European Journal of Agronomy 11 (1999) 145–155

Variation in N-recovery of winter wheat under Mediterranean

conditions studied with 15N-labelled fertilizers
C. Carranca a, *, A. de Varennes b, D.E. Rolston c
a Estação Agronómica Nacional, Av. República, 2784-505 NOVA OEIRAS, Oeiras, Portugal
b Instituto Superior de Agronomia, Tapada da Ajuda, 1300 Lisbon, Portugal
c University of California, Davis, CA, USA
Accepted 13 April 1999

Abstract

We studied the fate of 15N-labelled fertilizers applied to winter wheat (Triticum aestivum L., cv. Tejo) grown for
3 years on a Haplic Luvisol in southern Portugal. The nitrogen rate used was 60 kg ha−1 using ammonium nitrate as
basal dressing, and two top-dressings of 60 kg ha−1 of N each, either as urea or calcium nitrate. Microplots (PVC
cylinders with 30 cm diameter) with ceramic cups placed at the bottom were used, so that leachates could be collected
by hand pumping. A greater percentage of plant N was derived from the fertilizer in the driest year (year 1) than in
the other two years. However, biomass accumulation was smaller in that year and, therefore, plants took up less N.
As a result, the recovery of 15N-labelled fertilizer in the crop was in the range of 22–40%, with no significant
differences between years or treatments. Considerable amounts of N derived from fertilizer remained in the soil after
harvest (15 to 66% of total fertilizer N ). In years 1 and 2, the 15N in the soil derived from fertilizer was similar, and
not significantly affected by N form. In year 3, top-dressing with urea led to more of the 15N from fertilizer remaining
in the soil, compared with applying nitrate, and to greater values of total soil N, nitrate and extractable ammonium.
There was evidence of nitrate leaching in years 2 and 3, particularly after the first two N applications, during the
autumn–winter period, but not significantly affected by treatment. A significant amount of 15N remaining in the soil
was in 15 NH+ fixed in the clay fraction, especially in the surface layers and with urea application, suggesting that
4
ammonium fixation in montmorillonite was an important process for removing NH+ from solution and protection
4
from nitrification. N losses were similar (35 to 55%) in years 1 and 2 with both treatments, and in year 3 with
ammonium nitrate+calcium nitrate application. In year 3 the ammonium nitrate+urea treatment resulted in a
complete recovery of 15N applied. © 1999 Elsevier Science B.V. All rights reserved.

Keywords: Fixed 15NH ; Haplic Luvisol; 15N fertilizer use efficiency; 15N leaching; 15N recovery
4

1. Introduction unlabelled fertilizer, but to measure the recovery

Many field experiments have been carried out
to study N fertilization in wheat, and to assess the
efficiency of the different forms of N supply.
Fertilizer uptake by the crop can be estimated with
* Corresponding author. Tel.: +351-(01)-4416855;
fax: +351-(01)-4416011.
of fertilizer N in the soil and the N lost, a labelled
fertilizer has to be used. Environmental factors
influence plant growth, N uptake, and N losses.
Pilbeam (1996) assembled data from field experi-
ments on the fate of 15N-labelled fertilizers applied
to wheat grown in different parts of the world, but
no information was available from countries in
southern Europe. There are several publications

1161-0301/99/$ – see front matter © 1999 Elsevier Science B.V. All rights reserved.
PII: S1 1 6 1 -0 3 0 1 ( 9 9 ) 0 0 02 7 - 1
146 C. Carranca et al. / European Journal of Agronomy 11 (1999) 145–155
on the N uptake by wheat in Portugal and Spain,
but 15N was not used in these experiments
(Carvalho, 1987; Alves and Carvalho, 1988;
Milheiro and Pereira, 1988; Mompo and Armentia,
1992; Abreu et al., 1993; Carvalho and Basch,
1994).
We evaluated the fate of 15N-labelled fertilizers
applied to winter wheat grown under Mediterran-
ean conditions on a Haplic Luvisol at Elvas (south-
ern Portugal ) over a 3 year period, to determine
the recovery of fertilizer N in the crop and soil at
harvest. We also looked for evidence of nitrate
leaching and ammonium fixation.
2. Materials and methods
Experiments were established in 1991–92, 1992–
93, and 1993–94 at Elvas, in southern Portugal,
on a Haplic Luvisol (FAO/UNESCO, 1989), a
different area being used each year. Monthly mini-
mum air temperatures in the 3 years varied from
0–4°C (January) to 12–15°C (May–June) and
maximum temperatures from 12–14°C (January)
to 17–27°C (May). Except for the greater values
for maximum temperature during March in 1992
and 1994, the minimum and maximum temper-
atures did not differ significantly from one year to
another, and were not significantly different from
the 30-year average.
Total rainfall during the growing season was
291 mm, 307 mm, and 309 mm in years 1, 2 and 3
respectively. These values were smaller than the
median for the 30-year period. However, monthly
precipitation did not differ significantly from the
medians for the 30-year period. Soil water deficit
( Thorntwaite–Mather method ) was observed in
all years. In year 1 there was soil water deficit
Table 1
Some characteristics of the Haplic Luvisol at Elvasa
Year pH Organic C (g kg−1)
(H2O)
1 6.6 c 4.8 a
2 7.3 a 4.8 a
3 7.0 b 4.2 b
a Means in a column followed by the same letter do not differ significantly (P≤0.05)
from February to June. In year 2, the soil water
deficit occurred in January and from April to May.
In year 3, soil water deficit occurred in March and
April. The ratio rainfall–potential evapotranspira-
tion was 0.75, 1.20 and 1.07 in years 1, 2 and 3
respectively.
Every year, before sowing, 60 kg ha−1 of P O
2 5
and K O were applied as a basal dressing, and soil
2
samples were taken in either 10 or 20 cm
increments, to a depth of 60 cm, for physical and
chemical analyses. The texture of the soil was
sandy clay loam, and the clay content ranged from
16.4 to 27.6%. Montmorillonite was the dominant
clay mineral. After the basal fertilizer dressing the
area of soil used each year had different character-
istics ( Tables 1 and 4.). Organic C, total N and
fixed NH+ decreased with depth, but not pH, or
4
extractable NH+ and NO−.
4 3
Randomized blocks with 4 m2, with three repli-
cates and two N treatments (urea-15N or
Ca(15NO ) application) were used. For 15N
32
enrichment, PVC cylinders (microplots) were
pushed into the soil, with minimal soil disturbance.
The cylinders were 30 cm in diameter and 50 cm
long. About 1 cm of cylinder was left above the
soil surface to avoid water runoff. Below each
cylinder, three ceramic cups were placed at an
angle of about 45° to the vertical. The soil solution
moved into the ceramic cups in response to applica-
tion of vacuum to the porous cups by hand
pumping (<8×104 Pa).
Winter wheat (Triticum aestivum L., cv. Tejo)
was sown in November–December at a seeding
rate of 300 seeds per square metre in the cylinders
and surrounding plots. The rate for N fertilization
of winter wheat recommended in this region is
180 kg ha−1, for an expected yield of 4–5 t ha−1
(Milheiro and Pereira, 1988), in which two-thirds
Total N (g kg−1) Fixed NH+-N (mg kg−1)
4
0.67 a 95.2 b
0.58 b 108.9 a
0.55 b 102.4 a
 C. Carranca et al. / European Journal of Agronomy 11 (1999) 145–155
should be applied as a top-dressing. In accordance
with this, treatments consisted of a basal applica-
tion of 15NH 15NO (60 kg ha−1 of N, with a
4 3
4.91% double enriched with 15N in the microplots)
and either urea-15N or Ca(15NO ) as two top-
32
dressings (each of 60 kg ha−1 of N, with 4.90% or
3.64% enrichments with 15N in the microplots
respectively), applied at the end of February (mid-
tillering) and at the end of March (stem elonga-
tion). Plants were harvested at physiological matu-
rity (June in year 1, and May in years 2 and 3),
and separated into roots, straw and ears. For yield
evaluation, shoots were also removed from the
areas surrounding the cylinders and separated into
straw and ears.
The samples were oven dried at about 70°C for
48 h, weighed, finely ground, sieved and analysed
for total N and 15N enrichment by ANA-mass
spectrometry at the Seibersdorf Lab. ( Vienna,
Austria) or at the Scottish Crop Research Institute
(Dundee, Scotland).
After harvest, the cylinders were removed and
the soil separated into 10 cm slices. Below each
cylinder, at a depth of 50–70 cm, soil samples were
also taken. Total N and the 14N:15N ratio were
analysed by automatic combustion of soil samples
in Sn capsules, and ANA-mass spectrometry at
Berkeley ( USA). Fixed NH+ and the 14N:15N
4 2 and 3, probably due to a longer period of soil
ratio were determined by direct combustion in an
ANA-mass spectrometer at Berkeley ( USA), after
soil samples were extracted according to Silva and
Bremner (1966). Nitrate and extractable ammo-
nium (soluble plus exchangeable) were determined
by distillation after soil samples were extracted
with 2 M KCl.
Leachates were collected 1 or 2 days after it
rained, to allow the water to percolate down the
soil profile. Water samples were analysed for min-
eral N, and 15N enrichment of samples was deter-
mined by ANA-mass spectrometry at Seibersdorf
Laboratory ( Vienna, Austria) following separation
of NO− and NH+ by diffusion (Brooks et al.,
3 4
1989)
The percentage of 15N derived from the fertilizer
(%Ndff ) in the crop, remaining in the soil after
harvest or in the leachates, was calculated using
the atmospheric 15N abundance (0.3663% 15N
147
atom) as reference:
%Ndff=(15N atom excess /
plant, soil, leachate
15N atom excess )×100,
fertilizer
where %Ndff is the percent N derived from the
fertilizer, and
%N recovery =15N (kg ha−1 of N )/
plant, soil plant, soil
N rate (kg ha−1 of N )×100.
Plant, soil and leachate data were analysed
statistically after transforming the percentage
results through ‘arc sin’ and using the analysis of
variance (ANOVA) to estimate the effects of N
treatment, year, and depth. Means were compared
by least significant difference (LSD). Yields of
plants grown inside or outside of the cylinders
were compared by t-test.
3. Results and discussion
3.1. Crop response to N fertilizer
The year significantly affected most plant char-
acteristics, but treatment did not ( Table 2).
Biomass of the straw and total dry matter accumu-
lation were smaller in year 1 compared with years
water deficit in the first year.
The yield of plants grown in the cylinders did
not differ significantly from that of plants grown
in the outside areas, with unlabelled N fertilizer
(t=0.521). Thus, the 15N-enriched microplots were
representative of the whole plots.
N concentration and N removed in the straw
increased significantly from years 1 to 3. In the
ears and roots, the N concentration was greater in
year 3 than in other years. In consequence, the
total N taken up by the crop was smaller in year
1 compared with years 2 and 3. N absorption is
dependent on soil moisture (Boaretto et al., 1982).
Therefore, drought is the likely reason for the
small N uptake observed in year 1. The amounts
of N removed in years 2 and 3 (254 kg ha−1 of N
and 218 kg ha−1 of N respectively), were similar
to the values found by van Cleemput and Baert
(1984) for winter wheat grown on a clay soil in
 148

Table 2
Comparison of plant characteristics as affected by the year, and results of ANOVA to evaluate the effects of year and treatment on crop characteristicsa

Year Dry matter N concentration Removed N Ndff (%) Ndfs (%) Plant recovery of 15N
(g/cylinder) (g kg−1) fertilizer (%)

(g/cylinder) (kg ha−1)

Straw Ear Root Straw Ear Root Straw Ear Root Total Total Straw Ear Root Straw Ear Root Straw Ear Root Total

1 25.80b 25.14a 5.19b 5.20c 17.98b 4.90b 0.136c 0.451a 0.024c 0.611b 86.45b 42.0a 48.0a 38.0a 58.0c 52.0b 62.0b 4.5c 17.0a 0.7b 22.3a
2 76.80a 58.23a 17.44a 7.68b 20.52b 5.13b 0.566b 1.140a 0.087a 1.792a 253.66a 26.8c 29.3b 28.0b 73.2a 70.7a 72.0a 11.9b 25.9a 1.9a 39.6a
3 59.44a 28.03a 8.98b 13.50a 24.35a 7.56a 0.802a 0.673a 0.066b 1.541a 218.12a 32.3b 33.5b 31.3b 67.2b 66.5a 68.7a 20.4a 17.4a 1.6a 39.4a

Source of Dry matter Total N Removed N Ndff Ndfs Plant recovery

variation yield of fertilizer

Straw Ear Root Straw Ear Root Straw Ear Root Total Total Straw Ear Root Straw Ear Root Straw Ear Root Total

Year 34.11*** ns 46.84*** 185.43*** 17.62** 21.29** 187.13*** ns 130.79*** 11.61** 17.25** 22.31*** 6.25* 17.25** 22.31*** 6.25* 40.07*** ns 36.34*** ns
Treatment ns ns ns ns ns ns ns ns ns ns ns ns ns ns ns ns ns ns ns ns
Year× ns ns ns ns ns ns ns 6.07* ns ns 7.20* 9.73** ns 7.20* 9.73* ns ns ns ns ns
treatment

a Means in a column followed by the same letter are not significantly different (P≤0.05); ns, *, **, ***=F-values non-significant and significant at P≤0.05, P≤0.01,
C. Carranca et al. / European Journal of Agronomy 11 (1999) 145–155

and P≤0.001 respectively.

 C. Carranca et al. / European Journal of Agronomy 11 (1999) 145–155
Belgium, and fertilized with 180 kg ha−1 of N,
given as three dressings. The small amount of N
removed in year 1 (87 kg ha−1 of N ) was still
slightly greater than the average values of
60–80 kg ha−1 of N reported by dos Santos (1991)
and Carvalho (1987) for winter wheat grown in
Portugal.
The %Ndff in all plant parts was significantly
greater in year 1 compared with years 2 and 3
( Table 2). The values for %Ndff obtained in the 3
years are greater than those reported by Khanif
et al. (1983), van Cleemput and Baert (1984),
Bowen and Zapata (1991), and Sanaa et al. (1992),
probably due to the small initial content of mineral
N in the soil used in our experiments. Most of the
plant N was derived from the soil (%Ndfs)
( Table 2), as also reported by Sanaa et al. (1992)
and Isfan et al. (1994).
Recovery of 15N-labelled fertilizer in the crop
was in the range 22–40%, with no significant
differences between years or treatments. The roots
contained less than 2% of the applied N, as fre-
quently reported (Pilbeam, 1996).
Pilbeam (1996) suggested that there is a correla-
tion between the percentage recovery of 15N-
labelled fertilizer in the crop and soil, and the
precipitation–evaporation quotient for the site. In
humid environments more fertilizer would be reco-
vered in the crop than in the soil, and the opposite
would occur in dry environments. The explanation
presented was that in dry environments plants
would have greater root–shoot and C–N ratios.
The average value of 33.8% for recovery of 15N-
labelled fertilizer in the crop obtained in our
experiments is similar to values obtained in Prague
(Czech Republic) (Mouchová et al., 1996), and in
Washington ( USA) and Australia (Pilbeam, 1996).
This value is greater than average values obtained
in Syria and Israel (Pilbeam, 1996; Pilbeam et al.,
1997), but it is smaller than those from Belgium,
France, Germany and the United Kingdom
(Powlson et al., 1992; Pilbeam, 1996), with the
exception of an experiment with autumn-applied
fertilizer in the United Kingdom (Powlson et al.,
1986).
The correlation between recovery of 15N-
labelled fertilizer in the plant and the precipitation–
evaporation quotient does not appear to hold true
149
for different years at the same site. Our results
show that recovery of 15N-labelled fertilizer in the
crop was similar in the 3 years when the precipita-
tion–potential evapotranspiration ratios were very
different (0.75, 1.20 and 1.07 in years 1, 2 and 3
respectively). It is true that, in the driest year, the
recovery of 15N-labelled fertilizer was smaller in
some plant parts (straw and roots). However, this
resulted from a smaller amount of N removed in
the crop, since the %Ndff was greater in all plant
parts in this year, than in the other two.
3.2. Soil N
The amount of total N present in the soil at
harvest was significantly affected by year, treat-
ment, and depth ( Table 3). However, only in year
3 and with ammonium nitrate+urea application
was this value greater than the initial value
(Table 1), and significantly greater than the value
obtained with ammonium nitrate+calcium nitrate
application (Table 3).
The percentage of soil N derived from the
fertilizer (%N dff ) is a measure of the residual
soil
and downward movement of N in the soil as result
of fertilizer application. Most of the N derived
from the fertilizers was accumulated in the top
30 cm layer, though it was still detected down to
60–70 cm, especially at the end of years 2 and 3
(Table 3). A similar result was reported by Khanif
et al. (1983), Zapata and van Cleemput (1986)
and Isfan et al. (1994). The labelled N in the top
layers may not only derive from residual fertilizer,
but also from root debris and plant exudates
(Rolston et al., 1979; van Cleemput and Baert,
1984; Zapata and van Cleemput, 1986).
At harvest, the soil N derived from the 15N-
labelled fertilizer varied from 15 to 66% of total
fertilizer N. It was not significantly different in
years 1 and 2, independently of treatment, but in
year 3 the ammonium nitrate+urea application
led to a significantly higher recovery of 15N-
labelled fertilizer compared with the ammonium
nitrate+calcium nitrate treatment ( Table 3).
The amount of clay-fixed NH+ at the end of
4
the experiment was affected by year, treatment,
and depth ( Table 3). Clay-fixed ammonium was
always greater than 2 M KCl-extractable ammo-
150 C. Carranca et al. / European Journal of Agronomy 11 (1999) 145–155

Table 3
Residual N in the Haplic Luvisol, as affected by year, treatment, and depth, and results of ANOVA to evaluate the effects of year,
depth, and treatment on residual soil Na

Depth Total N (g kg−1) Fixed NH+-N (mg kg−1) Total N dff (%) 15N Recovery (%)
4 soil
(cm)
15NH 15NO 15NH 15NO 15NH 15NO 15NH 15NO 15NH 15NO 15NH 15NO 15NH 15NO 15NH 15NO
4 3 4 3 4 3 4 3 4 3 4 3 4 3 4 3
+urea-15N +Ca(15NO ) +urea-15N +Ca(15NO ) +urea-15N +Ca(15NO ) +Urea-15N +Ca(15NO )
32 32 32 32
Year 1
0–10 0.66 ADa 0.77 Aa 98.0 Aa 59.8 Ab 2.30 Aa 2.77 Aa 12.8 Aa 17.97 Aa
10–20 0.73 Aa 0.80 Aa 93.3 ABa 57.2 ABb 0.78 Ba 1.16 Ba 4.75 ABa 8.00 Ba
20–30 0.66 ADa 0.74 Aa 82.8ABa 73.5 Aa 0.58 Ba 0.52 Ba 3.19 Ba 3.21 Ba
30–40 0.54 BDa 0.69 Aa 71.2 Ba 57.8 ABa 0.44 Ba 0.44 Ba 1.70 Ba 2.19 Ba
40–50 0.46 BCa 0.50 Ba 64.2 Ba 50.6 ABa 0.34 Ba 0.25 Ba 1.10 Ba 0.90 Ba
50–70 0.34 Ca 0.46 Ba 72.7 Ba 48.6 Ba 0.24 Ba 0.19 Ba 0.90 Ba 0.60 Ba
Total 25.61 Ea 32.49 Da
Year 2
0–10 0.63 Aa 0 57 ACa 129.5 Aa 114.9 ABa 2.52 Aa 1.16 Ab 13.23 Aa 5.51 Aa
10–20 0.60 ACa 0.58 Aa 133.0 Aa 127.2 Aa 1.06 Ba 1.06 Aa 5.30 ABa 5.12 Aa
20–30 0.62 ACa 0.50 ABa 112.6 ABa 124.8 Aa 0.57 Ba 0.43 Aa 2.95 Ba 1.79 Aa
30–40 0.53 ABa 0.46 ABa 113.2 ABa 116.1 ACa 0.52 Ba 0.28 Aa 1.99 Ba 0.93 Aa
40–50 0.47 BCa 0.43 BCa 89.3 Ba 88.7 Ba 0.81 Ba 0.23 Aa 2.21 Ba 0.63 Aa
50–70 0.44 Ba 0.41 Ba 90.4 Ba 91.6 BCa 0.81 Ba 0.24 Aa 2.57 Ba 0.71 Aa
Total 28.25 Ea 14.69 Da
Year 3
0-–10 0.88 Aa 0.45 Bb 148.2 Aa 119.0 Ab 3.72 Aa 2.65 Aa 27.28 Aa 9.94 Ab
10–20 0.79 Aa 0.63 Ab 115.5 Ba 105.0 ACa 1.86 Ba 1.33 Ba 12.25 Ba 5.15 Aa
20–30 0.86 Aa 0.54 ABb 109.9 Ba 109.7 Aa 1.71 BDa 0.77 Ba 12.26 Ba 3.47 Aa
30–40 0.84 Aa 0.48 ABb 114.3 Ba 108.5 Aa 1.31 BCa 0.88 Ba 7.95 BCa 3.05 Aa
40–50 0.77 Aa 0.53 ABb 99.2 Ba 81.7 BCa 0.64 CDa 0.52 Ba 3.56 Ca 1.99 Aa
50–60 0.62 Ba 0.48 ABa 99.3 Ba 73.5 Ba 0.54 Ca 0.57 Ba 2.42 Ca 1.98 Aa
Total 65.72 Da 25.58 Db

Source of Total N Fixed NH+-N Total N dff 15N Recovery

4 soil
variation

Year (A) 20.9*** 74.76*** 7.7** 6.6**

Treatment (B) 19.5*** 16.54*** 4.8* 6.4*
Depth (C ) 16.9*** 13.89*** 23.8*** 18.2***
A×B 37.2*** 4.22* 2.3 ns 5.9**
A×C 2.1* ns 0.8 ns 0.7 ns
B×C 1.1 ns ns 0.4 ns 1.0 ns
A×B×C 0.9 ns ns 0.6 ns 1.2 ns

a In each year and for each soil characteristic, means in a row followed by the same letter or in a column followed by the same
capital letter did not differ significantly (P≤0.05); ns, *, **, ***=F-values non-significant and significant at P≤0.05, P≤0.01 and
P≤0.001 respectively.

nium. There was less NH+ fixed in the soil after
4
the first crop than in years 2 and 3, and the fixed
NH+ was also less at the end of year 1 than at
4
the beginning of the experiment (Table 1). In year
2 the amount of fixed NH+ was similar before
4
and after the crop, and did not differ between
treatment. In year 3, NH+ fixation was more
4
relevant with ammonium nitrate+urea application
than with ammonium nitrate+calcium nitrate
treatment, though there was no significant differ-
 C. Carranca et al. / European Journal of Agronomy 11 (1999) 145–155 151

ence between the amounts present before and after
the crop ( Tables 1 and 3). The fixed NH+ at the
4
beginning of the experiment in year 1 was smaller
than in years 2 and 3, probably due to the preced-
ing crop (a cereal ), with a consequent greater
uptake of soil N. In years 2 and 3 the preceding
crops ( legumes) may have increased the N content
in the soils. The fixed NH+ decreased significantly
4 3
with depth ( Table 3), independently of treatment.
The decrease with soil depth was already observed
before the crop was grown and may have been
accentuated by the surface incorporation of fertil-
izer N.
The amounts of NO− and extractable NH+
3 4
were affected by the year and treatment, but not
by depth (Table 4). In year 1 the extractable soil
NH+ at harvest in the ammonium nitrate+
4 4
calcium nitrate treatment was significantly lower
than at the beginning of the experiment, and
significantly lower than values in years 2 and 3. In
the urea treatment, extractable soil NH+ increased
4 4
significantly from years 1 to 3. In year 1 it was
smaller than the value at the beginning of the
experiment, but the opposite was true in years 2
and 3. Soil NO− at harvest was not significantly
3
different in the 3 years following ammonium
nitrate+calcium nitrate application, though in
year 2 the NO− content was significantly lower at
3
the beginning of the experiment. With ammonium
nitrate+urea application the soil NO− at harvest
was greater in year 3 compared with years 1 and
2. In year 3 the soil NO− at harvest was also
3
greater than at the beginning of the experiment,
but it was smaller in year 2. As with total soil N,
only in year 3 did the ammonium nitrate+urea
application result in greater levels of nitrate and
extractable NH+ at harvest, compared with
4
ammonium nitrate+calcium nitrate application.
Some NH+ fixation occurred from 15N-labelled
fertilizer ( Table 5) especially in the surface layers
and with ammonium nitrate+urea application.
We could find no data in the literature on the
NH+ fixation from 15N-labelled fertilizer in the
presence of wheat. It is argued that recently fixed

Table 4
Levels of soil-extractable NH+ and NO− for the whole depths at sowing and harvesting time, and results of ANOVA to evaluate
4 3
the effects of year, treatment, and depth on inorganic-Na

Inorganic-N Year Initial Treatment at harvest

15NH 15NO +urea-15N 15NH 15NO +Ca(15NO )

4 3 4 3 32
NH+-N (mg kg−1) 1 6.6 Aa 3.4 Cb 1.9 Bb
4
2 4.2 Bb 6.7 Ba 5.0 Aab
3 5.6 ABb 13.1 Aa 5.8 Ab
NO−-N (mg kg−1) 1 3.0 Ca 2.7 Ba 2.7 Aa
3
2 17.2 Aa 1.8 Bb 3.9 Ab
3 5.8 Bb 13.5 Aa 3.7 Ab

Source of variation NH+-N NO−-N

4 3
Year (A) 18.25*** 50.27***
Treatment (B) 9.26** 7.42**
Depth (C ) 0.50 ns 2.65 ns
A×B 4.64* 9.33***
A×C 0.45 ns 1.12 ns
B×C 0.16 ns 0.50 ns
A×B×C 0.28 ns 0.22 ns

a For each soil characteristic, means in a row followed by the same letter or in a column followed by the same capital letter are
not significantly different (P≤0.05); ns, *, **, ***=F-values non-significant and significant at P≤0.05, P≤0.01 and P≤0.001
respectively.
152 C. Carranca et al. / European Journal of Agronomy 11 (1999) 145–155

Table 5
Fixed NH+-15N as affected by treatment and depth, and ANOVA to evaluate the effects of year, depth, and treatment on fixed
4
NH+-15Na
4
Depth (cm) NH+-15N dff (%) NH+-15N recovery (mg kg−1 of N )
4 4
15NH 15NO 15NH 15NO 15NH 15NO 15NH 15NO
4 3 4 3 4 3 4 3
+urea-15N +Ca(15NO ) +urea-15N +Ca(15NO )
32 32
0–10 4.963 Aa 1.561 Ab 7.260 Aa 1.676 Ab
10–20 0.595 Ba 0.000 Ba 0.642 Ba 0.000 Ba
20–30 0.014 Ba 0.000 Ba 0.015 Ba 0.000 Ba
30–40 0.025 Ba 0.000 Ba 0.018 Ba 0.000 Ba
40–50 0.000 Ba 0.000 Ba 0.000 Ba 0.000 Ba
50–60/70 0.045 Ba 0.000 Ba 0.041 Ba 0.000 Ba
Total 5.642 a 1.561 b 7.976 a 1.676 b

Source of variation NH+-15N dff NH+-15N recovery

4 4
Years (A) ns ns
Depth (B) 10.73*** 16.98***
Treatment (C ) 4.32* 6.35*
A×B 2.20* ns
A×C ns ns
B×C 2.86* 3.60**
A×B×C ns ns

a For the same soil characteristic, means in a row followed by the same letter or in a column followed by the same capital letter
do not differ significantly (P≤0.05); ns, *, **, ***=F-values non-significant and significant at P≤0.05, P≤0.01 and P≤0.001
respectively.

NH+ can simply replace native NH+, without a
4 4
change in the total amount of fixed NH+
4
( Kowalenko and Ross, 1980). Our results do not
contradict this statement, since in years 2 and 3
there was no net increase in the amount of fixed
NH+. Fixed NH+ even decreased in year 1, either
4 4
due to absorption by the crop (Scherer and
Weimer, 1994) or by transformations in the soil.
However, the fact that there were differences in
the amounts of fixed NH+ when ammonium
4
nitrate+urea or ammonium nitrate+calcium
nitrate were used in year 3 suggests that the
capacity to fix NH+ by the 2:1 clay mineral
4
(montmorillonite) is an important process for
removing NH+ from solution, and to protect
4
against nitrification. According to Scherer (1993),
the ability of montmorillonite to fix NH+ ions
4
under wet conditions is negligible. If fixation
occurs it should be attributed to the presence of
zones with vermiculite or illite structures.
We could not evaluate how much of the N
remaining in the soil at harvest would be available
to the following crop, because the plots had to be
disturbed to collect the soil samples. Important
losses over the summer can only be due to ammo-
nia volatilization. Since the majority of the NH+
4
present was fixed in the clay minerals and minerali-
zation will be slow because of the soil water deficit,
losses were likely to be negligible. Farmers choose
the sowing date for wheat in the autumn according
to the amount of rainfall and, therefore, the major-
ity of N remaining in the soil at harvest would
probably be available for the next crop. The basal
dressing would then probably not be needed, and
this would in turn lead to smaller losses through
leaching during the first stages of plant growth.
This subject deserves further investigation.
3.3. N in the leachate
In year 1 no leachates could be collected owing
to the limited rainfall. Leaching of fertilizer N
 C. Carranca et al. / European Journal of Agronomy 11 (1999) 145–155
below the rooting zone occurred in years 2 and 3,
mostly after the autumn application of fertilizer
( Table 6). Similarly, Mompo and Armentia (1992)
in Spain, and Mouchová et al. (1996) in Prague
found that most of the N applied to wheat at
sowing was leached during the winter. Under these
conditions, it is advisable to apply N fertilizer only
late in the winter or in spring (Zapata and van
Cleemput, 1986; Mompo and Armentia, 1992).
Losses of NO− by leaching did not differ signifi-
3 clayey soils (Zapata and van Cleemput, 1986;
cantly with treatment in year 2, but were signifi-
cantly higher in year 3 with ammonium
nitrate+urea treatment. The %Ndff in leached
nitrate was not significantly different in both years
and treatments, with an average value of 6.3%.
Ammonium was detected in the leachate, but
only at very low levels (2.3 mg l−1 of NH+-N ),
4
and a very small percentage was derived from
labelled fertilizer (0.3%). It was not affected by
year, treatment or sampling time. Zhu et al. (1996)
did not detect NH+ in the leachate water, when
4
urea was applied to wheat in similar soils.
The infiltration rate of water in Vertisols and
Luvisols is low (0.04–0.09 cm h−1), and leaching
movement is not considered important in such
soils (Sigunga and Janssen, 1996). Therefore, it
can be assumed that N losses were mainly due to
bypass flow. Important losses of nitrate through
cracks in clayey soils were observed by White
(1987), Goss et al. (1993), Strong (1995) and Zhu
et al. (1996).
3.4. Unaccounted 15N
Nitrogen can be lost from soil mainly via ammo-
nia volatilization, denitrification, nitrification and
Table 6
Comparison of means of nitrate in the leachate by LSDa
Year Sampling date
2 92/12/23
93/02/12
93/03/15
3 94/01/10
94/03/03
a Means in a row followed by the same letter or in a column followed by the same capital letter do not differ significantly at
P≤0.05; nd=not determined owing to lack of leachate.
153
leaching. Part of the 15N-labelled fertilizer was lost
by leaching, especially in autumn–winter in years
2 and 3. Although leachates could not be collected
in year 1, N derived from fertilizer was present in
the 50–70 cm soil layer, indicating that downward
movement of fertilizer N occurred even in this year.
Losses by denitrification may also have occurred
during temporary periods of oxygen deficiency in
the microsites when the soils were very wet in such
Carvalho, 1987; van Cleemput et al., 1987). The
use of ammonium nitrate as a basal dressing may
have contributed to these losses.
The water deficit observed from February until
the end of the season in year 1, and the relatively
high air temperatures, especially during grain for-
mation, could explain the low N recovery by plants
due to NH volatilization following urea applica-
3
tion (Boaretto et al., 1982; Carvalho, 1987).
Ammonia volatilization from the aerial parts of
wheat at maturity, caused by hydrolysis of proteins
due to the relatively high air temperatures, can be
another source of N loss. According to Peoples
et al. (1995), about 20 kg ha−1 of N can be lost
from wheat in this way.
As a result of these losses, from 0 to 55% of
the 15N-labelled fertilizer could not be accounted
for at harvest. N losses were similar in years 1 and
2, with both treatments, and in year 3 with ammo-
nium nitrate+calcium nitrate application (in the
range 35–55%). In year 3 the treatment with
ammonium nitrate+urea resulted in the recovery
of all the 15N applied. The values for unaccounted
N (0–99 kg ha−1 of N ), though important, were
NO−-N (mg l-1)
3
15NH 15NO +urea-15N 15NH 15NO +Ca(15NO )
4 3 4 3 32
29.30 Ba 19.42 Ba
28.42 Ba 30.53 ABa
40.12 Ba 11.69 Ba
159.25 Aa 76.06 Ab
nd 14.53 B
154 C. Carranca et al. / European Journal of Agronomy 11 (1999) 145–155
generally smaller than the N losses estimated by
Carvalho (1987) for wheat grown in Portugal
(94–120 kg ha−1 of N ). They are also smaller than
the percentage average loss of 15N-labelled fertiliz-
ers reported from Syria (Pilbeam et al., 1997) and
Tunisia (Sanaa et al., 1992).
4. Conclusions
The recovery of 15N-labelled fertilizer by rain-
fed wheat grown under Mediterranean conditions
was similar to the urea or calcium nitrate top-
dressings. The level of 15N from labelled fertilizer
that remained in the soil at harvest was similar
with both top-dressings in the driest and wettest
years. When rainfall matched potential evapo-
transpiration the ammonium nitrate+urea appli-
cation led to a great recovery of 15N from labelled
fertilizer in the soil and greater values of total soil
N, NO− and extractable NH+. There was evidence
3 4
of NO− leaching in the two wettest years, indepen-
3
dently of the form of N fertilizer, particularly after
the first two N applications, during the autumn
and winter periods. Therefore, whenever poss-
ible, autumn applications of N fertilizer should
be avoided in order to reduce N leaching.
Ammonium-15N was fixed in the clay mineral,
especially in the surface layers (by the surface
incorporation of fertilizer N ) and with ammonium
nitrate+urea application. This process is impor-
tant for removing NH+ from soil solution, and to
4 Khanif, Y.M., van Cleemput, O., Baert, L., 1983. Fate of field-
protect against nitrification.
Acknowledgements
Financial support from the International
Atomic Energy Agency ( Vienna, Austria), grant
no. POR/5/003, is gratefully acknowledged.
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