Field Crops Research 101 (2007) 160–171

www.elsevier.com/locate/fcr

Wheat grain yield, phosphorus uptake and soil phosphorus fraction

after 23 years of annual fertilizer application to an Andosol
Shigeru Takahashi *, Muhuddin R. Anwar
Integrated Soil Fertility Management Research Team, National Agricultural Research Center, 3-1-1 Kannondai, Tsukuba, Ibaraki 305-8666, Japan
Received 15 December 2005; received in revised form 2 November 2006; accepted 3 November 2006

Abstract
A field experiment was conducted on an Andosol to evaluate wheat (Triticum aestivum L.) yield, P and N uptake and soil P fraction after long-
term fertilization (no fertilizer, NPK, NP, NK and PK treatments). Application rates of N, P and K fertilizers were 100, 65 and 83 kg ha1 year1 by
ammonium sulfate, superphosphate, and potassium chloride, respectively. Phosphorus fertilization was critical for grain yield since the NK
treatment did not increase yield compared with no fertilizer treatment. Agronomic efficiency of P was greater than agronomic efficiency of N,
although apparent recovery of P and N were 17 and 53%, respectively. Combination application of fertilizer P and N resulted in the greatest grain
yield over 23-year cultivation. Interaction impact on grain yield between P and N ranged from 71 to 109%, and was greater than the values for
cereals in the earlier works. The N/P ratios of wheat decreased by P application and increased by N application. The N/P ratios in NPK and NP
treatments were higher than the values attaining maximum yield for cereal crops reported by other works.
Increase in soil available P in the treatments with P application was modest after 23-year fertilization. Total inorganic P (Pi), Ca-Pi + Al-Pi + Fe-
Pi, increased in the treatments with P application at 0–15 cm. Total Pi was greater at 0–15 cm depth than at 30–50 cm depth. Although apparent
recovery of fertilizer P (Ca-Pi as superphosphate) was less than 20%, soil Ca-Pi was very low even in the treatments with P application. This meant
that unutilized fertilizer P did not remain in the form of Ca-Pi. In contrast to inorganic P, there was no significant difference in total organic P (Po),
Ca-Po + Al-Po + Fe-Po, among the treatments and soil depths. Regardless of fertilizer treatments, Al-Pi was the predominant form at 0–15 cm depth
and Al-Pi concentrations were similar to Fe-Pi concentrations at 30–50 cm depth. On the other hand, Fe-Po was greater than Al-Po at 0–15 cm
depth. Difference in inorganic P at 0–15 cm depth demonstrated that unutilized fertilizer P was transformed mainly to Al-Pi followed by Fe-Pi.
However, wheat seemed to absorb P from Al-Pi and Fe-Pi modestly.
# 2006 Elsevier B.V. All rights reserved.

Keywords: Fractionation; Long-term experiment; Phosphorus; Wheat

1. Introduction conducted on Andosols on changes in major and trace elements

Andosols cover more than 50% of upland areas in Japan
(Hirata et al., 1999). Phosphorus is one of the limiting nutrients
in Andosols which have high P fixing capacity due to their high
Al and Fe oxide concentrations. Thus, large amounts of P
fertilizer are applied to Andosol fields continuously. This may
be the reason for the high P/N ratio of fertilizer consumption in
Japan (Aulakh and Malhi, 2005).
Long-term experiments provide one of the means to measure
sustainable management systems in agriculture (Rasmussen
et al., 1998). Several long-term fertilizer experiments have been
* Corresponding author. Tel.: +81 298 38 8829; fax: +81 298 38 8837.
E-mail address: shigeru@affrc.go.jp (S. Takahashi).
0378-4290/$ – see front matter # 2006 Elsevier B.V. All rights reserved.
doi:10.1016/j.fcr.2006.11.003
in soil (Hirata et al., 1999; Takeda et al., 2005), soil organic C
(Mathews et al., 2001; Gudmundsson et al., 2004) and N
leaching (Nakamura et al., 2004). Few long-term fertilizer
experiments on wheat in Andosols are available.
The low efficiency of plant P uptake is the main problem
associated with P application. Nitrogen is also the key element
in achieving consistently high yield in wheat. The interaction of
N with P can be termed the single most important nutrient
interaction of practical significance (Aulakh and Malhi, 2005).
Recently, Sadras (2006) investigated the N/P ratio of cereals
(n = 759) and indicated over 40% of crops attaining maximum
yield had N/P ratio in a relatively narrow range between 4 and 6.
When P input from fertilizer exceeds P output in crop, P
accumulation in the soil gradually increase over time (Kuo
et al., 2005). The information obtained from P fractionation
 S. Takahashi, M.R. Anwar / Field Crops Research 101 (2007) 160–171
schemes has been used to estimate the fate of applied P and the
relationship between forms of P and plant P nutrition (Ryan
et al., 1985; Furlani et al., 1987; Cox et al., 1997). Thus, for
developing long-term P management strategies, it is important
to ascertain the forms and characteristics of P remaining in the
soil after repeated fertilizer P additions in an agro-ecosystem.
The method developed by Chang and Jackson (1957) has
been widely used for investigation of the soil P. Another
sequential P fractionation method was developed by Sekiya
(1983) for Andosols. Otani and Ae (1997) reported that Sekiya
method (Sekiya, 1983) seemed to give better information than
Chang and Jackson method (1957) about P status in Andosols in
terms of plant P availability. The original or modified Hedley
methods are other widely used methods of P fractionation
(Hedley et al., 1982; Sui et al., 1999; Saleque et al., 2004; He
et al., 2004; Kuo et al., 2005; Verma et al., 2005). In the Hedley
methods, Al- and Fe-bound P (Al-P and Fe-P) are found in the
same fraction. Otani et al. (1996) indicated that organic acids in
root exudates have a significant role for soil P utilization and the
solubility of inorganic Al-P and Fe-P in some organic acids was
substantially different. Hirata et al. (1999) reported continuous
increase in inorganic Al-P along with a decease in inorganic Fe-
P by continuous application of farmyard manure and inorganic
P fertilizer for 9 years. Therefore, inorganic Al- and Fe-P
should be determined separately.
McKenzie et al. (1992) and Schmidt et al. (1996) reported
that fertilizer P and manure applied annually for many years
resulted in accumulation of inorganic P and organic P fraction
in soil. However, Campbell et al. (1986) did not observe a
significant change in the soil organic P fraction when manures
were applied once every 3 years, while labile inorganic P levels
were increased. Several studies demonstrated that organic P
plays an important role in P cycling and plant nutrition
(Sharpley and Smith, 1985; Beck and Sanchez, 1994; Selles
et al., 1999). In Andosols, organic P can account for 30–70% of
soil total P (Amano, 1981). However, few studies have been
made to investigate the individual inorganic P and organic P in
Andosols by the long-term fertilizer applications (Otani and
Ae, 1997).
The objectives of the present study were: (1) to investigate
the wheat yield response to long-term fertilization, (2) to
evaluate P and N uptake and their interaction, and (3) to
investigate soil P fractions after long-term fertilization.
2. Materials and methods
2.1. Experimental site, design and treatments
A long-term field experiment has been conducted since 1981
at NARC, Tsukuba, Japan (368010 N, 1408010 E) on a low-humic
Andosol (Typic Hapludands). Winter wheat (Triticum aestivum
L.) was grown except for 1991/1992, 1992/1993 and 1996/
1997. Oat (Avena sativa L.) was grown in 1991/1992 and 1992/
1993, and winter rye (Secale cereale L.) was grown in 1996/
1997.
The experiment had five fertilizer treatments including no
fertilizer (no F), NP, NK, PK and NPK. The N, P, and K mean
161
nitrogen, phosphorus, and potassium, respectively. Each
treatment had two replications. The plot area was 63 m2 and
arranged by a randomized block design. Application rates of N,
P and K fertilizers were 100, 65 and 83 kg ha1 year1 by
ammonium sulfate, superphosphate, and potassium chloride,
respectively. All P and K fertilizers were applied basal prior to
sowing. Seventy kg ha1 of the N fertilizer was basal applied
prior to sowing and 30 kg ha1 of N was top-dressed at stem
elongation stage. Between 1981 and 2000, soils were plowed
till 20–25 cm by disk plow and then basal fertilizers were
incorporated into 15 cm by rotary tiller. After 2001, basal
fertilizers were incorporated into 15 cm by rotary tiller without
plowing. Stubble was incorporated into the field, while straw
was removed from the field. In this study, we focused on recent
three seasons (2001/2002, 2002/2003 and 2003/2004). Wheat
(cv. Norin61) was sown at 60 kg seeds ha1 (18 cm row
spacing) on 30 October 2001, 7 November 2002 and 5
November 2003, and harvested (2 m by 4 rows) on 5 June 2002,
10 June 2003 and 3 June 2004, respectively. Historical grain
yield data were cited from the literatures by Plant Nutrition
Diagnosis Laboratory, NARC (PNDL, 1991, 1994, 1996,
1998).
2.2. Plant sampling and analysis
At physiological maturity, grain yields were reported at
125 g kg1 moisture content and the plant materials (grain and
straw) were subsampled for colorimetrical N and P analysis by
an Autoanalyzer (TRACCS 800, Bran + Luebbe) after the
digestion with H2O2–H2SO4.
Apparent recovery of P (ARP) and of N (ARN) in NPK
treatment were calculated as follows:
P uptake in NPK treatment
 P uptake in NK treatment
ARP ð%Þ ¼ 100  :
P fertilizer applied
N uptake in NPK treatment
 N uptake in PK treatment
ARN ð%Þ ¼ 100  :
N fertilizer applied
Agronomic efficiency of P (AEP) and of N (AEN) in NPK
treatment were calculated as follows:
Grain yield in NPK treatment
 Grain yield in NK treatment
AEP ¼
P fertilizer applied
Grain yield in NPK treatment
 Grain yield in PK treatment
AEN ¼
N fertilizer applied
Harvest index (HI), phosphorus harvest index (PHI), and
nitrogen harvest index (NHI) were calculated as follows:
Grain yield at dry matter basis
HI ¼ :
Aboveground dry matter production
P uptake in grain
PHI ¼ :
Aboveground P uptake
162 S. Takahashi, M.R. Anwar / Field Crops Research 101 (2007) 160–171

N uptake in grain
NHI ¼ :
Aboveground N uptake

The ratio of N to P (N/P) was expressed as a weight basis,

namely N (g kg1)/P (g kg1), not mol basis.

2.3. Soil sampling and analysis

Soil sampling was conducted in June 2002 from 0 to 15 cm

depth and in October 2004 from three depths (0–15, 15–30 and
30–50 cm). Six soil cores (ca. 3 cm diameter) were collected in
each plot and mixed to form a composite sample from that plot.
The samples were air dried and ground to pass a 2-mm sieve. The
soil samples collected in 2002 were used for determination of
organic C and total N. Other soil analyses were conducted using
the soil samples collected in 2004. Organic C and total N were
determined by dry combustion with a NC-Analyzer (model NC-
95A, Sumica Chemical Analysis Serves). Available N was
determined by hot KCl extraction method (Gianello and
Bremner, 1986). Ammonium N in the KCl solution was
determined by colorimetrically by an Autoanalyzer (TRACCS
800, Bran + Luebbe). Available soil P was determined using the
Truog P method (Truog, 1930) and Bray2 P method modified by
Shoji et al. (1964). Phosphorus in the solution was determined
colorimetrically by the Murphy and Riley method (1962).
Phosphorus fractionation was conducted as follows (Sekiya,
1983): a soil sample (400 mg) was extracted with 40 mL of Fig. 1. Organic C and N concentrations at 0–15 cm depth. NS, not significant at
0.44 M CH3COOH for 2 h. The soil suspension was p = 0.05.
centrifuged, and the supernatant was decanted. The soil in
the tube was washed twice with 20 mL of 1 M NH4Cl and both significant difference in organic C and total N concentrations
washing solutions were added to the extract of CH3COOH (Ca- among the treatments (Fig. 1).
P fraction). After the extraction of Ca-P, the soil was extracted Available P concentrations at 0–15 cm depth by Truog and
with 40 mL of 1 M NH4F (pH 7) for 1 h. The suspension was Bray2 methods were greater in the treatments with P
centrifuged, and the supernatant (Al-P fraction) was collected. application than the treatments without P application
Then, the soil was washed twice with 20 mL of saturated NaCl, (Table 1). At 30–50 cm depth, available P concentrations in
which was discarded. The soil was then extracted with 40 mL the NP and PK treatments were greater than the no F treatment,
of 0.1 M NaOH for 17 h, centrifuged, and the supernatant (Fe-P
fraction) was collected. Table 1
Total P in Ca-, Al-, and Fe-P fractions were determined after Soil available P and available N concentrations
the digestion with H2O2–H2SO4. Inorganic P in each fraction Treatment Depth (cm) Truog P Bray2 P Available N
was referred to Ca-Pi, Al-Pi and Fe-Pi and organic P in each
NPK 0–15 11.4 b 30.7 b 6.7 NS
fraction was referred to Ca-Po, Al-Po and Fe-Po. Organic P in NP 0–15 14.0 bc 30.4 b 6.9
these fractions was calculated by subtracting inorganic P from PK 0–15 15.1 c 33.3 b 7.5
total P. Phosphorus concentrations in the solutions were NK 0–15 5.2 a 11.7 a 6.2
determined by the Murphy and Riley procedure (1962). No F 0–15 5.6 a 11.4 a 7.6
NPK 15–30 6.8 a 24.1 b 6.4 ab
2.4. Data analyses NP 15–30 8.3 c 23.0 b 7.1 c
PK 15–30 8.1 c 27.3 b 7.1 c
NK 15–30 5.8 ab 13.4 a 6.1 a
Differences in each measurement were examined by ANOVA,
No F 15–30 5.4 a 11.5 a 6.7 bc
with means separation by least significant difference at p = 0.05.
NPK 30–50 4.9 a 12.5 a 5.9 NS
NP 30–50 6.8 b 19.9 b 5.9
3. Results and discussion PK 30–50 6.1 b 20.0 b 5.5
NK 30–50 4.9 a 12.0 a 4.9
3.1. Soil fertility No F 30–50 5.0 a 12.4 a 5.6
Truog P, Bray2 P and available N (mg kg1). Data with same letter in a column
Soil organic C and total N concentrations at 0–15 cm depth at the same depth are not significantly different at p = 0.05. NS, not significant at
tended to increase in the NPK treatment, but there was no p = 0.05.
 S. Takahashi, M.R. Anwar / Field Crops Research 101 (2007) 160–171
while there was no significant difference in available P
concentrations between the NPK and no F treatments. The
greatest values for Truog P and Bray2 P concentrations were 15
and 33 mg kg1 in the PK treatment at 0–15 cm depth,
respectively. Usually Truog P method has been used as the soil
P test for upland soils in Japan and recommended values are
more than 44 mg kg1. The proper range of the Bray2 P
concentration is considered to be between 80 and 260 mg kg1
for paddy soils (Kogano, 1984; Komoto, 1984). Thus, increase
in available P in the treatments with fertilizer P was modest
after 23 years and available P was severely deficient in all
treatments.
Available N concentrations at 0–15 cm depth were slightly
higher than at 30–50 cm depth. Difference in available N
concentrations among the treatments at the same soil depths
was only a few.
3.2. Grain yield and yield component
It seemed that there was no increasing or decreasing trend of
grain yield during the experiment in each treatment (Fig. 2).
Among recent three seasons, grain yield was the highest in
2001/2002 and the lowest in 2002/2003. The NK treatment had
no yield increasing effect, while the PK treatment had a
significant effect of yield increase. Thus, most important single
element should be P in this study. This was in line with the
observation by Sinha et al. (1973) in wheat. In severe P deficient
soils such as the present study, application of N alone will
produce only a small increase in yield, much below the
potential (Aulakh and Malhi, 2005). The AEP ranged from 53
to 60 and AEN from 15 to 19 (Fig. 3). The AEP in this study was
higher than the values reported by Holford and Doyle (1993)
and Singh et al. (2005), and similar to values in diammonium
Fig. 2. Changes in grain yield during the long-term experiment and comparison
of grain yield among the treatments (mean of 2001/2002, 2002/2003 and 2003/
2004) and year (mean of five treatments). Data with same letter are not
significantly different at p = 0.05. Oat was cropped in 1991/1992 and 1992/
1993, and winter rye was cropped in 1996/1997. Yield data in 2000/2001 are not
available.
163
phosphate treatment by Dwivedi et al. (2004). The AEN was
similar (Doyle and Holford, 1993; Singh et al., 2005) or higher
than earlier reports (Delogu et al., 1998; López-Bellido and
López-Bellido, 2001; Kutra and Aksomaitiene, 2003; Singh
and Agarwal, 2004).
Comparing grain yield between the NPK and NP treatments,
K application seemed to have little effect on grain yield in this
study. Assuming no difference in wheat growth between the no
F treatment and (virtual) K treatment, interaction impact on
grain yield defined by Aulakh and Malhi (2005) can be
calculated as follows:
Interaction impact ð%Þ on grain yield
100  ½Grain yield response in NPK treatment
 ðGrain yield response in PK treatment
þ Grain yield response in NK treatmentÞ
¼ :
Grain yield response in PK treatment
þ Grain yield response in NK treatment
where grain yield response in the fertilized treatment = grain
yield in the treatment  grain yield in the no F treatment.
Interaction impact on grain yield ranged from 71 to 109%.
These values were high compared with the interaction impact
on grain yield between P and N in cereals reported by Aulakh
and Malhi (2005).
In the treatments without P application, 1000 grain weight
decreased compared with the treatments with P application
(Fig. 4). The 1000 grain weight was higher in 2001/2002 than in
2002/2003. These results were in line with the results in grain
yields. The HI in the treatments with greater yield tended to be
high compared with HI in the treatments with lower yield
(Fig. 5). While the HI between the years was not in accordance
with the order of grain yield, namely HI in 2003/2004 was the
lowest. On the basis of a literature review, van Duivenbooden
et al. (1996) indicated the average wheat HI of 0.41 with a range
0.35–0.49 that was in the range of present study.
In the P-deficient soil, P fertilization was critical for wheat
grain yield and AEP was greater than AEN. Values of
interaction impact on grain yield between P and N were greater
than the reported values in cereals. Combination application of
fertilizer P and N was definitely important for higher grain yield
over 23-year cultivation.
3.3. Nutrient uptake
Phosphorus and N uptake were in the following order: NPK
and NP treatments > PK treatment > NK and no F treatments
(Fig. 6). These trends in P and N uptake among the treatments
were the same as grain yield. In the NPK treatment, ARP
ranged from 16 to 17% and ARN from 45 to 59% (Fig. 3). The
ARP is often low and rarely exceeds 20% (Damodar Reddy
et al., 1999). This was also the case in the present study.
Although ARP was less than ARN, absorbed P contributed to
the grain yield effectively judging from AEP and AEN.
Mean values of PHI in each treatment ranged from 0.87 to
0.91 (Fig. 5). Although there was significant difference in PHI
among the treatments and year, the difference was small and the
164 S. Takahashi, M.R. Anwar / Field Crops Research 101 (2007) 160–171
Fig. 3. Agronomic efficiency of P (AEP) and N (AEN), and apparent recovery of phosphorus (ARP) and nitrogen (ARN), and in NPK treatment. NS, not significant at
p = 0.05.
effect of fertilizer P was not definite. Bauer et al. (1987)
reported that 94% of plant P at harvest was in the spike and
Clarke et al. (1990) found that 95% of spike P was in the grain.
From these two findings, PHI was estimated to be 0.9 (Clarke
et al., 1990). The present results were in line with this
estimation. Mean values of NHI in each treatment ranged from
Fig. 4. 1000 grain weight. Data with same letter are not significantly different at
p = 0.05. No data are available in 2003/2004.
0.75 to 0.80. There was no significant difference in NHI among
the treatments, while there was significant difference between
years. López-Bellido and López-Bellido (2001) reported NHI
was significantly influenced by N fertilizer rates and year.
Variations in NHI in the present study were narrow compared
with a range 0.71–0.78 (López-Bellido and López-Bellido,
2001) and 0.71–0.85 (Clarke et al., 1990). Greater PHI than
NHI regardless of the treatment was probably due to the higher
translocation efficiency for P than N (Clarke et al., 1990).
As expected from the relationships between PHI and HI, P
concentrations in grain were greater than those in straw (Fig. 7).
For grain, P concentrations in the treatments with P application
were greater than the treatments without P application and there
was no significant difference in P concentrations among the
years (Fig. 7). For straw, on the other hand, fertilizer treatment
had no significant influence, while P concentrations were
influenced by year. The N concentrations in grain were also
greater than those in straw (Fig. 8). For grain and straw, N
concentrations in the treatments with N application were
greater than the treatments without N application.
The N/P ratios were greater in the treatments without P
application than in the treatments with P application (Fig. 9).
There was no significant difference in N/P ratio among the year.
 S. Takahashi, M.R. Anwar / Field Crops Research 101 (2007) 160–171
Fig. 5. Harvest index (HI), phosphorus harvest index (PHI), and nitrogen harvest index (NHI). Data with same letter are not significantly different at p = 0.05. NS, not
significant at p = 0.05.
In natural species, seedlings of many different plant species
have similar N/P ratio when supplied with nutrients at free
access (Knecht and Göransson, 2004) and critical N/P ratio has
been concerned for detecting nutrient limitation (Koerselman
and Meuleman, 1996; Güsewell, 2004; Güsewell et al., 2003).
Agronomic studies of crop N/P ratios are scarce. Sadras (2006)
demonstrated that over 40% of cereal crops attaining maximum
yield had N/P ratio in a relatively narrow range between 4 and 6.
For cereals, van Duivenbooden et al. (1996) calculated an
optimum N/P  7 (P/N = 0.14). The N/P ratios of aboveground
part were 8.4 and 7.9 in NPK and NP treatments, respectively.
The N/P ratio in PK treatment was the lowest (6.5). The N/P
ratio is a direct function of N uptake and an inverse function of
P uptake. In the present study, soil available P status was very
poor and thus N/P ratio that gave the highest yield among the
treatments shifted to higher value.
The N/P ratio ranged from 5.8 to 9.9 in grain and from 13 to
22 in straw (Fig. 9). Aulakh and Malhi (2005) summarized that
the main cereal crops, such as wheat, rice, and corn, typically
have N/P ratios in both grain and straw in the narrow range of
4.2–6.7. This was not in accordance with the present results,
165
especially in straw. This was because that seeds generally have
higher P and N concentrations and lower N/P ratios than
vegetative structure (Güsewell, 2004).
Application rate of fertilizer P exceeded P uptake regardless
of the treatment. While, application rate of fertilizer N was
nearly same to N uptake in the NPK and NP treatments. Thus, N
balance should be match in both treatments. On the other hand,
exceed fertilizer P must be accumulated in the soil.
3.4. Soil phosphorus fraction
Total Pi (=Ca-Pi + Al-Pi + Fe-Pi) at 30–50 cm depth was
lower than the upper soils (Fig. 10). At 15–30 cm depth, disk
plowing till 20–25 cm should have influenced the P concentra-
tions. Total Pi was greater in the treatments with P application
than the treatment without P application at 0–30 cm depth. This
was true at 30–50 cm depth in the NP and PK treatments. We
have no explanation for the result at 30–50 cm depth in the NPK
treatment, but this may be related to the results of available P
concentrations at this depth. In contrast to total Pi, there was no
significant difference in total Po (=Ca-Po + Al-Po + Fe-Po)
166 S. Takahashi, M.R. Anwar / Field Crops Research 101 (2007) 160–171

Fig. 6. Aboveground P and N uptake. Data with same letter are not significantly different at p = 0.05. NS, not significant at p = 0.05.

among the treatments (Fig. 10). Thus, unutilized fertilizer P was
not accumulated in organic forms. Otani and Ae (1999) also
reported that organic P did not appear to increase by
immobilization of inorganic P in Andosols. There was no
significant difference in total Po among soil depth. This
suggested that total Po concentrations are stable till 50 cm was
stable regardless of fertilizer treatment. Judging from total Pi
and total Po, it seemed that there was no considerable leaching
of unutilized fertilizer P to 30–50 cm soil depth. At 0–15 cm
depth, total Pi was greater than total Po in the treatments with P
application and lower in the treatments without P application.
At 30–50 cm depth, organic forms were predominant in all
treatments.
Although Ca-Pi concentrations in the treatments with P
application at 0–15 cm depth were high compared with the
treatments without P application, the difference was small
(Fig. 11). There was no significant difference in Ca-Pi
concentrations at 15–30 and 30–50 cm depths among the

Fig. 7. Phosphorus concentrations in (A) grain and (B) straw. Data with same letter are not significantly different at p = 0.05. NS, not significant at p = 0.05.
 S. Takahashi, M.R. Anwar / Field Crops Research 101 (2007) 160–171 167

Fig. 8. Nitrogen concentrations in (A) grain and (B) straw. Data with same letter are not significantly different at p = 0.05. NS, not significant at p = 0.05.

Fig. 9. Ratio of N to P in (A) aboveground, (B) grain and (C) straw. Data with same letter are not significantly different at p = 0.05. NS, not significant at p = 0.05.
168 S. Takahashi, M.R. Anwar / Field Crops Research 101 (2007) 160–171

Fig. 10. Total Pi and total Po in soil. Data with same letter are not significantly different at p = 0.05. NS, not significant at p = 0.05. Total Pi = (Ca + Al + Fe)–Pi, total
Po = (Ca + Al + Fe)–Po.

Fig. 11. Soil inorganic phosphorus (Ca–Pi, Al–Pi and Fe–Pi). Data with same letter are not significantly different at p = 0.05. NS, not significant at p = 0.05.
 S. Takahashi, M.R. Anwar / Field Crops Research 101 (2007) 160–171
treatments. The Ca-Pi at 30–50 cm depth tended to show higher
values than the upper soils, but that difference was not
significant. The Ca-Pi accounted for about 1% of total Pi at 0–
30 cm depth and 2.0–3.9% of total Pi at 30–50 cm depth.
Although applied fertilizer P was Ca-Pi (superphosphate) and
ARP was less than 20%, unutilized fertilizer P did not remain in
the form of Ca-Pi.
The Al-Pi concentrations in the treatment with P application
were high compared with the treatments without P application
at 0–30 cm depth (Fig. 11). At 30–50 cm depth, Al-Pi was lower
than upper soils and the influence of P application was not clear.
The Al-Pi accounted for 62–69 and 47–54% of total Pi at 0–30
and 30–50 cm depths, respectively.
The Fe-Pi concentrations in the treatment with P application
were greater than the treatments without P application at 0–
30 cm depth, while there was no significant difference in Fe-Pi
concentrations at 30–50 cm depth (Fig. 11). At 30–50 cm
depth, Fe-Pi was lower than upper soil. The Fe-Pi accounted for
31–37 and 43–52% of total Pi at 0–30 and 30–50 cm depths,
respectively. This means that Al-Pi was the predominant form at
0–30 cm depth, while Al-Pi concentrations were similar to Fe-
Pi concentrations at 30–50 cm depth.
Fig. 12. Soil organic phosphorus (Ca–Po, Al–Po and Fe–Po). Data with same letter are not significantly different at p = 0.05. NS, not significant at p = 0.05.
169
Comparing the treatments with and without P application at
0–15 cm depth, increase in Al-Pi concentrations was greater
than Fe-Pi concentrations. Beck and Sanchez (1994) concluded
the NaOH-extractable inorganic P by the Hedley methods
served as a primary sink for fertilizer P added to soil in a
tropical Ultisol. This was also the case for continuous
fertilizations of soils with fertilizer P for 9 or more years
under some cropping systems (Richards et al., 1995; Motavalli
and Miles, 2002). The present study demonstrated that
unutilized fertilizer P was transformed mainly to (NH4F-
extractable) Al-Pi followed by (NaOH-extractable) Fe-Pi in the
Andosol.
Fertilizer treatments had no significant effect on Ca-Po, Al-
Po and Fe-Po concentrations at each soil depth (Fig. 12). The
Ca-Po accounted for 1.1–2.7% of total Po at 0–50 cm depth and
was very low compared with other two forms. The Al-Po was
greater at 30–50 cm depth than at 0–30 cm depth and the
reverse was true for Fe-Po. The Al-Po accounted for 35–48 and
47–53% of total Po at 0–30 and 30–50 cm depths, respectively.
The Fe-Po accounted for 50–63 and 44–51% of total Po at 0–30
and 30–50 cm depths, respectively. In contrast to inorganic P,
Fe-Po was greater than Al-Po at 0–30 cm depth.
170 S. Takahashi, M.R. Anwar / Field Crops Research 101 (2007) 160–171
Since organic P was not considered to be the sink of
unutilized fertilizer P and no significant difference in the
composition of organic P forms, P availability may largely
depend on the status of soil inorganic P. Minami et al. (1969)
found that correlation coefficient between P uptake by oat and
Al-Pi was higher than Ca-Pi and Fe-Pi in Andosols. Yamamoto
and Miyasato (1971) reported that more than 300 mg kg1 of
Al-Pi was necessary for better crop growth in an Andosol.
Osborne and Rengel (2002) reported that significant
genotypes variation was identified in wheat in the ability to
utilize P from Fe-Pi. Although Al-Pi was about 300 mg kg1
at 0–15 cm depth in the NK treatment, soil P status must have
limited wheat growth. Soil bulk density at 0–15 cm depth was
0.8 Mg m3. Assuming that mean P uptake of three seasons
(2.4 kg ha1 year1) in the NK treatment did not change
during the long-term experiment and wheat taken up P from 0
to 15 cm depth, P concentration at 0–15 cm depth in the NK
treatment should decrease 44 mg kg1 in 23 years that
corresponds to only 9% of total Pi. Thus, Al-Pi and Fe-Pi in
the NK treatment contributed to P uptake by wheat modestly.
This led to the idea that low availability of Al-Pi and Fe-Pi that
accumulated in the treatments with P application in the
present study.
4. Conclusion
The present study demonstrated how P deficit and
inorganic fertilization affected nutrient uptake by wheat
and soil P fraction in a long-term field experiment.
Application of P was critical for wheat growth and application
of N alone had little positive effect. Combination application
of fertilizer P and N resulted in the greatest grain yield over
23-year cultivation. Greater interaction impact on grain yield
between P and N was observed compared with the earlier
reports. The N/P ratio of aboveground part that gave the
greatest yield among the treatments was about 8 and higher
than the values attaining maximum yield for cereal crops
reported by other works.
Increase in soil available P in the treatments with P
application was modest after 23-year fertilization and available
P levels were far below the recommended level. The long-term
application of fertilizer P resulted in an accumulation of large
amounts of total Pi at 0–15 cm depth. In contrast to total Pi,
there was no significant difference in total Po among the
treatments. At 0–15 cm depth, Al-Pi was the predominant
inorganic form, while Fe-Po was greater than Al-Po. Applied
Ca-Pi as superphosphate was transformed mainly to Al-Pi
followed by Fe-Pi. However, wheat seemed to absorb P from
Al-Pi and Fe-Pi modestly.
Acknowledgements
We are indebted to Plant Nutrition Diagnosis Laboratory,
NARC for managing the long-term experimental field
by 2000. The authors would like to thank Ms. Y. Nemoto
and T. Komatsu for their assistance in the field and
laboratory.
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