C A N STOMATA PLAY A PART IN PROTECTING

PLANTS A G A I N S T A I R POLLUTANTS ?
T. A. MANSFIELD ~r, ONDREI MAJERNIK

Department of Biological Sciences, University of Lancaster, Great Britain

ABSTRACT

The role of stomata as ports of entry of pollutants into plants is discussed. Experimental evidence from several sources suggests that stomatal closure can help to
protect plants against pollution damage. The C02 concentration in the atmosphere
usually increases appreciably when toxic pollutants are present in dangerous amounts.
The closing response of stomata to small increases in C02 concentration is described,
and it is suggested that thi~ mechanism might be exploited to produce varieties of
plants more likely to survive in polluted atmospheres.
Experimental evidence is presented, showing that stomata respond, directly to
SO 2 in the atmosphere. In conditions of low relative humidity SO 2 suppresses
stomatal opening but, with a higher water content in the atmosphere, there is an
appreciable stimulation of opening. This latter effect is more likely to occur in
natural conditions and it will tend to increase the penetration of S02 into leaves.

In discussions of the effects of air pollutants on plants, stomata are usually briefly
mentioned as the probable ports of entry of gases and vapours into leaves, but,
considering the volume of published work in the entire field, it is surprising how
little is known about their true role, and, more especially, about the effects of
pollutants on the stomata themselves.
The cuticle covering the epidermis of leaves is not impermeable, as was once
thought. It is now known that many substances can both enter and leave the leaf
by passing through the cuticle. The rate of movement through the cuticle is,
however, generally slow since it offers a considerable resistance to diffusion.T h e
stomatal pores therefore provide the main means of communication with the
atmosphere, with the result that the guard cells regulate, to a large extent, what
goes in and out of the leaf. They are also the first cells to experience any pollutant
149
Environ. Pollut.(1) (1970) pp. 149--154--O Elsevier Publishing Company Ltd, England--Printed in Great Britain

150

T. A. MANSFIELD, ONDRFJ MAJERNIK

diffusing in from the atmosphere. If they react to the pollutant by losing turgor
so that the stomata close, the other cells inside the leaf will be offered some degree
of protection. However, if stomatal closure is permanent (that is, if the guard cells
are damaged and cannot regain turgor) the effect is likely to be serious: the subsequent growth of the plant will be affected because the main route for carbon
dioxide intake for photosynthesis wilt be blocked.
As long ago as 1921, Loftfield showed that lucerne (alfalfa) plants were only
susceptible to SO2 during the hours in which the stomata were open. Bobrov (1955)
observed that bands of damage on the young blades of P e a annua corresponded to
positions where the stomata had newly become functional. With other pollutants,
there has not been general agreement about whether stomatal closure prevents
damage. Also using Pea annua as a test plant, Juhren, Noble & Went (1957)
found that when the stomata were closed in low light intensity, the plants were
'quite resistant' to smog (air drawn from outside the laboratory in Los Angeles).
On the other hand, Dugger et al. (1962) reached the opposite conclusion,, that
phytotoxic components in smog (e.g. ozone) damaged bean plants (Phaseolus
vulgaris) whether the stomata were open or closed. They used fumigation chambers
and the toxic substances used were prepared specially for the investigation and
carefully controlled.
It is not difficult to see how data could have arisen leading to these two opposite
conclusions. The epidermis can be looked upon as constituting a resistance to
diffusion between the interior and exterior of the leaf. It is a variable resistance,
with a magnitude depending on the stomatal aperture, and it will control the rate
of movement of pollutants inwards along a concentration gradient. Unless the
magnitude of the resistance becomes infinite when the stomata are completely
closed, inward movement will still occur, but at a reduced rate. We know that/he
resistance does not become infinite, because the cuticle is slightly permeable, and
furthermore even when stomata are closed they are unlikely to provide a perfect
seal (also some stomata are often kept permanently open by dust particles jammed
in the pores). Consequently, pollutants will still enter the leaves, slowly, along the
concentration gradient, and if the plant is exposed to the pollutant for long enough,
or if the concentration outside the leaf is high enough, damage will still occur.
Whether stomatal closure gives the plant protection or not will therefore depend
on the amount of pollution, and on the duration of exposure.
If the stomata were to close in response to the pollutant, in the correct circumstances this would give the plant some degree of protection. The ideal situation
would be a reversible reaction of the stomata: closing when the pollutant is in the
atmosphere, opening at other times. There is a normal physiological response of
the stomata which puts this ideal situation well within the bounds of possibility.
It is well established that stomata show a considerable sensitivity to carbon
dioxide in the atmosphere. They begin to close when the concentration rises slightly
above the normal 300 ppm, and open widely when it falls below this figure. A lot

151

C A N STOMATA PLAY A PART 1N P R O T E C T I N G P L A N T S ?

of work has been published on this topic (see Meidner & Mansfield, 1968), and the
two studies outlined hereunder provide information especially relevant to the
present problem.
Heath & Russell (1954) studied in great detail the interaction between CO 2
concentration and light intensity on stomatal aperture in wheat. We have taken
some of their data and converted them to values which are of relevance to the
present discussion, namely to show how the resistance to gaseous diffusion through
the stomata is determined by CO2 concentration (Fig. 1). As the CO2 concentration

""

~o
0---.-~_ 0

~o

\.

.g
I

200

I
600

400
[CO,] in

I
800

ppm

Fig. 1. Effect of CO2 concentration on stomatal aperture of wheat at two light intensities:

0 , 8 Klux; O
(3, 2.7 Klux. Recalculated data from Heath & Russell (1954). For wheat
stomata the cube root of viscous conductance is directly proportional to diffusive conductance.
increased from zero, the resistance offered by the stomata to diffusion increased
considerably.
Gaastra (1959) obtained some data for turnip which give a particularly good
illustration of this response. He measured the effect of CO2 concentration on
transpiration rate, which is, of course, dependent on the diffusive capacity of the
stomata. In a light intensity of 4500 lux, transpiration fell to less than half as the
CO2 concentration was increased from 310 to 420 ppm. This light intensity is of
the same order of magnitude as that occurring on cloudy days in the UK, and the
increase in CO2 concentration is similar to that which occurs on such days in towns
as a result of pollutants being pumped into the atmosphere. Measurements of CO 2
concentrations in towns have not often been made over long periods, presumably
because there is little general interest in a non-toxic pollutant. Records have been
obtained in Paris, however, showing diurnal cycles of 340 ppm minimum and
maxima in the range 400-600 ppm, over the period December-April.
The concentration of CO2 tends to increase as the concentration of other pollutants increases; in fact the correlation is so good that some workers have found

152

T. A. MANSFIELD, ONDREJ MAJERNIK

that measurements of CO2 levels can provide an accurate guide to the concentrations of other, less readily measured, pollutants. Consequently, the stomatal
closure in response to the CO 2 should offer the plant some protection against the
dangerous pollutants.
Is there any possibility that this physiological response might be exploited to
provide plants more likely to survive in polluted atmospheres? The stomatal
closing reaction to CO2 seems to be of wide occurrence among plant species, though
the magnitude of the effect does vary and the stomata of some crops, such as tomato
and cotton, seem to be relatively insensitive to CO2 (Pallas, 1965; Hurd, 1969).
Among individuals of a particular species there is the usual statistical variation
that one expects in physiological responses, and it is reasonable to suppose that
with a suitable selection programme crop varieties could be produced with enhanced
stomatal responses to CO2. The possibilities here are completely unexplored, but
some aspects are being examined in our current research programme at Lancaster.
S t o m a t a l responses to S O 2

We are also investigating the direct response of the stomata to SO2 in the
atmosphere. The reaction of the stomata to the pollutant itself may well be a major
factor in determining a plant's resistance. Studies with broad bean have revealed
two kinds of stomatal reaction to SO2, which depend on the relative humidity of
the atmosphere. The effect which is probably of most significance occurs when the

darknes~

) light (10Klux)

~o-.~.....~ o~ e

c
o

8'
"u

2o,. 4

lo

~'.-, .~

1'o

1~

time

Fig. 2. Effectof treatment with 0.5 ppm SO2 on stomatal aperture of broad bean in light and
darkness. Relative humidity 42 % at 18C.
C), control; - , identical conditions plus
0-5 ppm SO2. Ordinate scale: total pressure minus pressure across the leaf in a resistance porometer,
indicating stomatai opening in arbitrary units (Allaway & Mansfield, 1969).

153

CAN STOMATA PLAY A PART IN PROTECTING PLANTS?

relative humidity is greater than 40% at 18C (water vapour pressure deficit less
than 7 mm Hg). It involves an opening of the stomata to much wider apertures
than normal (Fig. 2). This reaction is potentially very damaging, for wide open
stomata will permit rapid access of the pollutant to the interior of the leaf (Majernik
& Mansfield, 1970). When the water content of the atmosphere is lower (less than
40% R.H. at 18C) we have found the opposite response, namely closure of the
stomata in the presence of SO2 (Fig. 3).

C
r-

o o-.._.o._____o..._.,
o_.....o__....

~2
00

*0

6
time

10

12

Fig. 3. Effect of treatment with 1 ppm SO2 on stomatal aperture of broad bean in low relative
humidity (32% at 18C) during a 12 hour photoperiod. O - - O , control;
, identical
conditions plus 1 ppm SO2.
The existence of these two opposite responses to SO2 reveals a complicated
situation in the plant which we have not begun to understand at the cellular level.
It is not, however, necessary to have such understanding to appreciate the significance of these observations in relation to the question of plant survival. In Western
Europe the most serious SO2 pollution is at times when the relative humidity is
relatively high, and consequently it is the first response (Fig. 2) which is likely to
predominate. An important question to be answered in future research is whether
there is any possibility of preventing such abnormal opening of stomata. Abnormal
individuals with only semi-functional stomata, which in normal circumstances
would be less successful than the normal population, might be more suited to
survival in polluted atmospheres.
This discussion is intended to be applicable to situations in which plant growth
is severely restricted by SO 2 pollution, which usually applies only to major industrial
conurbations and areas adjacent to them. In such places the concentration of
SOz in the atmosphere varies over a wide range, and the most hazardous times for
plants are the occasional rises to high levels. Peak levels of SOz rarely rise above

154

T. A. MANSFIELD,ONDREJ MAJERNIK

1 ppm, and usually lie between 0.25 and 1 ppm. Our experimental data for SO 2
are applicable to this situation. Similarly, the concentration o f CO2 in the
atmosphere only increases significantly in large population centres. Thus the
data presented here may be relevant to the growth of plants in and a r o u n d
our towns, but, unless general pollution levels increase, they are not likely
to be applicable to crops on a wider scale. The problem o f growing a suitable
variety o f plants in our towns is not, however, an insignificant o n e - - n o one will
dispute that they are essential for an agreeable environment. The ultimate answer
to this problem .is, o f course, to reduce the a m o u n t of atmospheric pollution in
population centres. Some progress is being m a d e in this direction but there is still
much to be achieved, and, meanwhile, the vegetation continues to be damaged.
There is thus a case for a really determined research effort to see whether plants
can be produced that will grow reasonably well in these situations. Studies o f the
role o f stomata are but one facet of the overall problem, but the possibility that
they might contribute to a plant's 'self-defence' system is well wort.h our
consideration.

ACKNOWLEDGEMENT
The financial support o f the Natural Environment Research Council is gratefully
acknowledged.

REFERENCES

ALLAWAY,W. G. & MANSFIELD,T. A. (1969). 'Automated system for following stomatai behaviour
of plants in growth cabinets.' Can. J. Bot., 47, 1995-8.
BoBRov, R. A. (1955). 'The leaf structure of Poa annua with observations on its smog sensitivity
in Los Angeles County.' Am. 3". Bot., 42, 467-73.
DUGGER, W. M., TAYLOR,O. C., CARDIFF,E. & THOMPSON,C. R. (1962). 'Stomatal action in
plants as related to damage from photochemical oxidants.' PI. Physiol. Lancaster, 37, 487-91.
GAASTnA,P. (1959). 'Photosynthesis of crop plants as influenced by light, carbon dioxide, temperature and stomatal diffusion resistance.' Meded. LandbHoogesch. Wageningen., 59, 1-68.
HEATH,O. V. S. & RUSSELL,J. (1954). 'Studies in stomatal behaviour. VI. An investigation of the
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JUHREN,M., NOBLE,W. & WENT,F. W. (1957). 'The standardisation ofPoa annua as an indicator
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LoFrFIELD,J. V. G. (1921). 'The behaviour of stomata.' Pubis Carnegie Instn., 314, 1-104.
MAJERNIK, O. & MANSFIELD,T. A. 1970. 'Direct effect of SO2 pollution on the degree of
opening of stomata.' Nature, Lond. 227, 377-8
MEIDNER,H. & MANSFIELD,T. A. (1968). Physiology o f Stomata. London; McGraw-Hill.
PALLAS, J. E. (1965). 'Transpiration and stomatai opening with changes in the carbon dioxide
content of the air.' Science, N.Y., 147, 171-3.