Bioavailability

Effect of pH on bioavailability of Ni
for oats
a case study

General introduction

What is bioavailability?
Used in different disciplines
Various definitions
Soil Quality: nutrients/contaminants
in soil actually available to receptor
organisms (uptake)

The journey of a chemical

input

Adsorbed/
precipitated

Soil solution

B
i
o
m
e
m
b
r
a
n
e

Biological
response

physical-chemical reactions transport uptake relocation effects

Popular theories
Free Ion Activity Model (FIAM)
free metal ion activity is the parameter that
determines the effects

Bio-Ligand Model (BLM)

uptake is a result of competition for binding of
the chemical by all reactive components (abiotic
& biotic)

What processes/factors affect

bioavailability?
Abiotic--Physical chemical factors
soil composition, pH, structure, etc.

Biotic factors
species, root activity, cellular
compartment, etc.

Case study

Reading materials
Effects of soil characteristics on Ni
toxicity to oats
1. pH effects
2. Soil composition effects
This lecture focuses on the pH effects

Why pH?
pH: -log(H+)
wide range in soil
can be manipulated
affecting both the physical chemical
and the uptake process

pH effects on basic uptake

process
Water culture using nutrient
solution
avoid implication from soil

(almost) all Ni is free Ni2+ ion

Bioavailability, water culture

What factors might affect the uptake of
metal ions?
- Metal ion concentration??
- pH ??
- Other factors??

Plant growth and Ni concentration in

Water
pH 4
Concentrations
in mg/l Ni
0

0.25

0.5

shoot dry weight (g/plant)

EffectNiconc.onplantgrowth
1.4

pH4

1.2
1.0

3 week
2 week
1 week

0.8
0.6
0.4
0.2
0.0
0

50

100

[Ni-nutrient solution] (M)

150

shoot dry weight (g/plant)

EffectNiconc.onplantgrowth
1.4

pH6

1.2
1.0
0.8

3 week
2 week
1 week

0.6
0.4
0.2
0.0
0

50

100

[Ni-nutrient solution] (M)

150

pH effects on Ni toxicity in water

1 mg/liter Ni

pH 5

pH4pH5pH6pH7

pH effects
relative dry weight

120%
100%
80%

EC50

60%
40%
20%
0%

pH7 pH6

10

pH5

pH4

20

30

40

[Ni-nutrient solution] (M)

50

60

From effects to uptake

Growth is one of the plant responses to Ni
toxicity
In most cases, the extent of the
physiological responses is related to the
chemical concentration in tissue (uptake)
What are the effects on uptake?

Relationship between pH, Ni conc.

and uptake
[Ni-root] (logmmol/kg)

2.0

root

pH7
1.5

pH6
pH5
1.0

pH4
0.5
-0.5

0.0

0.5

1.0

1.5

2.0

[Ni-nutrient solution] (logM)

2.5

Relationship between pH, Ni conc.

and uptake
[Ni-shoot] (logmmol/kg)

2.0

shoot

1.5
1.0
0.5

pH7

0.0

pH4

-0.5
-1.0
-0.5

0.0

0.5

1.0

1.5

2.0

[Ni-nutrient solution] (logM)

2.5

pH dependent uptake
Note uptake scale is logarithmic
Low Ni concentration big pH effect
High Ni concentration very small pH effect
What could be the reason of the fact that the
pH dependence of the uptake vanishes at
[Ni]>1 mg/l ? See photograph.

Uptake equation
pH dependent uptake can be described using
linear equations
shoot:
log[Ni-shoot] = 1.15 + 0.65log[Ni2+-free] + 0.11 pH

Conclusions water culture

Ni at higher concentration is toxic
Ni is taken up by the roots and transported
to the shoots
Ni uptake is dependent on the pH and [Ni]
Higher pH, higher uptake, higher toxicity!

What would happen in soil?

Bioavailability of Ni in soil
What chemical forms (species) will Ni take
when added to soil? solid/solution?
Will pH affect Ni distribution among these
chemical forms?
If we assume pH effects on plant uptake
remain the same, What would be the final
pH effects on Ni toxicity in soil?

dry weith shoot (g/pot)

Ni conc. effect on plant growth in

soil
20

pH4.7
15

10

0
0

200

400

600

Ni in soil (mg/kg)

800

dry weight shoot (g/pot)

Ni conc. effect on plant growth in

soil
20

pH6.0
15

10

0
0

200

400

600

Ni in soil (mg/kg)

800

pH effects in soil
100mgNi/kgsoil

pH4.7pH5.2pH6.0pH6.8

pH effects in soil
based on total metal content
relative dry weight of shoot

125%
100%
75%

EC50

50%
pH5.2

25%

pH6.8

pH4.7

pH6.0

0%
0

10

[Ni-added] (mmol/kgsoil)

12

14

Why the pH effects differ from

those in water culture?
To understand it, we need to separate the
effects on soil from those on uptake
Uptake via soil solution
0.01 Molar CaCl2 extraction

EC50
Soil CaCl2, versus nutrient solution
12

40
30

10

CaCl2 extraction

30
20
20

soil

Nutrient solution

10
10

4
2

00

soil EC 50 (mmol/kgsoil)

solution
EC 50 (M)
EC50 (M)

40

0
3

6
pH

Reason for the difference??

Practical point of view, simple to have just
one EC50 independent of pH !!
Theoretical insight needed to explain the
difference

Ni speciation in CaCl2 extract

Free metal ion considered to be the
bioavailable species
Ni forms complexes with dissolved
organic matter: fulvic, humic acid etc.
Measuring free Ni with the DMT
technique

Relationship between Ni in CaCl2

versus free Ni2+
Log[Ni2+free]=0.76+1.09log[Ni2+CaCl2]0.14pH

Free Ni2+ concentration in CaCl2 extraction

decreases with increasing pH.

EC50
free Ni2+ in CaCl2 extract,
nutrient solution
25

Ni2+ CaCl2
extraction

EC50 (M)

20
15
10

Ni2+ nutrient
solution

5
0
3

pH

Adsorption isotherm
FromCaCl2extraction,wederived:
Log[Niadsorbed]=1.89+0.77log[NiCaCl2]+0.6pH(1)

RelationshipbetweenNiCaCl2andfreeNi2+:

Log[Ni2+free]=0.76+1.09log[Ni2+CaCl2]0.14pH(2)
(1)+(2)gives:
Log[Niadsorbed]=1.35+0.71log[Ni2+free]+0.70pH

Plantuptakeequation
fromnutrientsolutionexperiment
log[Nishoot]=1.15+0.65log[Ni2+free]+0.11pH(1)

Soil adsorption isotherm

From CaCl2 extraction
Log[Niadsorbed]=1.35+0.71log[Ni2+free]+0.70pH(2)

Conclusion soil adsorption versus

uptake
Both biotic uptake and binding to soil increase with
increasing pH
Bioavalibility in nutrient solution increases with
increasing pH
pH effect for soil is bigger than for uptake
Result is that the bioavailability when expressed on Ni
bound to soil (app. total Ni) decreases with increasing pH

Model approach to link soil metal

content to metal uptake
QMsoil=Ks(Me2+)ms(H+)as

(1)

QMbiota=Kb(Me2+)mb(H+)ab

(2)

(1)+(2)gives,assumingMe(total)=QMsoil

QMbiota=KbKs(mb/ms)QMtotal(mb/ms)(H+)(abas(mb/ms))

Outcome model
Log[Nishoot]=3.41+0.92log[Nisoil]0.53pH

Outcome model prediction of Ni uptake

in soil
log[Ni-shoot] mmol/kg

2.0

pH4.7

pH5.4
pH6.2

1.5

pH6.8

1.0
0.5
0.0
-0.5
-1.0
-1.5

-1.0

-0.5

0.0

0.5

1.0

log[Ni-soil] mmol/kg

1.5

2.0

What have we found about the

pH effects on Ni bioavailability?