Europ. J.

Agronomy 51 (2013) 25–33

Contents lists available at ScienceDirect

European Journal of Agronomy

journal homepage: www.elsevier.com/locate/eja

Plant stand, nodulation and seed yield in soybean as affected by

phosphate fertilizer placement, source and application method
Fernando Salvagiotti a,∗ , Mirian Barraco b , Damian Dignani c , Hector Sanchez d ,
Alfredo Bono e , Pedro Vallone f , Guillermo Gerster a , Carlos Galarza f ,
Jorgelina Montoya f , Vicente J. Gudelj f
a
Dep. Agronomía, EEA INTA Oliveros, Ruta 11 km 353 (C 2206), Santa Fe, Argentina
b
EEA INTA General Villegas, San Martin 26 (6230), General Villegas, Buenos Aires, Argentina
c
NOVA S.A. Ruta 9 km 373,9 – (2500) Cañada de Gómez, Santa Fe, Argentina
d
EEA INTA Famailla, Ruta Prov. 301 km 32 (4132), Famaillá, Tucumán, Argentina
e
EEA INTA Anguil, Ruta 5 km 580 (6326), La Pampa, Argentina
f
INTA Marcos Juárez, Ruta 12, (2580) Marcos Juárez, Córdoba Republica, Argentina

a r t i c l e i n f o a b s t r a c t

Article history: Establishing an optimum plant population at planting is the start-point for assuring seed yield in crops.
Received 9 April 2013 In addition to plant establishment, the onset of a fully functional nodule system is crucial for nitrogen
Received in revised form 23 June 2013 nutrition in soybean. Fertilizer application in close contact with seeds during planting operation would
Accepted 29 June 2013
lead to phytotoxicity and plant loss. The objectives of the present study were to study the effects of two
phosphate fertilizer sources: single super phosphate (SSP) and monoammonium phosphate (MAP) (i) at
Keywords:
increasing phosphorus rates applied with seed (WS) and (ii) using different fertilizer placement strategies
Phosphate fertilizer
on plant stand, seed yield and nodule onset. Two fertilization experiments in soybean (Experiments I
Phytotoxicity
Plant stand
and II) were performed in eleven sites in an ample area of the Argentinean soybean production area in
2005–2006 and 2006–2007 seasons (site-years, SY). Experiment I was designed for evaluating the effects
of two P fertilizer sources (Sou): SSP and MAP combined with three methods of fertilizer application
(Me): WS, side-banded (SB) or broadcast anticipated (BA) applying 16.2 kg P ha−1 . Broadcast application
was done at least 90 days before planting. Experiment II was planned for evaluating the above mentioned
fertilizer sources at increasing P rates (0, 5.4, 10.8 and 16.2 kg P ha−1 ) applied with seed (WS). Fertilizer
application with seed had less detrimental effect on plant stand achievement (PSA) when SSP was used as
compared with MAP. Reductions in PSA were directly related with clay content when using both fertilizer
source, but when using MAP, the decay in PSA was also negatively related to silt content. Broadcast as
well as side-banded application of both fertilizer sources had no damaging effects on PSA. Seed yield
response to fertilizer application was similar among different fertilizer placement strategies. In most
cases, reduction in PSA did not affect seed yield, suggesting a high capacity of soybean to compensate
plant loss. Nodule onset was severely affected by application of MAP with seed, but this was not translated
in a reduced seed yield.
© 2013 Elsevier B.V. All rights reserved.

1. Introduction combination of insecticide and herbicide (Hayes et al., 1979) or

Establishing an optimum plant population at planting is the
start-point for assuring seed yield in crops, since radiation cap-
ture is warranted early in the cycle (Andrade, 1995; Purcell
et al., 2002) also affecting water and nutrient uptake (Azam-Ali
et al., 1994). Several factors may reduce plant population estab-
lishment such as low seed vigor (Ellis, 1992), toxicity due to a
∗ Corresponding author. Tel.: +54 3476 498010; fax: +54 3476 498010.
E-mail addresses: fsalvagiotti@correo.inta.gov.ar, fsalvagiotti@hotmail.com
(F. Salvagiotti).
fertilizer toxicity (Bremner and Krogmeier, 1989). Seed yield sensi-
tivity to plant loss varies among crops, being of highly importance
in crops that develop few reproductive structures like maize, but of
less magnitude in crops that may compensate plant loss by devel-
oping compensatory strategies (e.g. tillering, branching). The linear
relationship between seed number per plant and plant growth rate
during the critical period found by Vega et al. (2001) shows the
larger plasticity in soybean for generating reproductive sinks in
comparison with other crops like sunflower or maize.
Soybean is the principal summer crop in Argentina (Wilcox,
2004). Seed yield reduction associated with nutrient constraints in
field conditions has been reported for sulfur (S) and phosphorus

1161-0301/$ – see front matter © 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.eja.2013.06.006
26 F. Salvagiotti et al. / Europ. J. Agronomy 51 (2013) 25–33

(P) (Gutierrez-Boem et al., 2007; Salvagiotti et al., 2004, 2012).

Phosphate fertilizer market has been dominated by diammonium
phosphate (DAP), monoammonium phosphate (MAP) or triple
super phosphate (SPT), all of them characterized by high-analysis
P content (more than 20% of P in its formulae). In the last years, sin-
gle superphosphate (SSP) market increased in Argentina, because
includes P and S in their grade. Since a lower P concentration is
found in SSP in comparison with MAP or DAP, larger fertilizer rates
are needed to apply in order to reach the same P level. All the
above mentioned P fertilizer sources have more than 85% of phos-
phate in soluble forms (Chien et al., 2011), but bioavailability of
phosphates will be related to soil pH and associated free Fe and
Al content (Sample et al., 1980). Most soils dedicated to agricul-
ture in Argentina showed moderately acid reaction (Alvarez et al.,
2009; Salvagiotti et al., 2012), and thus, low Fe or Al content that
will enhance available P from fertilizers. Varied results have been
reported with regards to agronomic efficiency using different P fer-
tilizer sources. Lu et al. (1987) observed a better performance of
SSP in combination with urea when related with DAP in calcareous
soils. Nandini Devi et al. (2013) in an acid soil showed similar agro-
nomic P use efficiency when comparing SSP and DAP. However, no
research has been made comparing MAP and SSP in Mollisols, the
main soils in the soybean production area in Argentina.
In general, fertilizer application in close contact with seeds
during planting operation would lead to phytotoxicity. Many
times soybean planters are not equipped with devices to separate
fertilizer from seeds, and phytotoxic effects are expected to hap-
pen. Processes that cause phytotoxicity depend on fertilizer type.
Ammonium phosphates fertilizers may alter seedling growth by
a direct effect of ammonium (Lowell and Tsao, 1986), but also
by modifying pH in the area surrounding the fertilizer granule,
that may have acidic (MAP) or alkaline reactions (Young et al.,
1985). On the other hand, reaction of fertilizers like SSP may cause
sharp decreases up to 1.5 in pH (Hedley and McLaughlin, 2005)
that may also affect seedling growth. In a study performed on
maize and wheat, Cummins and Parks (1961) showed less detri-
mental effect on plant germination of 20% superphosphate as
opposed to ammonium based fertilizers, when all were applied
with seed. Early studies have compared the effects of anhydrous
ammonia, concentrated superphosphates or ammonium sulphate
on plant emergence in wheat and maize (Allred and Ohlrogge, 1964;
Cummins and Parks, 1961; Olson and Dreier, 1956), but no compar- Fig. 1. Distribution of experimental sites in the soybean production area in
isons have been made between MAP and SSP in soybean plants. Argentina. Site numbers are described in Table 1.
The negative impact of fertilizer on plant emergence decreases
as soil moisture increases and the time that the seed and the fer-
tilizer granule are in close contact decreases (Chapin and Smith, a large proportion of bacteria from inoculants are expected to hap-
1960). Therefore it is expected less phytotoxic effects in heavy tex- pen (McDermott and Graham, 1989). In addition to reductions in
tured soils which have a greater water holding capacity, in addition plant stand, changes in pH or in ammonia concentration in the prox-
to a larger cation exchange capacity. Another option for avoiding imity of the fertilizer granules may alter plant nodulation (nodule
the application of fertilizer with seeds is broadcasting P fertilizer weight and/or number) when fertilizers are applied with seeds, but
before planting or in winter when soil is in fallow. In soils of the Mid- this issue has not been explored before.
west in USA, Bordoli and Mallarino (1998) showed no differences in In the present study we studied the effects of two phosphate
grain yield on maize when comparing broadcast and deep banded fertilizer sources (SSP and MAP) (i) at increasing phosphorus rates
application at planting. Also, Mallarino et al. (2009) observed in applied with seed (WS) and (ii) using different fertilizer placement
soybean similar responses in seed yield when applying P fertilizers strategies on plant stand, seed yield and nodule onset in soils with
in the fall and in the spring. different texture in the Argentinean soybean production area.
In addition to plant establishment, the onset of a fully functional
nodule system is crucial for nitrogen nutrition in soybean. Bradyrhi-
zobium japonicum specifically induce nodule formation in soybean 2. Materials and methods
and is the principal bacteria in commercial inoculants (Graham,
2008). Its functionality is important at around one month after 2.1. Generalities
planting, but the recognizing process between bacteria and the
host plant occurs early, in the first days after radicle emergence Two fertilization experiments in soybean (Experiments I and II)
(Zhang and Smith, 2002). Low pH, reduced soil moisture and low were performed in eleven sites in an ample area of the Argentinean
temperatures may alter this process (Sadowsky, 2005). These fac- soybean production area in 2005–2006 and 2006–2007 seasons (i.e.
tors may decrease the proportion of nodules in the crown, where 11 site-years, SY, Fig. 1).
 F. Salvagiotti et al. / Europ. J. Agronomy 51 (2013) 25–33 27

Table 1
Soil features and planned plant population in eleven site-years (SY).

SY Season Site Soil type Texture Organic C (g kg−1 ) pH Bray1 P (mg kg−1 ) S-SO4 (mg kg−1 ) PPP PNc

1 2005–2006 AG6 EH L 8.8 6.0 30 n.a. n.a. 10

2 2005–2006 GV6 TH L 10.2 6.2 10 9.3 46 41
3 2005–2006 MJ61 TA SiL 14.8 6.0 19 4.8 40 31
4 2005–2006 MJ62 TA SiL 14.8 6.0 19 4.8 40 29
5 2005–2006 SJ6 VA SiL 16.4 5.9 12 n.a. 46 43
6 2005–2006 LV6 EH SiL 10.7 6.5 13 8.6 46 26
7 2006–2007 GV7 TH L 10.5 6.1 11 n.a. 46 35
8 2006–2007 MJ71 TA SiL 13.2 6.2 18 3.3 48 36
9 2006–2007 MJ72 TA SiL 13.2 6.2 29 2.5 48 32
10 2006–2007 AG7 EH SaL 7.4 6.0 30 n.a. 39 22
11 2006–2007 LR7 TH SiL 12.8 6.3 8 n.a. 42 40

Soil type: TH, thaptoargic hapludol; TA, typical argiudol; VA, vertic argiudol; EH, entic haplustol.
Texture: L, loam; SiL, silty-loam; SaL, sandy-loam
PPP, planned plant population (seeds m−2 ); PNc , plant number achieved in the control (non-fertilized) treatment; n.a. = not available.

All experiments were performed under no tillage in soils that respective problems of hail and drought GV7 and AG6 were not
represented a large range of texture (Table 1). Experiment I was analyzed for seed yield.
designed for evaluating the effects of two P fertilizer sources
(Sou): single superphosphate (SSP) and monoammonium phos-
2.3. Data analysis
phate (MAP) combined with three methods of fertilizer application
(Me): WS, side-banded (SB) or broadcast anticipated (BA). In all
Analysis of variance was performed using the Proc Mixed pro-
cases a P rate of 16.2 kg P ha−1 was evaluated. Broadcast applica-
cedure in SAS (Littell et al., 1996). In both experiments, replication
tion was done at least 90 days before planting. Experiment I was
nested within SY was used as an error term for evaluating SY effect
performed in AG6, GV6, MJ61, SJ6, LV6, GV7, MJ71, AG7 and LR7.
(Gomez and Gomez, 1984). In case of interaction among factors
Experiment II was designed for evaluating the above mentioned fer-
the slice statement was used for detecting differences within each
tilizer sources at increasing P rates (0, 5.4, 10.8 and 16.2 kg P ha−1 )
interaction. Lineal and lineal–plateau functions were fitted to ana-
applied with seed (WS). Since SSP contains 11% S in its formulae,
lyze the effects of increasing P rates on PSA. Quadratic regression
treatments were equilibrated using calcium sulphate in order to
was used to analyze the effects of increasing P rates on seed yield.
provide the same amount of S in each treatment. Experiment II was
Agronomic efficiency (AE) of fertilizer P was estimated at a P rate
done in all SY. In both experiments, treatments were arranged in
when maximum seed yield response was determined. Partial factor
a randomized complete block design, with three replications. Each
productivity (PFP) was calculated as seed yield P rate−1 . Multiple
experimental unit consisted of 4–5 rows wide (0.52 m spacing) and
regression analysis was used in order to predict PSA as a function
10–20 m long. Planned plant population (PPP) was decided based
of P rate and soil variables: texture (clay and silt percentage), soil
on local recommendation for optimal growth in each SY.
organic matter content, soil pH and soil moisture. Variables that
were significant at 5% level were left in the final model.
2.2. Measurements

In each SY, a composite soil sample was taken at 20 cm depth 3. Results

before planting. Carbon content (Walkley and Black, 1934), phos-
phorus P Bray I (Bray and Kurtz, 1945), S-sulphate soil content
(Chesnin and Yien, 1950) and soil texture (i.e. clay, silt and sand
proportion) by pipet method (Gee and Bauder, 1986) was deter-
mined. Phosphorus P Bray is the recommended soil analysis to test
P fertility since most soils in the soybean production area Argentina
had a moderately acid reaction (Ferraris et al., 2002). Carbon con-
tent was converted to organic matter (OM) using a recuperation
factor of 0.58. An additional soil sample in the upper 7 cm discard-
ing the 2 upper cm was taken, in order to quantify soil moisture in
the area where fertilizer granule and seed were in contact.
At V3 stage (Fehr and Caviness, 1977), plant stand was deter-
mined in the two central rows of each plot. In each experimental
unit, plant stand achievement (PSA) was calculated as
PNt
PSA(%) = × 100
PNc
where PNt and PNc are plant number achieved in each treatment
and in the control (non-fertilized), respectively.
In SJ6, GV7 and MJ71, nodule weight and number were deter-
mined in R2 stage by detaching nodules from a 1 m long row plant
sample. Determinations were done in the control (non-fertilized)
and in treatments that received the largest P rate: SSP and MAP
applied with seed and site-banded MAP.
At harvest, seed yield was determined in each plot and adjusted
to a standard moisture content of 0.13 kg H2 O kg grain−1 . Due to
3.1. Soil features and climatic conditions
Experiments were performed in typical soils of the soybean
production area in Argentina, including loam, silty-loam and sandy-
loam textures with organic carbon content that ranged from 7.4 to
13.2 g C kg−1 . They all had a moderately acid reaction (pH values
between 5.9 and 6.5) and limestone content is considered negligi-
ble in these soils (SAGyP-INTA, 1990). Phosphorus content in the
upper 20 cm oscillated from 8 to 30 ppm (Table 1).
Rainfall between November and March, period in which soy-
bean was grown in all sites, oscillated between 382 mm in AG6 to
1415 mm in LV6. In all sites, actual rainfall showed values 14 to 81%
larger than historical records (Fig. 2). In general, mean maximum
and minimum temperature in the same period showed values equal
or slightly greater (1–6%) than historical records. In GV7, mean
(1)
minimum temperature was 10% larger than historical records.
3.2. Effects of method of fertilizer application on PSA and seed
yield (Experiment I)
Fertilizer source did not affect PSA but a significant SY × Me
interaction was observed (P < 0.01, Table 2). In all SY, fertilizer appli-
cation with seed severely reduced PSA with different magnitude,
producing plant losses that ranged from 20 to 48% (Fig. 3). In SB and
BA treatments, reductions in PSA were of less magnitude, from 0
28 F. Salvagiotti et al. / Europ. J. Agronomy 51 (2013) 25–33

Fig. 2. Actual and Historical decadal records for rainfall, maximum and minimum temperature in the 11 site-years. Historical record corresponds to climatic series data of
30 years. Arrows indicate planting date.

to 17%. In most cases (7 out 9 SY), SB and BA did not significantly
differ from control plots.
A significant SY × Me interaction was observed for seed yield
(P < 0.01, Table 2). Significant increase in seed yield in response to
P fertilizer addition was only observed in GV6, SJ6 and LR7 with a
respective increase of 23, 24 and 12% over the control non-fertilized
treatment (Fig. 4). In GV6 and LR7, no differences among meth-
ods were observed, but, in SJ6, broadcast anticipated application
significantly increased seed yield 8% over WS application (Fig. 4).
3.3. Effects of fertilizer rate applied with seed on PSA and seed
yield (Experiment II)
A significant SY × Sou × P interaction was observed in PSA
(P = 0.02; Table 3). The analysis of the SY × P interaction using
the slice statement showed that PSA decreased in all SY when
P rate increased except for MJ62 (Table 4). In the remaining SY,

Table 2
Principal effects and ANOVA for plant stand achievement (PSA) and seed yield (cor-
rected to 0.13 kg H2 O kg grain−1 ) as affected by two fertilizer sources (Sou): single
superphosphate (SSP) and monoammonium phosphate (MAP) and three methods of
application (Me): BA = broadcast 90 days before planting; SB = side-banded at plant-
ing; WS = applied with seed in 9 sites-years in 2005–2006 and 2006–2007 seasons
(SY). (Experiment I). P rate was 16.2 kg P ha−1 for both fertilizer sources.

PSA (%) Seed yield (kg ha−1 )

Control 100 3293

Sou
MAP 87 3494
SSP 90 3437
Me
BA 94 3567
SB 94 3545
WS 67 3458

P value

SY 0.02 <0.01
Me <0.01 <0.01
Fig. 3. Plant stand achievement in soybean in 9 site-years during 2006 and 2007
Sou 0.12 0.18
in response to fertilizer application method (Experiment I). Each column is the
Me × Sou 0.20 0.36
average of two fertilizer sources (monoammonium phosphate and single super
SY × Me <0.01 <0.01
phosphate). Lines in each column are the standard error of the mean. Asterisks indi-
SY × Sou 0.86 0.12
cate sites where fertilizer applied WS significantly reduced PSA respect to control
SY × Me × Sou 0.14 0.18
plot (P < 0.05).
 F. Salvagiotti et al. / Europ. J. Agronomy 51 (2013) 25–33 29

Table 3
Principal effects and ANOVA for plant stand achievement (PSA) and seed yield (cor-
rected to 0.13 kg H2 O kg grain−1 ) as affected by two fertilizer sources (Sou) applied
with seed (WS): single superphosphate (SSP) and monoammonium phosphate
(MAP) and four phosphorus (P) rates in 9 sites-years in 2005–2006 and 2006–2007
seasons (SY). (Experiment II).

PSA (%) Seed yield (kg ha−1 )

Sou MAP 80 3472

SSP 83 3466
P 0 100 3372
5.4 82 3535
10.8 74 3497
16.2 69 3471

P value

SY <0.01 <0.01
Sou <0.01 0.88
P <0.01 0.01
Sou × P 0.18 0.88
SY × Sou <0.01 0.03
SY × P <0.01 <0.01
SY × Sou × P 0.02 0.86
Fig. 4. Soybean seed yield in 7 site-years (SY) during 2006 and 2007 in response
to fertilizer application method (Experiment II). Each column is the average of
two fertilizer sources (monoammonium phosphate and single super phosphate).
Lines in each column are the standard error of the mean. Asterisks indicate sites with seed (Table 5). The relationship between PSA and P rate in
where response to fertilizer application (fertilized (WS + SB + BA) vs. control plot) the remaining sites was adequately fitted by a lineal–plateau func-
was significant (P < 0.10). AG6 and GV7 were not analyzed due to drought and hail,
tion (Fig. 5). The reduction in PSA at increasing P rate (parameter
respectively.
b) varied from 2.1 to 6.5% per unit of applied phosphorus. The P
rate at which the minimum PSA was attained ranged from 5.8 to
both fertilizer sources applied with seed reduced PSA, but different 11.8 kg P ha−1 (Table 5). A significant fertilizer source effect was
trends were observed in the relationship between P rate and PSA observed only in SJ6 and LR7. In both sites, MAP had a more detri-
(Fig. 5). mental effect on PSA as shown by parameter b. Application of MAP
In MJ61, MJ71 and MJ72 a lineal trend between PSA and P rate with seed decreased PSA 5.6 and 5.1% per unit of applied P in SJ6 and
was observed with no differences between fertilizer sources. Plant LR7, respectively. When using SSP, this decrease rate was 45 and
stand decreased at a between 1.8 and 2.1 plants/kg of P applied 60% lower than when applying MAP, respectively (Table 5). In both

Fig. 5. Plant stand achievement in response to P addition using two fertilizer sources: monoammonium phosphate (MAP) and single super phosphate (SSP) applied with
seed in 11 site-years (Experiment II). Parameters of the equations of each SY are shown in Table 5.
30 F. Salvagiotti et al. / Europ. J. Agronomy 51 (2013) 25–33

Table 4 Table 6
Probability values of the interactions Site-year × Source and Site-year × P rate as Mean values, analysis of variance and preplanned contrasts P values for nodule
separated by the slice statement in SAS (Experiment II). weight and nodule number per m−2 relative to a control non-fertilized and three
fertilizer allocation treatments (FT) in 3 site-years in 2005–06 and 2006–07 (SY).
P value
Means Nodule weight (g m−2 ) Nodule number per m−2
SY PSA Seed yield PSA Seed yield
SY × Soua SY × Pa SY
SJ6 7.59 1581
AG6 0.72 – <0.01 – GV7 4.60 1145
GV6 0.09 0.17 <0.01 <0.01 MJ71 2.95 346
MJ61 0.27 0.52 <0.01 0.99 SE 0.60 262
MJ62 0.74 0.89 0.96 0.63 FT
SJ6 <0.01 0.56 <0.01 <0.01 Control 4.52 1081
LV6 0.96 0.47 <0.01 0.68 MAP SB 6.17 1217
GV7 0.59 – <0.01 – SSP WS 5.28 929
MJ71 0.96 0.34 <0.01 0.32 MAP WS 4.21 871
MJ72 0.07 0.21 <0.01 0.07 SE 0.68 133
AG7 0.16 <0.01 <0.01 0.32
LR7 <0.01 0.32 <0.01 <0.01
Means P values
Italic values indicate P < 0.10.
a
SY <0.01 <0.01
Interaction.
FT 0.04 0.66
SY × FT 0.16 0.08
Table 5
Parameters of lineal and lineal-plateau regression equations between PSA and P Contrasts P values
rate in 10 site-years (SY) using two fertilizer sources: Monoammonium phosphate
(MAP) and single super phosphate (SSP). a = PSA when no fertilizer is applied; b = PSA Control vs MAP SB 0.03 0.32
decrease at incrementing P rate; c = P rate at which minimum PSA is reached. SSP WS vs MAP WS 0.13 0.67
MAP SB vs MAP WS <0.01 0.02
SY Sou Parameters r2 Predicted PSA at c
FT are monoammonium phosphate (MAP) applied side-banded (SB) and with seed
a b c (WS) at planting and single superphosphate (SSP) applied with seed at planting. P
rate was 16.2 kg P ha−1 for both fertilizer sources.
AG6 100 −4.3 6.8 0.86 71
Italic values indicate P < 0.10.
GV6 103 −4.3 6.7 0.86 72
MJ61 101 −1.8 – 0.93 a

MJ62a – – – – –
the rest of the SY (Table 4). The SY × P interaction showed a sig-
SJ6 MAP 102 −5.6 11.3 0.99 39
SSP 100 −3.1 10.6 0.99 66
nificant increase in seed yield in response to P addition in 3 SY
LV6 100 −3.1 8.1 0.85 75 (GV6, SJ6 and LR7) (Table 4). A quadratic response curve explained
GV7 100 −3.9 5.8 0.85 78 this response in these SY, showing the maximum seed yield at P
MJ71 101 −1.8 – 0.88 a
rates of 10.4, 11.2 and 14.6 kg P applied per ha for LR7, SJ6 and
MJ72 100 −2.1 – 0.91 a
GV6, respectively (data not shown). Maximum observed response
AG7 100 −6.5 6.1 0.96 61
LR7 MAP 104 −5.1 11.8 0.95 44 to P addition ranged between 247 and 375 kg ha−1 , determined at P
SSP 101 −2.1 10.8 0.97 79 rates that oscillated between 5.2 and 7.3 kg P ha−1 . Agronomic use
a
Parameters for MJ62are not shown because no significant regression could be
efficiency and PFP ranged from 34 to 70 and 312 to 794 kg grain kg−1
fitted. of applied P (Fig. 6, inset).

SY, fertilizer source did not affect parameter c, and thus, the P rate
at which minimum plant stand was achieved was ca. 11 kg P ha−1 .
At this P rate, in SJ6, PSA is expected to be 40% when MAP is applied,
showing a 39% reduction as compared with addition of SSP. A simi-
lar relative reduction in PSA (ca. 44%) is expected in LR7 in response
to source application (Table 5).
Soil organic matter, clay and silt content in addition to P rate
accounted for 46% of the variation in PSA, when MAP was applied
with seed. On the other hand, when SSP was applied the best equa-
tion only included P rate and clay content, and explained 51% of
variation in PSA. Soil pH and soil moisture had no significant con-
tribution in both equations. The equations for both fertilizer sources
were:
PSAMAP = 1.09 − 0.02 P rate + 0.025 clay − 0.13 OM − 0.005 silt(2)
PSASSP = 0.77 − 0.017 P rate + 0.01 clay
3.4. Effects of method of fertilizer application on nodule formation
Independently of site (i.e. no significant site × treatment inter-
action), side-banded applications of MAP significantly increased
nodule weight by 36% respect to the control treatment (P < 0.05,
Table 6). A significant reduction (ca. 32%) in nodule weight was
observed when MAP was applied WS when compared with SB
application. No significant differences (P > 0.10) were observed
when comparing banded application of MAP with SSP applications
with seed (Table 6).
In contrast, banded applications of MAP did not increase signif-
icantly nodule number respect to the control, and no significant
interaction was observed among site-years. However, fertilizer
applications WS (both SSP and MAP) significantly reduced 26%
nodule number respect to banded applications of MAP without
differences between fertilizer sources (Table 6).
(3)

In both cases, a positive contribution of clay content on PSA 4. Discussion

was observed. On the opposite, increasing P rate, silt content and
organic matter in soil showed a negative effect.
A significant SY × Sou and SY × P interaction were observed for
seed yield (Table 3). Analyzing the SY × Sou interaction as separated
by the slice statement, only a significant 18% decrease in the SSP
treatment respect to MAP was observed in AG7, however, in this
site no significant response to P fertilization was observed (data not
shown). No effect of fertilizer source on seed yield was observed in
Soybean increased seed yield in response to P fertilization in
sites that showed soil P content not above 12 mg kg−1 . Soil test
calibration studies in the area (Ferraris et al., 2002) showed a
soil P threshold of 13 mg kg−1 below which response to P fertil-
ization is most likely to occur. Maximum seed yield responses
to P addition in these sites were observed at P rates between
5 and 7 kg P ha−1 , with PFP larger than 300 kg seed P rate−1 and
 F. Salvagiotti et al. / Europ. J. Agronomy 51 (2013) 25–33
Fig. 6. Seed yield in response to P addition using two fertilizer sources (monoam-
monium phosphate (MAP) and single super phosphate (SSP)) in site-years where
response to P fertilization was significant (see Table 4). The inset shows predicted
maximum seed yield response (kg ha−1 ) and P rate (kg ha−1 ), Partial factor produc-
tivity of P (PFP, kg seed kg−1 P applied) and Agronomic use efficiency (AE, kg < 0.01
seed yield kg−1 of applied P) estimated at maximum seed yield response.
agronomic efficiency above 34 kg seed kg−1 of applied P fertilizer.
In soybean, Garcia and Salvagiotti (2009) showed values of PFP of
ca. 350 kg seed kg P rate−1 and AE values up to 24 kg seed kg−1 of
applied P fertilizer, which are smaller than PFP and AE observed in
the present study. It is important to note that Garcia and Salvagiotti
(2009) calculated AE and PFP at a fixed P rate (i.e. 10 kg P ha−1 ).
In the present study, increasing P rates were evaluated, and thus,
optimum P rate and maximum response at this P rate could be
31
calculated by deriving the quadratic function, giving a more precise
estimation of PFP and AE.
Response to anticipated P fertilization is related to sorption-
desorption characteristics of the soils, associated with clay content
and type, soil pH, presence of reactive oxides or soil redox status
(Sims and Pierzynski, 2005). Soils in the pampean region are domi-
nated by Mollisols, which have low P retention capacity (Boschetti
et al., 1998; Gutierrez Boem et al., 2008; Silva Rossi et al., 2013).
In line with these, the present study showed a similar response to
P addition when fertilizer was applied side-banded at planting or
broadcasted 90 days before planting, with both SSP and MAP. Sim-
ilar results were observed in soils with the same texture in the USA
Midwest (Bordoli and Mallarino, 1998; Mallarino et al., 2009).
In all sites fertilizer application with seed reduced PSA as com-
pared with site-banded applications for both fertilizer sources.
However, in responsive sites, no differences in seed yield were
observed among fertilization placement (WS vs. SB or WS vs. BA),
suggesting that soybean could compensate up to 48% of plant loss.
On the other hand, the lack of response to fertilizer application
when applied with seed in the non-responsive sites was not asso-
ciated with reductions in PSA, since the other fertilizer placement
treatments (SB or BA) did not show significant increases in seed
yield. Previous research showed that early plant loss (at around V3)
reduced seed yield up to 33% (Conley et al., 2008). Clapp and Small
(1970) showed sharp reductions in plant stand when granular fer-
tilizers were applied with seed accompanied by small increases in
seed yield. In contrast, Teigen and Vorst (1975) showed no changes
in seed yield when plant stand was reduced up to 50% (from an opti-
mal plant stand of 30 seeds m−2 ) in V7. In the present study, plant
losses at the highest fertilizer rate ranged from 1 to 48%, without
affecting seed yield, which ranged from 2360 to 4583 kg ha−1 . Pre-
vious studies showed that increases in the plant stand variability
(quantified as vegetative biomass production per plant), did not
affect seed yield per plant in soybean, implying a larger capacity
for sustaining seed yield under plant stand reductions as compared
with maize (Andrade and Abbate, 2005). Therefore, the magnitude
of plant loss due to fertilizer application with seed shown in the
present study was not enough to affect seed yield.
In most locations, reductions in PSA were independent of fertil-
izer source. However, in sites that showed significant differences
between fertilizer sources (SJ6 and LR7), SSP had a lower decay
rate in PSA as P rate increased than MAP. These observations sug-
gest that in these experiments, the effects of ammonium in the area
close to the seed had more impact on plant stand reduction than
a reduced pH, and thus, for the same recommended P rate a larger
PSA is expected when SSP is applied with seed. As reductions in PSA
did not decrease seed yield, both fertilizer sources increased seed
yield in a similar manner in the responsive sites. This was expected
since P in both fertilizer sources has a high water solubility that
ensures agronomic performance (Young et al., 1985).
Soil texture in addition to P rate played an important role in
determining PSA as evaluated by a multiple linear regression. Clay
content was positively associated with PSA, using both fertilizer
sources. Increasing clay content in soils may increase water holding
capacity and thus, reducing the negative effects of ammonia on
seedlings. Likewise, the equation for MAP was negatively related
to silt content. Truog et al. (1925) found severe reductions in plant
stand in sandy soils with low soil moisture content. Unexpectedly,
soil moisture was not included in both models.
Phosphorus addition increased both nodule number and weight
when MAP was applied side banded. Previous studies have shown
increases in both variables when fertilizer P was applied in dif-
ferent experimental systems (Abbasi et al., 2010; Waluyo and Lie,
2004). However, this positive effect was reverted when fertilizer
was applied with seed. Reductions in nodule number suggest a local
effect of applied fertilizer, may be associated with a poor bacterial
32 F. Salvagiotti et al. / Europ. J. Agronomy 51 (2013) 25–33
infection when bacteria–plant interaction happens. Saline stress
has shown reductions in nodule number in soybean (Singleton
and Bohlool, 1984), explained by a diminution of hair root growth
and deformation or reduction in isoflavone secretion that reduces
plant-bacteria communication (Muñoz et al., 2012). In pea, the
close proximity of a concentrated band of more than 40 kg N ha−1
applied as urea reduced more nodule number than nodule weight
(Clayton et al., 2004). These authors justified this reduction in nod-
ule number as a consequence of a high nitrate concentration that
constrained the nodulation process rather than a phytotoxic effect.
In the present study, N addition at the highest MAP rate was ca.
13 kg N ha−1 that may have not affected nodulation as suggested
by these authors. However, a likely local negative effect of ammo-
nia release from fertilizer when MAP was applied WS may have
reduced nodule number and weight. Application of SSP WS slightly
increased nodule weight but reductions in nodule number are dif-
ficult to explain in terms of toxicity. Nandini Devi et al. (2013)
showed increases in both nodule number and weight when SSP
similar to those obtained with DAP, but no phytotoxic effects were
observed in nodule attributes.
5. Conclusions
Fertilizer application with seed had less detrimental effect on
PSA when SSP was used as compared with MAP. Reductions in PSA
were directly related with clay content when using both fertilizer
source, but when using MAP, the decay in PSA was also negatively
related to silt content. Broadcast as well as side-banded applica-
tion of both fertilizer sources had no damaging effects on PSA. Seed
yield response to fertilizer application was similar between differ-
ent fertilizer placement strategies. In most cases, reduction in PSA
did not affect seed yield, suggesting a high capacity of soybean to
compensate plant loss. Finally, nodulation was severely affected
by application of MAP with seed, but this was not translated in a
reduced seed yield.
Acknowledgments
We thank Dr Nacira Muñoz (IFRGV, INTA) and Ing. Agr. Juan Mar-
tin Enrico (INTA Oliveros) for critical comments on the manuscript.
This work was supported by INTA (Argentine Institute of Agricul-
tural Research) and Proyecto Fertilizar.
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