1. Introduction
Black walnut (
Juglans nigra L.) is the highest value tree species in the continental United States [
1], and it typically occurs as a minor component of mixed deciduous forests in central and eastern parts of the U.S. [
2]. The demand for high quality hardwoods, such as black walnut, that are used in fine furniture, cabinetry, gunstocks, paneling, and specialty crafts has been increasing dramatically. Due to its high value, black walnut has been routinely harvested from natural forests, causing depletions in the supply of quality trees. As a result, there has been considerable interest in recent decades in the establishment and intensive management of black walnut in plantations [
3,
4]. Although only about 5585 ha of black walnut plantations exist in the U.S. (representing about 1% of total black walnut cubic foot volume in the U.S.) [
5], plantation area is rapidly increasing. In Indiana, which is centrally located within black walnut’s native range, 2400–3200 ha of trees are planted every year and black walnut represents about 18% of planted trees [
6].
Based on the high nutrient demand, rapid growth potential, and extraordinary value of black walnut timber, some industrial black walnut plantation managers in Indiana and Spain have been operationally fertilizing stands every year following planting with the intention of significantly promoting growth and reducing rotation ages [
7,
8,
9]. Although several studies have shown improved plantation productivity with fertilization in both black walnut and other stand types [
10,
11], others have demonstrated low fertilizer nitrogen use efficiency (FNUE) and low productivity [
12]. To address low FNUE and low productivity issues, various “enhanced-efficiency fertilizers” (EEF) have been developed to reduce N loss, increase N availability, improve fertilizer N recovery (FNR), and enhance plantation growth and yield [
11,
13]. The EEFs can be divided into slow release (SRN), controlled release (CRN), and stabilized (SNF) N fertilizers [
13]. The SRN products slowly release fertilizer N due to microbial decomposition [
14]. The CRN products have coatings around the fertilizer N to alter the rate, pattern, and duration of fertilizer N release [
14,
15]. The SNF products have compounds to inhibit rapid fertilizer N transformation to less stable forms [
16]. The different attributes of the various N containing EEF products increase the options for N fertilization under diverse conditions to help optimize plant N uptake and increase FNUE compared to urea.
However, although fertilization has been found to reduce nutrient limitations and improve plantation productivity and performance, it may also result in negative impacts such as stimulating the growth of non-target vegetation or increasing the offsite movement of nutrients through losses due to volatilization, runoff, or leaching. These factors and related uncertainties lead to the need to improve our understanding of fertilizer fate and use efficiency in plantation settings in order to limit nutrient losses and improve plantation productivity. The goal of this study was to characterize N uptake and partitioning within aboveground ecosystem components and to determine whether FNR was greater for EEFs compared to urea. To accomplish this, we compared fertilizer N uptake in mid-rotation black walnut plantations following fertilization for three urea-based EEF products, uncoated urea, and an unfertilized control to determine if there were differences among treatments within aboveground ecosystem components for: (1) N concentrations; (2) δ15N levels to quantify the incorporation of applied N; and (3) FNR and partitioning.
2. Materials and Methods
2.1. Site Description
In July 2010, eight non-contiguous areas were selected in a privately-owned mid-rotation black walnut plantation with a total area of about 50 ha (centered at approximately 40.32877° N, 86.70805° W) in Sheffield Township, Tippecanoe County, IN, USA. Slopes in the plantation were under 5% with the majority of soils underlying the plantation being well-drained to excessively drained. Soils under the plantations included Hononegah fine sandy loam (37.3% of the land area), Ockley silt loam (10.4% of the land area), Fincastle-Crosby complex (9.7% of the land area), Sleeth loam (8.2% of the land area), Cohoctah fine sandy loam (7.3% of the land area), Ouiatenon sandy loam (7.3% of the land area), Mahalasville silty clay loam (7.1% of the land area), and various minor components.
Each selected area (about 6 ha each) served as a block, with areas selected to spatially represent the entire plantation. The plantation was established in 1989 with a tree spacing of 6.9 m between rows and 3.0 m between trees within rows. Within each block, five dominant crop trees were selected for inclusion in the study. Crop trees were selected to spatially represent the entirety of the block, with sufficient space between trees to prevent cross-contamination from the application of 15N-enriched fertilizers. Crop trees selected for the study had an average diameter at breast height (±standard deviation) of 24.63 ± 2.69 cm, average total height of 15.19 ± 1.67 m, average live crown length of 8.73 ± 1.80 m, and the average live crown ratio was 0.58 ± 0.10. At the time of selection, each crop tree in each block was randomly assigned to one of five fertilizer treatments (described below).
The understory of the plantation was relatively homogeneous and was populated by a mixture of woody and herbaceous species, with the majority of understory species present being herbaceous. The main woody species among the competing vegetation was black raspberry (Rubus occidentalis L.), while the herbaceous species composition varied seasonally and consisted of various members of the Poaceae, Asteraceae, and Brassicaceae, with small numbers of individuals from other families. Historical control of competing vegetation in this stand prior to the study consisted mainly of mechanical control via mowing. The vegetation control history and methods in this stand, as well as the density and composition of vegetative competition observed, can be considered representative for black walnut plantations in this region.
2.2. Fertilizer Treatments and 15N Labeling
The five fertilizer treatments consisted of an unfertilized control, Agrotain Ultra (Agrotain), Arborite EC (Arborite), Agrium ESN (ESN), and urea. All fertilizers were enriched to 0.5 atom% 15N during manufacture, except for the unfertilized control treatment which served as both an isotopic and fertilization control throughout the study. All fertilizers were urea-based, with Agrotain, Arborite, and ESN serving as examples of urea-based enhanced-efficiency fertilizers (EEF) which employ different mechanisms for controlling release rates. The urea applied in the Agrotain Ultra treatment was an SNF coated with a urease inhibitor, N-(n-butyl) thiophosphoric triamide (NBPT), to reduce ammonia volatilization from applied urea (Koch Agronomic Services, LLC, Wichita, KS, USA). The urea applied in the Arborite EC treatment was an SRN coated with water-soluble boron and phosphate using a proprietary process in order to regulate rates of N release and loss from fertilizer granules (Weyerhaeuser Company, Seattle, DC, USA). The urea applied in the Agrium ESN treatment was a CRN contained within a polymer coating which moderates N release rates in proportion to soil temperature (Agrium, Inc., Calgary, AB, Canada). All fertilizer applications were surface broadcast by hand at a rate of 224 kg N ha−1 within a 100 m2 circular plot centered on each selected crop tree. Each fertilized tree received 22.4 kg N via its assigned fertilizer treatment except crop trees assigned to the control treatment. For logistical reasons, the experiment was installed and executed in two stages, with fertilizer treatments being applied to blocks 1–4 in May 2011 and to blocks 5–8 in May 2012. Initial pre-fertilization time zero sampling and post-fertilization sampling of crop tree and ecosystem components are described below.
2.3. Sample Collection
Black walnut crop tree leaves were collected from the middle one-third of the south side of each crop tree selected for inclusion in the study prior to application of fertilizer treatments and at intervals during the first growing season after fertilization. At each sampling time, 10 complete leaves (i.e., leaflets and rachis) were collected from each crop tree by hand using an aerial lift (Altec, Inc., Birmingham, AL, USA), with each leaf serving as a sub-sample. Initial pre-fertilization (time zero) leaves were collected in July of the year prior to fertilization (i.e., July 2010 for blocks 1–4 and July 2011 for blocks 5–8) due to the absence of crop tree leaves at the time of fertilization. After fertilization, leaves were collected from each black walnut crop tree as described above in June (one month after fertilization), July (two months after fertilization), and August (three months after fertilization) in the year of fertilization (i.e., 2011 for blocks 1–4 and 2012 for blocks 5–8). Upon collection, each leaf was placed in a labeled paper bag before placement in a cooler for immediate transport to the Forest Ecology, Soils, and Silviculture Laboratory (FESSL) in the Department of Forestry and Natural Resources at Purdue University (FESSL, West Lafayette, IN, USA) for processing.
Competing vegetation and forest floor litter (i.e., the Oi horizon) were collected from four 0.0625 m2 (25 cm × 25 cm) sub-sampling areas located approximately 2.82 m to the NE, SE, SW, and NW of each crop tree prior to application of fertilizer treatments and at intervals during the first year after fertilization. At each sampling time, all living vegetation emerging from the soil within each sub-sampling area was hand-cut at ground level and placed in a labeled paper bag before placement in a cooler for immediate transport to FESSL for processing. All forest floor litter of the Oi horizon was then collected from within each sub-sampling area and treated as described for the competing vegetation. Sub-sampling areas were randomly spatially shifted slightly at each sampling time so as to avoid re-sampling areas during the course of the study. Initial pre-fertilization (time zero) sampling of competing vegetation and litter occurred immediately prior to fertilization (i.e., May 2011 for blocks 1–4 and May 2012 for blocks 5–8). After fertilization, competing vegetation and litter were collected from sub-sampling areas surrounding each crop tree in June (one month after fertilization), July (two months after fertilization), August (three months after fertilization), November (six months after fertilization), and May of the following year (12 months after fertilization), with this sampling scheme initiated in 2011 for blocks 1–4 and in 2012 for blocks 5–8.
2.4. Sample Preparation and Analyses
Upon arrival at FESSL, leaf areas of all black walnut crop tree leaf sub-samples were immediately measured using an LI-3000 portable leaf area meter (LI-COR, Inc., Lincoln, NE, USA). Sub-samples of plant-based ecosystem components (i.e., black walnut crop tree leaves, competing vegetation, and litter) were air-dried to constant mass and dry masses were recorded before being ground in a Wiley mill (Thomas Scientific, Swedesboro, NJ, USA) to pass a 20-mesh screen. Subsequently, all ground sub-samples for each plant-based ecosystem component × fertilizer treatment × sampling time × block combination were composited, homogenized, and submitted to the Department of Forest Resources and Environmental Conservation at Virginia Tech for determination of N concentration and δ
15N using a coupled elemental analysis isotope ratio mass spectrometer (IsoPrime 100 EA-IRMS, Isoprime, Ltd., Manchester, UK; [
11]).
2.5. Calculations and Statistical Analyses
For all sampled ecosystem components (i.e., black walnut crop tree leaves, competing vegetation, and litter), all dependent variables measured and reported in
Table 1,
Table 2,
Table 3,
Table 4,
Table 5,
Table 6 and
Table 7 were directly quantified, except fertilizer N recovery (FNR). For estimation of FNR in leaves of black walnut crop trees, total leaf area contained within the canopy of each crop tree was estimated using the allometric relationships described by [
17]. The equation employed was:
where: A = the estimated total leaf area (m
2) of a given black walnut crop tree; DBH = the diameter at breast height (cm) of the black walnut crop tree; and LCR = the live crown ratio of the black walnut crop tree determined by dividing the length of live crown (m) of the black walnut crop tree by its total height (m). After total leaf areas were estimated for each black walnut crop tree, the FNR in leaves of each black walnut crop tree canopy was estimated as follows.
First, for leaves of each crop tree at each sampling time, leaf N content was calculated by multiplying the average dry mass of sub-sampled leaves by the measured N concentration (%, on a dry mass basis). Second, reported δ15N values for each crop tree leaf sample at each sampling time were mathematically reverted to 15N:14N values via simple rearrangement of the equation used for calculating δ15N. Third, total mass of 15N in each crop tree leaf sample was determined based upon the calculated leaf N content and 15N:14N of each sample. Fourth, the calculated total mass of 15N in each crop tree leaf sample was divided by the average area of leaf sub-samples for each crop tree at the corresponding sampling time to yield total mass of 15N per unit leaf area for each crop tree at each sampling time. Fifth, values for total mass of 15N per unit leaf area of unfertilized control trees were assumed to be representative of 15N natural abundance in crop tree leaves; consequently, these values were averaged for each sampling time and subtracted from the calculated total mass of 15N per unit leaf area for each fertilized crop tree at each sampling time to yield an estimate of the mass of fertilizer-derived 15N per unit leaf area for each fertilized crop tree at each sampling time. Sixth, the estimated mass of fertilizer-derived 15N per unit leaf area for each fertilized crop tree at each sampling time was multiplied by the estimated total leaf area of each fertilized crop tree to produce an estimate of the mass of fertilizer-derived 15N present in the canopy of each fertilized crop tree at each sampling time. Finally, the estimated mass of fertilizer-derived 15N present in the canopy of each fertilized crop tree at each sampling time was divided by the mass of 15N initially applied via each fertilizer treatment and expressed as a percentage to produce an estimate of FNR from the leaves of each fertilized black walnut crop tree at each sampling time.
Fertilizer N recovery from competing vegetation and litter were determined as follows. First, sample N contents were determined by multiplying the measured N concentration of each sample by the average dry mass of its vegetation or litter sub-samples collected from each 0.0625 m2 sub-sampling area. Second, reported δ15N values for each vegetation or litter sample at each sampling time were mathematically reverted to 15N:14N values via simple rearrangement of the equation used for calculating δ15N. Third, total mass of 15N in each vegetation or litter sample was determined based upon the calculated sample N content and 15N:14N of each sample. Fourth, values for total mass of 15N in each competing vegetation or litter sample collected at each sampling time from areas surrounding unfertilized control trees were assumed to be representative of 15N natural abundance in competing vegetation or litter, respectively, at each sampling time; consequently, these values were averaged for each ecosystem component at each sampling time and subtracted from the calculated total mass 15N per sample of vegetation and litter samples taken from areas surrounding fertilized crop trees at each sampling time to yield an estimate of the mass of fertilizer-derived 15N per sample for each competing vegetation or litter sample. Fifth, the resulting values were then divided by 0.0625 m2 and multiplied by 100 m2 to yield estimates of fertilizer-derived 15N contents of the competing vegetation and litter present within the 100 m2 area surrounding each crop tree to which fertilizer treatments were applied. Finally, the estimated mass of fertilizer-derived 15N present in the competing vegetation or litter of the 100 m2 areas surrounding each fertilized crop tree at each sampling time was divided by the mass of 15N initially applied via each fertilizer treatment and expressed as a percentage to produce an estimate of FNR from the competing vegetation and litter components surrounding each fertilized black walnut crop tree at each sampling time.
The experiment was designed and executed as a randomized complete block design (n = 8). Data for each response variable were analyzed using ANOVA (p < 0.05) followed by Tukey’s Multiple Pairwise Comparison (α = 0.05) to detect significant means separations within significant model factors (SAS 9.4, SAS Institute, Inc., Cary, NC, USA). Data for each ecosystem component (i.e., black walnut crop tree leaves, competing vegetation and litter) were analyzed separately, with fertilizer treatment, sampling time, and fertilizer treatment × sampling time included as model factors for each ecosystem component.
5. Conclusions
This study examined the short-term impacts and fates of urea-based EEFs compared with uncoated urea in a mid-rotation black walnut stand in Indiana, USA. Fertilizer N recovery did not vary by fertilizer treatment, but competing vegetation was a strong sink for applied N with the potential to negatively impact FNUE and growth of target trees. The low FNR rates observed in this study corroborate results of some studies but differ from others where more than 90% of applied N was recovered. Although not quantified, storage in woody tissues and soil compartments, volatilization, and leaching likely contributed to the low FNR rates observed in our study. This highlights the need for nutrient budget studies to comprehensively quantify native contributions and fate and transport of applied fertilizers in aboveground and belowground ecosystem components to enable accurate accounting of what translates to productivity and losses from the system. Only ESN exhibited a slow-release pattern, as indicated by lower δ15N values early in the season, which increased over time until 3 months after application before declining thereafter. Fertilization with Agrotain, Arborite, and Urea resulted in higher δ15N values in ecosystem components more rapidly after application, suggesting rapid increases in nutrient availability and little to no slow-release behavior. Based on patterns of N uptake by competing vegetation observed in this study, it appears that intensive plantation management practices where weed control is combined with fertilization could improve the availability of applied N for crop trees.