deviations (n=3). Four soil treatments: Control (no amendment), CLG (clay + lime + gypsum), CLG + NPK (clay + lime + gypsum + N, P, K fertilisers), and CLG + Compost (clay + lime + gypsum + compost).\u003c/p\u003e","description":"","filename":"floatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-7136714/v1/7fbf4a18bd3329eee41921bd.png"},{"id":87742946,"identity":"b8ebe330-861b-4df4-8764-a33d92289bc1","added_by":"auto","created_at":"2025-07-28 13:51:18","extension":"jpeg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":495342,"visible":true,"origin":"","legend":"\u003cp\u003eThe effects of four soil treatments on (a) pH (b) EC (c) NH\u003csub\u003e4\u003c/sub\u003e\u003csup\u003e+\u003c/sup\u003e-N, (d) NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e-\u003c/sup\u003e\u003csub\u003e \u003c/sub\u003e-N (e) DOC, and (f) MBC in the soil at wheat growth stage Z61 (beginning of anthesis). Different letters above bars indicate significant differences (p \u0026lt; 0.05) among treatments. Error bars are standard deviations (n=3). Four soil treatments: Control (no amendment), CLG (clay + lime + gypsum), CLG + NPK (clay + lime + gypsum + N, P, K fertilisers), and CLG + Compost (clay + lime + gypsum + compost). EC (electrical conductivity), NH\u003csub\u003e4\u003c/sub\u003e\u003csup\u003e+\u003c/sup\u003e-N (ammonium nitrogen), NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e—\u003c/sup\u003eN (nitrate nitrogen), DOC (dissolved organic carbon), MBC (microbial biomass carbon).\u003c/p\u003e","description":"","filename":"floatimage5.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7136714/v1/4dd48dac55fb638c4294857e.jpeg"},{"id":87742953,"identity":"27fc4783-b4c5-4cd9-9bf7-8b10211e3397","added_by":"auto","created_at":"2025-07-28 13:51:18","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":242741,"visible":true,"origin":"","legend":"\u003cp\u003eVenn diagrams showing (a) bacterial, (b) archaeal, and (c) fungal operational taxonomic units (OTUs) in the soil at wheat growth stage Z61 (beginning of anthesis) under different soil treatments. Colours represent different treatments. Numbers in overlapping regions shared OTUS among treatments; non-overlapping values indicate OTUs unique to each treatment. Four soil treatments: Control (no amendment), CLG (clay + lime + gypsum), CLG + NPK (clay + lime + gypsum + N, P, K fertilisers), and CLG + Compost (clay + lime + gypsum + compost).\u003c/p\u003e","description":"","filename":"floatimage6.png","url":"https://assets-eu.researchsquare.com/files/rs-7136714/v1/7152b7836f546b69a4fd9884.png"},{"id":87742947,"identity":"eb869ea9-9750-4657-970f-c1379743c1f2","added_by":"auto","created_at":"2025-07-28 13:51:18","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":14572,"visible":true,"origin":"","legend":"\u003cp\u003eNon-metric multi-dimensional scaling (NMDS) plots showing beta-diversity of (a) bacterial, (b) archaeal, and (c) fungal communities based on Bray–Curtis distances at wheat growth stage Z61 (beginning of anthesis) under four different soil treatments. Four soil treatments: Control (no amendment), CLG (clay + lime + gypsum), CLG + NPK (clay + lime + gypsum + N, P, K fertilisers), and CLG + Compost (clay + lime + gypsum + compost).\u003c/p\u003e","description":"","filename":"floatimage7.png","url":"https://assets-eu.researchsquare.com/files/rs-7136714/v1/dfd319e20a42a34ec112983f.png"},{"id":92390971,"identity":"55d5f846-8f47-455a-9f11-06d9402624f1","added_by":"auto","created_at":"2025-09-29 08:39:34","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":4763680,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7136714/v1/aa28c952-7939-47db-8729-636bf195d802.pdf"},{"id":87742950,"identity":"8a0ba092-a2e6-403b-98cf-1f63d2951867","added_by":"auto","created_at":"2025-07-28 13:51:18","extension":"docx","order_by":5,"title":"","display":"","copyAsset":false,"role":"supplement","size":230390,"visible":true,"origin":"","legend":"","description":"","filename":"Supplementaryfiguresandtables.docx","url":"https://assets-eu.researchsquare.com/files/rs-7136714/v1/7e1e5c9f7f2a0b50eaead8bc.docx"}],"financialInterests":"","formattedTitle":"Exploring the effect of soil amelioration on wheat growth, nutrition, and soil microbiomes in acidic sandy soil","fulltext":[{"header":"Introduction","content":"\u003cp\u003eWheat (\u003cem\u003eTriticum aestivum\u003c/em\u003e L.) is a major crop in Australia, significantly contributing to the nation\u0026rsquo;s economy. However, soil acidity significantly constrains productivity, particularly in the Western Australian grain belt region, where over 70% of topsoils and 50% of subsoils fall below recommended pH level 5.5 and 4.8, respectively (Gazey et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Acidic subsoils significantly impact wheat production, causing annual economic losses AU\u003cspan\u003e$\u003c/span\u003e1.6\u0026nbsp;billion (Herbert, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Petersen, \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). This decline in soil quality has broader implications for food security, biodiversity, and ecosystem function.\u003c/p\u003e\u003cp\u003eBoth organic and inorganic soil amendments are used to improve soil fertility. Common organic materials include manures, composts, plant residues, household waste, sewage sludge and biochar, while inorganic amendments include lime, gypsum, clay minerals. These materials can improve nutrient and water availability, stimulate microbial activity, and enhance plant nutrient absorption (Tejada et al., \u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Xu et al., \u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Lime, compost, and inorganic fertilisers have been widely studies for their capacity to enhance soil fertility and crop productivity (Marschner, \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2011\u003c/span\u003e), improve soil aggregation and bulk density, and optimise air\u0026ndash;water relations (Manirakiza \u0026amp; Şeker, \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Tejada et al., \u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e2009\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eLiming is a proven strategy for neutralising soil acidity, increasing pH and improving fertility (Orton et al., \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). It enhances the availability of key nutrients such as calcium (Ca) and magnesium (Mg) (Bossolani et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), both essential for plant and microbial development (Guo et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Jha et al., \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Liu et al., \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Liming reduces the toxicity of aluminium ions (Al\u003csup\u003e3+\u003c/sup\u003e) by replacing them with calcium ions (Ca\u003csup\u003e2+\u003c/sup\u003e), thereby raising soil pH to enhance nutrient availability (Paradelo et al., \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). In non-tillage systems, surface lime application is effective for ameliorating surface acidity (Carmeis Filho et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Joris et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Tiritan et al., \u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e2016\u003c/span\u003e), but less for acidic subsoil (Azam \u0026amp; Gazey, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eCombining deep tillage with lime incorporation has been shown to improve rooting depth, water use efficiency, and wheat grain yield by promoting root development (Scanlan et al., \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2014\u003c/span\u003e) and increasing tolerance to moisture stress during grain filling (Azam \u0026amp; Gazey, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Gypsum (CaSO\u003csub\u003e4\u003c/sub\u003e.2H\u003csub\u003e2\u003c/sub\u003eO), often applied with lime, helps displace excess sodium ions (Na\u003csup\u003e+\u003c/sup\u003e) in saline soils and enhances water infiltration while also supplying Ca (Oster, \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e1982\u003c/span\u003e). In Finland, gypsum application reduced phosphorus (P) runoff to surface waters by up to 50% (Ollikainen et al., \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Applying lime together with gypsum effectively reduces subsoil acidity particularly by facilitating the movement of basic cations into the subsoil (Tang et al., \u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e2013\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eHowever, lime alone may not fully restore degraded sandy soil prone to nutrient leaching (Hamza \u0026amp; Anderson, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2003\u003c/span\u003e). Organic amendments like compost and manure can improve soil structure, increase organic matter, and support microbial activity\u0026mdash;key factors for long-term soil health (Lal, \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). These amendments stabilise soil aggregates, enhance soil water-holding capacity, and slowly release nutrients, complementing the benefits of lime (Whalen et al., \u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e2000\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eCompost increases soil organic matter, improves physical properties such as infiltration, aeration and porosity, and supports microbial growth by increasing biomass, and biodiversity (Irshad et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Sharif et al., \u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). It also supports essential nutrients (N, P, K, Ca, and Mg) for plants to absorb through their roots (Irshad et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2013\u003c/span\u003e) and reduce soil erosion. In a US study, composted sandy clay loam retained 80% of simulated rainfall and reduced runoff by 60% (Faucette et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). Organic matter can also raise soil pH and reduced Al\u003csup\u003e3+\u003c/sup\u003e toxicity by enhancing buffering capacity and forming Al-chelates (Wang et al., \u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eClay amendment offers another option for improving sandy soils by increasing water and nutrient retention, crop growth and soil carbon content (Davenport et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Schapel et al., \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Ye et al., \u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Fine particles such as clay and silt contribute to better aggregate formation, higher nutrient availability, and improved soil organic carbon (SOC) stability (Bronick \u0026amp; Lal, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Feng et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Regelink et al., \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), and they also enhance cation exchange capacity (Chorom et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e1994\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eSoil microbial communities are crucial in nutrient cycling and soil function. Soil harbours a rich diversity of microbes (Azim, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), including bacteria, fungi, and algae, making up about 2% of SOC (Lakshmipathi et al., \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). These microorganisms help decompose organic matter, recycle nutrients, and support soil structure and function (Heikkinen et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Lazcano et al., \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). However, soil acidity can reduce microbial activity and diversity (Kuramae et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Navarrete et al., \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2013\u003c/span\u003e) by lowering carbon availability, restricting nutrient, and increasing toxic elements like aluminium (Al) and manganese (Mn) (Bowman et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Tian et al., \u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Amendments such as lime and compost can counter these effects by enhancing microbial biomass and diversity (Sun et al., \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Zhao et al., \u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) and, in turn, improving nutrient availability and plant uptake (Zhang et al., \u003cspan citationid=\"CR79\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eWhile many studies have examined individual amendments like lime, compost, or biochar, relatively few have explored the combined effects of multiple amendments on plant growth, soil health and microbial communities in sandy soil. Addressing this gap is essential for developing sustainable management practices for acidic sandy soil. The field experiment of \u0026ldquo;re-engineering soils\u0026rdquo; project aimed to address multiple soil constraints including acidity, compaction, poor water-holding capacity, water repellence, and low fertility\u0026mdash;through soil amelioration with lime, clay, gypsum, fertilisers, and organic matter to enhance water use efficiency of grain crops in Western Australia. This study investigates the effects of soil amelioration on wheat growth, nutrition and soil microbiomes through a glasshouse experiment. The specific objective of the study is to assess the effects of soil amendments on soil health, plant performances, and soil microbiomes. We hypothesise that combinations of different soil amendments will enhance wheat productivity by improving soil physiochemical properties and supporting diverse microbial interactions. These findings will inform sustainable management strategies to improve crop performance and soil health in acidic sandy soils.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cp\u003eSite description and soil collection\u003c/p\u003e\u003cp\u003eUndisturbed soil cores were collected from a field trial conducted by the Department of Primary Industries and Regional Development (DPIRD) as part of the re-engineering soil project in Bolgart, Western Australia (31\u0026deg;18\u0026rsquo;59.9\u0026rdquo;S, 116\u0026deg;34\u0026rsquo;37.5\u0026rdquo;E). The site had previously received following four soil treatments (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e\u003cb\u003e)\u003c/b\u003e to address multiple soil constraints. In total, 36 intact soil cores (4 treatments \u0026times; 3 replications \u0026times; 3 destructive harvests) were collected using cylindrical PVC pots (10 cm diameter, and 20 cm height) with sharpened bases, hydraulically press into the soil. Cores were transported to The University of Western Australia (UWA) for glasshouse experiment. Baseline soil chemical properties are detailed in Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e. The Bolgart region has a Mediterranean-type climate, characterised by hot, dry summer and cool, wet winters, with an average annual rainfall of 612 mm. Temperatures range from 10.5\u0026ndash;26.7\u0026deg;C. The soil was moderately acidic (topsoil pH 5.2, subsurface pH 4.6) with a loamy sand texture, and low to medium Al (~\u0026thinsp;6 mg kg\u003csup\u003e\u0026ndash;1\u003c/sup\u003e). The field was heavily compacted, with penetrometer resistance readings between 3 and 4 MPa (Azam et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2024\u003c/span\u003e) before the application of soil amendments.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eExperimental treatments applied to the top 10 cm of field soil\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"2\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTreatments\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eDescription\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eT\u003csub\u003e1\u003c/sub\u003e (Control)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eNo amendment.\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eT\u003csub\u003e2\u003c/sub\u003e (CLG)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eClay @ 110 t ha\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, Lime @ 1.5 t ha\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, Gypsum @ 1 t ha\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e.\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eT\u003csub\u003e3\u003c/sub\u003e (CLG\u0026thinsp;+\u0026thinsp;NPK)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eClay @110 t ha\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, Lime @ 1.5 t ha\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, Gypsum @ 1 t ha\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, Nitrogen (N) @ 40kg ha\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, Phosphorous (P) @12.5 kg ha\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e and Potassium (K) @15 kg ha\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e.\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eT\u003csub\u003e4\u003c/sub\u003e (CLG\u0026thinsp;+\u0026thinsp;Compost)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eClay@110 t ha\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, [email protected] t ha\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, Gypsum@1 t ha\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, Compost@42t ha\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e.\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eExperimental design and glasshouse set up\u003c/p\u003e\u003cp\u003eThe 36 PVC soil cores were arranged in a completely randomised design into the UWA glasshouse incorporating the same four treatments and three harvest stages, each replicated three times. Five wheat seeds (cv. Scepter) were sown per pot and thinned to two plants eight days after sowing. Moisture content was maintained at 75% of field capacity during germination and increased to 100% during active growth. Glasshouse temperatures were maintained at 20\u0026ndash;26\u0026deg;C, with relative humidity ranging from 34.9\u0026ndash;84.7%. Plants were grown from 1 May to 10 September 2024. Crop development stages were recorded following the Zadoks\u0026rsquo; decimal growth scale (Zadoks et al., \u003cspan citationid=\"CR78\" class=\"CitationRef\"\u003e1974\u003c/span\u003e), with sampling focused on three key stages Z23 (main stem and three tillers), Z61 (beginning of anthesis), and Z92 (Grain hard, not dented by thumbnail).\u003c/p\u003e\u003cp\u003ePlant growth and nutrient analysis\u003c/p\u003e\u003cp\u003ePlants were harvested at Z23, Z61 and Z92 (12 pots per stage). Measurements included plant height, tiller number, leaf area, chlorophyll content, and shoot and root dry weights. Chlorophyll content was assessed using a SPAD meter (SPAD-502, Spectrum\u0026reg; Tech, Inc., Ramsey, NJ, USA) on the youngest fully expanded flag leaves. Leaf area was determined using ImageJ software (Schneider et al., \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). Roots were washed under running tap water, and both roots and shoots were oven-dried at 70\u0026deg;C for four days to constant weight. At maturity, grain yield per plant and 1000-grain weight were also recorded. The 1000-grain weight was estimated by weighing 100 grains and multiplying by 10. Harvest index (HI) was calculated as:\u003c/p\u003e\u003cp\u003eHI = [grain yield/ (grain yield\u0026thinsp;+\u0026thinsp;shoot dry weight)] \u0026times; 100 (Metho \u0026amp; Hammes, \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2000\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eDried shoots and grains were ground using a mini grinder for nutrient analysis. Macronutrients such as Nitrogen (N), Phosphorous (P), Potassium (K), Calcium (Ca), Magnesium (Mg), Sulphur (S), and micronutrients like Zinc (Zn), Manganese (Mn), Copper (Cu), Iron (Fe) were measured. A 0.15 g subsample was digested using a 3:1 (v/v) mixture of HNO\u003csub\u003e3\u003c/sub\u003e and HClO\u003csub\u003e4\u003c/sub\u003e, following the method of (Simmons \u0026amp; Loneragan, \u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e1975\u003c/span\u003e). The digests were analysed via inductively coupled plasma optical emission spectroscopy (ICP-OES,Optima 5300 DV, USA). Each batch included blanks and two standard references materials for quality control. Total nitrogen (N) was analysed in shoot and grain using the Dumas method (Elementar vario MACRO CNS; Elementar, Germany) (Rayment \u0026amp; Lyons, \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2011\u003c/span\u003e), with 35 mg of ground material.\u003c/p\u003e\u003cp\u003eSoil chemical analysis\u003c/p\u003e\u003cp\u003eRhizosphere soil samples (three replicates per treatment) were collected at each harvest and stored at 4\u0026deg;C for later analysis. Analysed parameters included pH (in H\u003csub\u003e2\u003c/sub\u003eO and 0.01M CaCl\u003csub\u003e2\u003c/sub\u003e), electrical conductivity (EC), ammonium nitrogen (NH\u003csub\u003e4\u003c/sub\u003e\u003csup\u003e+\u003c/sup\u003e-N), nitrate nitrogen (NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e-N), dissolved organic carbon (DOC), and microbial biomass carbon (MBC). Soil pH and EC were measured with a 1:5 soil water ratio (Rayment \u0026amp; Lyons, \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2011\u003c/span\u003e), using a calibrated pH meter (Thermo Scientific Orion Star A111). NH\u003csub\u003e4\u003c/sub\u003e\u003csup\u003e+\u003c/sup\u003e-N and NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e-N were measured using standard colorimetric methods (Srivastava et al., \u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eSoil MBC was assessed using the chloroform fumigation-extraction method (Vance et al., \u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e1987\u003c/span\u003e). Soil samples were pre-incubated at 25\u0026deg;C for 7 days at 45% water-holding capacity. Moist sub-samples (10 g dry weight equivalent) were fumigated with alcohol-free chloroform (CHCl₃) for 24 hours, then extracted with 0.5 M potassium sulphate (pH 8.5). Non-fumigated sub-samples were extracted using the method of (Hargreaves et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2003\u003c/span\u003e) followed by shaking for one hour. Extracts were syringe filtered (0.45 \u0026micro;m, Filtropur S) and analysed using an automated total organic carbon analyser (Shimadzu, TOC-500, Tokyo, Japan). Data are expressed on an oven-dry basis (105\u0026deg;C, 24 h) and represent the mean of three replicates per treatment (n\u0026thinsp;=\u0026thinsp;3).\u003c/p\u003e\u003cp\u003eSoil microbiome analysis\u003c/p\u003e\u003cp\u003eRhizosheath soil\u0026mdash;soil tightly adhering to the roots (Pang et al., \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), was collected by gently shaking the roots for about 30 seconds. Soil samples were stored in sterile 20 mL tubes at \u0026minus;\u0026thinsp;20\u0026deg;C prior to DNA extraction and metagenomic sequencing.\u003c/p\u003e\u003cp\u003e\u003cem\u003eDNA extraction and quantification\u003c/em\u003e\u003c/p\u003e\u003cp\u003eDNA was extracted from 0.25 g of rhizosphere soil using the DNeasy PowerSoil Pro\u0026reg; DNA Isolation Kit (Qiagen, Germantown, MD, USA), following the manufacturer's protocol with a modified lysis step. Cell lysis was performed using a FastPrep bead-beating system (Bio-101, Vista, CA, USA) at 5.5 m/s for 30 seconds. Extracted DNA was stored at \u0026minus;\u0026thinsp;20\u0026deg;C until further analysis. DNA quality and concentration were assessed using Qubit Fluorometer (Thermo Fisher Scientific Inc., Waltham, MA) and NanoDrop spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA). DNA libraries were prepared using the Oxford Nanopore Technologies\u0026rsquo; Ligation Sequencing gDNA - Native Barcoding Kit (SQKNBD112.24) and sequenced using a DNA sequencer (MinION\u0026trade; Mk1B, Oxford Nanopore Technologies).\u003c/p\u003e\u003cp\u003e\u003cem\u003eSequencing data analysis\u003c/em\u003e\u003c/p\u003e\u003cp\u003eNanopore electrical signals were basecalled (in \u0026lsquo;sup\u0026rsquo; mode) and demultiplexed using Dorado (v0.7.2, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://github.com/nanoporetech/dorado\u003c/span\u003e\u003cspan address=\"https://github.com/nanoporetech/dorado\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). Reads were trimmed for adapter sequences, demultiplexed and filtered for quality reads (Q\u0026thinsp;\u0026gt;\u0026thinsp;10, length\u0026thinsp;\u0026gt;\u0026thinsp;200 bp) using NanoFilt (v2.8.0). High-quality reads were taxonomically classifid using Kraken2\u003csup\u003e1\u003c/sup\u003e (v2.1.2) with the \u0026ldquo;core nt\u0026rdquo; reference database \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://benlangmead.github.io/aws-indexes/k2\u003c/span\u003e\u003cspan address=\"https://benlangmead.github.io/aws-indexes/k2\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e, created on 12/28/2024). Kraken2 outputs were converted to BIOM format using kraken-biom (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://github.com/smdabdoub/kraken-biom\u003c/span\u003e\u003cspan address=\"https://github.com/smdabdoub/kraken-biom\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). Microbial diversity analyses and visualisations were conducted using the R packages phyloseq\u003csup\u003e2\u003c/sup\u003e, ggplot2\u003csup\u003e3\u003c/sup\u003e and VennDiagram\u003csup\u003e4\u003c/sup\u003e.\u003c/p\u003e\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003eStatistical analysis\u003c/h2\u003e\u003cp\u003eStatistical analyses were conducted separately for each harvest using Genstat and R (R Core Team, \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Normality of data checked prior to analysis. One-way analysis of variance (ANOVA) was used to compare plant growth and nutrient data across treatments. Tukey\u0026rsquo;s Honest Significant Difference test was used for post-hoc comparisons. Statistical significance was accepted at p\u0026thinsp;\u0026le;\u0026thinsp;0.05. Alpha diversity of microbial communities was analysed using permutational multivariate analysis of variance (PERMANOVA).\u003c/p\u003e\u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003ePlant growth\u003c/p\u003e\u003cp\u003eFigure \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e shows the growth performance of wheat (cv. Scepter) under four soil treatments at three key growth stages: (a) Z23, (b) Z61, and (c) Z92. Visual differences in plant height, leaf area, chlorophyll content, and shoot development were apparent among treatments (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Plants in amended soils appeared taller, more vigorous and greener than those in the control treatments, which exhibited poor growth, pale leaves, and reduced stature.\u003c/p\u003e\u003cp\u003eAt Z23 growth stage, significant differences in growth parameters were observed (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Although tiller number per plant did not differ significantly between treatments, all amended soils produced significantly taller plants with greater leaf area and significantly higher shoot and root dry weights than the control. Among treatments, The CLG\u0026thinsp;+\u0026thinsp;Compost had the highest chlorophyll content (SPAD value), followed by CLG and CLG\u0026thinsp;+\u0026thinsp;NPK.\u003c/p\u003e\u003cp\u003eTable S2 presents detailed physiological parameters at growth stage Z23. Soil amendments significantly enhanced all growth metrics\u0026mdash;except tiller number\u0026mdash;relative to the control. Plants in the CLG\u0026thinsp;+\u0026thinsp;Compost treatment reached an average height of 36.33 cm with maximum leaf area (118.7 cm\u0026sup2;), chlorophyll content (40.69), shoot dry weight (1.92 g plant\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) and root dry weight (0.807 g plant\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e). ANOVA confirmed significant treatments effects on plant height (p\u0026thinsp;=\u0026thinsp;0.0007), leaf area (p\u0026thinsp;=\u0026thinsp;0.002), chlorophyll content (p\u0026thinsp;=\u0026thinsp;0.023), shoot dry weight (p\u0026thinsp;=\u0026thinsp;0.016), and root dry weight (p\u0026thinsp;=\u0026thinsp;0.032). Tiller number did not vary significantly among treatments (p\u0026thinsp;=\u0026thinsp;0.142).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eAt Z61growth stage, similar trends were observed (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). While plant height and tiller number did not significantly differ from the control (Table S2), all amended treatments resulted in significantly larger leaf area (p\u0026thinsp;=\u0026thinsp;0.00015), higher chlorophyll content (p\u0026thinsp;=\u0026thinsp;0.024), and greater shoot and root dry weight (p\u0026thinsp;=\u0026thinsp;0.0134 and p\u0026thinsp;=\u0026thinsp;0.007 respectively).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eAt maturity (i.e., Z92 growth stage), the amended treatments significantly increased tiller numbers, shoot and root dry weights, 1000-grain weight, and grain yield compared to the control (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). The CLG\u0026thinsp;+\u0026thinsp;Compost treatment resulted in the highest yield metrics.\u003c/p\u003e\u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e summarises plant physiological and yield parameters at Z92 growth stage. Although plant height tended to increase across treatments\u0026mdash;from 68.33 cm in the control to 73.33 cm in CLG\u0026thinsp;+\u0026thinsp;Compost\u0026mdash;these differences were not statistically significant (p\u0026thinsp;=\u0026thinsp;0.079). However, all other parameters showed marked improvements with soil amendments. The CLG\u0026thinsp;+\u0026thinsp;Compost treatment achieved the highest tiller number (6.33; p\u0026thinsp;=\u0026thinsp;0.0025), shoot dry weight (9.40 g plant\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; p\u0026thinsp;=\u0026thinsp;0.013), and root dry weight (2.96 g plant\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; p\u0026thinsp;=\u0026thinsp;0.0041), all significantly greater than the control.\u003c/p\u003e\u003cp\u003eTreatment also significantly influenced grain yield (p\u0026thinsp;=\u0026thinsp;0.00016), with CLG\u0026thinsp;+\u0026thinsp;Compost producing the highest yield (7.70 g plant\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e), followed by CLG (6.99 g plant\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) and CLG\u0026thinsp;+\u0026thinsp;NPK (6.96 g plant\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e). All amended treatments significantly outperformed the control (5.99 g plant\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e). The CLG\u0026thinsp;+\u0026thinsp;Compost treatment also had the highest 1000-grain weight (48.96 g), again outperforming the control (42.49 g) (p\u0026thinsp;=\u0026thinsp;0.0082). Similarly, HI, a measure of grain production efficiency, was significantly higher in the amended soils (p\u0026thinsp;=\u0026thinsp;0.0025), with CLG\u0026thinsp;+\u0026thinsp;Compost recording the highest value (45.04%), followed by CLG (44.24%), CLG\u0026thinsp;+\u0026thinsp;NPK (44.07%), and the control (41.31%).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003ePlant physiological and yield parameters of wheat (cv. Scepter) at growth stage Z92 (grain hard, not dented by thumbnail) under four soil treatments.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"9\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eStage\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eTreatments\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"6\" nameend=\"c8\" namest=\"c3\"\u003e\u003cp\u003ePlant physiological and yield parameters\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003ePlant\u003c/p\u003e\u003cp\u003eheight\u003c/p\u003e\u003cp\u003e(cm)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eTiller number\u003c/p\u003e\u003cp\u003e(plant\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eGrain\u003c/p\u003e\u003cp\u003eyield\u003c/p\u003e\u003cp\u003e(g plant\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1000-grain\u003c/p\u003e\u003cp\u003eweight\u003c/p\u003e\u003cp\u003e(g)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eShoot dry\u003c/p\u003e\u003cp\u003eweight\u003c/p\u003e\u003cp\u003e(g plant\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eRoot dry\u003c/p\u003e\u003cp\u003eweight\u003c/p\u003e\u003cp\u003e(g plant\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eHarvest\u003c/p\u003e\u003cp\u003eindex\u003c/p\u003e\u003cp\u003e(%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e\u003cp\u003e\u003cb\u003eZ92\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eControl\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e68.33\u0026thinsp;\u0026plusmn;\u0026thinsp;2.31a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e4.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.021a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e42.49\u0026thinsp;\u0026plusmn;\u0026thinsp;1.12a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e8.52\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e41.31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.77a\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCLG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e71\u0026thinsp;\u0026plusmn;\u0026thinsp;1.0a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e46.75\u0026thinsp;\u0026plusmn;\u0026thinsp;1.23b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e8.82\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07ab\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2.73\u0026thinsp;\u0026plusmn;\u0026thinsp;0.27b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e44.24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.78b\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCLG\u0026thinsp;+\u0026thinsp;NPK\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e69.67\u0026thinsp;\u0026plusmn;\u0026thinsp;2.08a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.96\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e46.73\u0026thinsp;\u0026plusmn;\u0026thinsp;2.15b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e8.83\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12ab\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2.83\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e44.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06b\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCLG\u0026thinsp;+\u0026thinsp;Compost\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e73.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e7.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13c\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e48.96\u0026thinsp;\u0026plusmn;\u0026thinsp;0.42b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e9.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2.96\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e45.04\u0026thinsp;\u0026plusmn;\u0026thinsp;0.64b\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eP value\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e0.079\u003c/b\u003e\u003csup\u003e\u003cb\u003eNS\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e0.0025**\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e0.00016***\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003e0.0082**\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u003cb\u003e0.013*\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u003cb\u003e0.0041**\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e\u003cb\u003e0.0025**\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cem\u003eControl\u003c/em\u003e no amendment; \u003cem\u003eCLG\u003c/em\u003e clay\u0026thinsp;+\u0026thinsp;lime\u0026thinsp;+\u0026thinsp;gypsum; \u003cem\u003eCLG\u0026thinsp;+\u0026thinsp;NPK\u003c/em\u003e clay\u0026thinsp;+\u0026thinsp;lime\u0026thinsp;+\u0026thinsp;gypsum\u0026thinsp;+\u0026thinsp;N, P, K fertilisers; and \u003cem\u003eCLG\u0026thinsp;+\u0026thinsp;Compost\u003c/em\u003e clay\u0026thinsp;+\u0026thinsp;lime\u0026thinsp;+\u0026thinsp;gypsum\u0026thinsp;+\u0026thinsp;compost.\u003c/p\u003e\u003cp\u003eData are means\u0026thinsp;\u0026plusmn;\u0026thinsp;standard error (n\u0026thinsp;=\u0026thinsp;3), *** p\u0026thinsp;\u0026lt;\u0026thinsp;0.001, ** p\u0026thinsp;\u0026lt;\u0026thinsp;0.01, * p\u0026thinsp;\u0026lt;\u0026thinsp;0.05, NS - not significant. Different letters within a column indicate significant differences\u003c/p\u003e\u003cp\u003e(p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) among treatments.\u003c/p\u003e\u003cp\u003ePlant shoot and grain nutrient\u003c/p\u003e\u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e summarises the shoot nutrient concentrations at the vegetative (Z23) and anthesis (Z61) stages under the four soil treatments. Significant treatment effects were observed for several nutrients at Z23 growth stage. All amended treatments had significantly higher N levels than the control. The CLG\u0026thinsp;+\u0026thinsp;Compost treatment had the highest P (4.89 mg plant\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) and K (30.04 mg plant\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e), followed by CLG and CLG\u0026thinsp;+\u0026thinsp;NPK. The control had the lowest P (3.15 mg plant\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) and K (8.19 mg plant\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e). The CLG\u0026thinsp;+\u0026thinsp;Compost treatment also had the highest Ca and S levels, while Mg did not significantly differ across treatments (p\u0026thinsp;=\u0026thinsp;0.311). Among the micronutrients, CLG\u0026thinsp;+\u0026thinsp;Compost had significantly higher Zn levels (0.08 mg plant\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) than the control (0.05 mg plant\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; p\u0026thinsp;=\u0026thinsp;0.036), while Mn, Cu, and Fe did not significantly vary among treatments.\u003c/p\u003e\u003cp\u003eNutrient trends in plant shoot at Z61 growth stage, largely mirrored those at Z23 growth stage (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Nitrogen concentration remained highest in the CLG\u0026thinsp;+\u0026thinsp;Compost treatment (126.39 mg plant\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e), followed by CLG (121.26 mg plant\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) and CLG\u0026thinsp;+\u0026thinsp;NPK (117.26 mg plant\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e), all significantly higher than the control (107.10 mg plant\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e). The CLG\u0026thinsp;+\u0026thinsp;Compost treatment also had the highest P and K levels, followed by CLG and CLG\u0026thinsp;+\u0026thinsp;NPK, all significantly greater than the control. The amended treatments produced significantly higher Mg, Zn and Cu levels than the control (p\u0026thinsp;=\u0026thinsp;0.017), with no significant differences for S, Mn, and Fe.\u003c/p\u003e\u003cp\u003eStatistical analysis of the nutrient content in wheat grain revealed that all the treatments increased nutrient concentrations compared to the control, although the extent of enhancement varied between the nutrients and treatments (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Most of the nutrients showed highly significant differences due to the treatment effects. The CLG\u0026thinsp;+\u0026thinsp;Compost treatment consistently yielded the highest nutrient accumulation across all elements measured, followed by the CLG\u0026thinsp;+\u0026thinsp;NPK and CLG treatment. Macronutrients such as P, Ca, Zn exhibited very highly significant responses (p\u0026thinsp;=\u0026thinsp;0.00013, 0.00035, and 0.00012, respectively), indicating strong treatment impacts on their availability or uptake. N, K, S, and Mg also showed significant differences (p\u0026thinsp;=\u0026thinsp;0.0036, 0.0048, 0.0012, and 0.024, respectively). On the other hand, Mn and Cu showed no significant differences (p\u0026thinsp;=\u0026thinsp;0.06 and 0.455, respectively), suggesting that the treatment had little to no effect on these two micronutrients.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eNutrient content in wheat (cv. Scepter) shoots at growth stage Z23 (main stem and three tillers) and Z61 (beginning of anthesis) under four soil treatments. Macronutrients (N, P, K, Ca, Mg, S) and micronutrients (Zn, Mn, Cu, Fe) were measured.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"12\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c12\" colnum=\"12\"\u003e\u003c/div\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eStage\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eTreatments\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"10\" nameend=\"c12\" namest=\"c3\"\u003e\u003cp\u003ePlant shoot nutrient (mg plant\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eN\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eP\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eK\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eCa\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eMg\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eZn\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003eMn\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003eCu\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c12\"\u003e\u003cp\u003eFe\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e\u003cp\u003e\u003cb\u003eZ23\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eControl\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e22.62\u0026thinsp;\u0026plusmn;\u0026thinsp;2.47a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3.15\u0026thinsp;\u0026plusmn;\u0026thinsp;0.55a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e8.19\u0026thinsp;\u0026plusmn;\u0026thinsp;1.74a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e9.89\u0026thinsp;\u0026plusmn;\u0026thinsp;0.53a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e2.76\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.24a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e0.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e0.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e0.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c12\"\u003e\u003cp\u003e0.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.16a\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCLG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e31.64\u0026thinsp;\u0026plusmn;\u0026thinsp;2.32b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3.39\u0026thinsp;\u0026plusmn;\u0026thinsp;0.27a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e22.06\u0026thinsp;\u0026plusmn;\u0026thinsp;2.83b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e11.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.89b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e3.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.32a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e0.07a\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e0.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e0.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c12\"\u003e\u003cp\u003e0.29\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12a\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCLG\u0026thinsp;+\u0026thinsp;NPK\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e32.43\u0026thinsp;\u0026plusmn;\u0026thinsp;3.53b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.70ab\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e19.47\u0026thinsp;\u0026plusmn;\u0026thinsp;3.83b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e11.97\u0026thinsp;\u0026plusmn;\u0026thinsp;0.95b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e3.56\u0026thinsp;\u0026plusmn;\u0026thinsp;1.26a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e0.06a\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e0.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e0.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c12\"\u003e\u003cp\u003e0.36\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23a\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCLG\u0026thinsp;+\u0026thinsp;Compost\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e35.26\u0026thinsp;\u0026plusmn;\u0026thinsp;2.73b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e4.89\u0026thinsp;\u0026plusmn;\u0026thinsp;0.41b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e30.04\u0026thinsp;\u0026plusmn;\u0026thinsp;4.49c\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e14.80\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30c\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e3.85\u0026thinsp;\u0026plusmn;\u0026thinsp;0.38a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31c\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e0.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e0.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e0.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c12\"\u003e\u003cp\u003e0.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14a\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eP value\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e0.009**\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e0.021*\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e0.0003***\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003e0.0007***\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u003cb\u003e0.311NS\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u003cb\u003e1.47e-06***\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e\u003cb\u003e0.036*\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e0.650NS\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e\u003cb\u003e0.455NS\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c12\"\u003e\u003cp\u003e\u003cb\u003e0.709NS\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e\u003cp\u003e\u003cb\u003eZ61\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eControl\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e107.10\u0026thinsp;\u0026plusmn;\u0026thinsp;7.43a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e18.27\u0026thinsp;\u0026plusmn;\u0026thinsp;1.30a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e119.68\u0026thinsp;\u0026plusmn;\u0026thinsp;5.97a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e12.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.95a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e5.04\u0026thinsp;\u0026plusmn;\u0026thinsp;1.08a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e11.63\u0026thinsp;\u0026plusmn;\u0026thinsp;2.07a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e0.27\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e0.37\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e0.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c12\"\u003e\u003cp\u003e2.55\u0026thinsp;\u0026plusmn;\u0026thinsp;1.13a\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCLG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e121.26\u0026thinsp;\u0026plusmn;\u0026thinsp;2.94ab\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e22.62\u0026thinsp;\u0026plusmn;\u0026thinsp;1.94ab\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e151.25b\u0026thinsp;\u0026plusmn;\u0026thinsp;3.57c\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e20.72b\u0026thinsp;\u0026plusmn;\u0026thinsp;2.07c\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e7.49\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20ab\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e14.52\u0026thinsp;\u0026plusmn;\u0026thinsp;1.46a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e0.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06ab\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e0.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e0.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c12\"\u003e\u003cp\u003e1.74\u0026thinsp;\u0026plusmn;\u0026thinsp;0.55a\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCLG\u0026thinsp;+\u0026thinsp;NPK\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e117.26\u0026thinsp;\u0026plusmn;\u0026thinsp;5.56ab\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e21.04\u0026thinsp;\u0026plusmn;\u0026thinsp;1.33ab\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e142.79\u0026thinsp;\u0026plusmn;\u0026thinsp;4.84b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e18.29\u0026thinsp;\u0026plusmn;\u0026thinsp;2.20ab\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e6.35\u0026thinsp;\u0026plusmn;\u0026thinsp;1.15ab\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e12.85\u0026thinsp;\u0026plusmn;\u0026thinsp;0.95a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e0.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02ab\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e0.39\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e0.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c12\"\u003e\u003cp\u003e1.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21a\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCLG\u0026thinsp;+\u0026thinsp;Compost\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e126.39\u0026thinsp;\u0026plusmn;\u0026thinsp;1.45b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e25.95\u0026thinsp;\u0026plusmn;\u0026thinsp;1.48b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e155.79\u0026thinsp;\u0026plusmn;\u0026thinsp;3.82c\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e26.11\u0026thinsp;\u0026plusmn;\u0026thinsp;3.13c\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e8.75\u0026thinsp;\u0026plusmn;\u0026thinsp;0.89b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e15.25\u0026thinsp;\u0026plusmn;\u0026thinsp;1.81a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e0.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07c\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e0.49\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e0.023\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0ab\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c12\"\u003e\u003cp\u003e0.60\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12a\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eP value\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e0.023*\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e0.009**\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e0.0003***\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003e0.002**\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u003cb\u003e0.017*\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u003cb\u003e0.136\u003c/b\u003e\u003csup\u003e\u003cb\u003eNS\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e\u003cb\u003e0.038*\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e0.291\u003c/b\u003e\u003csup\u003e\u003cb\u003eNS\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e\u003cb\u003e0.016*\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c12\"\u003e\u003cp\u003e\u003cb\u003e0.073\u003c/b\u003e\u003csup\u003e\u003cb\u003eNS\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cem\u003eControl\u003c/em\u003e no amendment; \u003cem\u003eCLG\u003c/em\u003e clay\u0026thinsp;+\u0026thinsp;lime\u0026thinsp;+\u0026thinsp;gypsum; \u003cem\u003eCLG\u0026thinsp;+\u0026thinsp;NPK\u003c/em\u003e clay\u0026thinsp;+\u0026thinsp;lime\u0026thinsp;+\u0026thinsp;gypsum\u0026thinsp;+\u0026thinsp;N, P, K fertilisers; and \u003cem\u003eCLG\u0026thinsp;+\u0026thinsp;Compost\u003c/em\u003e clay\u0026thinsp;+\u0026thinsp;lime\u0026thinsp;+\u0026thinsp;gypsum\u0026thinsp;+\u0026thinsp;compost.\u003c/p\u003e\u003cp\u003eData are means\u0026thinsp;\u0026plusmn;\u0026thinsp;standard errors (n\u0026thinsp;=\u0026thinsp;3); *** p\u0026thinsp;\u0026lt;\u0026thinsp;0.001, ** p\u0026thinsp;\u0026lt;\u0026thinsp;0.01, * p\u0026thinsp;\u0026lt;\u0026thinsp;0.05, NS - not significant. Different letters within a column and stages indicate significant differences (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) among treatments.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eNutrient content in wheat (cv. Scepter) grain at growth stage Z92 (grain hard, not dented by thumbnail) under four soil treatments. Macronutrients (N, P, K, Ca, Mg, S) and micronutrients (Zn, Mn, Cu) were measured.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"12\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c12\" colnum=\"12\"\u003e\u003c/div\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eStage\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e\u003cp\u003eTreatments\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"9\" nameend=\"c11\" namest=\"c3\"\u003e\u003cp\u003eGrain nutrient (mg plant\u003csup\u003e\u0026ndash;1\u003c/sup\u003e)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"1\" nameend=\"c12\" namest=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eN\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eP\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eK\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eCa\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eMg\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eZn\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003eMn\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c12\" namest=\"c11\"\u003e\u003cp\u003eCu\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e\u003cp\u003e\u003cb\u003eZ92\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eControl\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e169.29\u0026thinsp;\u0026plusmn;\u0026thinsp;6.34a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e22.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.55a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e34.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.82a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e6.60\u0026thinsp;\u0026plusmn;\u0026thinsp;0.60a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e8.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.35a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e0.15\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e0.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c12\" namest=\"c11\"\u003e\u003cp\u003e0.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01a\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCLG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e208.7\u0026thinsp;\u0026plusmn;\u0026thinsp;8.41b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e26.80\u0026thinsp;\u0026plusmn;\u0026thinsp;0.41b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e38.20\u0026thinsp;\u0026plusmn;\u0026thinsp;3.35ab\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e7.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.45ab\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e9.43\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e0.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e0.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c12\" namest=\"c11\"\u003e\u003cp\u003e0.04\u0026thinsp;\u0026plusmn;\u0026thinsp;0.006a\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCLG\u0026thinsp;+\u0026thinsp;NPK\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e203.50\u0026thinsp;\u0026plusmn;\u0026thinsp;11.0b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e25.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.93b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e36.97\u0026thinsp;\u0026plusmn;\u0026thinsp;1.94a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.53\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e7.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.33ab\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e9.71\u0026thinsp;\u0026plusmn;\u0026thinsp;0.51bc\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e0.19\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02a\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e0.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03ab\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c12\" namest=\"c11\"\u003e\u003cp\u003e0.04\u0026thinsp;\u0026plusmn;\u0026thinsp;0.006a\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCLG\u0026thinsp;+\u0026thinsp;Compost\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e229.85\u0026thinsp;\u0026plusmn;\u0026thinsp;14.47b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e30.04\u0026thinsp;\u0026plusmn;\u0026thinsp;1.18c\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e42.56\u0026thinsp;\u0026plusmn;\u0026thinsp;1.66b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.80\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15c\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e8.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.37b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e10.89\u0026thinsp;\u0026plusmn;\u0026thinsp;0.54c\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e0.32\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03b\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e0.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02ab\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c12\" namest=\"c11\"\u003e\u003cp\u003e0.04\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00a\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eP value\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e0.0036**\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cb\u003e0.00013***\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003cb\u003e0.0048**\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u003cb\u003e0.00035***\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u003cb\u003e0.024*\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u003cb\u003e0.0012**\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e\u003cb\u003e0.00012***\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e\u003cb\u003e0.06\u003c/b\u003e\u003csup\u003e\u003cb\u003eNS\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"2\" nameend=\"c12\" namest=\"c11\"\u003e\u003cp\u003e\u003cb\u003e0.455\u003c/b\u003e\u003csup\u003e\u003cb\u003eNS\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cem\u003eControl\u003c/em\u003e no amendment; \u003cem\u003eCLG\u003c/em\u003e clay\u0026thinsp;+\u0026thinsp;lime\u0026thinsp;+\u0026thinsp;gypsum; \u003cem\u003eCLG\u0026thinsp;+\u0026thinsp;NPK\u003c/em\u003e clay\u0026thinsp;+\u0026thinsp;lime\u0026thinsp;+\u0026thinsp;gypsum\u0026thinsp;+\u0026thinsp;N, P, K fertilisers; and \u003cem\u003eCLG\u0026thinsp;+\u0026thinsp;Compost\u003c/em\u003e clay\u0026thinsp;+\u0026thinsp;lime\u0026thinsp;+\u0026thinsp;gypsum\u0026thinsp;+\u0026thinsp;compost.\u003c/p\u003e\u003cp\u003eData are means\u0026thinsp;\u0026plusmn;\u0026thinsp;standard errors (n\u0026thinsp;=\u0026thinsp;3); *** p\u0026thinsp;\u0026lt;\u0026thinsp;0.001, ** p\u0026thinsp;\u0026lt;\u0026thinsp;0.01, * p\u0026thinsp;\u0026lt;\u0026thinsp;0.05, NS - not significant. Different letters within a column and stages indicate significant differences (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) among treatments.\u003c/p\u003e\u003cp\u003eSoil chemical properties\u003c/p\u003e\u003cp\u003eThe organic and inorganic amendments significantly influenced properties at Z61 growth stage, including pH, EC, ammonium nitrogen (NH₄⁺-N), nitrate nitrogen (NO₃⁻-N), dissolved organic carbon (DOC), and MBC (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cem\u003eSoil pH and electrical conductivity (EC)\u003c/em\u003e\u003c/p\u003e\u003cp\u003eStatistical analysis of soil chemical properties (Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e) showed that the control had the lowest pH (7.1 in H\u003csub\u003e2\u003c/sub\u003eO), while the CLG\u0026thinsp;+\u0026thinsp;Compost treatment recorded the highest pH (7.91). Intermediate values were observed in the CLG (7.50) and CLG\u0026thinsp;+\u0026thinsp;NPK (7.62) treatments. Both CLG\u0026thinsp;+\u0026thinsp;NPK and CLG\u0026thinsp;+\u0026thinsp;Compost significantly increased soil EC (0.075 and 0.074 dS m⁻\u0026sup1;, respectively), whereas CLG (0.048 dS m⁻\u0026sup1; and 0.055 dS m⁻\u0026sup1;) was statistically similar to the control (0.055 dS m⁻\u0026sup1;).\u003c/p\u003e\u003cp\u003e\u003cem\u003eSoil NH\u003c/em\u003e\u003csub\u003e\u003cem\u003e4\u003c/em\u003e\u003c/sub\u003e\u003csup\u003e\u003cem\u003e+\u003c/em\u003e\u003c/sup\u003e\u003cem\u003e-N and NO\u003c/em\u003e\u003csub\u003e\u003cem\u003e3\u003c/em\u003e\u003c/sub\u003e\u003csup\u003e\u003cem\u003e\u0026minus;\u003c/em\u003e\u003c/sup\u003e\u003cem\u003e-N concentrations\u003c/em\u003e\u003c/p\u003e\u003cp\u003eSoil treatments significantly affected NH₄⁺-N and NO₃⁻-N concentrations. NH₄⁺-N levels remained unchanged in the control and CLG treatments (0.02 mg kg⁻\u0026sup1;), while CLG\u0026thinsp;+\u0026thinsp;Compost significantly increased NH₄⁺-N (0.047 mg kg⁻\u0026sup1;) compared to CLG\u0026thinsp;+\u0026thinsp;NPK (0.03 mg kg⁻\u0026sup1;). NO₃⁻-N levels also varied significantly (p\u0026thinsp;=\u0026thinsp;1.71\u0026times; 10\u003csup\u003e\u0026ndash;7\u003c/sup\u003e), with values increasing across treatments: control (0.61 mg kg⁻\u0026sup1;), CLG (0.86 mg kg⁻\u0026sup1;), CLG\u0026thinsp;+\u0026thinsp;NPK (0.82 mg kg⁻\u0026sup1;), and CLG\u0026thinsp;+\u0026thinsp;Compost (0.93 mg kg⁻\u0026sup1;).\u003c/p\u003e\u003cp\u003e\u003cem\u003eCO\u003c/em\u003e\u003csub\u003e\u003cem\u003e2\u003c/em\u003e\u003c/sub\u003e \u003cem\u003erespiration and soil microbial biomass carbon\u003c/em\u003e\u003c/p\u003e\u003cp\u003eThe CLG\u0026thinsp;+\u0026thinsp;Compost treatment recorded the highest CO\u003csub\u003e2\u003c/sub\u003e respiration rates (31.24 mg kg⁻\u0026sup1; C), followed by CLG\u0026thinsp;+\u0026thinsp;NPK (18.76 mg kg⁻\u0026sup1; C), both significantly greater than CLG (14.50 mg kg⁻\u0026sup1; C), and the control (14.69 mg kg⁻\u0026sup1; C). A similar trend was observed for MBC, where MBC was lowest in the control (9.56 mg C kg⁻\u0026sup1; soil) and significantly increased in CLG (14.15 mg C kg⁻\u0026sup1;), CLG\u0026thinsp;+\u0026thinsp;NPK (16.7 mg C kg⁻\u0026sup1;), and CLG\u0026thinsp;+\u0026thinsp;Compost (21.71 mg C kg⁻\u0026sup1;) (p\u0026thinsp;=\u0026thinsp;0.0003).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cem\u003eSoil microbial diversity\u003c/em\u003e\u003c/p\u003e\u003cp\u003eThe Venn diagrams in Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e illustrate the number of unique and shared operational taxonomic units (OTUs) of bacteria, archaea, and fungi across treatments, revealing distinct microbial community structures at Z61. Shared OTUs represent the core microbiome, while unique OTUs reflect treatment-specific effects. This study identified 6382 bacterial, 112 archaeal, and 314 fungal OTUs across the four treatments. Bacterial OTUs (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003ea) showed the greatest diversity and highest proportion of shared taxa compared to archaeal (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eb), and fungal communities (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003ec). The CLG\u0026thinsp;+\u0026thinsp;Compost treatment harboured the greatest number of unique OTUs, suggesting enhanced microbial diversity driven by organic matter and nutrient availability.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eMicrobial beat-diversity analysis (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003e) showed clear compositional shifts in microbial communities among treatments. Non-metric multi-dimensional scaling revealed that different soil amendments significantly altered bacterial (p\u0026thinsp;=\u0026thinsp;0.001) and archaeal (p\u0026thinsp;=\u0026thinsp;0.009) community structures. However, alpha-diversity metrics (Fig. S2) indicated that microbial richness and evenness were not significantly affected by the treatments (Tukey\u0026rsquo;s test, p\u0026thinsp;\u0026gt;\u0026thinsp;0.05).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eSoil acidification constrains plant growth and microbial activity, reducing crop productivity. Understanding how plants respond to the remediation of acidic soils\u0026mdash;particularly the role of rhizosphere microbial communities\u0026mdash;is vital, given the central role of microbial functioning in maintaining soil health (Zhao et al., \u003cspan citationid=\"CR81\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). This study evaluated the effect of various soil amendments on wheat growth, nutrient uptake, and rhizosphere microbial communities in acidic soil. The results indicate that applying soil amendments enhances wheat performance and soil microbial characteristics by improving microbial activity in the rhizosphere and modifying soil pH and other physiochemical properties.\u003c/p\u003e\u003cp\u003eThe CLG, CLG\u0026thinsp;+\u0026thinsp;NPK and CLG\u0026thinsp;+\u0026thinsp;Compost treatment significantly improved plant growth, nutrient concentrations, soil chemical attributes, and microbial activity, underscoring their potential for ameliorating acidic soils. These outcomes support previous findings on the benefits of organic and inorganic soil amendments for enhancing crop productivity and soil health (Chen et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Liu et al., \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Among the treatments, CLG\u0026thinsp;+\u0026thinsp;Compost had the strongest positive effect\u0026mdash;likely due to the labile organic carbon in compost, which supports microbial taxa associated with soil remediation (Benbi et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Li et al., \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eImproved plant growth and biomass accumulation\u003c/p\u003e\u003cp\u003eAll soil amendments enhanced wheat growth, with the CLG\u0026thinsp;+\u0026thinsp;Compost treatment yielding the most pronounced improvements. Across all growth stages, this treatment produced significantly greater shoot and root biomass, leaf area, and chlorophyll content than the control (Table S2). These results align with previous studies highlighting how organic amendments promote plant growth by improving soil structure, nutrient availability, and water retention (Agegnehu et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe increased plant biomass observed in the CLG and CLG\u0026thinsp;+\u0026thinsp;NPK treatments illustrates the contribution of inorganic amendments in supplying essential nutrients in readily available forms (Goyal et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e1999\u003c/span\u003e). However, the superior performance of CLG\u0026thinsp;+\u0026thinsp;Compost suggests that organic inputs offer additional benefits, such as enhanced soil organic matter, greater microbial activity, and improved nutrient uptake. These combined effects, including more dynamic rhizosphere microbial interactions likely explain the enhanced biomass accumulation (O'Connor et al., \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eEnhanced shoot nutrient uptake\u003c/p\u003e\u003cp\u003eNutrient analyses revealed significantly higher N, P, and K uptake in the soil amendment treatments\u0026mdash;particularly CLG\u0026thinsp;+\u0026thinsp;Compost\u0026mdash;at Z23 and Z61 growth stages. These macronutrients are essential for processes like photosynthesis, protein synthesis, and ATP production, contributing directly to increased growth and biomass (Marschner, \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). The elevated N content aligns with prior research indicating that organic amendments stimulate N mineralisation and improve N availability (Azeez \u0026amp; Van Averbeke, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Increased P and K levels (Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e) further support evidence that organic matter enhances nutrient retention and reduces nutrient losses (Adesemoye et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2009\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe increase in Zn content, especially in the CLG\u0026thinsp;+\u0026thinsp;Compost treatment, suggests improved micronutrient availability, which is essential for enzyme function and metabolic regulation. The lack of significant changes in Mg levels (Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e) could be due to competitive cation uptake, where Mg absorption is reduced in the presence of higher levels of other cations like Ca and K (Anderson et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), warranting further investigation.\u003c/p\u003e\u003cp\u003eEnhanced grain nutrient content\u003c/p\u003e\u003cp\u003eAll soil treatments significantly increased nutrient concentrations in wheat grain grown on acidified soils (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Notably, N, P, and K levels were significantly higher in grain from amended soils, suggesting improved nutrient supply and uptake. While CLG and CLG\u0026thinsp;+\u0026thinsp;NPK offered substantial improvements, CLG\u0026thinsp;+\u0026thinsp;Compost produced the greatest benefits, indicating that although inorganic amendments address immediate nutrient deficiencies, compost provides a more balanced nutrient profile and supports long-term soil health.\u003c/p\u003e\u003cp\u003eCompost improves soil structure and moisture retention while stimulating microbial activity and nutrient cycling, which are crucial for maintaining soil fertility (Ollikainen et al., \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). These properties likely contributed to the superior grain nutrient levels in CLG\u0026thinsp;+\u0026thinsp;Compost, consistent with findings that organic amendments enhance the rhizosphere environment, promote root development and enable more efficient nutrient absorption (Scanlan et al., \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Additionally, the significant increases in grain K, Ca, and Zn levels under CLG\u0026thinsp;+\u0026thinsp;Compost underscore its value in addressing micronutrient deficiencies common in acidic soils (Whitten, \u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). These findings also emphasise the importance of addressing surface and subsurface acidity for sustainable production, as surface-level treatments alone may be insufficient (Azam \u0026amp; Gazey, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIncreased grain yield and harvest index\u003c/p\u003e\u003cp\u003eAll soil treatments increased grain yield and HI, with the CLG\u0026thinsp;+\u0026thinsp;Compost treatment achieving the highest yield, supporting prior research showing that combining organic and inorganic amendments improves yield by enhancing soil fertility, nutrient availability, and water retention (O'Connor et al., \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). The increased HI, particularly in the CLG\u0026thinsp;+\u0026thinsp;NPK and CLG\u0026thinsp;+\u0026thinsp;Compost treatments, suggests a more efficient partitioning of biomass towards reproductive growth\u0026mdash;a key factor in crop productivity (Farooqi et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). (Gupta et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) also reported increased HI with integrated nutrient management strategies. The synergy between organic and inorganic inputs likely ensures vegetative and reproductive development towards reproductive growth\u0026mdash;a key factor in crop productivity (Farooqi et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). (Gupta et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) also reported increased HI with integrated nutrient management strategies. The synergy between organic and inorganic inputs likely ensures vegetative and reproductive development (Iqbal et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eEnhanced soil chemical properties and microbial biomass carbon\u003c/p\u003e\u003cp\u003eThe soil amendments significantly improved soil chemical properties, including pH and EC. The slight increase in pH in the CLG\u0026thinsp;+\u0026thinsp;NPK and CLG\u0026thinsp;+\u0026thinsp;Compost treatments indicates their capacity to buffer acidity, enhancing nutrient availability and supporting plant growth in acidic soils (Rengel, \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2003\u003c/span\u003e). Higher EC values suggest increased ion concentrations and nutrient availability, particularly for P (Chen et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Elevated NH\u003csub\u003e4\u003c/sub\u003e\u003csup\u003e+\u003c/sup\u003e-N levels and NO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e-N in the CLG\u0026thinsp;+\u0026thinsp;Compost and CLG\u0026thinsp;+\u0026thinsp;NPK treatments indicate enhanced N mineralisation and nitrification, consistent with previous findings showing that compost improves microbial activity and provides a slow-release N source (Azeez \u0026amp; Van Averbeke, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2010\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe significant increases in CO\u003csub\u003e2\u003c/sub\u003e respiration and MBC in CLG\u0026thinsp;+\u0026thinsp;Compost indicate enhanced organic matter turnover and a more biologically active soil ecosystem (Lal, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). These results are consistent with findings by (Owen et al., \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), who reported that organic amendments increase microbial abundance and activity by supplying labile organic carbon. The highest MBC observed in CLG\u0026thinsp;+\u0026thinsp;Compost confirms the positive influence on microbial communities and their vital role in nutrient cycling and organic matter decomposition (Lal et al., \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2004\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eShifted microbiome composition\u003c/p\u003e\u003cp\u003eBeta-diversity analysis revealed shifts in microbial community composition in response to soil amendments, as evidenced by treatment-specific taxa in Venn diagrams (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). Although alpha-diversity remained largely unchanged\u0026mdash;consistent with previous findings (Ali et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Siedt et al., \u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e2024\u003c/span\u003e)\u0026mdash;these results suggest that the amendments altered community composition without affecting overall microbial richness. However, the functional implications of these community shifts, warrant further investigation.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis study demonstrates the effectiveness of combining organic and inorganic amendments such as clay, lime, gypsum, NPK fertilisers, and compost\u0026mdash;in improving wheat growth, nutrient uptake, soil chemical properties, and rhizosphere microbiomes. While the CLG amendment effectively alleviated soil constraints, its overall benefits for soil health and agronomic value were significantly enhanced when combined with compost compared to NPK fertilizers. These findings enhance our understanding of how soil management practices influence soil health and microbial diversity, supporting of more sustainable farming systems in acidic sandy agroecosystems. The results suggest that optimising rhizosphere microbial communities through targeted soil amendments could offer a regenerative strategy to boost grain production and promote food security in the wheatbelt region of Western Australia. However, the findings are based on a single soil type, which may limit their applicability to other soil types and environmental conditions. Moreover, socioeconomic aspects\u0026mdash;such as the cost-effectiveness and labour demands of implementing these amendments, were not addressed. Future research should investigate the long-term impacts of these soil amendments, assess their effectiveness across diverse soil types and cropping systems, and evaluate their role in mitigating soil degradation and enhancing resilience under the increasing environmental stresses associated with climate change.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthors’ contribution:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eShompa Akter\u003c/strong\u003e\u003cstrong\u003e:\u0026nbsp;\u003c/strong\u003eConducted the glasshouse experiment, collected, analysed and interpreted the plant and soil data, collected soil samples for microbiome study, extracted DNA, prepared DNA library, and performed Oxford Nanopore Sequencing using MinION, drafted original manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMd Sultan Mia:\u0026nbsp;\u003c/strong\u003eConceived the original idea, developed the research concept, and designed the experimental framework,\u0026nbsp;supervised glasshouse experiment, data collection and curation, DNA extraction, library preparation, sequencing, and sequencing data acquisition from the MinION. Provided sequencing facilities, reviewed the drafts and final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eM. Asaduzzaman Prodhan:\u0026nbsp;\u003c/strong\u003eSupervised library preparation, MinION sequencing, and carried out the bioinformatics workflow for soil microbiome analysis, provided editorial comments.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGaus Azam:\u003c/strong\u003eConception and design,\u0026nbsp;supervised the experiment, provided with the experimental soils, and funding for the sequencing facilities, provided editorial comments.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eZakaria M. Solaiman:\u0026nbsp;\u003c/strong\u003eSupervised\u0026nbsp;the glasshouse experiment, plant \u0026amp; soil data collection and analysis,\u0026nbsp;critically discussed the results and substantially revised the manuscript.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eKadambot H.M. Siddique:\u0026nbsp;\u003c/strong\u003eSupervision, coordination, provided\u0026nbsp;funding for plant and soil analysis, discussed the results and discussion sections and substantially reviewed and edited the final manuscript\u0026nbsp;for important intellectual content.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding information\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDPIRD/GRDC funded project DAW1902_003RTX\u003c/p\u003e\n\u003cp\u003eUWA Institute of Agriculture\u003c/p\u003e\n\u003cp\u003eUWA School of Agriculture and Environment\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAuthors of this study are thankful to The UWA Institute of Agriculture and the UWA School of Agriculture and Environment for research support and facilities. We acknowledge “Soil Re-engineering Project” funded by the Department of Primary Industries and Regional Development (DPIRD), and the Grains Research and Development Corporation (GRDC) for research funding.Shompa Akther would like to express her sincere gratitude to “Australia Award Scholarship” funded by the Department of Foreign Affairs and Trade (DFAT) of the Australian Government for sponsor her to study at UWA. We thank Professor Nanthi Bolan for critical comments and suggestions on the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets that support the findings are available from the corresponding authors on reasonable request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAdesemoye A, Torbert H, Kloepper J (2009) Plant growth-promoting rhizobacteria allow reduced application rates of chemical fertilizers. 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Soil Biol Biochem 148:107863. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.soilbio.2020.107863\u003c/span\u003e\u003cspan address=\"10.1016/j.soilbio.2020.107863\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Soil amendments, Wheat yield, Nutrient content, Soil health, Soil microbiomes","lastPublishedDoi":"10.21203/rs.3.rs-7136714/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7136714/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cem\u003e\u003cstrong\u003eBackground and aims\u003c/strong\u003e\u003c/em\u003e Soil acidity poses a significant global challenge to soil health and the sustainability of agricultural production. In Western Australia’s grain belt, soil acidification—exacerbated by crop removal and the use of acidifying fertilisers—reduces land productivity. A combination of organic and inorganic soil amendments including lime, gypsum, clay, compost, and synthetic fertilisers could improve soil health and microbial function.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cstrong\u003eMethods\u003c/strong\u003e\u003c/em\u003e A glasshouse experiment was conducted using undisturbed soil cores amended three years earlier in a “re-engineering soils” field trial, which comprised four soil treatments: Control, CLG (clay, lime, gypsum), CLG + NPK (CLG + nitrogen, phosphorus, and potassium fertilisers), and CLG + Compost. The study followed a completely randomised design with three replications. Plant growth, nutrient uptake, and soil chemical and microbial properties were assessed at three key wheat growth stages, according to Zadoks’ scale: Z23, Z61, and Z92.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cstrong\u003eResults\u003c/strong\u003e\u003c/em\u003e These amendments improved plant growth, nutrient uptake, grain yield, and nutrient status, while also enhancing soil chemical properties, microbial biomass carbon (MBC), and microbial composition. CLG was the main amendment used to improve soil conditions, and its application with compost showed greater effectiveness compared to its use with NPK fertilisers.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003e\u003cstrong\u003eConclusion\u003c/strong\u003e\u003c/em\u003e This study offers insights into the benefits of organic and inorganic amendments for managing acidic sandy soil. The findings support the development of sustainable soil management strategies. Future work should explore the long-term impacts of these amendments across different soil types and cropping systems.\u003c/p\u003e","manuscriptTitle":"Exploring the effect of soil amelioration on wheat growth, nutrition, and soil microbiomes in acidic sandy soil","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-07-28 13:51:13","doi":"10.21203/rs.3.rs-7136714/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"61b33dc1-4676-49bf-a54e-a9d6a8060218","owner":[],"postedDate":"July 28th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-09-29T08:31:24+00:00","versionOfRecord":[],"versionCreatedAt":"2025-07-28 13:51:13","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7136714","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7136714","identity":"rs-7136714","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}