Soil Science and Plant Nutrition

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Fractionation of phosphorus in soils with different

geological and soil physicochemical properties in
southern Tanzania

Tomohiro Nishigaki, Soh Sugihara, Kazuki Kobayashi, Yohey Hashimoto,

Method Kilasara, Haruo Tanaka, Tetsuhiro Watanabe & Shinya Funakawa

To cite this article: Tomohiro Nishigaki, Soh Sugihara, Kazuki Kobayashi, Yohey Hashimoto,
Method Kilasara, Haruo Tanaka, Tetsuhiro Watanabe & Shinya Funakawa (2018) Fractionation
of phosphorus in soils with different geological and soil physicochemical properties in southern
Tanzania, Soil Science and Plant Nutrition, 64:3, 291-299, DOI: 10.1080/00380768.2018.1436406

To link to this article: https://doi.org/10.1080/00380768.2018.1436406

Published online: 07 Feb 2018.

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SOIL SCIENCE AND PLANT NUTRITION
2018, VOL. 64, NO. 3, 291–299
https://doi.org/10.1080/00380768.2018.1436406

ORIGINAL ARTICLE

Fractionation of phosphorus in soils with different geological and soil

physicochemical properties in southern Tanzania
Tomohiro Nishigakia,b, Soh Sugihara c, Kazuki Kobayashid, Yohey Hashimotoe, Method Kilasaraf, Haruo Tanakac,
Tetsuhiro Watanabea and Shinya Funakawaa
a
Graduate School of Agriculture, Kyoto University, Kyoto, Japan; bGraduate School of Agriculture, Tokyo University of Agriculture and Technology,
Fuchu-shi, Tokyo, Japan; cInstitute of Agriculture, Tokyo University of Agriculture and Technology, Fuchu-shi, Tokyo, Japan; dGraduate School of
Bio-Applications and Systems Engineering (BASE), Tokyo University of Agriculture and Technology, Koganei-shi, Tokyo, Japan; eDepartment of Bio-
applications and Systems Engineering (BASE), Tokyo University of Agriculture and Technology, Koganei-shi, Tokyo, Japan; fCollege of Agriculture,
Sokoine University of Agriculture, Morogoro, Tanzania

ABSTRACT ARTICLE HISTORY

Soil phosphorus (P) forms have been practically defined as chemically fractionated pools. A knowledge of Received 31 August 2017
the abundance and diversity of P forms in soil, and the factors affecting them, will lead to better soil Accepted 31 January 2018
management. However, little is known about the differences in P forms among soils with different KEY WORDS
geological properties in tropical Africa. The aim of this study was to investigate the P forms in soils with Extractable Al; extractable
different physicochemical properties formed under different geological conditions in southern Tanzania Fe; sequential fractionation;
and to identify the factors affecting the P forms in these soils. In total, 37 surface soil samples were collected soil fertility; Sub-Saharan
from three geological groups; the plutonic (mainly granite) rock (PL) group, the sedimentary and meta- Africa
morphic rock (SM) group, and the volcanic ash (V) group. Soil P was sequentially extracted by NH4Cl, NH4F,
NaHCO3, NaOH + NaCl, and HCl, and inorganic (Pi) and organic P (Po) in each fraction were determined. The
lowest total P was in the PL group (average, 360 mg P kg-1) because of the high sand content. Iron (Fe)-P
(NaOH-Pi) was the major form in this group, accounting for 8.4% of total P. In the SM group (average total P,
860 mg P kg-1), Fe-P was the major form in most, accounting for 7.8% of total P. Soils in the SM group
occasionally had high calcium (Ca)-P due to application of chemical fertilizer at the collection site. The V
group had the highest total P (average, 1600 mg P kg-1) and its major P form was Ca-P, which was possibly
derived from primary minerals (i.e., apatite), accounting for 14% of total P. In addition, the high oxalate-
extractable Al possibly caused the accumulation of Al-P in the V group. Oxalate-extractable Fe generally
increased with increasing Fe-Pi, while oxalate-extractable Al increased with increasing organic P and Al-Pi in
soils in all three geological groups. These results demonstrate that the soil P forms differ greatly among sites
in southern Tanzania with different geological conditions and associated soil properties.

1. Introduction P forms in highly weathered soils common in tropical areas, while

Ca-P would be the main form in younger soils which include
Soil phosphorus (P) is one of the most important nutrients
apatite derived from parent materials (Walker and Syers 1976). In
controlling plant biomass production in tropical agroecosys-
addition, the application of chemical fertilizers such as superpho-
tems where highly weathered soils are common. These soils
sphate could increase the amount of Ca-P in surface soils in
effectively adsorb and occlude P, in many cases leading to P
arable lands (Wright 2009). The accumulation of soil organic
limitation (Tiessen et al. 1992). The soil characteristics signifi-
matter in surface layers could enhance organic P accumulation
cantly affect the type of P forms, which is chemically defined,
in forest soils, while the lack of soil organic matter in sandy or
and P bioavailability and stability (Wright 2009).
cropland soils may lead to less organic P (Tiessen et al. 1992).
The dominant form of P in soils differs depending on the
Several methods have been developed and used to study the
parent materials and the soil physicochemical properties. Many
forms, amounts, and dynamics of soil P (Chang and Jackson
conditions, including land-use conditions, affect soil formation.
1957; Hedley et al. 1982; Tiessen and Moir 1993; Kuo 1996). The
Many studies have reported on the transformation of P during
sequential fractionation method of soil P is based on the differ-
pedogenesis (Walker and Syers 1976; Tiessen et al. 1984;
ent solubilities of P forms in various extracts and is widely used to
Schlesinger et al. 1998); Calcium- (Ca-) P as primary minerals
determine P availability in soils. The distribution of P within
was the most dominant form at the beginning stage of pedo-
chemically defined pools, such as labile, inorganic, and organic
genesis, and the P released during weathering transformed into
fractions, provides an indication of the potential stability of P in
labile pools including soil organic matter and then into less-labile
soil, which differs depending on geological conditions and the
pools associated with the aluminum- (Al-) and iron- (Fe-) (hydr)
associated soil properties. Hedley et al. (1982) proposed a
oxides. Therefore, it is likely that Al-P and Fe-P would be the main
method to differentiate labile to less-labile forms using resin,

CONTACT Soh Sugihara sohs@cc.tuat.ac.jp Graduate School of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu-shi,
Tokyo 183-8509, Japan
The supplemental data for this article can be accessed here
© 2018 Japanese Society of Soil Science and Plant Nutrition
292 T. NISHIGAKI ET AL.
NaHCO3, NaOH, and HCl, but this method does not separate Al-P
and Fe-P. It has been applied in many studies to estimate P
availability to plants. Chang and Jackson (1957) and Kuo (1996)
devised a soil P fractionation method using NH4F as the extrac-
tant to separate Al-P from Fe-P, which led to a better under-
standing of the P transformation process during pedogenesis.
Abdala et al. (2015) simplified the method and used it to analyze
the effect of long-term manure application on P accumulation in
highly weathered agricultural soils in southern Brazil. These frac-
tionation methods provide the information about the availability
and stability of P that have led to the current soil P conditions
and allow the appropriate assessment of available P based on
the P forms in soils.
A few studies have assessed fractionated P and available P in
soils on a regional or country scale in tropical Africa. Azeez et al.
(2013) and Auxtero et al. (2013) reported on available P in a
range of Nigerian and Angolan soils, and evaluated different
soil P extraction tests. Nwoke et al. (2003) evaluated P fractions
and adsorption characteristics in relation to soil properties of
West African savanna soils. Funakawa et al. (2012) surveyed 95
soil samples collected throughout Tanzania to assess soil ferti-
lity and factors affecting it and found high levels of total P and
available P (measured by the Bray method) in the southern
volcanic mountain range. In Tanzania, there are a range of soil
types that have developed from many types of geological
parent materials on different time scales and under various
environmental conditions as affected by the Great Rift Valley.
In southern Tanzania, ejecta from volcanoes was carried mainly
southeastward by winds and deposited as tephra layers over
the geomorphologically stable plateau of the Southern
Highlands (Msanya et al. 2007; Fontijn et al. 2010). This volcanic
Figure 1. Map of site locations in southern Tanzania.
ejecta resulted in potentially high soil fertility, which is related
to high soil organic matter in this area (Funakawa and Kilasara
2017). The dominant P form is Ca-P as primary minerals, or Al-P
derived from the highly active Al in volcanic soils around the
volcanoes. On the other hand, the P form fixed by Al- and Fe-
(hydr)oxides in soils is dominant in the highly weathered soils
distributed around the volcanic area in southern Tanzania. To
date, however, there have been no studies on the differences in
P forms among soils formed under various geological condi-
tions and associated soil properties in tropical Africa.
The overall aim of this study, therefore, was to analyze the
abundance and diversity of soil P forms, including Al-P, Fe-P, Ca-
P, and organic P, in relation to the geological conditions and the
soil physicochemical properties on a regional scale. This informa-
tion can be used to improve management to benefit agricultural
production, soil quality, and water quality. The specific aims of
this study were as follows: (1) to extract the P forms from soil
samples using the sequential extraction method, and (2) to
identify the factors contributing to the types of P in soils formed
under different geological conditions and associated soil physi-
cochemical properties in southern Tanzania.
2. Material and methods
2.1. Study area and soil sampling
The soil samples used in this study were mainly collected from
arable lands and woodlands in Mbeya and Iringa regions,
southern Tanzania, between 32 39′ 18.1′′ E and 36 3′ 35.2′′ E
and between 7 40′ 14.9′′ S and 9 36′ 28.6′′ S (Fig. 1). The soil
samples were collected from the most intensive agricultural

production area in Tanzania, as well as the northern part
around Mt. Kilimanjaro, with cool temperatures and relatively
high precipitation. Soil samples were collected from the sur-
face layer at 37 sites.
The elevation of the sites ranged from 480 to 2730 m above
sea level. The mean annual precipitation ranges from 500 to
>2000 mm and is highest around Lake Nyasa. There are a
variety of geological parent materials in southern Tanzania
as affected by geological activities in the Great Rift Valley.
The studied soil samples were classified into three major
geological groups based on their parent materials according
to the geological map of Tanzania and field observations; the
plutonic rock (mainly granite) group (PL; No. 1–9), the sedi-
mentary and metamorphic rock group (SM; No. 10–12, 22,
24–25, 30–31, 34–37), and the volcanic ash group (V; No.
13–21, 23, 26–29, 32–33). Although No. 29 and No. 33 were
located in the PL group according to the geological map, they
were classified into the V group based on their soil physico-
chemical properties and field observations.
At each site, soil samples from surface layers were collected
from a pedon on a flat or gently sloping surface without signs
of water erosion. Only soil sample No. 34 contained gravel.
The collected soil samples were air-dried, sieved to pass
through a 2-mm mesh screen, and stored at ambient tempera-
ture until analysis.
2.2. Basic soil physicochemical properties
Soil pH was determined in both deionized water (H2O) and
1 M KCl at a soil-to-solution ratio of 1:5, and electric conduc-
tivity (EC) was determined in deionized water at a soil-to-
solution ratio of 1:5. Soil particle size distribution was deter-
mined by the wet-sieving and pipet method (Gee and Or
2002). Extractable Al and Fe contents were determined by
the dithionite-citrate method (Ald and Fed), the acid ammo-
nium oxalate method (Alo and Feo), and the sodium pyropho-
sphate method (Alp and Fep), as described by Courchesne and
Turmel (2008). Total carbon (C) and nitrogen (N) were quanti-
fied by the dry combustion method with an NC analyzer (Vario
Max CHN, Elementar, Hanau, Germany).
2.3. Total phosphorus and sequential chemical
fractionation of phosphorus
Soils were digested with 60% HClO4 following Kuo (1996), and
total P (TP) was determined by the molybdovanadate method
(Kitson and Mellon 1944). Soil P was sequentially fractionated
using a chemical fractionation method as described by Kuo
(1996) and modified by Abdala et al. (2015). Sieved soil (2 g)
was added to a 50-ml centrifuge tube and sequentially
extracted by adding 40 ml of each extractant solution. The
extractant solutions were added in the following order: 1 M
NH4Cl, 0.5 M NH4F (pH 8.2), 0.5 M NaHCO3 (pH 8.5), 0.1 M
NaOH + 1 M NaCl, and 1 M HCl. In the two first extractions, the
centrifuge tube was shaken for 1 h at 100 rpm, and in the
subsequent three extractions the tube was shaken for 18 h at
100 rpm. After each extraction, the tubes were centrifuged at
2500 g for 20 min, and the supernatant was filtered (5C,
ADVANTEC) and then collected to measure the concentration
SOIL SCIENCE AND PLANT NUTRITION 293
of inorganic P (Pi) using the ascorbic acid method (Murphy
and Riley 1962) after adjusting the pH of the solution. Soluble
and loosely bound P was extracted with NH4Cl; Al-P was
extracted by NH4F; easily mineralizable organic P was
extracted by NaHCO3; Fe-P was extracted by NaOH; and Ca-P
was extracted by HCl (Abdala et al. 2015). The TP in each
fraction, except the HCl fraction, was digested with ammo-
nium persulfate in an autoclave at 103.5 kPa and 120°C for
60 min, and the concentration was determined by the method
of Murphy and Riley (1962). The organic P (Po) was calculated
as the difference between total P and Pi. The difference
between TP and the sum of inorganic and organic P extracted
by the sequential fractionation was defined as residual P.
2.4. Available phosphorus
To evaluate the relationship between available P and the soil P
forms determined by the above sequential method, available
P was evaluated by two different methods; the alkaline bicar-
bonate method of Olsen et al. (1954) and the acid ammonium
fluoride extraction of Bray and Kurtz (1945), with
modifications.
Olsen P was determined using an extraction method fol-
lowing Kuo (1996). Sieved soil (2 g), half a teaspoon of black
carbon, and 40 mL 0.5 M NaHCO3 (pH 8.5) solution were
added into a 125-ml Erlenmeyer flask and shaken for 30 min.
The mixture was filtered and the extract was used to deter-
mine the P concentration using the ascorbic acid method
(Murphy and Riley 1962) after adjusting the pH of the solution
with 2.5 M H2SO4.
Bray P was extracted from the soil using the modified Bray
No. 2 method following Nanzyo (1997). Soil (1 g) and 20 ml
extractant (0.03 M NH4F and 0.1 M HCl) were added to a 50-ml
centrifuge tube and shaken for 1 min. The mixture was fil-
tered, and the extract was used to determine the P concentra-
tion by the method of Murphy and Riley (1962).
2.5. Data analysis
All statistical analyses were performed with SYSTAT 11
(SYSTAT Inc., Richmond, CA, USA). All data are expressed on
a dry-weight basis. The Shapiro–Wilk test was used to test
whether data were normally distributed. Multiple comparisons
were conducted using Tukey–Kramer test. Pearson’s correla-
tion analysis was used to test the relationships between soil
physicochemical properties and total P, available P, P contents
in the sequential fractionation method, and total C. In all
analyses, p < 0.05 indicated statistical significance.
To evaluate the contribution of P forms to available P,
forward-stepwise regression analysis was applied with P frac-
tions extracted by the sequential fractionation method as the
explanatory variable, and available P (Olsen P and Bray P) as
the objective variable. Prior to the regression analysis, the
inorganic P fraction extracted by NaHCO3 was excluded from
the explanatory variables to avoid multicollinearity that had a
strong positive correlation with NH4F extraction (r = 0.98,
P < 0.001) in all soils and in each geological group. On the
other hand, we did not exclude either NH4F-Pi or HCl-Pi
despite of their significant positive correlation with each
294 T. NISHIGAKI ET AL.

other because they were extracted by the different solutions
with contrasting pH and they had a different slope of the
regression line in each geological group.
3. Results
3.1. Soil characteristics
The soil physicochemical properties differed among the three
geological groups (Table 1). The soil texture was generally
coarse in the PL group, which had a high sand content. The
soil texture was generally fine in the SM group, which had a
high clay content. The highest contents of total C, total N, total
P, Alo, Alp, and Feo were in the V group followed by the SM
group. The highest Fed content was in the SM group. Total C
content was clearly and strongly correlated with the Alp con-
tent (r = 0.94, p < 0.001) and Alo content (r = 0.79, p < 0.001) in
all soils, but weakly correlated with clay + silt (r = 0.34,
p = 0.04) and Feo contents (r = 0.48, p = 0.003).
The Feo/Fed indicates the degree of crystallinity of Fe oxi-
des (Lair et al. 2009). The Feo/Fed values were high in the V
group (range, 0.3–0.8) indicating that this group has the
stronger effects of volcanic materials than the other geological
groups (Table 1). A low Feo/Fed ratio generally indicates the
dominance of crystalline Fe forms, mainly goethite and hema-
tite in the tropical savanna (Agbenin 2003).
3.2. Total phosphorus and fractionated phosphorus
The amount of TP ranged from 53 to 4900 mg P kg-1 among
the soils (mean, 1100 mg P kg-1). As shown in Table 1, the V
group had the highest TP among the geological groups, fol-
lowed by the SM group.
The P content in each fraction and its ratio to TP are
summarized in Table 2. The sum of Pi and Po in the NH4Cl
fraction accounted for only 0.4% of TP for all soils. The sum of
Pi and Po in the NH4F, NaHCO3, and NaOH + NaCl fractions
accounted for 13%, 5.5%, and 12% of the TP, respectively, for
all soils. The HCl-Pi content ranged from 0.8 to 1700 mg P kg-1
among the soils, corresponding to 0.3% to 44% of TP. Residual
P accounted for 60% of TP averaged over all soil samples,
ranging from 21% to 87% of TP in individual samples. The
lowest residual P content was in the PL group, and it was
strongly correlated with Fed (r = 0.51, p = 0.001) in all soils. The
total inorganic P (TPi) content in all the fractions ranged from
10 to 3500 mg P kg-1, accounting for 6.3–70% of TP, while the
total organic P (TPo) content ranged from 16 to 360 mg P kg-1,
accounting for 3.1–31% of TP. The highest ratio of TPo to TP
was in the PL group, followed by the V group.
In every fraction, the lowest Pi and Po contents were in the
PL group (Table 2). The NH4Cl-Pi and NaOH + NaCl-Pi in the SM
group were comparable to those in the V group. The contents
of other fractions (NH4F-Pi, NaHCO3-Pi, and HCl-Pi) were much
higher in the V group than in the PL and SM groups. There
was a significant positive correlation between NH4F-Pi and
HCl-Pi (r = 0.92, p < 0.001) in all soils. There was a significant
positive correlation between NH4F-Pi and Alo in the soils with
Alo + 0.5 Feo of less than 20 g kg-1, which is used as criteria for
andic soil properties (Soil Survey Staff 2014), in all the groups
(r = 0.67, p < 0.001), but lower correlation in all soils (r = 0.41,
p = 0.01). In addition, Feo but not Alo was significantly corre-
lated with NaOH + NaCl-Pi in every geological group. The HCl-
Pi and its ratio to TP increased with Feo/Fed only in the V
group (Fig. 2). The NaOH + NaCl-Pi increased with Feo/Fed in
each subgroup in the SM group (Fig. 3): The samples in the SM
group were divided in those from the northern part (Nos.
10–12, 22, 24, and 30) and those from the southernmost part
(Nos. 31, and 34–37) of the study area.
3.3. Available phosphorus
The Bray P content ranged from 3.8 to 450 mg P kg-1 in all the
soils, accounting for 0.5–33% of TP, whereas the Olsen P
content ranged from 2.8 to 43 mg P kg-1, accounting for
0.2–9.6% of TP. The P contents extracted by these two meth-
ods were significantly correlated with each other (r = 0.87,

Table 1. Physicochemical properties of soil samples collected in southern Tanzania.

All soils (N = 37) PL (N = 9) SM (N = 12) V (N = 16)
Property Mean (range) Mean (range) Mean (range) Mean (range)
pH (H2O) 5.9 (4.8–7.1) 5.8 a (4.9–6.6) 5.9 a (4.8–7.1) 6.0 a (4.9–7.0)
pH (KCl) 4.8 (4.1–6.4) 4.8 a (4.2–5.7) 4.8 a (4.1–6.4) 4.8 a (4.3–5.7)
EC, mS m−1 2.0 (0.2–9.6) 0.9 a (0.4–2.0) 1.9 a (0.2–9.3) 2.7 a (0.7–9.6)
Sand, % 51 (17–84) 67 a (44–84) 47 b (18–72) 44 b (17–78)
Silt, % 20 (1.7–39) 8 c (1.7–16) 17 b (4.3–36) 30 a (15–39)
Clay, % 29 (6.3–70) 25 a (8.3–47) 36 a (14–70) 26 a (6.3–44)
TC, g C kg−1 33 (3.2–140) 9.8 b (3.2–27) 26 ab (5.0–90) 51 a (12–140)
TN, g N kg−1 2.5 (0.3–9.3) 0.8 b (0.3–1.9) 1.9 b (0.5–6.5) 3.9 a (0.8–9.3)
TP, mg P kg−1 1100 (53–4900) 360 b (53–860) 1100 ab (350–3500) 1600 a (390–4900)
Ald, g Al kg−1 7.6 (0.0–45) 2.5 a (0.3–7.0) 6.5 a (0.0–18) 11 a (0.1–45)
Alo, g Al kg−1 11 (0.2–57) 1.2 b (0.2–2.6) 5.3 b (0.7–14) 21 a (2.7–57)
Alp, g Al kg−1 2.9 (0.6–11) 1.0 b (0.6–1.8) 2.3 ab (0.6–8.6) 4.4 a (0.6–11)
Fed, g Fe kg−1 22 (0.8–85) 11 b (0.8–25) 36 a (9.6–85) 17 b (4.3–36)
Feo, g Fe kg−1 5.3 (0.1–22) 0.5 c (0.1–1.5) 4.4 b (0.3–7.9) 8.7 a (3.2–22)
Fep, g Fe kg−1 0.8 (0.1–2.1) 0.4 b (0.1–1.0) 0.9 a (0.1–2.1) 1.0 a (0.2–2.0)
Alo + 0.5 Feo, g kg−1 14 (0.2–63) 1.4 b (0.2–3.4) 7.5 b (1.4–17) 25 a (6.2–63)
Feo/Fed 0.3 (0.0–0.8) 0.1 b (0.0–0.3) 0.1 b (0.1–0.3) 0.6 a (0.3–0.8)
PL: plutonic rock group; SM: sedimentary and metamorphic rock group; V: volcanic ash group; EC: electronic conductivity; TC: total carbon; TN: total nitrogen; TP:
total phosphorus; Ald: Al extracted by dithionite-citrate; Alo: Al extracted by acid ammonium oxalate; Alp: Al extracted by sodium pyrophosphate; Fed: Fe extracted
by dithionite-citrate; Feo: Fe extracted by acid ammonium oxalate; Fep: Fe extracted by sodium pyrophosphate.
Different letters indicate significant differences (p < 0.05) among geological groups.
 SOIL SCIENCE AND PLANT NUTRITION 295

0.9 a (0.1) 2.1 a (0.2) 3.0 a (0.2) 190 a (11) 100 a (6.3) 300 a (18) 96 a (5.5) 25 a (2.0) 120 a (7.5) 83 a (6.6) 47 a (4.2) 130 a (11) 270 a (14) 790 a (49) 640 a (38) 180 a (13)
1.8 a (0.2) 1.9 a (0.2) 3.7 a (0.4) 50 b (3.7) 34 b (3.3) 84 b (7.0) 32 a (2.6) 13 b (1.3) 45 ab (3.8) 86 a (7.8) 36 ab (4.1) 120 a (12) 80 a (7.0) 730 a (70) 250 ab (21) 84 b (8.9)
39 b (17)
TPo

0.4 a (0.2) 0.9 a (0.4) 1.3 a (0.6) 11 b (4.3) 16 b (6.9) 26 b (11) 6.4 a (2.4) 4.3 b (1.9) 11 b (4.3) 24 a (8.4) 18 b (7.6) 41 a (16) 3.0 a (1.5) 280 a (66) 44 b (17)
TPi
Residual
HCl
Pi

Figure 2. Relationship between inorganic phosphorus extracted by HCl in

Pi + Po

sequential extraction method (HCl-Pi) and iron crystallinity (Feo/Fed).

NaOH + NaCl

p < 0.001). The content of Bray P in the V group was 8.7 and
1.8 times as high as that in the PL and SM groups, respectively,
Po

while the content of Olsen P in the V group was 2.3 and 1.5
PL: plutonic rock group; SM: sedimentary and metamorphic rock group; V: volcanic ash group; Pi: inorganic phosphorus; Po: organic phosphorus.

times as high as in the respective groups, although there was

no statistical difference among all the geological groups. The
Pi

ratio of P content extracted by the Bray method to TPi was the

highest in the PL group followed by the SM group, while the
mg P kg−1 (% of total P)

ratio to TP was not significantly different among all the geo-

Pi + Po

logical groups (Table 3). The ratio of P content extracted by

the Olsen method to TP, as well as that to TPi, were highest in
the PL group (Table 3). A summary of forward stepwise ana-
NaHCO3

lysis of available P and fractionated P data is shown in Table 4.

Po

The NH4F, NH4Cl, and HCl fractions were selected as the

independent variables for Bray P, while the NH4F, NaOH,
NH4Cl, and HCl fractions were selected as the independent
Pi

variables for Olsen P.

Different letters correspond to significant differences (p < 0.05) among the geological groups.
Pi + Po

4. Discussion
4.1. Effect of geological conditions and associated soil
physicochemical properties on Ca-P and Fe-P in soils
NH4F
Po

Iron plays a substantial role in recalcitrant P accumulation, as

Table 2. Sequentially fractionated phosphorus in three geological groups.

we found a strong positive correlation between residual P and

Fed (r = 0.51, p = 0.001) across all soils. However, for some soils
Pi

in the V group, there was a positive correlation between

residual P and extractable Al (Ald, Alo and Alp) rather than
Fed. Chang and Jackson (1957) showed the predominance of
Pi + Po

Fe-P in the residual fraction, which is first extractable by

reductive extraction after the NH4F, NaOH, and H2SO4 treat-
ments. Redel et al. (2016) reported a high positive correlation
NH4Cl

between residual P and Feo, although they did not measure

Po

Fed in their study.

The high TP content and the high ratio of HCl-Pi (Ca-P) to
TP in the V group suggest that the soils had been weakly
Pi

weathered, supported by their high Feo/Fed. Primary apatite is

possibly predominant in the V group (Walker and Syers 1976).
Geological group

The amount of HCl-Pi and its ratio to TP increased with Feo/Fed

only in the V group (Fig. 2). The high Feo/Fed and Alo + 0.5 Feo
resulted from the strong influence of volcanic materials, lead-
ing to high Ca-P in the V group. On the other hand, the high
SM
PL

Ca-P in the PL and SM groups is likely due to external factors,

296 T. NISHIGAKI ET AL.

Figure 3. Relationship between inorganic phosphorus extracted by NaOH and NaCl in sequential extraction method (NaOH + NaCl-Pi) and iron crystallinity (Feo/Fed).
Samples in volcanic ash group were divided into four groups based on distance from caldera and content of Al and Fe extracted by acid ammonium oxalate (Alo + 0.5 Feo): samples near
Rungwe caldera were within 15 km in north-west direction or 30 km in south-east direction (because volcanic material was distributed southeastward by wind), and high Alo + 0.5 Feo is
above 20 g kg-1. Samples in sedimentary and metamorphic rock group were divided into two groups: northern part (<9 17′ 00′′S) and southern part of study area.

Table 3. Available phosphorus in three geological groups. In the SM group, a few samples (Nos. 10, 34, 35, and 37)
Bray P Olsen P had high HCl-Pi. The high HCl-Pi values in Nos. 10 and 37 could
P content / TP / TPi P content / TP / TPi be attributed to chemical fertilizer, as both sites were used as
Geological group mg P kg−1 % mg P kg−1 %
arable land, while the high value in Nos. 34 and 35 may have
PL 13 a 5.7 a 32 a 5.9 a 3.5 a 19 a
resulted from primary Ca-P in parent materials containing
SM 62 a 5.4 a 22 ab 9.1 a 0.9 b 6.3 b apatite in the surface layer. In the PL group, the ratio of HCl-
V 110 a 8.0 a 18 b 14 a 1.1 b 3.2 b Pi to TP was low, indicating that there was little effect of
PL: plutonic rock group; SM: sedimentary and metamorphic rock group; V: chemical fertilizer, probably because fertilizer was readily lost
volcanic ash group; TP: total phosphorus; TPi: total inorganic phosphorus.
by leaching from the sandy soils (Sugihara et al. 2012).
Different letters indicate significant differences (p < 0.05) among the geological
groups. The Feo content was significantly correlated with
NaOH + NaCl-Pi in every geological group while the Alo con-
tent was not. Thus, the NaOH + NaCl-Pi in the soil samples in
such as the application of chemical fertilizer and the specific this study was primarily associated with Feo, namely Fe-P,
arid environment associated with the geomorphological con- consistent with the results of previous studies (e.g., Kuo
ditions. The samples in the V group were clearly divided into 1996). The decrease in NaOH + NaCl-Pi with decreasing Feo/
those with low Feo/Fed (< 0.6), which were collected from sites Fed in each geological subgroup in the SM group (Fig. 3)
relatively far from the center of the recently erupted Rungwe suggested that the Fe-P content decreased during the weath-
caldera (Nos. 13–18, 21, 29, and 33), and those with high Feo/ ering process, as indicated by the crystallization of Fe in soils.
Fed that were collected from sites near the caldera (Nos. 19, However, there was no correlation between Fe crystallinity
20, 23, 26–28, and 32). This indicated that the soils in the outer (Feo/Fed) and NaOH + NaCl-Pi in the PL and V groups. This
areas had developed from relatively old parent materials. The could be attributed to the low Feo content in the PL group. In
samples from the inner area were distributed from the caldera addition, the re-adsorption of the extracted P to active Al and
southeastward, consistent with the distribution of volcanic Fe in soils probably occurred during the extraction in the V
materials as reported elsewhere (Msanya et al. 2007; Fontijn group. The correlation coefficient between Feo and
et al. 2010). NaOH + NaCl-Pi for the soils except those with high Alo + 0.5

Table 4. Forward stepwise analysis of available phosphorus and fractionated inorganic phosphorus.
Regression
Adjusted R square coefficient Standard error Standard regression coefficient Partial correlation coefficient F value
Bray P 0.89
Constant term −6.7 7.7 −6.7 0.76
NH4F-Pi 0.86 0.11 1.4 0.81 65
NH4Cl-Pi 23 2.5 0.64 0.85 87
HCl-Pi −0.24 0.06 −0.66 −0.55 14
Olsen P 0.80
Constant term 3.2 0.88 3.2 13
NH4F-Pi 0.05 0.01 1.0 0.56 15
NaOH-Pi 0.04 0.01 0.39 0.58 16
NH4Cl-Pi 0.98 0.27 0.34 0.54 13
HCl-Pi −0.01 0.01 −0.49 −0.34 4.1

Feo (≥20 g kg-1) (r = 0.69, p < 0.001) increased from that for all
the soils (r = 0.53, p = 0.001). Thus, the high Alo + 0.5 Feo
(≥20 g kg-1) likely caused the underestimation of
NaOH + NaCl-Pi, which was also observed for NH4F-Pi as dis-
cussed later. The re-adsorbed P was probably accounted to be
in the following fractions, HCl-Pi and residual P.
4.2. Effects of active Al on accumulation of Al-P and
organic P in soils
The amount of active Al (Alo) is an important factor in P
accumulation (Satti et al. 2007). In the soils with low Alo +
0.5 Feo (< 20 g kg-1) in all the geological groups, NH4F-Pi
increased with increasing Alo (r = 0.67, p < 0.001), suggesting
the contribution of Alo to the accumulation of possibly Al-P.
However, there was no correlation between NH4F-Pi and Alo in
all soils including those with high Alo + 0.5 Feo (≥20 g kg-1).
This was probably because of the re-adsorption of the
extracted P to a lot of active Al and Fe in soils during NH4F
extraction, as well as NaOH + NaCl extraction already dis-
cussed above, leading to an underestimation of NH4F-Pi.
On the other hand, NH4F-Pi should consist of not only Al-P
but also Ca-P since the fluoride ion reacts with both Al and Ca,
especially in the soils rich in Ca-P, such as those in the V group
(Syers et al. 1972). As we detected a significant positive corre-
lation between NH4F-Pi (assumed to extract Al-P and a part of
Ca-P) and HCl-Pi (assumed to extract mainly Ca-P) (r = 0.92,
p < 0.001), the extracted Ca-P in NH4F-Pi increased with
increasing HCl-Pi. Thus, we evaluated the relative abundance
of Al-P in NH4F-Pi by comparing the Alo content and the ratio
of NH4F-Pi to the sum of NH4F-Pi and HCl-Pi (Fig. 4). The higher
this ratio, the stronger the influence of Al-P and the weaker
the influence of Ca-P in NH4F-Pi. The ratio increased with
increasing Alo in the SM group and some of the samples
with low Alo + 0.5 Feo (<20 g kg-1) in the V group (Fig. 4).
Combined with the fact that Alo is significantly correlated with
NH4F-Pi, it is indicated that Alo played a significant role in the
Figure 4. Relationship between relative abundance of Al-associated inorganic
phosphorus in NH4F-Pi fraction and Al extracted by acid ammonium oxalate
(Alo).
Samples in volcanic ash group were divided into two groups based on content of Al and Fe
extracted by acid ammonium oxalate (Alo + 0.5 Feo): high Alo + 0.5 Feo is above 20 g kg-1.
SOIL SCIENCE AND PLANT NUTRITION 297
accumulation of Al-P in the soils with low Alo + 0.5 Feo.
However, the ratio began to decrease in some samples with
high Alo + 0.5 Feo (≥20 g kg-1), which likely reflected the
underestimation of NH4F-Pi as already discussed above. The
ratio in the PL group was generally high regardless of the Alo
content because of its low HCl-Pi (Table 2).
It is notable that active Al (Alo) and Al associated with humus
(Alp) significantly contributed to the increase in total C content and
the amount of TPo in all soils, even in apparently nonvolcanic soils
with low active Al content. This is consistent with the results of
Percival et al. (2000), who reported that Alp, even at low concentra-
tions, was the principal active Al controlling the degree of soil
organic matter stabilization and accumulation, both in volcanic
and crystalline clays in New Zealand. Redel et al. (2016) also
reported that Alo played a significant role in promoting organic P
accumulation in volcanic soils in Chile. It seems, however, that TPo
reached a peak in the soils with high Alo (>14 g kg-1), which
corresponds to those with high Alo + 0.5 Feo (≥20 g kg-1), in the
V group (Fig. 5). This finding suggests that these V soils are under-
saturated with organic matter and thus TPo. In the PL group,
organic P would play an important role to supply available P
because the extracted organic P was comparable to extracted
inorganic P. Tiessen et al. (1992) and Nwoke et al. (2003) also
suggested that P availability in tropical soils that have not been
supplemented with mineral P fertilizer is largely dependent on the
NaOH-extractable organic P in the Hedley sequential extraction
procedure.
4.3. Contribution of P forms to available P in soils
The highest extraction ratio of both Bray P and Olsen P to TPi in the
PL group indicated a high ratio of loosely bound P in TPi. This is
probably because extraction was enhanced by the sandy texture
and lower TPi in PL soils than in V and SM soils. Although the
contents of Pi extracted by the Bray method and the Olsen method
generally showed a strong positive correlation, the result of the
stepwise regression analysis of available P data showed that the
Figure 5. Relationship between total organic phosphorus extracted by sequen-
tial extraction method (TPo) and Al extracted by acid ammonium oxalate (Alo).
298 T. NISHIGAKI ET AL.
Figure 6. Relationship between Bray P and sum of inorganic phosphorus
extracted by NH4F and HCl (NH4F-Pi + HCl-Pi).
Alo: Al extracted by acid ammonium oxalate; Feo: Fe extracted by acid ammonium oxalate.
various fractionated P pools made different contributions to avail-
able P, as defined by the two methods (Table 4).
The NH4F, NH4Cl, and HCl fractions were selected as the
explanatory variables for Bray P. The low pH and the fluoride
ion used in the Bray method extracted the same inorganic P
pools with that extracted by the NH4F and HCl steps in the
fractionation method. However, as shown in Fig. 6, the con-
tent of Pi extracted by Bray P was much less than the sum of
NH4F-Pi and HCl-Pi when soils had high Alo + 0.5 Feo (≥20 g kg-
1
), possibly because of re-adsorption during extraction in the
Bray method. As discussed above, NH4F-Pi in the soils with
high Alo would also be underestimated compared with that in
soils with low Alo, even though a higher concentration of the
extracts was used for the sequential extraction. These results
highlight the difficulty in completely extracting Al-P using
NH4F solution from soils with high active Al and Fe contents.
The NH4F, NaOH, NH4Cl, and HCl fractions were selected as
the explanatory variables for Olsen P, since all of these fractions
contributed to Olsen P. In the PL group, the predominance of
NaOH + NaCl-Pi was highlighted by its proportion of TPi (54%),
higher than that in the V and SM groups (13% and 33%, respec-
tively). Therefore, our results suggested that the Olsen method,
which takes all the P forms into account to evaluate available P,
is more widely applicable than the Bray method to assess avail-
able P in southern Tanzania where soil P forms differ greatly
among sites with different geological conditions. Further
research is required to assess the relationship between the P
content measured by these methods and plant growth.
5. Conclusions
The P forms in surface soils varied among sites in southern
Tanzania depending on the geological conditions and asso-
ciated soil physicochemical properties. In the PL group, the TP
content was low because of the sandy soil texture and Fe-P was
predominant. In the SM group, Fe-P was the major P form, but
some samples had a high Ca-P content due to the application of
chemical fertilizers. Soils in the V group had high TP and Ca-P,
which may have been derived from primary apatite, and high
Alo possibly caused the accumulation of Al-P. The Feo and Fed
were generally responsible for the high Fe-P (NaOH-Pi) and
residual P, respectively, while Alo contributed to the accumula-
tion of organic P and Al-P in soils in all three geological groups.
The abundance and diversity of soil P forms varied greatly
among sites with different geological conditions and associated
soil properties in southern Tanzania. Appropriate land manage-
ment on a regional scale should be established based on this
information on the availability and stability of soil P.
Acknowledgments
We thank Professor E. Marwa, and staff of Sokoine University of Agriculture
for technical support in the field and laboratory experiments in Tanzania.
Funding
This work was supported by grants from the Kyoto University Education
and Research Foundation, Japanese Society for the Promotion of Science
(JSPS) Grants-in-aid for Scientific Research (KAKENHI) Nos. 24255019,
24228007, and 17H06171, and a JSPS Research Fellow Grant (No.
14J06313)..
ORCID
Soh Sugihara http://orcid.org/0000-0002-5874-8957
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