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D. L6pez-Hermindezand M. Nifio
Laboratorio de Estudios Ambientales, IZT. Facultad de Ciencias UCV, Apdo. 47058, Caracas 1041-
A, Venezuela
(Received November 28, 1991; accepted after revision June 10, 1992)
ABSTRACT
L6pez-Hern~mdez, D. and Nifio, M., 1993. Phosphorus mineralization during laboratory incubation
in soils derived from different textured parent materials. In: L. Brussaard and M.J. Kooistra (Edi-
tors), Int. Workshop on Methods of Research on Soil Structure/Soil Biota Interrelationships. Geo-
derma, 56: 527-537.
The dynamics of phosphorus in soils derived from different textured parent materials were studied.
Soils weathered from the three parent materials were Haploborolls but different in texture and solum
depth. Microbial phosphorus by hexanol fumigation, and mineralizationrates of organic P (Po) were
assessed with a 32p dilution technique and P was fractionated using a modification of the method of
Hedley et al.
Microbial P and Po mineralization rates were significantly affected by parent materials. The high
rates of mineralization for the fine-textured soils correspond well with their higher content in sub-
strates able to mineralize e.g. total carbon, Po and microbial Pi. The residual P (e.g. more stable Pi
and Po forms) is the principal P fraction in the analyzed Mollisols.
INTRODUCTION
The amounts and forms of soil phosphorus (P) vary with soil-forming fac-
tors (Walker, 1965; Smeck, 1973). Moreover, Walker (1965) emphasized
the role of P as perhaps the key element in pedogenesis because of its great
ecological significance. More recently Cole and Heil (1981 ) have discussed
the influence of P concentration on the tendency of C and N to accumulate
during pedogenesis.
Precipitation inputs of P are generally below 1 kg ha-1 yr-1 in both tem-
perate (Likens et al., 1977) and tropical (L6pez-Hern,'lndez, 1991 ) ecosys-
METHODS
Site description
Soils from the summit position of three toposequences were used in this
study. The sites are located in southwestern North Dakota, and have been the
subject of intensive studies on erosion, organic carbon and nitrogen changes
resulting from cultivation (Aguilar, 1984; Schimel et al., 1985; Schimel, 1986 ).
The toposequences were on three different parent materials with varying tex-
tures (sandstone, siltstone and shale) with grassland as the native plant com-
ponent. The siltstone site has never been fertilized; the other sites have re-
ceived about 10 kg N ha -t yr -t (Schimel et al., 1985) The soil of all three
sites were classified as Typic Haploborolls (Soil Survey Staff, 1975 ). Selected
soil properties are shown in Table 1.
PHOSPHORUS MINERALIZATIONDURING LABORATORYINCUBATION 529
TABLE 1
FieM sampling
Soil samples for incubations were collected in August 1984. Three 7.5 cm
diameter, l 0 cm deep cores were taken from each site. The samples were kept
in a cooler with ice and returned to Fort Collins, Colorado for analysis.
Incubations
Fifty grams < 2 m m samples of dry soil were placed into Nalgene PMP
wide mouth 125 ml jars, wetted to 50% field capacity with distilled water and
incubated with 1 ml of garden soil solution (E.T. Elliott, pers. commun., 1987)
to restore biological activity. The wetter samples were left to incubate for 4
days. After that period, 2 ml 32p-labelled 10-5M KH2PO4 carrier with an ac-
tivity of approximately 40/zCi m l - l was added to the soil in order to obtain
80% of field capacity and the system was mixed thoroughly with a glass rod
to give a distribution of 32p in the soil as uniform as possible. The labelled
soils were maintained in an incubation chamber designed to measure CO2-C
evolution and at a constant room temperature (25 °C). Three aliquots from
each sample were destructively sampled at days 0, 2, 7, 14, 21 and 51. On
each sampling day the individual systems were thoroughly mixed to distrib-
ute the 32p evenly and sub-samples were taken for subsequent phosphorus,
activity and water contents analysis.
Isotope analysis
On each sampling date, triplicate samples from each site were taken in 100
ml centrifuge tubes and shaken for 1 h in a reciprocal shaker with 40 ml of de-
ionized water plus 0.5 g of bagged anion exchange resin (Dowex 1 * 80-50
> 30 mesh) in a bicarbonate form.
After the shaking period the resin bag was removed and Pi was recovered
by exchanging with 30 ml ofa 0.5M HC1 solution for 1 h. Aliquots of the resin
recovered were analyzed for Pi by Murphy and Riley (1962) reagent and for
32pi by Cerenkov liquid scintillation counting. Isotopic exchangeable phos-
530 D. LOPEZ-HERN,~iJqDEZ AND M. NIIT/O
sol.dE1 (a)
[ SAMPLE AJ
(b)
RESIN Pi 16h
RESIDUE + S U P E R N A T A N T
FOR Pi AND Po
WATER SOLUBLE
Pi AND Po
RESIDUE
Pi AND Po
RESIDUE
phorus (IEP) was determined after decay and humidity corrections at each
sampling date by using the isotope dilution technique (White, 1976) accord-
ing to the formula:
total activity added (cpm g-1 soil)
IEP -
specific activity of resin extract (cpm #g-~ P)
Resins have been shown to adsorb practically only inorganic phosphorus
(Hedley et al., 1982), therefore during the incubation period any increase in
IEP is a direct consequence of the release of Pi by microbial activity, since
exchange of 3zp with inorganic mineral forms appears to contribute little to
IEP in the studied soils (Sweet, 1981 ).
Rates of Po mineralization were calculated from the difference between IEP
values at the start (0 day) and IEP values at a given incubation period.
Phosphorusfractionation
At the last sampling date (day 51 ) a modified version of the Hedley et al.
(1982) fractionation of soil P was performed (Fig. 1). In the procedure, a
sequential extraction removed labile Pi and Po first, followed by the more
stable forms. The most biologically labile Pi was extracted first with anion
exchange resin (Amer et al., 1955 ). A duplicate sample of the soil was treated
with hexanol (McLaughlin et al., 1986) and then extracted with the resin.
The extra Pi and Po extracted (Hexanol-Resin P minus Resin P) corresponds
to part of the microbial P present in the soil (Fig. la). The suspension was
flocculated with 1.6 g of NaCI, the clear solution contains mainly Po desorbed
from the soil surface.
The resin extraction was followed by sodium hydroxide to remove Po as-
sociated with humic compounds (Batsula and Krivonosova, 1973; Fares et
al., 1974; Bowman and Cole, 1978 ) and Pi more strongly bound to Fe and A1
compounds (L6pez-Hern~lndez and Burnham, 1974, Burnham and L6pez-
Hern~indez, 1982). An acid extractant (1M HCI) removed mainly apatite-
type minerals (Williams et al., 1971 ). Residual P was not analyzed, a sepa-
rate total P analysis by NaOH fusion was performed, however.
Total P (Pt) in the neutralized extracts were determined by acidified so-
dium persulfate oxidation (Environmental Protection Agency, 1971 ) and Po
calculated as the difference between Pt and Pi-
RESULTS
Rates of mineralization
"--" SANDSTONE
IO0- -- -- SHALE
SfLTSTONE
T
50-
f ~ y
O.
2r I 511
Doys
P fractionation
The shale parent material site gave the highest value in resin Pi ( 7.1 #g g- ] )
as compared with the other two sites (3.9 and 4.5 for sandstone and siltstone,
respectively) (Table 2 ). Microbial Pi was also present in appreciable amounts
in the fine materials ( 14.3 and 10.3 #g g - ~for the siltstone and shale, respec-
tively), whereas the coarse-textured sandstone site yielded a lower value ( 7.3
/zg g- ] ). As expected after resin extraction the amount of P~ remaining in
solution was low (0.63-0.80 gg g - ] ) in all the three sites. On the other hand,
Po remaining in solution was, however, important although the values did not
differ significantly between sites (Table 2 ). Inorganic P extracted with NaOH
was also significantly higher in the siltstone than in the sandstone and shale.
PHOSPHORUSMINERALIZATIONDURINGLABORATORYINCUBATION 533
TABLE 2
Phosphorus fractions (mg kg- t ) from the sequential extraction. Standard deviation in parentheses
A similar result was found for NaOH extracted Po. HCl-extracted Pi w a s high
for the sandstone and shale sites, and significantly differs from the siltstone
site.
Finally, total P as determined by NaOH fusion as well organic P for the
fine-textured parent material (shale and siltstone) exceeded the value of the
coarse particle-size sandstone (Table 1 ). Substracting the sum of the frac-
tions from total P, residual P is calculated. It appears that residual P which
corresponds to the more stable Pi and Po forms, is the principal P fraction in
all the analyzed Mollisols; residual P fraction was higher in the shale (414
#g-~) when compared with the sandstone and siltstone with 314 and 359/zg
g- 1, respectively.
DISCUSSION
TABLE 3
IEP values at the beginning (To) and 51 days of the incubation period. % Po mineralized at 14 days
as compared with 51 days
are higher than the theoretical value (0.13 mg kg- 1 day- 1 ) found by Harri-
son (1982a) for a woodland in the Lake District in England. The data, how-
ever, cannot be compared directly because the method used by Harrison
( 1982b ) included a shorter incubation period. Harrison's experiments were
performed using an incubation temperature of 13°C, the experiments re-
ported here were done at a higher temperature (25°C) and perhaps a more
optimal moisture content. Temperature is very well known to affect the rate
of Po mineralization (Thompson and Black, 1947; Ruiz and Lrpez-Hermin-
dez, 1977).
The high rate of mineralization for the fine-textured soils corresponds well
with their higher content in substrates able to mineralize e.g. total carbon,
organic P and microbial Pi (Tables 1 and 2 ).
O'Halloran et al. (1985, 1987) studied P fractionation in a spatial varia-
bility design study. They reported a decrease in resin Pi and NaOH Pi with
increasing sand content. The results presented here are not entirely compa-
rable with O'Halloran's results because of the reduced number of samples
used. The information confirms, however, the results in the case of resin Pi,
but not for NaOH extractable Pi. When comparing the information obtained
for HC1-Pi non significant differences were found amoung the different tex-
tured parent materials. O'Halloran et al. ( 1985 ), on the contrary, reported a
positive correlation for HC1-P~ and sand content.
Schimel (1986) measured net gross N mineralization and CO2 evolution
during the first 4 days of incubation in soils derived from the same parent
materials. He concluded that net N mineralization was similar in all sites;
however, the gross N mineralization, and particularly the CO2 evolution, was
higher in the fine-textured materials than in the coarse-textured sandstone.
The results presented by Schimel (1986) for N and C coincide with the infor-
mation presented here for Po mineralization rates. CO2 evolution curves (to
be discussed in another paper) during a 28 d incubation period resembles
those obtained for isotopic exchangeable P, indicating that the increase in the
exchangeable P fraction (IEP) is a direct consequence of the release of Pi by
microbial activity.
PHOSPHORUS MINERALIZATION DURING LABORATORY INCUBATION 535
CONCLUSIONS
Our results for soils with the same taxonomic clasification (Typic Haplo-
borolls) but differing in textural characteristics show that higher rates of mi-
neralization occurred in the fine-textured parent material soils compared with
the sandstone derived soil. The high rate of mineralization for the fine-tex-
tured soils corresponds well with their higher content in organic P and micro-
bial Pi. The residual P (e.g. stable Pi and Po forms) is the principal P fraction
in the analyzed Haploborolls.
ACKNOWLEDGEMENTS
C.V. Cole and E.T. Elliott contributed to the genesis of this study and pro-
vided the soil samples and laboratory facilities. This research was supported
by NSF-BSR-810528 l, the Great Plains Agroecosystems Project. Work re-
ported in this manuscript was done while the senior author was at NREL,
Fort Collins, CO, on sabbatical leave from Universidad Central de Venezuela.
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