The experimental field was located in Kuuma, Jokioinen in Southwestern Finland (60.22 oN, 24.78 oE, 110 m a.s.l.). The field has been drained and cultivated at least from the end of the 19th century and it has a subsurface drainage system. Prior to the beginning of this experiment, the field was in monoculture with spring cereals, conventionally cultivated and ploughed for several years.
[FIGURE 1 HERE]
The experimental site was placed between the subsurface drainage pipes so that each plot (8 x 18 m = 144 m2) had hydrologically as similar conditions as possible (Fig. 1). The experimental setup was a fully randomized plot experiment in four blocks. Originally, the field was set up with four treatments: half of the CT and NT plots were sown with a cover crop (Italian ryegrass, Lolium multiflorum). However, the ryegrass did not manage (mean yield 410 ± 300 kg dry matter ha− 1) and as it did not cause any differences in the results between treatments, we decided to divide the plots into two treatments only (CT and NT) resulting in eight replicate plots per treatment in the data analysis. Our cover crop yields were clearly below the mean Italian ryegrass biomass of 2.2 Mg dry matter ha− 1 observed in boreal climatic conditions (Känkänen 2019) or the < 1 Mg ha− 1 biomass estimated necessary for carbon sequestration (Blanco-Canqui et al. (2020)). Despite not considering it in the statistical comparisons, the cover crop leaf area index was included in the models and calculations for GHG fluxes.
The whole field was ploughed in the spring 2017, and the experiment was set up on May 24th, 2018. The CT treatment was ploughed in spring 2018 (May 24th, 2018), spring 2019 (May 20th, 2019), autumn 2019 (Nov 18th, 2019) and autumn 2020 (Nov 30th, 2020). The sowing dates were June 14th, 2018, May 22nd, 2019 and June 12th, 2020.
The experimental field was growing oat (Avena sativa) in 2018, barley (Hordeum vulgare) in 2019 and oat in 2020. The sowing density was 500 seeds m− 2 for both barley and oat. The plots were fertilized each year with Yara Mila Y3 fertilizer which contains 26% nitrogen (N), 3% phosphorus (P) and 8% potassium (K). The applied amount was 260 kg ha− 1 each year which corresponds to N rate of about 60 kg per hectare. The fertiliser was given once a year at the time of sowing. To control weeds during the experiment, herbicides such as Premium Classic (12 kg ha− 1), Starane (0.25 l ha− 1), Refine Super SX (0.25 kg ha− 1) and Primus (0.006 l ha− 1) were applied annually as a one-time treatment according to the user manual.
Ploughing, sowing, plant protection, fertilizing and harvesting were done using typical farming machinery. Sowing was performed using VM300SK direct sowing machine (Vieskan Metalli Oy, Finland) in 2019–2020 and Väderstad 300-400C/S (Väderstad Ab, Sweden) in 2018. The CT plots were mouldboard ploughed to the depth of 20 cm. Plot yields were determined in the middle of the plots with an experimental combine harvester (Sampo Rosenlew SR2010) in Sep 18th, 2018, Sep 26th, 2019 and Sep 22th, 2020. Biomass outside of these areas was harvested with a field-scale combine harvester.
The peat layer was deep (over 2 m), and its organic carbon content was about 25% in the topsoil 0–20 cm, 29% in the 20–40 cm layer, and 35% in the 40–60 cm layer. The topsoil was sampled in May 2018 (except density and porosity), and its properties are presented in Table 1. Soil conductivity and acidity were determined using the ISO 11265 and ISO 10390 methods, respectively. Content of nutrients was analysed as in Vuorinen and Mäkitie (1955) and that of soil carbon and nitrogen using the dry combustion method (Leco TruMac CN Determinator, Leco Corp. St. Joseph, MI, USA). Soil core samples for dry bulk density and porosity (diameter 5 cm) were taken from the surface layer (0-17.5 cm) of each plot in Oct 2020 using the Kopec corer, and the samples were dried at 37°C.
Closed opaque chambers were used to measure ecosystem respiration and N2O and CH4 fluxes. In each plot, a 60 cm x 60 cm steel collar was installed to the depth of 10–15 cm. An aluminium chamber (height 40 cm) mounted at the top of the collar was sealed with water in the groove of the upper edge of the collar. Steel extensions were used when the height of the crop exceeded the height of the chamber. The measurements were done during daytime between 10 am and 2 pm approximately biweekly, or monthly in the winter. The chambers were closed for 30 minutes, and four 20 ml gas samples were taken with a 60-ml plastic syringe to pre-vacuumed vials (Exetainer, Labco Limited, UK) in 10-minute intervals starting immediately after closing. Prior to sampling, the syringe was pumped a few times to mix the air in the chamber. The samples were analysed with a gas chromatograph (Agilent 7890 Agilent Technologies, Inc., Wilmington, DE, USA) equipped with flame ionizer and electron capture detectors, and a nickel catalyst for converting CO2 to CH4. The gas chromatograph had a 2 ml sample loop and a backflush system for separating water from the sample and flushing the precolumn between the runs. The precolumn and analytical columns consisted of 1.8 and 3 m long steel columns, respectively, packed with 80/100 mesh Hayesep Q (Supelco Inc., Bellefonte, PA, USA). Nitrogen was used as the carrier gas and a standard gas mixture of known concentration of CO2, N2O and CH4 was used for a calibration curve with seven concentration points. An autosampler (222 XL Liquid handler, Gilson Medical Electronics, France) fed the samples to the loop of the gas chromatograph. A linear regression model was fitted to calculate gas concentrations and the ideal gas law was used to solve the flux rate for every enclosure. The criterion of R2 > 0.9 for CO2 was used as the acceptance limit for all gases.
A transparent chamber (60x60x60 cm) made of polycarbonate plexiglass was used to measure crop NEE approximately biweekly during the growing season. The chamber was equipped with a Vaisala GMP-343 probe and temperature and humidity sensor (Vaisala Oy, Vantaa, Finland) and two fans for mixing the air during the measurement. One or two layers of a white fabric shroud and one blackout curtain were used to control the amount of light entering the chamber (approximately 100, 50, 25, 0% of ambient radiation). The measurements were done in the same collars as the opaque chamber measurements. Four measurements with different amount of entering light were taken from each subplot in order to cover a large range of light conditions during one measurement round. Each measurement took one minute with a five second sampling rate. The chamber was flushed after each measurement to reconstitute ambient CO2 and air humidity contents. A 10 second lag time was applied after closing and before starting the measurement to exclude the deadband when the flux was not yet stabilised. Clear sky conditions were preferred to avoid problems related with changing cloudiness and to achieve the widest possible range of available light. On the hottest summer days, freezer blocks were used to cool the chamber air. The temperature change inside the chamber was less than 1.5 degrees which was also used as a criterion for data filtering.
Due to the short chamber enclosure time, the change of gas concentration during the chamber enclosure was mostly linear. The primary measurement results as parts per million (ppm) unit were converted to g m− 2 h− 1 in accordance with the ideal gas law using the measured temperature inside the chamber.
If the snow cover was > 20 cm, a concentration snow gradient method Maljanen et al. (2003a) was used to determine the GHG fluxes. A probe made of a steel pipe (Ø 3 mm), with a three-way valve and a plastic syringe, was used to sample 15 ml of air just above the snow cover, in the bottom of the snow cover and at one depth in between in tree replicates per plot. The gas was stored in the pre-vacuumed vials and the concentrations were determined gas chromatographically as with the opaque chamber samples. The fluxes were calculated using the equation based on Fick’s law as in Maljanen et al. (2003a).
Measurements for bare soil respiration were made in 7/2019–5/2021. We installed one steel air ventilation pipe 27 cm in diameter and 30 cm in length to the depth of 5–10 cm in each of the 16 subplots. All green vegetation was removed within the chamber area. For the measurements, the cylinders were closed with a cover equipped with a CO2 sensor (GMP-343; Vaisala Oyj, Vantaa, Finland) and a small fan. One measurement lasted for one minute with a five second sampling rate, and was taken about once in a week or two. The measurement frequency was raised to determine the effect of ploughing on soil respiration in autumn 2020 when the respiration was measured 0, 30, 90 and 210 minutes after ploughing as well as once per day in the next two days. The respiration chamber method showed higher values and more deviation compared to the large opaque chamber method particularly during the winter period in 2020–2021, and soil respiration was not modelled on an annual basis due to poor performance of the empiric soil respiration model. However, data from the period 7/2019–9/2020 were used to estimate the total soil respiration in the growing seasons of 2019 and 2020. The additional carbon loss caused by ploughing was also included in the estimate of total annual respiration in 2020 in 2019 when determining the NEE.
Photosynthetically active radiation (PAR) was continuously measured at the edge of the field by LI-190 quantum PAR sensor (LI-COR, Lincoln, Nebraska, USA) with a one minute sampling rate. Soil temperature was measured at the depth of 10 cm (changed to depth of 5 cm in May 2020 to achieve better response of CO2 to air temperature) in CT and NT plots (one plot per treatment) with Elcolog sensors (Elcoplast Oy, Tampere, Finland). The air temperature, precipitation and radiation data were taken from the weather station of Finnish Meteorological Institute (FMI) located about 10 kilometres from the site. Gaps in the measured PAR data were filled with global radiation data from FMI corrected using the ratio of global radiation and the measured PAR.
Leaf area index (LAI) was measured usually at the same time with the transparent chamber measurements with a portable LAI meter (SunScan; Delta-T Devices Ltd, Cambridge, United Kingdom). Because the measured LAI corresponds poorly to the actual photosynthetically active biomass in the late growing season due to ripening of cereals, we selected the highest measured LAI values for the beginning of August and assumed that LAI is zero in the end of August when the measured GP was negligible. LAI of the cover crop in the respective treatments was set to zero after ploughing in the CT plots and at mid-December in the NT plots. LAI values > 3 were set to 3 as they were assumed not to affect GP due to saturation of the reflectance (Aparicio et al. 2000).
NEE (net ecosystem exchange) consists of GP (gross photosynthesis) and ER (ecosystem respiration) and thus GP was estimated for each NEE measurement by (Eq. 1),
where the full darkened transparent chamber measurement result (ER) is subtracted from the light-dependent flux (NEE) measured during the same day. ER and GP were modelled using nonlinear regression (fitnlm function in MATLAB 2019b) for all 16 plots and all years separately summing up to 96 models in total. Modelling was done for the period from May to April. Empiric models were used for ER as in (Lohila et al. 2003) and for GP as in (Kandel et al. 2013). Instead of the phytomass indices used in the above publications, we used LAI to describe the stage of the crop growth. Air temperature and PAR were assumed to be the same for all plots, whereas we used the measured LAI for each plot distinctly and the soil temperature from either CT or NT plots.
We used the following equation defined by Long et al. (1993) for GP to estimate empirical coefficients (Amax and k):
$$GP=\frac{{A}_{Max}*PAR}{k+PAR}*LAI{*T}_{Scale}$$
1
where PAR is the measured photosynthetically active radiation, LAI is the measured leaf area index, Amax is the asymptotic maximum, and k is a half-saturation value. TScale represents the temperature sensitivity of photosynthesis and follows the equation performed by (Raich, Rastetter et al. 1991):
$${\text{T}}_{Scale}=\frac{(T-{T}_{min})(T-{T}_{max})}{\left(T-{T}_{\text{m}\text{i}\text{n}}\right)\left(T-{T}_{max}\right)-{(T-{T}_{opt})}^{2}}$$
2
where T is the measured temperature, photosynthetically active minimum temperature Tmin is -2 ˚C, maximum Tmax is 40 ˚C and the optimum is 20 ˚C as in (Kandel et al. 2013).
ER consists of autotrophic (Rauto) i.e. plant respiration and heterotrophic (Rhetero) i.e. soil respiration (Lloyd and Taylor 1994):
$$ER= {R}_{hetero}+{R}_{auto}$$
3
$${R}_{hetero}={R0}_{s}*\text{e}\text{x}\text{p}\left({E0}_{s}\right(\frac{1}{56.02}-\frac{1}{{T}_{soil}+46.02}\left)\right)$$
4
$${R}_{auto}=LAI*{R0}_{p}*\text{e}\text{x}\text{p}\left({b}_{d}\right(\frac{1}{10+273}-\frac{1}{{T}_{air}+273})$$
5
, where Tsoil is the measured soil temperature, LAI is the measured leaf area index, Tair is the measured air temperature, R0s is soil respiration at the reference temperature 10 ˚C, R0p is plant respiration at the reference temperature at 10 ˚C, E0s is ecosystem sensitivity set to 308 and bd was the temperature dependence of dark respiration set to 5000 as in (Lohila, Aurela et al. 2003).
The empirical coefficients (R0p and bd) were estimated with a nonlinear regression model similarly as in the case of GP. Hourly timeseries of GP and ER were predicted with the above equations using the modelled parameters and hourly timeseries of the field measurements. Missing values of LAI and soil temperature were acquired by interpolation. Gaps in soil temperature during sowing, harvesting or ploughing were filled with averaged air temperature using daily moving mean (correlation between soil and averaged temperature was R2 = 0.8 in the full dataset). The NEE was calculated by subtracting the modelled GP from the modelled ER. ER was estimated using data from both opaque chambers and darkened transparent chambers. Annual fluxes were computed as integral of the hourly fluxes with a trapezoidal method (trapz function in MATLAB 2019b). Net ecosystem carbon balance consists of NEE and the amount of carbon removed in yield. The carbon content of the crop yield was assumed to be 45% (Jensen et al. 2005).
Hourly soil respiration was modelled for each plot for the period of May 22, 2019 – Sep 22, 2020 based on the soil respiration measurements and Eq. 4. Total effluxes were counted for growing seasons (between sowing and harvesting) of 2019 and 2020.
Data cleaning and analysis
The flux measurements were visually examined, and obvious outliers and data points from the beginning of the measurement were removed if the flux was clearly not yet stabilized. For transparent chamber measurements, the criteria R2 > 0.9 for the fitted linear assumption of flux measurements would exclude a large amount of data, especially with small CO2 change, leading to a biased dataset. Therefore we decided to add the criterion Sxy < 2.3 g m− 2 h− 1 as in Kutzbach et al. (2007) (Sxy is the standard deviation of the residuals and 2.3 g m− 2 h− 1 is the 95% percentile of values). Data points containing gaps or possibly erroneous content (due to changing cloudiness etc.) in the background values were marked for a later decision about removal.
In the modelling phase, fitted values were examined visually, and marked outliers and other clear outliers were removed to avoid distortion. In 2018, 34 out of 427 GP values and 21 of 366 ER values were removed. In 2019, 66 out of 537 GP values and 5 of 467 ER values were removed. In the dataset of 2020, 86 out of 568 GP values and 23 of 499 ER values were removed. Average correlations of models were 0.83, 0.78 and 0.92 for ER, and 0.86, 0.91 and 0.80 for GP in 2018, 2019 and 2020, respectively. Measured versus modelled values of GP and ER are shown by years in Fig. 2. All models for soil respiration were accepted although data were highly variable and the average R2 of the models was 0.47 ± 0.24.
The coefficients for converting N2O and CH4 into CO2 equivalents were 273 and 27, respectively (Forster et al. 2021).
The statistical comparisons of the daily or annual flux values or soil properties and yields between the treatments were made by using one-way ANOVA procedure in the MATLAB version R2019b. Normal distribution of the variables was investigated using histograms and homogeneity of the variances using the Levene test. The difference was considered statistically significant when P < 0.05.