How Do PLANTROOTSACQUIRE

MINERALNUTRIENTS?
CHEMICALPROCESSES
INVOLVED
IN THE RHIZOSPHERE
P. Hinsinger*
Faculty of Agriculture
University of Western Australia
Nedlands, Western Australia 6907
Australia

I. Introduction
11. Definition of the Rhizosphere
111. Root-Induced Changes of Ionic Concentrations in the Rhizosphere
N. Root-Induced Changes of Rhizosphere p H
V. Root-Induced Changes of Redox Conditions in the Rhizosphere
VI. Root-Induced Complexation of Metals in the Rhizosphere
VII. Other Interactions Involving Root Exudates
VIII. Conclusion
References

I. INTRODUCTION

The acquisition of mineral nutrients by plant roots is a broader concept than the
uptake, which is the transfer of nutrients from the outer medium into the root. In-
deed, the acquisition of nutrients also encompasses the many processes occurring
in the immediate environment of the root as a consequence of its activity and in-
fluencing the dynamics of nutrients in the so-called rhizosphere prior to being ul-
timately taken up by the plant. Following Clarkson (1985), I should still “suggest
that many of the intriguing processes that occur in the root-soil interface merit a
more purposeful and integrated investigation, especially by plant physiologists,
than they have received,” and I would add “by soil scientists too.”
*Present address: Institut National de la Recherche Agronomique, UFRA de Science du Sol,
F-34060 Montpellier Cedex 2, France

22s
Advnnrcr in Agronomy,Volume 64
Copyright 0 1998 by Academic Press. All rights of reproduction in any form reserved
0065-2113/98 $25.00
226 P. HINSINGER

Most plant physiologists have addressed plant nutrition exclusively for hydro-
ponic conditions. Consequently, they have concentrated their research on the sole
uptake processes. Similarly, most soil scientists have studied the dynamics of min-
eral nutrients in soils in the absence of plants. Even though these reductionist ap-
proaches have increased our knowledge of the mechanisms involved in nutrient
dynamics, their major limitation has been to ignore, or at least underestimate, the
importance of the interactions between plant roots and soil constituents for min-
eral nutrition.
Nevertheless, evidence of the profound chemical changes that occur in the rhi-
zosphere has accumulated, as reviewed by Marschner et af. (1986), Darrah (1993),
and Hinsinger (1994). Chemical conditions in the rhizosphere can thus be drasti-
cally different from those in bulk soil. This should alter the quality of the diagno-
sis of nutrient bioavailability, which is commonly deduced from the analysis of
bulk samples of soil. A better prediction of nutrient bioavailability and mineral nu-
trition would thus take into account the chemical state and physical extent of the
rhizosphere. These factors are either a direct effect of root activity itself or an in-
direct effect of the roots, i.e., the effect of the root-stimulated rhizosphere mi-
croflora. Even though the latter effect may be of prime importance, especially for
major nutrients such as N and P,it will not be considered in this review, which con-
centrates on the direct effects of plant roots themselves. Some major groups of mi-
croorganisms occurring in the rhizosphere are root symbionts, such as N-fixing
bacteria and mycorrhizal fungi (Curl and Truelove, 1986). Their role in supplying
especially N and P to plants has been extensively reviewed elsewhere (e.g., Quis-
pel, 1983; Bolan, 1991; Marschner and Dell, 1994). The present review aims at
highlighting the processes involved in the acquisition of mineral nutrients from
soil by plant roots, with a particular emphasis on the chemical interactions occur-
ring in the rhizosphere as a direct result of root activity.

II. DEFINITION OF THE RHIZOSPHERE

Since the early work of Hiltner (1904), who first used the term rhizosphere to
describe the stimulation of microbial biomass and activity in the soil surrounding
plant roots, the concept behind the word has received several more-or-less loose
definitions (Curl and Truelove, 1986). It is now best defined as the volume of soil
influenced by root activity. According to this broad definition, the size of the rhi-
zosphere may extend well beyond the first millimeters of soil adhering to plant
roots when one considers the uptake of water and mobile nutrients, notably nitrate,
or the release of volatile compounds (Darrah, 1993). For the poorly mobile nutri-
ents such as phosphate, the extension of the rhizosphere is often limited to less than
1 mm (Hubel and Beck, 1993). In addition, the spatial extension of the rhizosphere
varies widely for any single nutrient, as shown in Table I for phosphorus.
 Table I
Spatial Extension of Phosphous Depletion in the Rhizosphere of Various Species"

Plant properties Bulk soil properties Rhizosphere properties

Age Sand Clay OrganicC P P-depletion

Species (days) (mgg-') (mg g - ' ) (mg g-') pH (mg kg-') Analytical method zone (mm) Source

Maize 5 770 80 20 5.7 158" A' 0.4 Kraus et al., 1987

6 100 210 22 5.6 6' A' 0.7 Hubel and Beck, 1993
5 850 65 - 5.8 42c AJ 1.9 Hendriks er al., 1981
Clover 10 170 210 46 7.4 36od Organic pd 0.8 Tarafdar and Jungk, 1987
Wheat 15 170 210 46 7.4 360" Organic pd 1.5 Tarafdar and Jungk. 1987
Ryegrass 10 - 90 23 7.4 347' HCI' 1.0-3.5 Gahoonia et a[., 1992a
10 - 500 5 5.7 400' HCI' 1.8-2.3 Gahoonia et al., 1992a
Rape 23 680 150 19 6.7 28Y H,SO,f I .6-3.0 Gahoonia and Nielsen, 1992
398 NaOH8 1.7-2.4 Gahoonia and Nielsen, 1992
20h KHC0,h 4.0 Gahoonia and Nielsen. 1992
7 sandy loam 6.9 27oh A' >4.0 Bhat and Nye, 1973
Onion 49 silty clay loam 7.3 2oJ A' 2.0 Owusu-Bennoah and Wild, 1979

"Modified from Hiibel and Beck, 1993, with kind permission from Kluwer Academic Publishers.
"Exchangeable P.
'Water-soluble P (Paauw, 1971).
dOrganic P (Tarafdar and Jungk, 1987).
'4M HC1-extractable P.
f6M H,SO,-extractable P in a sequential procedure (Gahoonia and Nielsen, 1992).
g0.lM NaOH-extractable P in a sequential procedure (Gahoonia and Nielsen, 1992).
h0.5M KHC0,-extractable P in a sequential procedure (Gahoonia and Nielsen, 1992).
'Determined by 33P-autoradiography.
'NaHC0,-extractable P.
228 P. HINSINGER

The variations in the extension of the rhizosphere largely depend on the physi-
cal properties of soil that influence the transfer of ionic and molecular compounds
and the resulting shape of their concentration gradients in the soil. Particle size,
soil structure, and water content, among others, are critical physical properties of
soil that determine the geometry of the rhizosphere (Kuchenbuch and Jungk, 1984;
Nye, 1986). Plant roots can alter some of these properties-for example, roots al-
ter soil structure by decreasing soil porosity in the rhizosphere as a consequence
of their radial growth (Dexter, 1987; Bruand ef al., 1996). They can also dramati-
cally decrease the water content in the rhizosphere as a direct consequence of wa-
ter uptake (Hamza and Aylmore, 1992). They may thereby directly influence the
extension of the rhizosphere and the dynamics of all nutrients.
The spatial extension of the rhizosphere also varies with plant species (Table I).
This partly relates to plant species varying in their strategies to further extend their
rhizosphere through the development of root hairs (Drew and Nye, 1969) and in-
directly through mycorrhizal symbiosis (Clarkson, 1985; Bolan, 1991). Li et al.
(1991) showed that P, which supposedly is taken up within several millimetres
from the root surface (Table I), can be acquired from a distance of up to 12 cm by
the hyphae of mycorrhizal fungi. This distance corresponds to about the maximum
spread of external hyphae of a range of vesicular-arbuscular mycorrhizal fungi, as
shown by Jakobsen et al. (1992). Plant species also differ in their rooting patterns.
Some species develop original features such as proteoid roots, which result in large
variations in the surface area of the soil-root interface and in the total volume of
rhizosphere soil used to acquire nutrients (Bowen, 1980; Lamont, 1982; Clarkson,
1985; Dinkelaker el al., 1995). Even though such considerations on the geometry
of the rhizosphere are critical for a quantitative approach to plant nutrition at the
rhizosphere level (Darrah, 1993) and even more so at the whole-plant level (Clark-
son, 1985;Barber, 1995),they will not be further discussed here. This chapter con-
centrates on the qualitative aspects of the chemical processes involved in the ac-
quisition of nutrients from rhizosphere soil.

III. ROOT-INDUCED CHANGES OF IONIC

CONCENTRATIONS IN THE RHIZOSPHERE

The uptake of water and nutrients, which is the major function of plant roots,
results in either the accumulation or depletion of all the ions contained in the soil
solution in the rhizosphere. This process occurs both for mineral nutrients and for
other, nonessential elements (e.g., Si) and possibly for toxic elements. The nature
and intensity of the changes in ionic concentrations depend on the correspondence
of the requirements of the plant and the supply by the soil. Nutrients such as Ca or
Mg, which usually occur at large concentrations in the soil solution (Fried and
 HOW DO PLANT ROOTS ACQUIRE MINERAL NUTRIENTS? 229

Table I1
Estimated Contribution of Root Interception, Mass-Flow, and Diffusion to the Mineral
Nutrition of a Field-GrownMaize Yielding 9500-kg Grain per Hectare‘

Process (kg ha-’)

Mineral nutrient Uptake Interception” Mass flow Diffusiond

Cations
Ca 40 60 I50 0
Mg 45 15 100 0
K I95 4 35 156
Anions
S 22 1 65 0
N 190 2 I50 38
P 40 I 2 31

“From S. A. Barber, 1995,“Soil Nutrient Bioavailability: AMechanistic Approach,” 2nd. ed. Copy-
right 1995 by John Wiley and Sons, Inc.; adapted by permission of John Wiley and Sons, Inc.
”The amount of nutrients supplied by root interception was calculated assuming that growing roots
would intercept 1% of the available nutrients in the soil.
T h e amount of nutrients supplied by mass flow was calculated from measured average soil solution
concentrations of nutrients in the bulk soil multiplied by an estimated water consumption of 2500 m3
per hectare.
dThe contribution of diffusion to plant uptake was obtained by difference.

Shapiro, 1961),may be transferred by mass flow to the root-soil solution interface

at a greater flux than required by the root (Table 11). It is the same for sulphate,
among major nutrients that are taken up as anions. These ions may thus accumu-
late in the rhizosphere, as shown for Ca and Mg by Youssef and Chino (1987) and
Lorenz et al. (1994). In calcareous soils the accumulation of Ca can generate cal-
cium carbonate precipitates around roots (Jaillard, 1985), a process that may im-
pair plant growth through lime-induced chlorosis. Similarly, when Ca and sulphate
accumulate concurrently in the rhizosphere, precipitation of calcium sulphate
(gypsum) can occur (Malzer and Barber, 1975; Jungk, 1996).
In saline soils, Na and CI that occur in large concentrations in the soil solution
can accumulate in the rhizosphere and reach much greater levels of concentration
due to water intake by plant roots (Sinha and Singh, 1974,1976; Marschner, 1995).
Such accumulation of Na and CI near the roots can severely impair plant growth
and mineral nutrition, even for soils in which salinity would not be considered ex-
cessive due to its acceptable level in bulk soil conditions as derived from soil paste
measurements. In addition, the resulting increase in osmotic potential (in absolute
value) in the rhizosphere (Hamza and Aylmore, 1992) can restrict water uptake by
roots, causing species that are not adapted to saline conditions to rapidly wilt and
2 30 P. HINSINGER

ultimately die. Hamza and Aylmore (1992) showed that for high soil-water con-
tent, the accumulation of salt does not increase exponentially near plant roots as
one might predict because of an important back-diffusion of the solutes to the bulk
soil. The processes of accumulating ions and salts in the rhizosphere are expected
to be enhanced by an increased water uptake rate, as shown by Sinha and Singh
(1974, 1976) for Na and C1. The effect on soil osmotic potential will in turn de-
press the water transpiration rate so that complex interrelationships between solute
accumulation and water uptake will be established (Hamza and Aylmore, 1992;
Stirzaker and Passioura, 1996).
Conversely, nutrients commonly occurring at low concentrations in the soil so-
lution, such as K and even more so P (Fried and Shapiro, 1961), are transferred by
the mass-flow process in amounts insufficient to meet the requirements of the plant
(Table 11). Their uptake thus results in a decrease in their concentration in the soil
solution near plant roots; this depletion then generates a concentration gradient and
diffusion of ions toward the roots (e.g., see Lewis and Quirk, 1967; Farr et al.,
1969; Kraus et al., 1987). Claassen and Jungk (1982) have estimated that the K
concentration in the soil solution in the rhizosphere of maize may thereby be de-
creased from several hundred to only 2-3 kmol per litre (Fig. 1).
Such a severe decrease in K concentration results not only in the diffusion of K
toward the root but also in profound consequences for K dynamics. According to
the mass-action law applied to ion exchange, it shifts the equilibrium of adsorp-
tion-desorption of K toward an enhanced desorption, leading to a depletion of
exchangeable K (Claassen and Jungk, 1982; Kuchenbuch and Jungk, 1982;

K in the soil solution

(PM)
800

600

400

200 - - - - - - - - -
0 . I I I i

0 2 4 6 8
Distance from the roots (mm)
Figure 1 Profile of K concentration in the soil solution as a function of distance from the root sur-
face of 2.5-day-old maize seedlings grown in two different soils. Concentrations of K near roots were
decreased to as low as 2-3 )LM(modified from Claassen and Jungk, 1982, with kind permission from
Wiley-VCH Verlag GmbH and Professor N. Claassen).
 HOW DO PLANT ROOTS ACQUIRE MINERAL NUTRIENTS? 23 1

AK (cmol kg'soil)
nonexch. K exch. K

I.

0.4 4 I I I I 1

0 2 4 6 8 10

Distance from the roots (mm)

Figure 2 Profile of change in exchangeable and nonexchangeable K in the rhizosphere of rape
grown for 7 days o n a loess-derived soil. Exchangeable K was depleted up to about 8 mm from the
roots. Nonexchangeable K as measured by substracting exchangeable K to HCI-extractable K was
mostly depleted within less than 2 mm from the root surface. By comparing the amount ofexchange-
able K depleted from the rhizosphere with the amount of K taken up by rape plants, it was shown that
total nonexchangeable K contributed a major proportion (71%) of K uptake (modified from Kuchen-
buch and Jungk, 1984, with kind permission from Wiley-VCH Verlag GmbH and Professor A. Jungk).

Wehrmann and Coldewey-Zum Eschenhoff, 1986; Niebes et al., 1993) and even-
tually to a depletion of nonexchangeable K (Kuchenbuch and Jungk, 1984;Jungk
and Claassen, 1986), as shown in Fig. 2. This root-induced release of nonex-
changeable K contributed up to 80% of the uptake of the plants in soils where the
release of nonexchangeable K would have been expected to be negligible when
considering the concentration of K in the bulk soil solution (Kuchenbuch and
Jungk, 1984; Niebes et al., 1993). Indeed, the release of nonexchangeable K from
interlayer sites in phyllosicates, which contribute a large proportion of soil K in
many soils, requires the occurrence of very small concentrations of K in the soil
solution (Sparks, 1987; Fanning et al., 1989)due to the high affinity of these sites
for K. Concentrationsof K as measured in the bulk soil solution of many soils and
especially in intensively cultivated soils are usually much above these critical con-
centrations. Thus, only negligeable amounts of nonexchangeable K are generally
expected to be released, even though many field experiments have revealed that
nonexchangeable K can contribute to a significant and sometimes major propor-
tion of K removal by crops (Bertsch and Thomas, 1985; QuCmener, 1986; Bosc,
1988). But when taking into account the peculiar conditions occurring in the rhi-
zosphere such as the extremely small K concentration reported by Claassen and
232 P. HINSINGER

Ryegrass Rape

0
I .
I

2
ij

Figure 3 X-ray diffraction spectra of mineral material obtained from the rhizosphere of Italian
ryegrass and rape as a function of cropping duration. The only source of K for plants was interlayer,
nonexchangeable K supplied as phlogopite, which exhibits a typical peak at 1.0 nm. No mineralogical
transformation of phlogopite was detectable in the control without plants (data not shown) and in the
rhizosphere of ryegrass after 2 days. Conversely, the appearance of a second peak at 1.4 nm after 2-3
days for both species indicated the transformation of phlogopite into a vermiculite clay mineral, which
accompanied the release of interlayer K due to K uptake by plant roots. After 16-32 days of cropping,
the vermiculitization of phlogopite was almost complete, suggesting that plant roots can induce severe
weathering of soil minerals due to chemical interactions occurring in the rhizosphere (modified from
Hinsinger and Jaillard, 1993, and Hinsinger etal., 1993, with kind permission from Blackwell Science
Ltd.).

Jungk (1982, fig. l), one can explain this apparent contradiction. By decreasing K
concentration below the critical value required for K release to occur, plant roots
can mobilize significant amounts of nonexchangeable K. This has been clearly es-
tablished when supplying ryegrass with a K-bearing phyllosilicate as the sole
 HOW DO PLANT ROOTS ACQUIRE MINERAL NUTRIENTS? 2 3 3

source of K, exclusively as nonexchangeable, interlayer K, and measuring the up-

take of K by ryegrass and the concurrent depletion of solution K in the rhizosphere
(Hinsinger and Jaillard, 1993). This root-induced release of interlayer K resulted
in a concomitant mineralogical transformation of the K-bearing phyllosilicate in
the rhizosphere (Fig. 3) within only a few days of cropping (Hinsinger etal., 1992;
Hinsinger and Jaillard, 1993). Similar mineralogical transformations were report-
ed from field plots by comparing bulk soil mineralogy with the mineralogical com-
position of soil sampled in the rhizosphere of maize roots (Kodama et al., 1994).
Plant roots are thus able to induce a substantial weathering of soil minerals as a di-
rect consequenceof the high flux of K uptake and of the subsequent, steep decrease
in K concentration occurring in their rhizosphere (Hinsinger and Jaillard, 1993).
Similar depletion of exchangeable and nonexchangeable ammonium is very
likely to occur given that its chemical behavior is close to that of K. Wehrmann
and Coldewey-Zum Eschenhoff (1 986) and Trofymow et al. (1987) evidenced a
severe depletion of exchangeable ammonium in the rhizosphere. Mengel el al.
(1990) showed the ability of plants to promote the release of fixed, nonexchange-
able ammonium in their rhizosphere. Species such as Italian ryegrass and oilseed
rape were particularly efficient at depleting nonexchangeable ammonium in the
first 2-3 mm of the rhizosphere (Scherer and Ahrens, 1996),which agrees with the
peculiar ability of these species to deplete nonexchangeable K (Kuchenbuch and
Jungk, 1984; Jungk and Claassen, 1986; Hinsinger et al., 1992; Hinsinger et al.,
1993).The sink effect of the absorbing roots, and the consequent shift of the cation
exchange equilibria, is the driving force of the root-induced release of nonex-
changeable ammonium, as it is for K (Hinsinger and Jaillard, 1993). Fixed am-
monium may thereby contribute a major proportion of N nutrition to crops (Men-
gel and Scherer 1981 ;Keerthisinghe et d., 1985; Baethgen and Alley, 1987).Once
again, the peculiar chemical conditions of the rhizosphere lead to reassessing the
relative importance of the various processes involved in nutrient dynamics.
The depletion of P in the rhizosphere of various species has also been shown in
different soils by numerous authors (Lewis and Quirk, 1967; Bhat et al., 1976;
Kraus et al., 1987;Steffens, 1987;Gahoonia er al., 1992a;Kirk and Saleque, 1995;
Saleque and Kirk, 1995; Hinsinger and Gilkes, 1996) (Fig. 4A). As in soil K, a se-
vere decrease in soil P in the rhizosphere may cause a shift in the adsorption-de-
sorption and dissolution-precipitation equilibria involved in the dynamics of soil
P. However, due to the poor reversibility of P sorption onto soil constituents (Bar-
row, 1983; Parfitt, 1978) and to the low solubility of the various phosphate min-
erals occurring in soils (Lindsay et d.,1989),very small solution P concentrations
must be reached for these phenomena to proceed to a significant extent. Such crit-
ical P concentrations might then be too low for sustaining adequate growth of
plants. For some species, the external P requirement, i.e., solution P concentration
required for near-maximum plant growth, can be as low as 1-5 pM (Asher and
Loneragan, 1967;Fohse et al., 1988) or even 10 times lower for P-efficient species
2 34 P. HINSINGER

NaOH-P (pg P g-' soil)

A
400 r
m Q

loo
0 '
t I 1

40 r
B

30

20

10

0
0 5 10
Distance from roots (mm)
Figure 4 Profile of NaOH extractable P(Na0H-P) in the rhizosphere of ryegrass grown on an alu-
mina sand with P supplied (A) as P sorbed onto alumina or (B) as phosphate rock. The profiles ob-
tained in the rhizosphere of ryegrass grown for 14 days with either alumina P or phosphate rock P as
the sole source of P are compared with profiles obtained in the absence of plants (control). A distinct
depletion of NaOH-P is detectable up to 2 mm from the root surface when P is supplied as P sorbed
onto alumina (A). Conversely, when Pis supplied as phosphate rock (B),an accumulation of NaOH-P
occurs in the rhizosphere of ryegrass, which is maximal at about 1-1.5 mrn from the roots. This in-
crease of NaOH-P indicates a root-induced dissolution of phosphate rock occurring at a rate faster than
that of P uptake (modified from Hinsinger and Gilkes, 1996, with kind permission from Blackwell Sci-
ence Ltd.).

such as perennial ryegrass (Breeze ef al., 1984).Other species, including such veg-
etable crops as lettuce, tomato, and potato, require much higher P concentrations
for achieving maximum growth (Asher and Loneragan, 1967; Fox, 1981). Thus,
 HOW DO PLANT ROOTS ACQUIRE MINERAL NUTRIENTS? 23 5

plant species vary widely in their ability to cope with low soil-Pconcentrations. In
addition to such considerations, the rate of desorption of soil P or the rate of dis-
solution of P-bearing soil constituents would need to be larger than fluxes of P up-
take to prevent any growth restriction. As pointed out by Darrah (1993), we still
lack knowledge about the kinetics of those reactions involved in P dynamics, so
the amount of P mobilized by plant roots from the rhizosphere is hardly pre-
dictable.
Surprisingly, though, some studies suggest that P may sometimes accumulate in
the rhizosphere rather than be depleted (Brewster et al., 1976; Hiibel and Beck,
1993; Hinsinger and Gilkes, 1995; Saleque and Kirk, 1995; Hinsinger and Gilkes,
1996, 1997) (Fig. 4B). Ruiz ( I 992) and Ruiz and Arvieu ( 1 992) calculated a four-
fold increase in solution P concentration at the root-mineral interface of oilseed
rape seedlings supplied with hydroxyapatite as the sole source of P. They con-
cluded that the roots induced a dissolution of hydroxyapatite at a faster rate than
that of P uptake, leading to the relative accumulation of P in the rhizosphere. They
suggested that roots acted either through proton excretion or Ca uptake, as can be

Ca,(PO,),OH + 7 H,Of -
deduced from the following dissolution equation of hydroxyapatite:
5 Ca2+ + 3 H,PO, + 8 H,O
Similar conclusions were drawn by Hinsinger and Gilkes (1995, 1996, 1997) to
(1)

explain the increase in NaOH-extractable P and concomitant dissolution of phos-

phate rock (carbonate apatite) that occurred in the rhizosphere of various species
(Fig. 4B). They showed for ryegrass that proton excretion was likely to be the dri-
ving force of this root-induced dissolution of phosphate rock occumng at a faster
rate than that of P uptake. Kirk and Saleque (1995) calculated that, whereas total
soil P was depleted to about 50% of its initial value in the vicinity of rice roots
growing in a flooded soil at various rates of P fertilization, a 5-20-fold increase in
solution P concentration occurred at 2 4 mm from the root surface. These results
agree with a model proposed by Nye (1983) for the diffusion of two interacting
solutes in the rhizosphere. In this model-for the diffusion of protons away from
the roots and the diffusion of P dissolved from a Ca-phosphate source toward the
roots (due to differential coefficients of diffusion of the two solutes)-Pcan be ex-
pected to be depleted in the immediate vicinity of plant roots and to accumulate
away from the roots, This would occur in the soil zone where dissolution due to
proton excretion is faster than the diffusion of P toward the roots due to P uptake.
It would result in some dissolved P diffusing away from the roots rather than to-
ward the roots (Nye, 1983; Kirk and Saleque, 1995). The profile of isotopically ex-
changeable P obtained by Hiibel and Beck (1993) in the rhizosphere of maize also
agrees with this model (Fig. 5). Hubel and Beck attributed the accumulation of P
peaking at about 0.5 mm away from the roots to the mobilization of some organ-
ic soil P due to phosphatase excreted by plant roots or to an artifact (P contained
in the root hairs). For mineral nutrients of which dynamics are not simply the re-
236 P. HINSINGER

lsotoplcally exch. P
(mg dm")

1
80

60

40

20

0
1
0 1 2 3
Distance from the root (mm)
Figure 5 Profile of isotopically exchangeable P around a primary root of maize grown for 4 days
in 33P-phosphate-labeled soil. Isotopically exchangeable P peaks at about 0.5 m m from the roots, in-
dicating that both depletion near plant roots and accumulation further away from the roots combine in
the rhizosphere. The accumulation of isotopically exchangeable P can be interpreted either as an arti-
fact (P included in root hairs, which extend up to about 0.5 mm) or as the result of root-induced mobi-
lization of poorly mobile P, such as organic P due to phosphatase excretion (modified from Hiibel and
Beck, 1993, with kind permission tiom Kluwer Academic Publishers).

sult of an ion exchange reaction between soil solution and one single pool of soil
solid constituent and subsequent diffusion toward the root, multiple processes can
interact and render difficult the task of predicting rhizosphere conditions and con-
sequent plant uptake.
Nevertheless, these studies suggest that the uptake of mineral nutrients not only
is the ultimate stage of the acquisition process at the root-soil interface but also
results itself in severe changes in ionic concentrations that can then shift the equi-
libria of adsorption-desorption or dissolution-precipitation involved in the dy-
namics of nutrients in the soil. The uptake may thus be regarded as a crucial and
active (in the broad sense) stage of the acquisition process. However, when con-
sidering the depletion of a nutrient in the rhizosphere, estimating its actual bene-
fit for the plant is still difficult. Although nutrient depletion may result in the mo-
bilization of less spatially or chemically accessible forms of the depleted nutrient
as shown for K, it should result as well in a decrease in the uptake rate according
to the Michaelis-Menten relationship between uptake flux and external concen-
tration (Epstein and Hagen, 1952). This holds true unless concentration levels
reached in the rhizosphere are maintained above the level required for maximum
growth (external requirement). Because of the lack of quantification of the differ-
ent processes involved here, it is difficult to state if these phenomena may be re-
 HOW DO PLANT ROOTS ACQUIRE MINERAL. NUTRIENTS? 237

garded as efficient strategies of mineral nutrition (Darrah, 1993), although some

work has clearly shown for P that the plant species that exhibit the lower external
Prequirements are among the most efficient in acquiring soil P (Fohse e l al., 1988).
Undoubtfully these root-induced changes of ionic concentrations deserve further
consideration as major components of the whole process of nutrient acquisition by
higher plants.

N.ROOT-INDUCED CHANGES OF RHIZOSPHERE pH

The uptake of nutrients can also affect soil pH. Indeed, as nutrients are taken up
as ions, the differential rates of uptake of cations and anions result in an imbalance
of positive and negative charges entering the root cells. When an excess of cations
over anions are taken up, the plant root compensates by releasing excess positive
charges as protons, thereby resulting in acidification of the rhizosphere (Nye,
1981; Romheld, 1986; Haynes, 1990). Conversely, when an excess of anions over
cations are taken up by the root, the excess negative charges are released as hy-
droxyls or bicarbonate ions, leading to alkalinization of the rhizosphere. The cor-
responding changes in pH are the most documented of the root-mediated chemi-
cal changes occurring in the rhizosphere. This is partly due to the development of
simple methods for measuring rhizosphere pH, notably, the dye indicator-agar
technique (Marschner et al., 1982).
Since N is the most demanded mineral nutrient for numerous plants (Mengel
and Kirkby, 1987; Marschner, 1995) and since it can be taken up by the plant as
an anion (as NO;), as a cation (as NH:), or as a molecule (as NJ, it has often been
reported to play a major role in the overall cation-anion balance of the plant (Nye,
1981; Romheld, 1986). Indeed, many studies have shown that plants supplied with
ammonium acidify their rhizosphere, whereas they alkalinize it when supplied
with nitrate (Riley and Barber, 1971; Jarvis and Robson, 1983; Marschner and
Romheld, 1983; Weinberger and Yee, 1984; Romheld, 1986; Gahoonia et al.,
1992a). In addition, legumes relying on symbiotic N, fixation have been shown to
acidify their rhizosphere to compensate for an excess of cations being taken up
(Jarvis and Robson, 1983; Romheld, 1986). This is, however, an oversimplified
picture of the process, since nitrate-fed plants such as oilseed rape, chickpea, or
lupins have been repeatedly shown to excrete protons and acidify their rhizosphere
(Grinsted et al., 1982; Marschner and Romheld, 1983; Loss et al., 1993; Hinsinger
and Gilkes, 1995, 1996). Some studies have also shown that the various parts of a
single root exposed to identical, external conditions can behave differently, lead-
ing to localized processes of acidification and alkalinization (Weisenseel er al.,
1979; Marschner et al., 1982; Marschner and Rornheld 1983; Haussling et al.,
1985; Jaillard et al., 1996) (Fig. 6).
238 P. HINSINGER

Figure 6 Map of pH values as obtained around the roots of a 7-day-old seedling of maize ac-
cording to the videodensitometry technique of Jaillard er al. (1996).The image was obtained 120 min
after embedding the roots in an agarose sheet containing bromocresol green as a pH dye indicator and
KNO, I mM, which had been adjusted at an initial pH of 4.60 by adding HCI. The image was acquired
with a scanning video camera and was then computed using image-analysis software. The pH map
shows that various parts of the roots behave differently. While the apical region is excreting hydroxyl
equivalents, resulting in an increased rhizosphere pH, the basal parts of the roots are excreting protons,
especially in the zones of emergence and elongation of laterals, at 2&30 and 80-100 mm respective-
ly, from the root tip (modified from Jaillard er aL, 1996. with kind permission from Kluwer Academic
Publishers).

It is now largely accepted that pH changes in the rhizosphere essentially origi-

nate from the imbalance of anions and cations taken up by plants (Nye, 1981;
Haynes, 1990). Compared with the corresponding excretion of protons, the con-
tribution to rhizosphere acidification of other processes, such as root respiration
and organic acid exudation, has not been much studied. According to Nye (1986),
the respired CO, may contribute a significant proportion of rhizosphere acidifica-
tion only in alkaline and calcareous soil conditions and/or when its diffusion is im-
paired (as in waterlogging, for instance). The abundance of calcareous soils in tem-
perate regions of the world suggests that the contribution of this phenomenon to
rhizosphere acidification would require more thorough investigations. The exuda-
tion of organic acids has occasionally been reported to contribute to pH changes
around roots of P-deficient seedlings of oilseed rape (e.g., Hoffland, 1992). Pe-
tersen and Bottger (1991) estimated that organic acids excreted by maize roots
contributed to less than 0.3% of rhizosphere acidification. Bearing in mind that the
common organic acids that can be excreted in the rhizosphere are dissociated in
the pH conditions of the cytoplasm (Hedley et al., 1982a; Nye, 1986; Haynes,
1990; Jones and Darrah, 1994), they should thus be released as organic anions and
not be regarded as responsible per se for an acidification of the rhizosphere. Nev-
ertheless, their release should be taken into account in the overall balance of
cations and anions crossing the plasmalemma (e.g., Dinkelaker er af.,1989), which
finally determines the net excretion of protons or hydroxyl equivalents. Whatever
 HOW DO PLANT ROOTS ACQUIRE MNERAL NUTRIENTS? 239

the origin of pH changes, modifications of up to 1-2 pH units have been commonly

reported in the rhizosphere of diverse species ( e g , see Riley and Barber, 1971;
Marschner and Romheld, 1983).
Soil pH is known to be a critical factor influencing many chemical reactions in
the soil environment (Mengel and Kirkby, 1987). For instance, the dynamics of
various forms of inorganic Pare strongly pH-dependent, with dissolution of P from
crystalline and sorption complexes and speciation of P in solution being strongly
dependent on the pH of soil solution (Murrmann and Peech, 1969; Barrow, 1984;
Lindsay et al., 1989). In soils of moderate to high pH, some P reacts with Ca ions
to form various sparingly soluble calcium phosphates such as octocalcium phos-
phate or hydroxyapatite (Arvieu, 1980; Freeman and Rowell, 198l), which require
a supply of protons to dissolve and release P (see Eq. 1). According to the mass
action law, the excretion of protons by plant roots should shift this equilibrium re-
action to the right, thereby enhancing the dissolution of hydroxyapatite (Kha-
sawneh and Doll, 1978; Kirk and Nye, 1986). Indeed, the ability of some species,
such as buckwheat, oilseed rape, and various legumes, to utilize P when supplied
as a phosphate rock (i.e., a carbonate apatite that also requires protons to dissolve)
is related to their capacity to excrete protons (Aguilar and van Diest, 1981; Bekele
et al., 1983; Ruiz, 1992; Hinsinger and Gilkes, 1995 and 1997). Riley and Barber
(1971) and Gahoonia et al. (1992a,b) showed that soybean and ryegrass fed with
ammonium were more efficient for mobilizing soil P in some soils than when fed
with nitrate. Similar conclusions were drawn by Hinsinger and Gilkes (1996) for
ryegrass supplied with a phosphate rock as the sole source of P. These studies thus
suggest that proton excretion occurring especially when N is supplied as ammo-
nium might improve Pnutrition by enhancing the dissolution of some forms of in-
organic P, most likely Ca-bound P, in the rhizosphere. In flooded soils where adapt-
ed plants, such as lowland rice, are expected to rely solely on ammonium (because
nitrate is reduced to ammonium as a result of the ambient reducing conditions),
root-induced solubilization of acid-soluble soil phosphates has been shown to con-
tribute a substantial proportion of P uptake (Kirk and Saleque, 1995; Saleque and
Kirk, 1995). These authors showed, however, that in this particular case the root-
induced dissolution of soil P was only partly due to proton excretion by plant roots
(see Section V). In addition, a stimulation of proton excretion has been reported
for P-deficient species such as oilseed rape (Grinsted er al., 1982; Moorby et al.,
1988; Ruiz, 1992). In this respect proton excretion by plant roots may thus be re-
garded as an adaptative strategy for P acquisition.
As for P, the concentration of Fe in soil solution is severely decreased when pH
increases, reaching a minimum for pH ranging from 7.4 to 8.5, due to the pH-de-
pendent solubility of iron oxyhydroxides (Lindsay, 1974; Lindsay, 1979; Schwert-
mann, 1991). Considering the solubility diagram of these Fe-bearing minerals in
oxidizing conditions (Lindsay, 1974), the activity of total soluble Fe in soil solu-
tion decreases from lO-'Mat pH 3.5 down to lo-" Mfor pH 8.5.Aroot-induced
240 P. HINSINGER

decrease in rhizosphere pH would thus increase the activity of Fe in the soil solu-
tion by up to several orders of magnitude. Indeed, Oertli and Opoku (1974) showed
that an enhanced proton excretion by maize roots, as obtained in response to a large
K supply and consequent excess of uptake of cations over anions, resulted in an
improved mobilization of Fe from a synthetic ferric hydroxide. In addition, many
species have been shown to respond to Fe-deficiency by acidifying their rhizos-
phere (Romheld et al., 1984; Marschner et al., 1986, 1989). Nevertheless, in the
pH range commonly found in soils, Fe activity is always below M (see pre-
ceding discussion), which is the value required for many plants to meet their Fe
requirements. Such a value is attained only for soil pH of about 3 (Lindsay, 1974).
The extent to which proton excretion is capable of supplying a sufficient amount
of Fe to roots for adequate plant growth is thus questionable unless very high flux-
es of proton excretion occur at the soil-root interface.
Considerable proton effluxes have indeed been encountered in the rhizosphere.
Romheld et al. ( 1 984) reported that the roots of Fe-deficient sunflower excreted an
average of about 5.6 pmol H+ hour-' g-' fresh weight of root, whereas Fe-ade-
quate plants released small amounts of hydroxyl equivalents. They found that lo-
cally the proton efflux could be as high as 28 pmol H+ hour-' g-' fresh weight
of root, especially near apical root zones, which were the preferred sites of excre-
tion. Proton effluxes of the same order of magnitude have been reported for roots
of oilseed rape (Jaillard, 1987; Ruiz, 1992). Jaillard (1985, 1987) has shown that
plant roots were able to grow in compacted, highly calcareous soils by dissolving
the surrounding calcium carbonate due to a large flux of Ca uptake and to a con-
sequent, large proton efflux. The dissolved calcium carbonate (calcite) was shown
to reprecipitate subsequently into the vacuole of root cells (Jaillard et af., 1991) to
form calcified roots, which can constitute up to 25%of the total calcium carbon-
ate present in some calcareous soils under natural grasslands (Jaillard, 1984). Jail-
lard (1987) showed in short-term experiments in controlled conditions that living
roots of oilseed rape were able to precipitate calcite into their cells within only a
few hours. Hinsinger et af. (1993) have shown that oilseed rape was also able to
mobilize nonexchangeable Mg by dissolving a Mg-bearing phyllosilicate as a re-
sult of the severe pH decrease that its roots induced in the rhizosphere (Table 111).
Conversely, ryegrass grown in identical conditions proved unable to mobilize any
significant amount of Mg (Hinsinger and Jaillard, 1993), as a consequence of the
high pH that it maintained in its rhizosphere (Table 111).
The preceding examples address the case of acidification of the rhizosphere as
a profitable strategy for acquiring mineral nutrients. However, alkalinization of the
rhizosphere is likely to be as widespread as acidification, or even more so, as in-
ferred by Nye ( I 986), considering that nitrate is the prominent source of N for non-
legume plants in most field conditions. In addition, the excretion of protons should
not be regarded as a universal solution to the problems encountered by plants while
acquiring nutrients. Rhizosphere acidification can have detrimental effects on root
 HOW DO PLANT ROOTS ACQUIRE MINERAL NUTRIENTS? 241

Table 111
Rhizosphere pH and Mobilizationof Mg (expressed as g kg-' of applied Mg)
as a Function of Time in the Cropping ExperimenLa
~

Time (days) 0 4 8 16 32

Rhizosphere pH rape 7.03 6.56 5.55 4.22 4.27

ryegrass 7.03 7.22 8.09 7.48 7.69
Amounts of Mg mobilized rape - -1 3 15 21
ryegrass - -1 0 2 3

UMgwas supplied as a Mg-bearing phyllosilicate (phlogopite). The Mg cations contained in this

mineral constitute the silicate framework and are thus nonexchangeable. Their release requires a dis-
solution of the phyllosilicate, which can occur in acid conditions such as those encountered in the rhi-
zosphere of rape after 1 6 3 2 days of cropping.

growth and mineral nutrition when soil pH is eventually decreased to very low val-
ues. Gahoonia ( 1993) showed, for instance, that ryegrass fed with ammonium de-
creased its rhizosphere pH to 4.4, which is a pH value prone to aluminium toxic-
ity (Kinraide, 1991). Indeed, Gahoonia (1993) measured a concurrent increase in
extractable Al in the rhizosphere of ryegrass. Thus, it is clear that plant roots should
not always acidify their rhizosphere and particularly not when growing in already
acid soils so as to prevent increased risks of aluminium, manganese, or even pro-
ton toxicity (Marschner, 1995). Some researchers have shown that species grow-
ing naturally in very acid soils, such as Norway spruce for instance, rather alka-
linize their rhizosphere (Marschner et al., 1991). Youssef and Chino (1989) have
shown that some plant species can increase rhizosphere pH in acid conditions and
decrease it in neutral or alkaline conditions, revealing the capacity of plants to
adapt to adverse soil conditions. Another major limitation of acid soils for plant
growth is related to deficiencies in various mineral nutrients, especially P defi-
ciency (Marschner, 1995). In acid soils, phosphate ions are indeed strongly sorbed
on various soil minerals and especially Fe- and Al-oxyhydroxides; the charge of
these minerals being pH dependent. Since the desorption of phosphate ions from
these minerals involves ligand exchange (Parfitt, 1978), hydroxyls or bicarbonate
ions excreted by plant roots may desorb some phosphate and render it available to
the plant, as suggested by Gahoonia et al. (1 992a). Gahoonia et al. (1992a) showed
that when increasing their rhizosphere pH, roots of ryegrass fed with nitrate were
more efficient at desorbing P in an Fe-oxyhydroxide-rich soil (oxisol) than when
fed with ammonium. Improved P nutrition can thus result from either root-induced
decrease or an increase in rhizosphere pH, depending on the dominant forms of in-
organic P present in the soil (Gahoonia et al., 1992a).
Whether root-mediated pH changes of the rhizosphere should be regarded as an
adaptative strategy of nutrient acquisition or not, there is no doubt that the actual
2 42 P. HINSINGER

pH in the rhizosphere should be taken into account rather than the pH of the bulk
soil when considering nutrient dynamics. Furthermore, the actual pH may be a
poor indicator of the real effect exerted by the root on its environment. Consider-
ing that the activity of protons and hydroxyls is influenced by the total solute con-
tent of the rhizosphere solution and that a large part of protons or hydroxyl equiv-
alents produced by the roots may be consumed in diverse adsorption-desorption
or dissolution-precipitation reactions with soil minerals, the consequences of the
excretion activity of the roots may extend much beyond what is indicated by a di-
rect measurement of the resulting pH in the rhizosphere. For instance, for nitrate-
fed ryegrass and subterranean clover grown in an artificial soil with phosphate rock
(carbonate apatite) as the sole source of P,Hinsinger and Gilkes (1996) found that
up to 20-25% of the applied phosphate rock dissolved in the rhizosphere, where-
as the pH decreased only minimally (<O. 1 pH unit). Considering that the dissolu-
tion of apatite minerals is assumed to consume protons (see Eq. (1) for hydroxya-
patite) they calculated, according to the dissolution equation of the carbonate
apatite used, that 16-19 Fmoles of protons had been consumed to account for the
root-induced dissolution of phosphate rock. Taking into account the proton-buffer-
ing capacity of the soil used, this should have led to an increase of about 0.3-0.4
pH units in the rhizosphere of both species. These results thus indicate that rye-
grass and clover excreted slightly more protons than required for dissolving phos-
phate rock and much more than predicted from the pH change in the rhizosphere
(Hinsinger and Gilkes, 1996). The critical factor influencing the acquisition by
plant roots of mineral nutrients such as P is thus the actual flux of protons of hy-
droxyl equivalents that are excreted at the soil-root interface rather than the re-
sulting pH change.

V. ROOT-INDUCED CHANGES OF REDOX

CONDITIONS IN THE RHIZOSPHERE

The redox conditions of a soil, which can be described by either the redox po-
tential (Eh) or the pe (negative log of the activity of electrons; with Eh (mV) =
59.2 pe), is an environmental parameter of critical importance for the dynamics of
those elements that can occur at different oxidation states in soils. Among miner-
al nutrients, this is particularly the case for Fe and Mn.
As previously pointed out, because of the low solubility of Fe-bearing minerals
such as iron oxyhydroxides and iron oxides, the activity of soluble Fe species in
soils is commonly much below the activity required for adequate plant growth
(Lindsay, 1974). Besides its strong dependency on pH, the solubility of iron oxy-
hydroxides and iron oxides is very much dependent on redox conditions. When
oxidant conditions are prevailing, which is the case in most soils as long as the
 HOW DO PLANT ROOTS ACQUIRE MINERAL NUTRIENTS? 243

transfer of gases is not impaired and PO, is about atmospheric PO,, the activity of
Fe2+ and other Fe" species is less than the activity of Fe3+and other Fe"' species
at the usual pH values found in the soils, except for alkaline soils (Lindsay, 1979).
However, when reduction occurs and pe decreases, Fe" species rapidly become
dominant over Fe"' species; the ratio Fe2+-Fe3+ increases 10-fold for a decrease
in pe by one unit (Lindsay, 1979). The reduction of iron-hydroxide can be de-

Fe(OH), + 3 H30+ + e-
from which the following can be deduced:
-
scribed by the following equation (Lindsay, 1979):
Fe2+ + 6 H,O, (2)

log(Fe2+) = 15.74 - (pe + pH) - 2 pH (3)

In oxidant conditions, when PO, equals atmospheric PO,, pe + pH = 20.61;
whereas it can decrease to as low as 2 when reducing conditions are prevailing
(Linday, 1979). Under such circumstances the activity of Fe" species can increase
by several orders of magnitude. Reduction processes can thus be very efficient for
enabling higher plants to meet their Fe nutritional demand. Root respiration con-
sumes 0, and may thus decrease PO, and pe, thereby increasing the activity of Fe
in the soil solution. However, PO, must be decreased to very low values before a
significant increase occurs in the solubility of iron oxyhydroxides and iron oxides.
This can occur only when soil physical conditions are impairing the transfer of
gasses to a great extent, i.e., in anoxic conditions that are altogether unfavorable
for adequate plant growth (discussed later).
The reducing activity of plant roots can also be attributed to the release of re-
ducing compounds such as phenolics (caffeic acid) or aliphatic acids (malic acid)
in the rhizosphere (Brown and Ambler, 1973; Romheld and Marschner, 1983). Al-
though this process has been suspected to be of ecological importance, it has been
shown to be too restricted to account for the reducing capacity of most plant
species (Romheld and Marschner, 1983; Bienfait et al., 1983). For bean, for in-
stance, Bienfait et al. (1983) found that the release of reductant in the rhizosphere
contributed about 14% of the total amount of Fe reduced by roots of Fe-sufficient
plants and less than 2% for Fe-deficient plants. They also showed that the release
of reductant less than doubled for Fe-deficient compared with Fe-sufficient bean
plants, whereas the reducing activity of the roots increased 15-fold. This reducing
activity, which was found to be located at the root surface rather than being re-
leased into the rhizosphere, was attributed to a plasmalemma-bound reductase
sytem (Chaney et al., 1972; Bienfait et al., 1983; Romheld and Marschner, 1983).
This suggests that the corresponding root-induced change in redox potential is
likely to be spatially restricted to the root-soil interface and not to extend into the
rhizosphere. The efficiency of such an enzymatic process for Fe reduction in cal-
careous soils can, however, be questionned: it is certainly of little significance if
not accompanied by a concomitant acidification of the rhizosphere, since this re-
244 P. HINSINGER

a b l e IV
Effect of Fe Deficiency on Rhizwphere pH and on the Amount of Fe Reduction
and Fe Uptake by a Chlorosis-ResistantCultivar of Chickpep

Fe reduction Fe uptake
(nmol Fe g-’ root (nmol Fe g - ’ root
Fe status Rhizosphere pH fresh wt. hour-’) fresh wt. hour-’)

Fe-sufficient 6.0 200 0.39

Fe-deficient 3.9 1100 7.58

“Modified from Marschner, 1990.

ductase system has been found to be inhibited at pH above 7 (Marschner ef al.,

1989). Nevertheless, numerous species respond to Fe deficiency both by an en-
hanced proton excretion and by an increase in the reducing capabilities of their
roots, showing a remarkable example of “cooperative strategy” (Romheld and
Marschner, 1983). This is illustrated for chickpea in Table IV. The stimulation by
Fe deficiency of the reducing activity andor proton excretion by plant roots has
been described as “strategy I” for Fe acquisition by nongraminaceous species
(Marschner et al., 1986).
The reduction of Fell1to Fe” as related to the reducing activity of the roots is
thus a major adaptative strategy of dicotyledonous species and nongrass mono-
cotyledonous species for coping with the poor solubility of iron oxyhydroxides
and iron oxides in soils (Chaney ef al., 1972; Marschner et al., 1989). The reduc-
ing activity of roots is likely to play an equally important role in Mn nutrition: The
reduction of MnO, has been observed in the rhizosphere of different species (Godo
and Reisenauer, 1980; Gardner et al., 1982). Uren (1981) has clearly established
with axenic-grown sunflower plants that the roots were themselves directly re-
sponsible for the reduction of manganic Mn, even though the mechanism has not
been fully elucidated.
When anoxic, reducing ambient conditions are prevailing in the bulk soil, as in
waterlogged soils, the activity of Fe” species can dramatically increase, as de-
duced from Eq. (2). Ferrous Fe, and also manganous Mn and sulfides, can there-
by reach phytotoxic levels that, together with the lack of 0,, may preclude plant
growth (Drew, 1988). To cope with such conditions, the roots of rice and other
plants naturally growing in submerged areas have developed an anatomical adap-
tation in their cortical tissue called aerenchyma: The translocation of 0, from the
shoots to the roots through the aerenchyma not only allows root cells to respire but
also results in a leakage of excess 0, in the rhizosphere (Armstrong, 1967; Ando
et al., 1983). This excess of 0, released by rice roots then diffuses in the rhizo-
sphere, leading to a significant increase in Eh (Fig. 7) up to a few millimetres away
from the roots (Trolldenier, 1988; Flessa and Fischer, 1992). The diffusion of 0,
 HOW DO PLANT ROOTS ACQUIRE MINERAL, NUTRIENTS? 245

Eh (mV)

Submerged soil

-200

-400
1 -iment
I I

0 5 10
Distance from the root (mm)
Figure 7 Profile of redox potential in the rhizosphere of 5-week-old rice grown in a submerged
soil and in river sediment. Redox potential as measured by redox microelectrodes decreased signifi-
cantly due to oxygen leakage from the primary root of rice, leading to a root-induced oxidation of the
rhizosphere detectable up to 1-3 mm from the roots (modified from Flessa and Fischer, 1992, with kind
permission from KIuwer Academic Publishers).

in the rhizosphere of roots of lowland rice thereafter results in an oxidation of fer-

rous Fe, as revealed by the decrease of Fe" concentrationand concomitant increase
of Fe"' concentration measured within a few millimeters from the root surface
(Fig. 8) in a flooded soil (Begg et al., 1994; Saleque and Kirk, 1995).
A further consequence of this oxidizing effect of rice roots is the precipitation
of ferric Fe as iron oxyhydroxides (Chen er al., 1980), which contributes the red-
brownish discolorationthat is clearly visible at the surface of rice roots after flood-
ing of the soil. Chen et al. (1980) identified iron coatings around rice roots as be-
ing mainly formed of goethite and lepidocrocite. Iron coatings have been reported
around roots of other plant species growing in submerged soils, e g , slash pine
(Fisher and Stone, 1991). The deposition of iron oxyhydroxide precipitates has
also been described to occur inside the root cortex in cell walls and intercellular
spaces of rice (Green and Etherington, 1977). The increase in redox potential in
the rhizosphere and subsequent immobilizationof ferrous Fe by oxidation and pre-
cipitation as iron oxyhydroxide by plant roots can prevent the uptake of toxic
amounts of ferrous Fe (Green and Etherington, 1977).
In addition, the oxidation of Fe" with 0, produces 2 mol of protons per mole
of Fe, according to Eq. (3) and the following equation (Ahmad and Nye, 1990):
4 F e 2 + + 0 , + 18H20*4Fe(OH),+8H,0+ (4)
The root-induced oxidation of ferrous Fe is thus expected to result in an acidifica-
tion of the rhizosphere. This has been shown by Begg et al. (1994) and Saleque
and Kirk (1995) for rice grown in a flooded soil (Fig. 8). These researchers showed
2 46 I? HINSINGER

Fe (mmol kg-' soil) PH

250
.* * * r7
200 t6
150

0 t2
0 5 10 15 20
Distance from the roots (mm)
Figure 8 Profile of Fell and Fell' concentration and of pH in the rhizosphere of 2 I -day-old low-
land rice grown in a flooded soil for 10 days. The Fe" concentration decreased up to 4 mm from the
roots, whereas Fe"l concentration steeply increased near rice roots, indicating that root-induced oxi-
dation of Fe occurred due to oxygen leakage into the rhizosphere. The larger extent of the depletion
zone of Fe" relative to the spread of accumulation of Fe"' is related to the larger mobility of Fe" com-
pared with Fe"'. Simultaneously, a steep decrease in soil pH was encountered, which is partly attrih-
uted to soil acidification resulting from the oxidation of Fe" according to Eq. (4) and to protons ex-
creted by rice roots to compensate for an excess of cation uptake (modified from Begg ef al., 1994).

that part of the strong decrease in pH that they measured in the rhizosphere of rice
was also due to proton excretion by rice roots that were relying exclusively on am-
monium as the soil-N source considering the reducing conditions of the bulk soil.
Kirk and Saleque (1995) and Saleque and Kirk (1995) also showed that the acid-
ification of the rhizosphere of lowland rice was responsible for the solubilization
of substantial amounts of soil P. In other words, root-induced changes in redox po-
tential not only influence the dynamics of elements with varying states of oxida-
tion such as Fe and Mn, but due to the concomitant change of pH that they impose
on the rhizosphere they can alter the availability of other nutrients, as evidenced
here for P (Kirk and Saleque, 1995; Saleque and Kirk, 1995). Nevertheless, the
major benefit of root-induced oxidation to the plant is the detoxification of the root
environment through a decrease in concentration of ferrous Fe and possibly
manganous Mn as well (Marschner, 1995).
The increase in Eh occurring in the rhizosphere of lowland rice in waterlogged
conditions and the consequent precipitation of iron oxyhydroxides around the
roots is probably the best-known evidence of an extensive change in redox condi-
 HOW DO PLANT ROOTS ACQUIRE MINERAL NUTRIENTS? 247

tions as induced by plant roots. In a similar way manganese oxides have been re-
ported to sometimes precipitate in the rhizosphere of lowland rice (Bacha and
Hossner, 1977).

VI. ROOT-INDUCED COMPLEXATION OF METALS

IN THE RHIZOSPHERE

The release of organic compounds by plant roots is a well-known phenomenon

that is at the origin of the stimulation of soil microflora in the immediate environ-
ment of the roots; i.e., the “rhizosphere effect” that was first defined as such by
Hiltner in 1904. It is now largely accepted that up to 20-30% of the total C as-
similated by higher plants is released in the rhizosphere as diverse exudates, in-
cluding respired CO, (Merckx et al., 1986a; Helal and Sauerbeck, 1989). These
exudates, however, remain difficult to quantify because of their rapid microbial
degradation (Uren and Reisenauer, 1988), and because similar metabolites are re-
leased by soil microorganisms.
Nevertheless, some root exudates exhibit complexing or chelating properties
with respect to metallic ions. Among those, numerous organic anions, which are
the conjugated bases of organic acids, have been reported to play a role in the mo-
bilization or immobilization of mineral nutrients or undesirable elements, as re-
lated to their complexing properties (Jones and Darrah, 1994). In addition, re-
searchers have shown that the exudation of organic acids by plant roots increases
as a response to nutrient deficiencies (Kraffczyk et al., 1984; Lipton et al., 1987;
Jones and Darrah, 1995), which might suggest their possible implication in the ac-
quisition of mineral nutrients from the rhizosphere.
Citrate, for instance, has been shown to be released in considerable amounts by
proteoid roots of white lupin (Gardner et al., 1982; Dinkelaker et al.,1989; Gerke
et al., 1994) and Proteaceae of the Banksia genus (Grierson, 1992; Dinkelaker et
al., 1995). Gardner et al. ( 1983)proposed that citrate excreted by the proteoid roots
of white lupin may play a major role in the dissolution of iron phosphates, through
the formation of a ferric hydroxy-phosphate complex. These researchers suggest-
ed that this complex might be of prime importance for the acquisition of P and Fe
by white lupin roots. This mechanism may partly explain the peculiar ability of
white lupin and presumably of most native Proteaceae species to cope with the
very poor P status of many Australian soils (Bowen, 1980). Furthermore, the en-
hanced formation of proteoid roots in P-deficient lupin (Marschner et d., 1986)
and the enhanced exudation of citrate in the rhizosphere of alfalfa and rape as a re-
sponse to P deficiency (Lipton et al., 1987; Hoffland et al., 1989, 1992) suggest
that they may be considered as adaptative strategies for the acquisition of nutri-
ents such as P. Indeed, the solubility of soil P has been shown to be influenced by
2 48 P. HINSINGER

organic anions such as citrate (Jones and Darrah, 1994). Staunton and Leprince
( 1996) showed that, compared with acetate, tartrate, salicylate, and oxalate, citrate
was the most efficient organic anion for increasing the proportion of phosphate in
soil solution. They showed that solution phosphate increased by a factor of two to
three for citrate concentrations ranging from 0.1 to 1 mM (Staunton and Leprince,
1994). Gerke (1994) reported a 20-fold increase in phosphate desorption from a
soil on addition of 50 pnol citrate g-I soil, which corresponds to the tremendous
concentration of citrate that Dinkelaker et al. (1989) found in the rhizosphere of
proteoid roots of white lupin.
While Parfitt (1979) and Grimal et al. (1995) privileged the hypothesis that or-
ganic anions excreted by ryegrass and maize may be involved in ligand exchange
reaction with phosphate ions sorbed onto iron oxyhydroxide (goethite) surfaces,
Bolan et al. (1994) proposed that the major effect of organic acids involved in the
release of soil P was related to A1 being complexed and to the subsequent solubi-
lization of P-A1 compounds. Indeed, they showed that addition to soil of various
organic acids commonly found in the rhizosphere resulted in decreased P sorption
and that organic acids extracted more soil P according to their ability to form sta-
ble complexes with Al (log K,,). Among the range of organic acids investigated,
oxalic and citric acids had the highest log K,, and had the largest effect on P up-
take and plant growth of ryegrass (Bolan et al., 1994). Ae et al. (1990) proposed
a similar mechanism to explain the peculiar ability of pigeon pea to take up P in a
P-deficient alfisol from India for which a major proportion of soil P was Fe-bound
P. They suggested that the roots of pigeon pea were more efficient than the roots
of other species due to excretion of organic acids that complexed Fe and resulted
in releasing Fe-bound P. Nevertheless, they found that pigeon pea had less citrate,
malate, malonate, and succinate in its root exudates than other less P-efficient
species such as soybean. Piscidic acid and its derivatives were identified as the pe-
culiar root exudates of pigeon pea that explained its ability to use Fe-bound P,
which was found to be almost unavailable to the other crops studied (Ae et al.,
1990).
The complexation of diverse micronutrients such as Co, Cu, Mn, and Zn
(Merckx et al., 1986b; Mench et al., 1987) and undesirable heavy metals such as
Cd (Mench and Martin, 1991) and Pb (Mench et al., 1987) has been shown to oc-
cur in the rhizosphere as a consequence of root exudation. Even tnough the root
exudates directly responsible for the complexing of these metals have not been
identified, speculations about the role of simple organic acids are supported by the
results of Mench and Martin (199 1). In addition, Gardner et al. ( 1982) and Dinke-
laker et al. ( 1 989) found an increase in amounts of available micronutrients such
as Fe, Mn, and Zn in the rhizosphere sampled near proteoid roots of white lupin,
which were also evidenced as root zones responsible for intense excretion of cit-
rate. Dinkelaker et al. (1989) estimated that a considerable amount of citrate was
excreted per plant, i.e., about 5.5 mmol per plant, which represented about 23% of
 HOW DO PLANT ROOTS ACQUIRE MINERAL NUTRIENTS? 249

Table V
Amounts of Citrate Excreted and of DTPA-ExtractableFe, Mn, and Zn in the Rhizosphere
of White Lupin Relative to Bulk Soil (mean value % standard deviationpb

Citrate Fe Mn Zn
(kmol g-I soil) (pmol kg-’ soil) (pmol kg-I soil) (pmol kg-’ soil)

Bulk soil - 34 ? 6 4428 2.8 ? 0.4

Rhizosphere soil 47.7 2 7.2 251 ? 43 222 ? 23 16.8 -t 2.4

“Modified from Dinkelaker et a/.. 1989, with kind permission from Blackwell Science Ltd.
hRhizosphere soil was sampled around proteoid rootlets of white lupin grown in a calcareous soil
for 13 weeks. DTPA (diethylen-triamin-penta acetic acid) extraction gives an estimate of available Fe,
Mn, and Zn.
“Not detectable (i.e. < 0.05 pmol g - ’ soil).

the dry weight of the whole plant. Citrate was excreted mostly around proteoid
rootlets where DTPA-extractable Fe, Mn, and Zn were found to increase-about
seven-, five- and six-fold, respectively, relative to bulk soil (Table V). Since a de-
crease in soil pH from 7.5 to 4.8 was also recorded in the rhizosphere of proteoid
roots, and since some reduction was likely to occur as well (Gardner et al., 1982),
it was not possible to definitely state that the exudation of citrate by proteoid roots
of white lupins contributed a major proportion of the mobilization of soil Fe, Mn,
and Zn (Dinkelaker et al., 1989). Jones et al. (1996a) recently demonstrated that,
except for alkaline pH conditions, citrate and malate might be responsible for a
substantial dissolution of iron hydroxide. They calculated that at rates of root ex-
udation reported in the literature, citrate and malate may thereby satisfy a signifi-
cant proportion of the Fe demand of plants. Considering, however, that simple or-
ganates such as citrate, oxalate, and malate, which are commonly reported as
important exudates in axenic-grown plants, might be rapidly metabolized by rhi-
zosphere microflora, their effective role in complexation processes in natural en-
vironments still remains questionnable (Darrah, 1991 ; Jones et d . , 1996b).
Treeby et al. ( 1989) reported that other root exudates such as the so-called phy-
tosiderophores can complex micronutrients such as Cu, Mn, and Zn. Mench and
Fargues (1995) reported that phytosiderophores produced by roots of an Fe-effi-
cient oat cultivar might also be involved in the mobilization of undesirable heavy
metals such as Cd and Ni from sludge-contaminated soils.
Phytosiderophores have been defined by Takagi et al. (1984) as a group of root
exudates exhibiting strong complexing properties with respect to ferric Fe (Taka-
gi, 1976) and identified as nonproteinogenic amino acids, such as mugineic acid
and its derivatives. In this respect, they are analogues of microbial siderophores,
which are literaly “iron bearers.” Literature on this topic has been extensively re-
viewed by Romheld and Marschner (1986a), Marschner ef al. (1989), and
250 P. HINSINGER

Phytosiderophore release
(pino1 g-l root d1 w
lo 1
8-
6-
4-
2-
0 -
Barley Wheat Oat Maize Sorghum

Figure 9 Amount of phytosiderophores(PS) released by roots of Fe-sufficient ( f Fe) and Fe-de-

ficient (- Fe) seedlings of Graminaceae species differing in their tolerance to lime-induced chlorosis.
The most chlorosis-resistant species, such as barley and wheat, exhibited the highest rates of PS release
and largest response to Fe-deficiency (modified from Romheld and Marschner. 1990, with kind per-
mission from Kluwer Academic Publishers).

Romheld (1991). The synthesis and release of phytosiderophores in the rhizo-

sphere are stimulated by Fe deficiency (Romheld, 1991) and have been described
as “strategy 11” for Fe acquisition, as developed exclusively by graminaceous
species (Marschner et al., 1986). Among Graminaceae, species differ widely in
their ability to produce phytosiderophores, both quantitatively (Fig. 9) and quali-
tatively. Most remarkably, among the range of graminaceous species studied by
Marschner et al. (1 989), the enhancement of the release of phytosiderophores by
Fe deficiency was reported to increase accordingly to the resistance of the species
to lime-induced chlorosis (Fig. 9).
The efficiency of phytosiderophores for complexing Fe as a function of the com-
petition with other metals or other complexing substances has been discussed by
Romheld (1991). Romheld and Marschner (1986b, 1990) showed that, compared
with Fe supplied as a microbial siderophore-Fe complex, a much larger uptake of
Fe was achieved by Fe-deficient barley plants when supplied with a phytosidero-
phore-Fe complex, although both phyto- and microbial siderophores were re-
sponsible for a similar amount of soil Fe being mobilized (Table VI). This prefer-
ential uptake of Fe-phytosiderophore relative to other complexes (more than three
orders of magnitude for uptake of Fe-HMA versus Fe-DFOB in Table VI) is re-
lated to the occurrence of a specific system of uptake of the undissociated Fe-phy-
tosiderophore complexes (Romheld and Marschner, 1986b; W i r h ef al., 1994).
This partly explains the efficiency of strategy I1 and justifies the terminology of
 HOW DO PLANT ROOTS ACQUIRE MINERAL NUTRIENTS? 251

Table VI
Rates of Fe Mobilization from a Calcareous Soil for a Phytosiderophoreand a Microbial
Siderophore and Rates of Uptake of Fe by Fe-Deficient Barley Plants
When Supplied with Fe as Fe-HMA or Fe-DFOB“.”

Fe mobilization Fe uptake
(nmol Fe g -’ (nmol Fe g-’ root
soil hour-’) dry weight hour-’)

Phytosiderophore (HMA) I .91 864.0

Microbial siderophore (DFOB) 1.60 0.3

HMA, hydroxymugineic acid; DFOB, fenioxamine B, or desfeml.

“Modified from Romheld and Mmchner, 1990, with kind permission from KluwerAcademic Publishers.
*he rates of Fe mobilization were obtained with applied concentrations of HMA and DFOB of
lo-’ M.

phytosiderophore, in spite of the ability of these substances to complex metals oth-

er than Fe (Treeby er al., 1989).
Again, a major limitation of the efficiency of phytosiderophores is their likeli-
hood to be degraded by rhizosphere microbes (Takagi et al., 1988; W i r h er al.,
1993). However, as inferred by Romheld (1991), it seems that the production of
phytosiderophores is sufficient to meet Fe requirements, at least in chlorosis-re-
sistant species such as barley, because phytosiderophores are released at high rates
that are both spatially and temporally confined. Indeed, Takagi et al. (1984) re-
ported that the release of phytosiderophores is a rythmic phenomenon restricted to
a period of 2-8 hours after the onset of daylight. In addition, Marschner et al.
( 1987) found for Fe-deficient barley that the release of phytosiderophores was
maximal immediately behind the root apex (Fig. 10).These characteristics of the
excretion of phytosiderophores are certainly beneficial to the acquisition of Fe, ac-
cording to the results deduced from the model put forth by Darrah (199 1). In this
model, Darrah (1991) predicted that short-term exudation at a high concentration
of a chelating compound such as phytosiderophores, which would be localized be-
hind the root tip, would lead to a more efficient acquisition of metal nutrients such
as Fe than a persistent exudation at a lower concentration, which would be uni-
formly distributed along the root. This holds true particularly when assuming that
rhizosphere microbial biomass is minimal behind the root tip, as evidenced by
Uren and Reisenauer ( 1988) and W i r h er al. ( 1993); in this case the site for max-
imal exudation coincides with the site for minimal potential degradation of exu-
dates by rhizosphere microflora.
The biosynthesis and excretion of complexing substances such as phy-
tosiderophores thus appear to be a “sophisticated” strategy developed by grami-
naceous species for coping with the low solubility of naturally occurring, Fe-bear-
252 P. HINSINGER

Phytosiderophorerelease
(pmol g" root DW per 4h)

100 1

80 -
60 -
40 -
20 -
0 1 'c
-- - I
I

1
0
1

0 5 10 15
Distance from root apex (cm)
Figure 10 Flux of phytosiderophore (PS) release as a function of the distance from root apex
(along the root) for Fe-sufficient (+ Fe) and Fe-deficient (- Fe) 15-day-old barley plants. Phytosidero-
phores were collected over a 4-hour period starting 2 hours after the onset of the light period. Vertical
bars indicate standard deviations (modified from Marschner et al., 1987).

ing secondary minerals (iron oxides) and for acquiring soil Fe. Studying the ex-
perimental weathering of a basalt rock containing Fe essentially as primary min-
erals (olivine and pyriboles), Femandes Barros and Hinsinger (1994) showed that
only minor amounts of Fe were released into the leaching solution in the absence
of plants due to the low solubility of Fe-bearing minerals in oxidizing conditions.
Conversely, in the presence of plants, they reported considerable amounts of Fe
released by the basalt and taken up by the seedlings of the various species studied
and above all by maize. Although the release of phytosiderophores was not mea-
sured in this experiment, it is likely that the mobilization of Fe from the primary
Fe-bearing minerals contained in the basalt was related to phytosiderophores ex-
creted by this graminaceous species. These results suggest that in addition to be-
ing of prime importance for plant nutrition, root exudates such as phytosidero-
phores may play a significant role in mineral weathering and pedogenesis
(Femandes Barros and Hinsinger, 1994) as already shown for numerous organic
substances excreted by soil microorganisms (Robert and Berthelin, 1986).
As previously mentioned, most root exudates act through an enhanced dissolu-
tion of the metal-bearing compound, increased release of the metal, and subse-
quent increase in its mobility toward the root. In some instances, however, the
 HOW DO PLANT ROOTS ACQUIRE MINERAL NUTRIENTS? 253

metalkxudate complexes can precipitate in the rhizosphere, resulting in an im-

mobilization of metals, as can occur for Ca in calcareous soils (Dinkelaker et al.,
1989) or A1 in acid soils (Dinkelaker etal., 1993; Jones and Darrah, 1994). As sug-
gested by Dinkelaker et al. (1993), the root-induced complexation of A1 due to the
release of complexing exudates into the rhizosphere of Norway spruce may part-
ly explain the tolerance of this species to A1 toxicity. Hue et al. (1986) clearly
showed that organic acids can detoxify A1 in acid soils. In their experiments, the
addition of organic acids alleviated the alteration of root elongation of cotton
seedlings, which normally occurs at concentrations of monomeric A1 above 1 IJ.M
in the bulk soil solution. Hue et al. (1986) showed that the most efficient A1 detox-
ifying organic acids were those that formed the most stable complexes with Al, no-
tably citric and oxalic acids, and to a lesser extent tartaric, malic, and malonic
acids. This work and that of others (e.g., Pellet et al., 1995) suggests that these root
exudates, which are commonly found in the rhizosphere, may contribute to the
adaptation of plants to A1 toxicity in acid soils.

Vn. OTHER INTERACTIONS INVOLVING

ROOT EXUDATES

Other very specifically oriented substances are produced by plant roots, such as
phosphatase and phytase ectoenzymes, that may play a major role in the catalytic
hydrolysis of organic P and in the subsequent acquisition of soil P (Tarafdar and
Jungk, 1987; Findenegg and Nelemans, 1993). This phenomenon might be criti-
cal, since 2040% of total soil Pis present as organic P (Mengel and Kirkby, 1987).
Tarafdar and Jungk ( I 987) showed a 3- to 10-fold increase in acid phosphatase ac-
tivity in the rhizosphere of onion, oilseed rape, clover, and wheat relative to bulk
soil and up to about a 3-fold increase in activity for alkaline phosphatase. An in-
crease in phosphatase activity in rhizosphere relative to bulk soil was also report-
ed for oilseed rape (Hedley er al., 1982b), maize (Dinkelaker and Marschner,
1992), lupins (Adams and Pate, 1992), and forest tree species such as Norway
spruce (Haussling and Marschner, 1989). In the work of Tarafdar and Jungk (1987)
the diverse phosphatases excreted in the vicinity of plant roots were likely to be
responsible for the concomitant, significant mobilization of soil organic P that was
indicated by the depletion of organic P in the rhizosphere (see Table I). Because
they concurrently measured an increase in both fungal and bacterial biomasses in
the rhizosphere relative to bulk soil, they could not definitively conclude whether
these diverse rhizosphere phosphatases were plant-borne or of microbial origin.
Experiments with axenic plants, however, have established that plant-borne phos-
phatases are excreted into the rhizosphere in the absence of microorganisms
(Amann and Amberger, 1989; Grimal et al., 1992) and may account for a sub-
254 P. HIN'SINGER

stantial part of the increase in phosphatase activity near plant roots (McLachlan,
1980). In addition, the excretion of phosphatase by plant roots has been shown to
be stimulated by P deficiency (McLachlan, 1980; Amann and Amberger, 1989;
Grimal et al., 1992), suggesting that the release of these enzymes should be con-
sidered as an adaptative response of the P-deficient plant. However, in addition to
their possible degradation by microbes, the competition of these enzymes with mi-
crobial phosphatases and their possible inactivation by adsorption onto soil reac-
tive components such as clay minerals (Quiquampoix et al., 1995; Leprince and
Quiquampoix, 1996) bring into question their effective role in soil environments.
Further work is needed in this area, since in many soils organic Pcontributes a ma-
jor proportion of soil P.
Major root exudates are the so-called mucilage-a gelatinous material made of
high-molecular-weight polysaccharides (Curl and Truelove, 1986). Polyuronic
acids that are well known for their important role in the cation exchange capacity
of the root cell walls account for a large proportion of this mucilagenous exudate.
The consequent exchange properties of mucilage explain their ability to bind
heavy metals such as Pb and Cd or micronutrients such as Cu and Zn (Morel et al.,
1986; Mench et al., 1987). In acid soils, Al can similarly be detoxified by a mas-
sive adsorption on mucilage (Horst et al., 1982). In addition to these binding prop-
erties of polyuronic sites in mucilage with respect to metal cations, polyuronate
ions may help desorb some anions, such as phosphate ions sorbed on soil miner-
als as shown for polygalacturonate by Nagarajah et al. (1970). Such a process
agrees with the findings of Grimal er al. (1995). They showed that mucilage ex-
creted by axenic-grown plants was sorbed on goethite, whereas phosphate was
desorbed from goethite in the rhizosphere of maize. Many other benefits have been
attributed to mucilage, including their role in establishing a better contact between
the roots and the porous soil matrix (Uren and Reisenauer, 1988), thereby im-
proving the transfer of water and mineral nutrients to the roots.

VIII. CONCLUSION

The rhizosphere, i.e., the volume of soil that is influenced by root activities, can
exhibit drastically different conditions compared with the bulk soil. Since the rhi-
zosphere conditions are those that are encountered by plant roots, understanding
them is critical to improving our knowledge of root functioning and plant nutri-
tion. The rhizosphere was once recognized only for its singular microbiology.
However, over the last two or three decades, evidence has accumulated that severe
changes in chemical conditions relative to the bulk soil are a major trait of the rhi-
zosphere. This review has concentrated on those peculiar modifications of chem-
ical conditions that are occurring in the rhizosphere as a direct consequence of the
 HOW DO PLANT ROOTS ACQUIRE MINERAL NUTRIENTS? 255

activity of plant roots. Obviously, some of these, such as many changes in ionic
concentrations and pH, are due simply to the uptake activity of the root. Although
changes in pH have received considerable attention, modifications of ionic con-
centrations certainly have to be considered as equally important and universally
widespread features of rhizosphere chemistry. Indeed, ionic concentrations and pH
are critical parameters that control many chemical reactions occumng at the
root-soil interface. Root-induced changes of these conditions will therefore influ-
ence the dynamics of many nutrients in the rhizosphere and ultimately their ac-
quisition by plant roots. The uptake of nutrients thus operates as a major driving
force in nutrient acquisition. It should, however, no longer be solely considered as
the ultimate mechanism involved in plant nutrition. One has to bear in mind that
nutrient uptake has a major effect on the chemical conditions occumng in the rhi-
zosphere, which will reciprocally determine the uptake activity of the root.
As evidenced mostly over the last decade, in addition to these interactions, oth-
er chemical processes, such as redox reactions, complexation, or enzymatic catal-
yses, can take place in the rhizosphere as a direct consequence of the exudation of
more or less specifically oriented metabolites produced by plant roots. The exu-
dation of organic acids and enzymes, for instance, can contribute a significant pro-
portion of the supply of major nutrients such as P to plant roots. Similarly, the ex-
udation of phytosiderophores by roots of grass species plays a major role in the
acquisition of poorly mobile micronutrients such as Fe and many other metals. A
better understanding of these peculiar chemical processes occurring at the
root-soil interface is thus a prerequisite for more accurately predicting the nutri-
tion needs of plants and the risks of undesirable micropollutants such as heavy met-
als entering the food chain.
Many of the aforementioned processes seem to be induced or stimulated in re-
sponse to nutrient deficiencies. This suggests that they may be regarded as strate-
gies of plant nutrition that evolved among higher plants to overcome adverse soil
chemical conditions (Marschner, 1995). Whether these processes can be consid-
ered as such, it should be borne in mind that the acquisition of mineral nutrients
not only relies on these diverse chemical processes but is largely influenced by (1)
the colonization of the soil by the root system and (2) the physical properties of
the intimate contact between the roots and the solid, liquid, and gaseous phases of
the soil. Considerable progress has been made in improving our knowledge of root
growth and rooting patterns (architecture of the root systems). In comparison, only
a limited amount of scientific data is relevant to the physical dimension of root-soil
interactions occurring in the rhizosphere. Thus, further investigations are needed
in this area.
In this chapter, chemical processes that occur in the rhizosphere as a direct con-
sequence of root activity were addressed. However, other processes that signifi-
cantly contribute to plant nutrition occur as a result of rhizosphere microflora. This
phenomenon can be regarded as an indirect effect of plant roots, since the activi-
256 P. HINSINGER

ty of microorganisms in the rhizosphere is largely supported by root exudation of

C compounds. Although this “rhizosphere effect” has been studied over almost a
century, many questions remain, especially regarding its actual benefit for plant
nutrition and plant growth. For instance, rhizosphere microorganisms are likely to
rapidly degrade those exudates-such as organic anions, phytosiderophores, and
enzymes-that are supposed to assist the plant in acquiring some mineral nutri-
ents. They also compete with plant roots for mineral nutrients. Rhizosphere mi-
croflora can thus have a detrimental effect on plant nutrition. The energetic cost of
rhizosphere microflora has been particularly addressed in the case of symbiotic
rhizosphere microorganisms such as N,-fixing bacteria and mycorrhizal fungi.
Nevertheless, mycorrhized plants often have a better P status than do nonmycor-
rhized plants, and over 95% of plant species are indeed mycorrhized. More inter-
estingly, some species that are never mycorrhized, such as oilseed rape and white
lupin, among crops, and many members of the Proteaceae family, among wild
species (Harley and Harley, 1987; Brundrett and Abbott, 1991), have been report-
ed as being some of the most efficient species for mobilizing soil P. This is attrib-
uted to their peculiar ability to excrete considerable amounts of protons and/or or-
ganic anions, such as citrate in particular. One may thus question whether these
root-induced chemical processes evolved in these species to compensate for the
lack of mycorrhizal support in P acquisition.Whatever the answer, the occurrence
of such plant species suggests that some rhizosphere characteristics may be worth
taking into account in plant breeding programs.
In today’s world, where conventional, intensive agricultural practices are being
challenged for both economic and environmental reasons, we should no longer
breed crops and pasture species that give a maximum yield under optimal grow-
ing conditions.From the plant-nutritionview point, this practice assumes that such
optimal conditions can be achieved with an adequate, and most often massive, use
of fertilizers. Sustainableagriculture, however, requires moderate consumption of
fertilizers. In this perspective, we should aim instead at selecting those species and
varieties that can most efficiently cope with a range of nonoptimal soil conditions.
A prerequisite to incorporating such considerations into our breeding programs is
a better understanding of the actual, combined effect on nutrient acquisition of the
various processes that occur in the rhizosphere. New experimental tools derived
from molecular biology, such as using mutants and genetic manipulations,will cer-
tainly help in ascertaining the relative contribution of the numerous mechanisms
that are involved. Moreover, using mathematical models of the combined phe-
nomena involved in the process of mineral nutrient acquisition will also help us to
improve our understanding of plant nutrition. For this purpose, as pointed out by
Darrah (1993), a more integrative and quantitative approach of rhizosphere
processes is indeed required. This is a fundamentalprerequisite to managing plant
nutrition in agricultural, forested, and natural environments.
 HOW DO PLANT ROOTS ACQUIRE MINERAL NUTRIENTS? 257

ACKNOWLEDGMENTS

This chapter is dedicated to the memory of Professor Horst Marschner, who contributed much to our
understanding of the chemical processes involved in the rhizosphere. I also thank Professor R. J. Gilkes,
Dr. J. C. Arvieu, and Dr. B. Jaillard for their comments on an earlier version of this chapter.

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