The soil used in the experiment was Podzol derived from loamy sand (Table 1). It was taken in early spring from a 0- to 30-cm-depth layer of a cultivated field (N 50.42947; E 22.60175). The soil was sieved through a 4-mm mesh to remove any debris. The soil size fractions were determined by a Mastersizer 2000 (Malvern, UK) laser diffractometer according to the procedure described by Ryżak and Bieganowski (2011). The apple pomace was the leftover of production of apple juice. It was stored in the freezer at a temperature of − 24 (± 2) °C until the beginning of the experimental procedures. The total phosphorus and potassium contents were determined colorimetrically and by flame photometry, respectively; the total N was measured according to the Kjeldahl method. The specific properties of both the soil and apple pomace are presented in Table 1.
Table 1 Selected properties of apple pomace and the soil Experiment I: Impact of AP on Artificial Soil Aggregates
The soil collected from the field was moistened to a field capacity, and nutrients were added in doses corresponding to N (NH4NO3) 30 kg ha−1, P (KH2PO4) 60 kg ha−1, K (K2SO4) 100 kg ha−1, and Mg (MgSO4 × 7 H2O) 25 kg ha−1. Unmodified apple pomace was added in the dose corresponding to 20 t of fresh mass per hectare (assuming that AP was mixed within the 0–10-cm top soil layer, 16.7 g of AP was added per 1 kg of soil). The artificial aggregates were formed from soil with and without AP using a 6- and 10-mm-thick acrylic glass plate with drilled 6- and 10-mm-diameter holes, according to the method proposed by Józefaciuk and Czachor (2014). The specified mass of the soil was manually pressed into the holes of the plates. Then, the cylindrical aggregates with 6 and 10 mm diameters with (AP+) and without apple pomace (AP−) were removed from the plates. The mass of the aggregates was checked to ensure uniform density (1.65 ± 0.1 Mg m−3).
Immediately after formation, 4 large (10 mm) or 16 small (6 mm) aggregates were carefully placed in 120-ml glass flasks. The total dry mass of the soil aggregates in the flasks ranged from approx. 4.6 to 5.4 g for the smaller and larger aggregates, respectively. The flasks were then airtight-closed for 2 h with a rubber cork prior to the first measurements of the cumulative CO2 concentration in the headspace. The concentration of CO2 was measured in 2 ml of gas sampled using a syringe on Shimadzu GC 14A equipped with a thermal conductivity detector (TCD). After sampling of the headspace gas, the flasks were covered with aluminum foil, in which approximately 10 small holes were made with a puncture needle. The holes allowed gas exchange at a limited rate of soil drying. Unavoidable water losses were corrected by daily watering to the initial weight using a pipette. The concentration of CO2 was repeatedly measured on the 1st, 2nd, 4th, 7th, and 21st day after the formation of the aggregates. Two hours before each measurement of the CO2 concentration, the flasks were airtight-closed with a rubber plug. Cumulative CO2 emission was used to assess the respiration rate of the soil aggregates (mg CO2-C kg−1 dry soil h−1). The aggregates used for the respiration and soil N measurements were stored in the dark at 24 °C day (14 h) and 18 °C night temperatures.
The impact of AP on soil N was evaluated by measurements of NO3−-N and NH4+-N in the soil aggregates. Twenty-four hours and 14 days after preparation, 20-g aggregates were frozen and kept at a temperature of − 24 (± 2) C prior to the measurements of the NO3−-N and NH4+-N concentrations. The concentration of NO3−-N and NH4+-N was determined in soil extracts (0.01 MCaCl2) using a flow-type spectrophotometric analyzer FIA-Star 5010 (Foss Tecator, Sweden).
Two sizes of artificially formed soil aggregates (6 and 10 mm) prepared according to the procedure described above were used for measurements of aggregate tensile strength, water and ethanol wetting rates, and the repellency index (RI). The measurements were performed after incubation for 90 days at room temperature (22 (± 2) °C). The water (Qw) and ethanol (Qe) wetting rates of the initially air-dried soil aggregate fractions were derived from the steady-state flow measurements conducted using an infiltration device (Lipiec et al. 2006). The rate of flow from the tube filled with either water or ethanol through the measured aggregate as a function of time was taken as a measure of the water wetting rate. The repellency index was the ratio of ethanol (not influenced by repellency) and water wetting rates calculated using a formula proposed by Leeds-Harrison et al. (1994). The RI (hydrophobicity) of the soil was taken as a ratio of Qw and Qe. Due to the very rapid disintegration, the ethanol wetting rate and RI were not measured for the smaller aggregates.
Crushing strength tests were performed using ZwickLine Testing Machine Z5.0 (ZwickRoell Testing Systems GmbH, Germany). The aggregates were crushed between two steel plates at the speed of 5 mm min−1, and the first peak of force recorded was used to calculate tensile strength.
Experiment II: Plant Response to AP
Faba bean (Vicia faba L.) cv. Granit and spring wheat (Triticum aestivum L.) cv. Kandela were used to study the impact of the apple pomace on plant growth. The faba bean seeds were noninoculated with Rhizobium. The plants were grown in cylinders with an inner diameter of 10 cm and a height of 40 cm filled with soil characterized by a moderate bulk density of 1.5 Mg m−3. The apple pomace was uniformly mixed with soil from the 0- to 10-cm layer in a dose corresponding to 20 t of fresh mass per ha. The soil was fertilized according to the crop demands to obtain 3 and 5 t of faba bean and spring wheat grain yields per hectare, respectively. Therefore, the doses of N, P, K, Mg, and S applied to the soil with faba bean were the same as in experiment I with the soil aggregates, and the dose of N in the soil with spring wheat was increased to 60 kg ha−1 (NH4NO3). The initial number of three seedlings of faba bean per pot was reduced to one soon after germination. In the spring wheat variant, the initial number of approx. 10 seedlings per pot was reduced to four after germination. The different number of plants per pot yielded similar plant biomass at the end of the experiment.
The conditions during the growth of both crops were as follows: day and night temperatures 24 °C (14 h) and 18 °C, photosynthetic active radiation 150 μmol photons m−2 s−1, and relative air humidity 60%. Both plant species were grown for 63 days and subjected to the following treatments: (1) C AP−: optimum soil water availability without addition of apple pomace; (2) C AP+: optimum soil water availability with addition of apple pomace; (3) D AP−: mild water deficit without addition of apple pomace; (4) D AP+: mild water deficit with addition of apple pomace. The soil water potential was maintained at the optimum level corresponding to field capacity at the soil water potential of − 33 kPa throughout the experimental period. Soil in the treatments with water deficit had an optimum water potential during the first 14 days after sowing; then, water deficit conditions were induced to maintain the soil water potential in the range from − 80 to − 60 kPa until cutting the plants at the end of the experiment. Specific soil water availability was maintained by watering the experimental cylinders to the specified weight every 24 h.
Leaf chlorophyll content was measured at day 63 using a Chlorophyll Content Meter CCM-300 (Opti-Sciences, Inc., USA) in the middle section of fully expanded leaves.
Leaf gas exchange was measured every fifth day starting from the beginning of limited watering. The measurement was conducted using a GFS3000 gas-exchange system equipped with a standard measuring head 3100-S (Walz GmbH, Germany). The conditions in the head chamber with the leaf were the same as in the growth chamber, and the CO2 concentration during the measurements was 400 ppm.
Leaf relative water content (RWC) was determined at day 63 in the first fully developed leaves. Excised leaves were immediately weighed to obtain fresh weight (FW), rehydrated in distilled water for 24 h, and weighed again to determine turgid weight (TW). Dry weight (DW) was determined after drying at 105 °C to constant mass. Relative water content was calculated using the equation: RWC = (FW − DW)/(TW − DW) • 100.
The presented data are the mean and standard error values (unless indicated otherwise) of the measurements of the plant or soil material (exact n is specified under the figures). Statistical analysis of the results was performed using confidence tests with the ANOVA analysis of variance (STATISTICA 13, StatSoft Inc.). Normality was evaluated using residual analysis (for n ≤ 4); in other cases (n > 4), the Shapiro-Wilk test was used. The means were compared with the use of the HSD Tukey test at p < 0.05.