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Absorption of boron by plant roots

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c 1997 Kluwer Academic Publishers. Printed in the Netherlands.

Chapter 4

Absorption of boron by plant roots

Hening Hu and Patrick H. Brown

Department of Pomology, University of California, Davis, CA 95616, USA

Abstract

Experimental evidence suggests that B uptake is the result of the passive assimilation of undissociated boric acid.
Boron uptake by a particular species should, therefore, be primarily determined by the B concentration in the
soil solution and the rate of water uptake by the plant. This simple explanation of B uptake, however, does not
adequately explain field observations where dramatic differences in B concentrations are observed between species,
even when these species are grown under similar environmental conditions. The apparent contradiction between
experimental results and in field observations, suggests, that B uptake is determined by factors that are as yet
unknown. In the following, we discuss experimental and field observations as they relate to B uptake and discuss
the mechanisms that may be involved in determining B uptake in diverse species.

Introduction results demonstrate that species can differ significant-

ly in their rate of B accumulation even when grown
Boron is an essential micronutrient for higher plants, under identical environmental conditions. This contra-
and B deficiency results in rapid inhibition of plant diction cannot be explained adequately by our current
growth. The rapid and specific inhibition of plant understanding of B absorption mechanisms.
growth that occurs upon removal of B is a consequence In this chapter we review current information on
of two important features of B physiology: the specific the mechanism of B absorption by plants and describe
structural role B plays in the cell wall (Hu and Brown, the biotic and abiotic factors that influence this pro-
1994; Loomis and Durst, 1992; Matoh, Chapter 5) and cess. This information is then discussed in light of
the limited mobility of B in the majority of species observed differences in B absorption amongst culti-
(Brown and Shelp, Chapter 7; Oertli and Richardson, vars and species.
1970). As a result of its critical role in expanding tis-
sues and its limited mobility, B has to be supplied
continually throughout the life of the plant, usually Mechanisms of boron absorption
through the root. For this reason, knowledge of the
physiology of B absorption is essential. Properties of boric acid
Boron is absorbed from soil solution by roots main-
ly as the undissociated boric acid. Theoretical consid- Boric acid is a very weak acid in aqueous solution:
erations predict that undissociated boric acid should be its activity as an acid appears to be related to OH
membrane permeable; in agreement with this, recent acceptance by the B(OH)3 rather than to H+ donation
experimental results suggest that B absorption is a pas- according to the following reaction:
sive, non-metabolic process. Based solely upon these
conclusions, it is predicted that B absorption is pri-
B(OH)3 + 2H2 O = B(OH)4 + H3 O
+ (pKa 9:25)
marily determined by B concentration in the uptake
medium and the transpiration rate of the plant. How-
Thus, in neutral or slightly acid soils, common-
ever, extensive field observations and experimental
ly occurring in plant growth environments, B exists
 E-mail: [email protected] mainly as undissociated boric acid (Raven, 1980).

*141783*
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50

Boric acid is soluble in water and has an ether– type of chain linkage, the permeability of water can
water partition coefficient of 0.035. Raven (1980) differ by up to 3-fold from one kind of lipid vesicles to
computed the permeability of plant cell membranes another.
to B(OH)3 on the basis of the ‘lipid solution’ mecha- In summary, it is clear that calculations of B fluxes
nism. Using the linear relationship between PM3=2 (P = based solely on theoretical lipid permeability are inad-
permeability coefficient, M = mol. wt) and the ether– equate to predict B absorption by plants. Genotypes
water partition coefficient, the computed value of P within a species could differ several fold in B perme-
for B(OH)3 is 810 6 cm s 1 . In another approach ability (see “the paradox of boron uptake”), which may
(Raven, 1980), using the linear relationship between have a physiologically important effect on B uptake by
log PM1=2 and N (the number of potential H-bonding roots.
groups, N = 2 for –OH, and N = 6 for B(OH)3 ), the
estimated P for B(OH)3 is 410 6 cm s 1 . This the- Role of boron complex formation in boron absorption
oretical calculation led Raven (1980) to suggest that
the permeability coefficient is high enough to account Another important property of boric acid that will influ-
for the measured magnitude of boric acid fluxes across ence B absorption is its ability to form cis-diol com-
many plant cell membranes. The use of active trans- plexes with a variety of organic molecules. For exam-
port of boric acid to maintain B distribution across a ple, in celery (Apium graveolens), B present in the cell
membrane away from thermodynamic equilibrium is sap and phloem exudates occurs almost entirely as the
consequently likely to be energetically expensive. On B–mannitol complex. No free boric acid was detected.
this basis, Raven (1980) predicted that B absorption by The reaction of boric acid with mannitol is described
plant roots would be a passive process. in the following reaction (Hu et al., 1997):
The conclusions made by Raven (1980) though
sound, may be over simplified. The ‘high’ cost of main-
taining a non-equilibrium concentration of an element, H3 BO3 + 2C6 O6 H14 =
mannitol
does not preclude the possibility of active absorption.
[(C6 O6 H12 )B(C6 O6 H12 )]
+ + 2H2 O
+H3 O
Protons, for example, have an estimated membrane [mannitol B mannitol ]
permeability of 10 3 to 10 4 cm s 1 (Dencher et al.,
1986), which is 100 to 1000 times more permeable The equilibrium constant for the reaction is 10 4 .
than boric acid. However, active H+ transport occurs The formation of borate–diol complexes has a much
in all living systems (Briskin and Reynolds-Niesman, lower pKa than that of borate. The pKa of B–mannitol
1991). complex (about 5.2) is four units lower than that of
Additionally, calculations of membrane permeabil- free boric acid (9.25) (Raven, 1980), strongly suggest-
ity based upon ether:water partition coefficients are ing that at normal B supply free boric acid concentra-
undoubtedly over simplified and may be misleading. tions inside the cytoplasm of celery (pH about 6.2–6.5)
For example, urea has a similar molecular wt (60) would approach zero. This situation is true for celery,
to boric acid (62) and is also uncharged at neutral in which mannitol is present at millimolar concentra-
pH. Finkelstein (1976) reported Purea in an artificial tions, and would also be true for other species where
membrane to be 0.6–4.010 6 cm s 1 , while Lee- B-chelating diols are present at millimolar or greater
Standelmann et al. (1991) calculated Purea in bar- concentration.
ley (Hordeum vulgare) to be 2.3–2.610 5 cm s 1 . There are a great number of biological compounds
Hence, membrane permeability to urea in barley was that can form complexes with B, both in the cytoplasm
found to be an order of magnitude greater than in and in the cell wall. Compounds capable of com-
an artificial membrane. Similarly, the permeability of plexing with boric acid include sugars, their deriva-
water can vary significantly between membranes of tives (sugar alcohols), phenols, organic acids, and
different composition and cannot be adequately pre- some polymers (Boeseken, 1949; Raven, 1980). Com-
dicted from theoretical considerations alone. Jansen mon examples are sorbitol, mannitol, glycerol, ribose,
and Blume (1995) measured the water permeability of apiose, nicotinamide-adenine-dinucleotide (NAD) and
lipid vesicles composed of a variety of different phos- fructose (Loomis and Durst, 1992; Makkee et al.,
pholipids with different head groups and fatty acyl 1985). Low molecular weight B-complexes with man-
chains. Depending on the nature of the head group, on nitol, sorbitol and fructose have been recently isolated
the chemical structure of the side chains and on the and characterised from plant tissue (Hu et al., 1997;

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 51

Penn et al., 1997). In addition, B complexes with concentration regardless of pH. In a time study from
rhamnogalacturonan-II in the cell wall have been char- 1 to 24 h, they did not find B accumulation within the
acterised (Ishii and Matsunaga, 1996; Kobayashi et al., root, nor did they find B accumulation against a con-
1996; O’Neill et al., 1996). centration gradient in solution B concentration ranges
The presence of significant amounts of B- from 0.0185 to 7.39 mM. Within 3 to 4 h of transfer to
complexing compounds in plant cells would have a CaSO4 solution, approximately 95% of the B, initial-
significant effect on B uptake if uptake is solely deter- ly absorbed by the root tissue was lost to the CaSO4
mined by the free boric acid concentration gradient solution. At pH 6.0, changes in substrate temperature
across the membrane. Raven (1980) suggested that the (from 10 to 30  C), salt composition (Ca or K salt) or
total B content of the tissue can greatly exceed the free addition of KCN and 2,4-dinitrophenol (DNP) failed to
boric acid concentration in the bathing solution due to exert any influence on B absorption. These results led
the formation of B complexes, so that the demonstra- Bingham et al. (1970) to conclude that B absorption is
tion of a net B influx into a tissue, in which the total a physical, non-metabolic process acting in response to
internal B concentration exceeds that of external boric B concentration gradient and that undissociated boric
acid concentration is not adequate to show that active acid is favoured for absorption.
transport is occurring. Wilders and Neales (1971) used sliced disks from
The uncharged nature of boric acid, the relatively carrot (Daucus carota) and red beet (Beta vulgaris)
high (though poorly defined) membrane permeability storage roots suspended in 0.1 m M boric acid solutions
of boric acid, combined with its propensity to form for absorption or desorption studies. They reported that
complexes both in the cell wall and in the cytoplasm disks of carrot and red beet absorb B to an equilibrium
greatly complicates the study of B absorption. Classic internal concentration which is greater than that of the
analysis of elemental uptake based upon determination external solution and that this accumulation of B in the
of transmembrane electrochemical gradients, analysis tissues is inhibited by anoxia, by DNP and also by low
of absorption kinetics and measurement of electrical temperature, suggesting active uptake. On the other
changes upon ion uptake do not readily apply to studies hand, B content of storage tissues, after a period of
of B. Nevertheless, important insights into the mecha- absorption in 0.1 m M boric acid solution, was rapidly
nism of B absorption can be gained from a review of and almost completely desorbed into a B free solution,
existing experimental data. suggesting a passive, diffusion type process. There-
fore, Wilders and Neales (1971) concluded that there
Is boron absorption active or passive? are two components of B uptake: a passive diffusion
of B(OH)3 and an active transport for B(OH)4 ion.
Over the last 30 years this question has been addressed Wilders and Neales (1971) based their conclusion
by many research groups with varied and conflicting of active uptake on 1) the observation that internal B
results. The reason is partly due to the characteristic concentrations were greater than the external B con-
permeability of boric acid across cell membranes and centrations as a result of the transport of the B(OH)4
the significant effects of cis-diol complex formation ion; and 2) that uptake was inhibited by anoxia and
which adds considerable complexity to the interpre- DNP. Their conclusion of active absorption based upon
tation of uptake experiments. Several of the studies the occurrence of higher internal B concentrations is
of B absorption were conducted using unrealistically not, however, supported by their data. The passive
high B concentrations with highly variable absorption membrane permeability of B(OH)3 (suggested by the
or desorption periods, and in only rare cases has the authors, and proposed by Raven, 1980) predicts that
compartmentation of B within the root been adequately B can diffuse into both internal and external solution.
considered. In other words, the internal B concentration would be
Bingham et al. (1970) conducted one of the first equilibrated with the external B concentration. From
detailed studies of B absorption using excised roots of the desorption data of Wilders and Neales (1971), we
barley. With a B concentration of 0.93 mM and a 4 h can calculate that B(OH)3 passively crosses the mem-
absorption period, a strong pH dependency in the alka- brane at a rate of at least 0.1 mol h 1 g 1 fresh wt,
line range for B absorption was found. Analysis of the while the B accumulation rate was more than 0.1 mol
pH effect on the proportion of B(OH)3 and B(OH)4 h 1 g 1 fresh wt. This means that the cells had to pump
in the absorption solution reveals that B absorption by B internally at a rate of at least 0.2 mol h 1 g 1 fresh
barley roots was primarily contingent upon the B(OH)3 wt to obtain a net accumulation of 0.1 mol h 1 g 1

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52

(1976), because the barley roots used were only rinsed

for 30 s or 1 min after B absorption. As a result, much
B remaining in the water free space and free space
in the cell wall may have been wrongly considered to
be absorbed B. However, it is not a simple matter to
determine what is an appropriate time for B desorption.
For example, if active uptake is the primary mode of
B accumulation, then the desorption period should be
long enough to remove all the non-specifically bound B
from the free wall space, e.g. 30 min (Bowen and Nis-
sen, 1976). On the other hand, if passive B uptake pre-
dominates, then the desorption should be short enough
to avoid passive desorption of B across the membrane,
while still removing the majority of the apoplastic B.
This is clearly an unattainable task for a highly per-
meable compound such as boric acid. Unfortunately,
the significance of the desorption period has not been
adequately considered in most studies of B absorption.
Figure 1. Relative uptake of boron as a function of solution pH. Bowen and Nissen (1977) used excised roots from
Uptake at pH 6 = 100% at each supply concentration. Solid line: per- 5 to 7 day old barley seedlings in their short term
centage of undissociated H3 BO3 . Adapted from Oertli and Grgurevic
(1975). (hours) B absorption studies. Within the concentration
range used, 0.2 – 5 mM, they did not find B accu-
mulation against a concentration gradient after 5 h in
fresh wt. It is extremely doubtful, that given the lim- the absorption solution. However, when they studied B
ited surface area provided by the storage root disks absorption as a function of the external B concentration
used, that B(OH)4 could be pumped into the cells at a over the range of 0.05 – 13.3 mM, and plotted the data
rate of 0.2 mol h 1 g 1 fresh wt. This is particularly as a double reciprocal plot, they observed that the data
improbable given the external B(OH)4 concentration could be described by a series of straight lines (phas-
of 32 nM (0.03% of 0.1 mM boric acid at pH 5.7) es) separated by sharp transitions or ‘jumps’, which
utilised by the authors. It seems clear that Wilders occurred at B concentrations of 0.16, 0.20, 0.47, 0.55
and Neales (1971) conclusion of active uptake was the and 8 mM, respectively. Based on these results, they
result of insufficient tissue rinsing and may also have concluded that B absorption in barley is an active trans-
been influenced by the relatively high concentrations port process since it is compatible with the concept of
of B-binding ligands in carrot (mannitol) and fructose multiphasic absorption mechanisms outlined by Nis-
in sugar beet (Beta vulgaris). sen (1974). To further support their argument, Bowen
Oertli and Grgurevic (1975) used excised root sec- and Nissen (1977) reported that B uptake from 1 mM
tions of 6 day old barley seedlings and studied the pH B solution was strongly dependent upon temperature
effect on B absorption. Changes in pH from 6 to 10 and over 3 – 47  C range, and also that B uptake from 1
B concentration from 0.093 to 0.93 m M suggested that mM B solution was inhibited by metabolic inhibitors,
B uptake and the final B contents increased nearly lin- including 0.05 mM DNP, 0.05 mM NaN3, 1 mM NaCN,
early with increasing B supply. The relative B uptake 5 mM arsenate and 0.05 mM dicoumarol.
(uptake at pH 6 = 100%) at all supply levels decreased A major concern with Bowen and Nissen’s (1977)
with increasing pH similar to the decrease of the frac- study is that the B concentrations used were much
tion of undissociated B(OH)3 (Figure 1). At pH 6, final higher (0.05 – 13.3 mM) than is physiologically rele-
B concentrations in roots and treatment solutions were vant for barley. It has been reported that barley plants
similar. Based on these results, they concluded that B have a low B requirement (Hu et al., 1996; Loomis and
in root tissues and external solution tends to approach Durst, 1992), while sensitive barley cultivars develop
a diffusion equilibrium which is governed by the pro- B toxicity symptoms after 11 days growth at solution
portion of B(OH)3 vs B(OH)4 in the system. B concentrations of 0.2 mM, and moderately resistant
The work of Bingham et al. (1970) and Oertli and cultivars developed toxicity 17 days after exposure to
Grgurevic (1975) was criticised by Bowen and Nissen 0.5 mM B (Nable, 1988). Bowen and Nissen (1977)

plso4del.tex; 1/10/1997; 19:13; v.7; p.4

 53

reported that B absorption is linear over a wide range of

B concentrations (0–0.2 mM or 0.2–10 mM) in cultured
tobacco (Nicotiana tabacum) cells and in the roots of
sunflower (Helianthus annuus) and squash (Cucurbita
pepo). There was no indication of saturation kinetics
and B absorption was not inhibited by DNP or KCN.
Although B absorption did respond to temperature in
cultured tobacco cells, absorption was not inhibited
by very low (2  C) temperature, and no inhibition of
uptake was reached at temperatures as high as 47  C
(Figure 2). The temperature effect was not typical of
temperature dependent kinetics, and the Q10 was less
than 2. On the basis of these results it was concluded
that shifts in B absorption with temperature were asso-
ciated with changes in membrane conformation (and
hence permeability) and the effects of temperature on
the relative activity of B molecules in solution.
In these experiments, 11 B incorporated during pre-
treatment growth was not displaced by 10 B in the
Figure 2. Influence of temperature on the uptake of 10 B into 11 B
precultured intact tobacco cells. Cells were placed in 0.1 mM 10 B uptake solution even in freeze-thaw killed cells, sug-
uptake medium for 30 min at various temperatures then transferred gesting that B can be incorporated into some non-
to B free desorption solution for 30 min at 5  C. Values are means exchangeable complexes within the cell (Brown and
 SE. Adapted from Brown and Hu (1994).
Hu, 1994). In desorption studies with cultured tobacco
cells, after a 30 min uptake period in either 0.1 or 10
mM10 B solution, it was found that B desorption was
observed transitions in B uptake at concentrations of almost complete within 1 min of transfer to B free solu-
0.47, 0.55 and 8 mM, which are well in excess of tion. This rapid desorption of B probably represents
the concentrations that are required for optimal bar- B within apoplastic spaces. Further loss of recently
ley growth. Therefore, it is extremely unlikely that acquired 10B was very limited. Based on these findings,
the uptake kinetics described by Bowen and Nissen Brown and Hu (1994) concluded that B absorption is
(1977) are of any physiological relevance. The tran- a non-metabolic process, which is controlled, in large,
sitions observed by Bowen and Nissen (1977) may by the formation of non-exchangeable B-complexes in
be better explained by the saturation of the B-binding the cytoplasm and cell wall.
capacity of various cellular compartments. In summary, an unequivocal conclusion about B
Thellier et al. (1979) conducted a B(OH)3 flux uptake mechanism can not be made at this time. The
study using isotopes of 10 B and 11 B in duckweed (Lem- reason is partly a result of the diverse plant materials
na minor). After a period in 10 B uptake solutions, and B concentrations used by different authors, the
plants were placed in either 11 B boric acid or pure failure to distinguish B compartmentation and the fail-
water to determine efflux kinetics. The results sug- ure to adequately assess B complex formation. It is
gested that four compartments existed within the cell, clear, however, that there is little or no sound evidence
i.e. free space, cytoplasm, vacuole and the cell wall. supporting active B absorption. Evidence to support a
Each of these compartments contained B concentra- non-metabolic process for B absorption, is somewhat
tions much higher than that of the external solution more convincing though far from conclusive. We pro-
(0.16 mM). They did not interpret the occurrence of pose that B absorption can be best explained as a pas-
higher internal than external B concentrations as evi- sive diffusion of free boric acid into the cell, followed
dence of active absorption of B, but attributed this to by a rapid formation of B-complexes within the cyto-
the formation of borate mono- or di-ester within the plasm and the cell wall. The decline in free boric acid
various compartments and subsequent exchange of B within the cell by the formation of B-complexes allows
isotopes. further absorption of B from the external solution and
Using stable B isotopes and inductively coupled results in tissue B concentration that can greatly exceed
plasma-mass spectrometry, Brown and Hu (1994) the free boric acid concentration in the uptake solution.

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54

Factors affecting boron absorption water consumption as well as B uptake, and that the
uptake of water and B was reduced 50% at high humid-
Boron absorption by plant roots is affected by various ity in comparison to low humidity. However, the B to
environmental factors, both in the soil and non-soil water uptake ratio remained constant (about 3.4 g
environment. Important factors influencing B adsorp- B mL 1 water at 0.93 mM external B) irrespective
tion from solution include the initial B content of the of humidity or temperature treatment. Temperature
soil, the pH of the soil, the type of exchangeable ions increased water and B absorption significantly at con-
present in the soil solution, the amounts and types of stant relative humidity (as a consequence of differences
minerals in the soil, the soil organic matter content, in vapour pressure deficits). For example, at an external
the wetting and drying cycles, and the water to soil B concentration of 0.05 mM, B absorption was about
ratio (Goldberg, Chapter 3; Keren et al., 1985). All 0.2 g per barley seedling at a temperature of 10  C.
these factors affect B absorption by plants. In coarse When temperature increased to 27  C, B absorption
textured, low organic matter, and high pH soils, low B was about 0.65 g per barley seedling. However, the
availability to the plants would be expected. ratio of B absorption to water uptake remained constant
Among the soil factors, pH is most important in at 0.39 g B mL 1 water. On the other hand, Oertli
affecting B absorption by plants (Gupta, 1979). In gen- (1994) found that the ratio of B absorption to water
eral, plant B absorption is decreased with the increase uptake did not correspond to the external B concentra-
in soil pH because of two reasons. Firstly, below pH tion. For example, at external B concentration of 0.05,
7, B(OH)3 predominates and since the affinity of soil 0.23 and 0.93 mM, the B to water uptake ratio was
clay for this species is relatively low, the amount of 0.39, 1.07 and 3.46 g B mL 1 water, respectively.
B adsorption by soil is small. As the pH increases, Forno et al. (1979) reported that two cassava (Mani-
B(OH)4 concentration relative to B(OH)3 increases, hot sp.) cultivars growing in nutrient solution devel-
and as a consequence of the relatively strong affinity oped B deficient symptoms when growing at root tem-
of B(OH)4 for clay minerals, the amount of adsorbed B perature of 18  C; their shoot B concentration ranged
increases (Keren and Bingham, 1985). In other words, from 9.7 to 11.9 mg B kg 1 dry wt. However, when
with the increase in soil pH, B availability to the root these plants were grown at 23  C root temperature,
decreases. For more information about reactions of B they developed much less severe B deficiency symp-
with soils, the reader is referred to Chapter 3 in this toms and symptoms appeared only towards the end of
volume. Secondly, Oertli and Grgurevic (1975) found the experiment. These plants accumulated 17 – 19 mg
that the absorption of B by solution grown barley roots B kg 1 dry wt in the shoot. Plants were symptom free at
decreased with the increase of solution pH, this corre- root temperatures of 28 – 33  C and they accumulated
sponded to the decrease in the fraction of undissociated 42 – 55 mg B kg 1 dry wt in the shoot.
B(OH)3 , demonstrating that B(OH)3 is the form of B An exception to the temperature effect on B absorp-
available to plants. tion described above was reported by Nable et al.
In non-soil environments, the transpiration rate (1990). These authors reported that nutrient solution
seems to be the most important factor governing B temperature had little effect on tissue B concentration
absorption. As a result, relative humidity, temperature within cultivars of barley. For example, at 15 M B
and light intensity will all alter B absorption. For exam- solution, B absorption in ‘Sahara 3771’ was the same
ple, Bowen (1972) made a comprehensive study about (20 mg B kg 1 dry wt) at 5 and 25  C. At 1 mM B in
the environment factors on B absorption and found solution, an increase in temperature from 5 to 25  C
that in sugarcane (Saccharum officinarum) seedlings, resulted in an increase of B absorption from 80 to 90 mg
increasing light intensity increased B absorption, while B kg 1 . However, different genotypes exhibited dif-
decreasing relative humidity increased B absorption. ferences in B accumulation.
Similarly, an increase in air temperature increased The reported environmental effects on B absorption
total B absorption even though the relative humidi- can largely be explained by changes in transpiration
ty was held constant. When other environment con- (Raven, 1980). Reduced humidity increases transpira-
ditions were constant, an increase in root temperature tion, resulting in an increase in B absorption. Increased
increased B absorption. temperature creates water vapour deficits, which will
Similar information was reported by Oertli (1994) also increase transpiration, resulting in an increase in
in barley seedlings. He found that increasing duration B uptake. Illumination has the same effect as increased
and intensity of illumination significantly increased temperature or reduced relative humidity, and as a

plso4del.tex; 1/10/1997; 19:13; v.7; p.6

 55

result, transpiration increases and B uptake increas- Firstly, B uptake may be partially under metabol-
es. However, the observed increases in B uptake with ic control. For example, Bowen and Nissen (1977)
the increase in root temperature can not be explained reported that active uptake accounts for less than 10%
by transpiration effects. No simple relationship exists of the total B accumulated by barley roots. Alternative-
between transpiration rates and whole plant B uptake. ly, there may be an active B exclusion mechanism. In
For example, in wheat (Triticum aestivum) cultivars, this regard, the extent of active uptake may be differ-
water use efficiency ranged from 3.1 to 4.0 g dry wt ent from species to species or from genotype to geno-
kg 1 water. The less than 30% difference in transpira- type. This mechanism may explain the differences in B
tion rate within cultivars of one species can not account absorption among different species (genotypes) and it
for the 400–700% difference in B uptake (Nable, could also account for the apparently conflicting results
1988). Further, genotype differences in resistance to obtained when metabolic inhibitors are used. Howev-
B toxicity and B absorption are known to occur in er, in view of the discussion in “Mechanisms of boron
roots assayed without the shoot attached and B uptake absorption”, we conclude there is little support for this
in these roots has been frequently observed demon- concept.
strating that transpiration is not essential for uptake Alternatively, one plausible explanation is that the
(Huang and Graham, 1990). exudation of B complexing agents into the rhizosphere
restricts B uptake from the soil. The extent to which B-
binding exudates are produced may be species depen-
The paradox of boron uptake dent. Organic compounds containing cis-diols, as well
as some amino acids or some organic acids can com-
The preponderance of available experimental evidence plex with B. Because the resultant B-complex is gen-
suggests that B uptake is a passive process, the rate of erally an anion, it would have a greatly reduced mem-
which is influenced by passive diffusion of B through brane permeability in comparison to uncharged boric
the membrane, the formation of B complexes within acid, and hence absorption would be reduced. Many
the cell and plant water fluxes. If these assumptions compounds present in root exudates, particularly the
are true then one would predict that B uptake would be pectin-rich polysaccharides that are abundant in plant
very closely correlated with water use and that species mucilage, would be capable of forming stable B com-
grown in like environments supplied with non-limiting plexes. These complexes could realistically reduce B
water would have very similar B uptake rates. Never- uptake. However, one might expect that these exudates
theless, there is clear evidence that B uptake differs would be saturated by B at high external B supply
dramatically between species and even within geno- and once saturated, uptake should show no difference
types of a single species. For example, Nable (1988) among genotypes. Even at the very high B application
reported that in solution culture experiments, the B rates (6.4 mM) used by Nable et al. (1990), this did
concentration or total B content in all organs of five not appear to be the case. Also, one would expect that
barley and six wheat cultivars differ dramatically. The experiments conducted in solution culture (in which
cultivars resistant to B toxicity always accumulated rhizosphere exudates would be diluted in the culture
less B than the sensitive cultivars. When grown in 5 medium) would result in diminished genotype differ-
mM B solution, barley cultivars ‘Sahara 3763’ and ences. In the relatively long term studies of Nable et
‘Schooner’ accumulated 112 and 710 mg B kg 1 dry al. (1990) (see also Nable, 1988; Paull et al., 1988)
wt in the youngest expanded leaf blade, respectively, a this did not appear to be true. On the other hand, geno-
more than 6-fold difference in B accumulation between type differences in long term B uptake studies have not
genotypes. These differences in B uptake cannot be been confirmed in short term solution culture experi-
explained through differences in water use, which has ments (Garnett et al., 1993). Nevertheless, this hypoth-
less than 1-fold difference among the cultivars. This esis cannot be entirely excluded without more specific
apparent contradiction between passive B uptake and information on the amount and localisation of these
significant differences among the genotypes is difficult putative B-binding exudates.
to reconcile but is of fundamental importance to studies Thirdly, root B-adsorption capacity may differ
of B nutrition. Several mechanisms have been postu- significantly and hence may affect B uptake. Tana-
lated to explain this apparent paradox (Nable, 1988; ka (1967) observed a 3- to 10-fold difference in the
Nable et al., 1990). adsorption capacity of roots between dicots and gram-
inaceous monocots with a much smaller difference

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56

within the dicots or within monocots. Yamauchi (1971) ences. Evidence to support this is provided in the work
demonstrated a highly significant correlation between of Garnett et al. (1993) who reported that in short term
‘protopectin’ (old terminology for water insoluble par- B uptake studies (4 h), the wheat cultivar ‘Halberd’
ent pectic substance) and 0.5 N HCl soluble but water (resistant to B toxicity) did not differ from ‘W1 MMC’
insoluble B in 33 species across 13 families. Hu et al. (sensitive to B toxicity) in B accumulation. In longer
(1996) reported that among the fourteen species stud- term studies (days) these two varieties did show sig-
ied, there is significant correlation between species B nificant difference in B accumulation. The results of
requirement and cell wall pectin. In general, more B Jenkin et al. (1993) may therefore reflect short term
is required by dicots than graminaceous monocots pre- effects and may not be relevant to long term B accumu-
sumably because of the higher pectin content in the lation. At present, insufficient information is available
dicots. Recently, Matoh et al. (1996) demonstrated to determine if there is any relationship between root
a high correlation between cell wall B and 2-keto-3- cell wall structure and B uptake.
deoxy sugars in the high molecular weight fragments If B uptake is a purely passive process, then the dif-
of cell walls. In combination, these results suggest ferences in B accumulation may be due to differences
that differences in the cell wall B requirement exist in membrane permeability to B. Membrane permeabil-
among species and that these differences might affect ity to urea, water and protons may all vary significantly
B uptake. This observation is not adequate, however, depending upon membrane composition (see “Proper-
to explain the observed differences in B uptake and is ties of boric acid”). Boric acid has a lipid solubili-
not consistent with results from uptake experiments. ty of the same order as urea and hence small differ-
For example, Hu et al. (1996) report a less than 2- ences in membrane composition (head groups, fatty
fold difference in cell wall B binding capacity between acyl chains, etc.) between genotypes may affect B
various monocot species even though differences in B uptake. If passive membrane permeation is the prima-
uptake can be as great as 4- to 7-fold between these ry means of B uptake, then B uptake would be linear
same species (Nable, 1988). Further, if the cell wall B- with increasing concentration, and would not be influ-
binding capacity is the key determinant of the observed enced by metabolic inhibitors. This basically agrees
differences in B uptake between species, then once the with existing information (see “Is boron adsorption
cell wall binding capacity is saturated, uptake should active or passive?”. If species and genotypes had differ-
be the same for all species. There is no indication that ent membrane permeability coefficients for boric acid,
this is the case. they would also exhibit a linear response to increasing
Fourthly, physical barriers related to root cell wall B supply. However, the slope of this response line may
structure may determine B uptake rates. Some evi- differ and the resulting B absorption rate would differ.
dence in support of this was provided by Jenkin et al. The genotype comparisons of Nable et al. (1990) can
(1993) who investigated B uptake in protoplasts of two be suitably explained in this manner.
barley varieties with stable B isotope techniques. The Similarly, small differences in the relative perme-
two barley varieties used were known to have signif- ability of membranes to B relative to water would
icant differences in B accumulation on a whole plant alter the ratio of B accumulated to water transpired,
and long term basis, with the B toxicity resistant culti- though the total amount of water transpired would
var ‘Sahara 3771’ accumulating much less B than the still directly affect B uptake. This would adequate-
sensitive cultivar ‘Stirling’. However, in the absence ly explain the observed relationship between water
of the cell wall, the two varieties did not show any uptake and B accumulation while accommodating the
significant difference in B accumulation (Jenkin et al., observed genotype differences. It is also possible that
1993). There are three possible explanations for this differences in membrane permeability to B may not
result: (1), the difference in B accumulation between be sufficient to influence whole plant B uptake during
these two cultivars is determined by the cell wall and, short term uptake studies, these differences may only
once the cell wall is removed, the cultivar difference become apparent after longer B uptake periods.
disappears; (2), since the membrane integrity was not Though differences in membrane permeability
assessed in the protoplasts used by Jenkin et al. (1993), could theoretically result in the observed differences
genotype difference might be overshadowed by mem- in B uptake between species (or genotypes), it should
brane leakiness as a consequence of the protoplast iso- be noted that this has not been experimentally verified.
lation technique; or (3), the duration of the uptake Further, we are unaware of any evidence of genotyp-
experiment was too short to determine species differ- ic differences in membrane composition resulting in

plso4del.tex; 1/10/1997; 19:13; v.7; p.8

 57

the differential membrane permeability of any solute. Bowen J E and Nissen P 1977 Boron uptake by excised barley roots
Finally, experimental evidence suggests that genotypic II. Characteristics and kinetics of active uptake. Physiol. Plant.
41, 109–115.
differences in B uptake are associated with relatively Briskin D P and Reynolds-Niesman I 1991 Determination of
few genes (Nable et al., 1990 and references therein), H+ /ATP stoichiometry for the plasma membrane H+ -ATPase
it would be intriguing to determine if these genes code from red beet (Beta vulgaris L.) storage tissue. Plant Physiol. 95,
for subtle differences in membrane composition. 242–250.
Brown P H, and Hu H 1994 Boron uptake by sunflower, squash and
Further research to determine the mechanism of cultured tobacco cells. Physiol. Plant. 91, 435–441.
genotype and species differences in B absorption is Dencher N A, Burghaus P A and Grzesiek S 1986 Determination of
clearly required. the net proton-hydroxide ion permeability across vesicular lipid
bilayers and membrane proteins by optical probes. Methods in
Enzymology 127, 746–760.
Finkelstein A 1976 Water and nonelectrolyte permeability of lipid
Conclusions and future directions bilayer membranes. J. Gen. Physiol. 68, 127–135.
Forno D A, Asher C J and Edwards D G 1979 Boron nutrition of
Boron uptake in higher plants is probably a passive 
cassava, and boron temperature interaction. Field Crops Res.
2, 265–279.
process acting in response to external boric acid con- Garnett T P, Tester M A and Nable R O 1993 The control of boron
centration, membrane permeability, internal complex accumulation by two genotypes of wheat. In Plant nutrition – from
formation and transpiration rates. Species differences genetic engineering to field practice. Ed. N J Barrow. pp 397–400.
Kluwer Academic Publishers, Dordrecht, The Netherlands.
in B uptake rate cannot be adequately explained on the
Gupta U C 1979 Boron nutrition of crops. Adv. Agron. 31, 273–307.
basis of current knowledge but may result from differ- Hu H and Brown P H 1994 Localisation of boron in cell walls of
ences in membrane permeability, B-complex forma- squash and tobacco and its association with pectin. Evidence for
tion inside or outside the root or some as yet unidenti- a structural role of boron in the cell wall. Plant Physiol. 105,
681–689.
fied mechanism. Hu H, Brown P H and Labavitch J M 1996 Species variability in
Future studies on B absorption should include con- boron requirement is correlated with cell wall pectin. J. Exp. Bot.
sideration of both short and long term uptake periods, 47, 227–232.
must be conducted at realistic B concentrations and Hu H, Penn S G, Lebrilla C B and Brown P H 1997 Isolation and
characterisation of soluble B-complexes in higher plants. The
should always consider the impact of B-complex for- mechanism of phloem mobility of boron. Plant Physiol. 113,
mation in experimental design. Experimental verifica- 649–655.
tion of the theoretical membrane permeability coef- Huang C and Graham R D 1990 Resistance of wheat genotypes to
ficient proposed by Raven (1980) remains a priority boron toxicity is expressed at cellular level. Plant and Soil 126,
295–300.
and may ultimately allow permeability to be compared Ishii T and Matsunaga T 1996 Isolation and characterisation of a
between species. The recent identification of closely boron-rhamnogalacturonan-II complex from cell walls of sugar
related genotypes differing significantly in B absorp- beet pulp. Carbohydr. Res. 284, 1–9.
Jansen M and Blume A 1995 A comparative study of diffusive and
tion rates has provided invaluable experimental mate-
osmotic water permeation across bilayers composed of phospho-
rial. These genotypes hold great potential for providing lipids with different head groups and fatty acyl chains. Biophys-
the physiological and genetic information necessary to ical J. 68, 997–1008.
determine the mechanism of B uptake in plants. Jenkin M, Hu H, Brown P, Graham R, Lance R and Sparrow D
1993 Investigations of boron uptake at the cellular level. In Plant
nutrition – from genetic engineering to field practice. Ed. N J
Barrow. pp 157–160. Kluwer Academic Publishers, Dordrecht,
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