سجاد
PHOTOSYNTHESIS
IN RELATION TO STOMATAL FREQUENCYAND DISTRIBUTION
R. 0
. FREELAND(WITH
ONE FIGURE)Received
May 4, 1948Introduction
The
concept that stomates play an all-important role in controlling thediffusion
of carbon dioxide into leaves during photosynthesis seems to bequite generally accepted by teachers and authors
of botany texts. In aplant physiology
text by MAXIMOV (6) may be found the following, "Thatthe carbon dioxide
enters the leaf mainly through the stomates may beshown by
a simple experiment. If on a certain portion of the leaf thestomata are coated
with vaseline, and the leaf is then exposed to light andafterward
treated with iodine, the blue color reaction will be observed onlyin
those portions where the stomata remained open." Also in the text byMEYER and ANDERSON
(7) one finds "Critical experiments have shown,however,
that the proportion of this gas (CO2) entering the leaves by thisroute
(directly through epidermis) is relatively small, and that practicallyall of the carbon dioxide
entering leaves diffuses in through the stomates."Similar
statements may be found in many other botanical books.It seems that this concept
stems mainly from the research of Blackmanand his contemporaries. BLACKMAN
(1) in a review of the research regardingthis problem concludes
that "Under normal conditions, practicallythe sole
pathway for carbon dioxide into or out of the leaf is by the stomata."Further
support and impetus were given to this idea by the classicalexperiments
of BROWN and ESCOMBE (2) which demonstrated andpartially
explained the enormous diffusive capacity of small pores andstomates.
More recently MASKELL (5) working with Prunus lauro-cerasusand
NUTMAN (9) studying Coffea arabica have reported that photosynthesisin these
plants is directly related to stomatal movement.On the other hand considerable evidence has
appeared in the literaturewhich indicates that the
rate of diffusion of carbon dioxide through epidermalcells of
leaves may be quite appreciable and should not be minimizedor
ignored. Blackman mentions that Boussigault and Barthelmvreported
data to the effect that the carbon dioxide exchange during "assimilation"was
independent of stomatal distribution. MITCHELL (8)found that
the leaves of tomato and Pelargonium absorbed carbon dioxideand accumulated
carbohydrates in appreciable quantities although thestomates appeared to be closed.
Furthermore, the amount of carbon dioxideabsorbed
by the leaves in which the stomates appeared to be closed wasapproximately equal
to the amount absorbed by the same leaves when595
PLANT PHYSIOLOGY
the stomates were open. HEINICKE and CHILDERS (4) and SCHNEIDER and
CHILDERS (10) in their extensive research with apple trees, found many
occasions when the rate of photosynthesis was quite high even though
thestomates were
closed. In this laboratory, many measurements of the apparentphotosynthesis from the upper and lower surfaces of leaves of various
plants have been made. As often as not the results have shown little
or no
direct correlation between photosynthesis and stomatal distribution.Some
of these data will be presented in this paper.Experimental procedure
The plants to
be included in this report are yellow-green coleus, Coleusblumei;
avocado, Persea americana; poinsettia, Euphorbia pulcherrima;4-
--<FIG. 1. Glass cups clamped on the top and bottom of a leaf. Description in text.
rubber
plant, Ficus elastica; begonia, Begonia sp.?; Rhoeo discolor; bean,Phaseolus
vulgaris (var. stringless green pod) ; tobacco, Nicotiana tabacum,and
geranium, Pelacrgonium zonale. These plants were selected because ofvariation in stomatal
distribution and thickness of cuticle. Six of thespecies
have stomates in the lower epidermis while the other three have596
FREELAND: STOMATAL FREQUENCY
stomates in both the upper and lower surfaces of their
leaves as shown intable I. The
amount or thickness of cuticle progresses from very little insuch plants
as coleus and tobacco to much in poinsettia and rubber plant.Apparent photosynthesis,
in terms of milligrams of carbon dioxide used,was
measured from the upper and lower surfaces of the leaves. Glass cupsof the design shown in figure
1 were clamped opposite each other on severalleaves of the experimental plant. These cups each had a
diameter of 4 cm.and
a depth of 8 cm. A little grafting wax applied around the rim of eachcup and
warmed slightly at the time the cup was applied to the leaf providedan
air-tight seal. Natural air passed through these cups to absorptiontowers
containing 0.1N KOH for carbon dioxide determination afterthe method of HEINICKE and
CHILDERS (4). Using flowmeters and constantsuction,
FREELAND (3), the rates of air flow were so regulated as toprevent excessive depletion of the carbon dioxide of the air and extreme
temperature rise in the cups.
A rate of air flow of about 0.5 cu. ft. perhour was adequate. Experimental plants were placed near a laboratory
window
in direct sunlight where on hot days they were shaded with oneor
two layers of cheesecloth. No other attempt was made to control thetemperature in the cups or around the plants. Temperatures were recorded
several times during the course of each experiment using thermocouples
and a
Leeds and Northrup thermopotentiometer. Little or no differencein temperature was
ever found in the cups attached to the oppositesides
of a leaf. The duration of each experiment was four hours.To
test the accuracy of the apparatus and methods, a large number ofdeterminations
of the carbon dioxide content of the air in the laboratoryhave been made. From these measurements it has been found that the milligrams
of carbon dioxide per cubic foot of air can be determined with a
standard deviation of 0.2.
Results and
discussionThe experimental results obtained for the
various plants are summarizedin table I. The data presented for each plant were obtained by
averaging
the results from three or more determinations made upon differentdays.
An examination of the data for those plants which have stomates only
in the lower
epidermis provides a basis for evaluating the importance ofstomates
in the absorption of carbon dioxide during photosynthesis. Forexample, in coleus, avocado, and
begonia, carbon dioxide diffused throughthe upper
epidermis in very significant amounts. In no case was the rateof
diffusion of carbon dioxide through the upper epidermis alone as greatas
it was through the lower epidermis plus stomates. However, from theknown structure
of leaves, it would appear to be a reasonable assumptionthat carbon dioxide could
diffuse through the lower epidermal cells, excludingstomates,
at a rate approximately equal to that through the upperepidermis.
On this basis the apparent photosynthesis calculated for the597
PLANT
PHYSIOLOGYtotal leaf epidermis,
excluding stomates, would really be twice the amountspresented for the upper epidermis in table
I. Applying this principle tocoleus,
as an example, the apparent photosynthesis in mg. of CO2 used perhour
per square decimeter would be 6.2 through epidermal cells alone and7.0
through stomates alone. The data for the other plants in this group,poinsettia, rubber plant, and Rhoeo
discolor, indicate that they belong in aseparate
class. In these plants there is little or no diffusion of carbondioxide
through the epidermal cells and the stomates are the primary pathwaythrough
which carbon dioxide diffuses during photosynthesis. A partialexplanation
for the difference between these two groups of plants withrespect
to carbon dioxide exchange through the astomatous epidermis maybe found in the thickness of the
epidermis or cuticle. There probably areother factors
as indicated by the data for begonia, which has a rather thickcuticle.
At any rate the data for these plants, having stomates in theTABLE I
SUMMARY OF THE DETERMINATIONS OF
APPARENT PHOTOSYNTHESIS IN TERMS OF THEAMOUNT OF CARBON DIOXIDE WHICH DIFFUSED INTO THE LEAF THROUGH THE ITPPER
AND LOWER EPIDERMIS
MG. OF
CO2/DM.2/HR. STOMATES/CM.2PLANT
UPPER LOWER UPPER LOWER
COLEUS
... .. 3.1 10.1 0 11,000AVOCADO
.... 4.1 5.9 0 15,200POINSETTIA
0.6 4.1 0 13,000RUBBER
PLANT ......* 2.3 0 17,000BEGONIA
2... 2.2 3.3 0 3,400RHOEO
DISCOLOR * 3.5 0 1,700BEAN
3.8 1,000 7,000TOBACCO
3.1 3.6 3,000 3,000GERANIUM
.... 1.3 3.7 2,300 15,000*
Not significant.lower
surface of the leaves, indicate that the statement of Blackman andothers that the
stomates are practically the sole pathway for carbon dioxideinto and
out of leaves under normal conditions is not true in many cases.The data
confirm the conclusions of Mitchell, Heinicke, and others to theeffect that
some plants may carry on a rather high rate of photosynthesiswith the
stomates closed.The
results for bean, toba2co, and geranium which have stomates in boththe
upper and lower epidermis of the leaves must be examined from a differentpoint
of view. For these plants the amounts of apparent photosynthesisrelated
to stomatal diffusion and epidermal diffusion respectivelycannot
be separated. In tobacco there is a direct correlation betweenstomatal
frequency and apparent photosynthesis for the upper and lowerside of the leaves. As indicated above
there is no way of determining howmuch,
if any, of the carbon dioxide exchange was due to epidermal diffusionexclusive of
stomates. For geranium the degree of correlation is598
FREELAND: STOMATAL FREQUENCY
much less and for bean it is probably
nonexistent. Therefore, one mustconclude
from these data that for those plants having stomates in both theupper
and lower epidermis of their leaves the diffusion of carbon dioxideinto the leaves
during photosynthesis may or may not show a direct correlationwith
stomatal frequency and distribution. It seems probable thatin
some plants, at least, some other factor (or factors) other than stomatalfrequency
and distribution plays a considerable role in determining thepathway of carbon
dioxide exchange during photosynthesis.Summary
1.
This is a report of measurements made to determine the relative significanceof
stomates versus the epidermis, exclusive of stomates, as routesfollowed by carbon dioxide which diffuses
into leaves during photosynthesis.2.
Plants were selected for the experiments to obtain wide variations instomatal frequency and
distribution, and thickness of cuticle.3. For those plants having
stomates only on one side of their leaves,considerable
variation occurred with respect to the absorption of carbondioxide
through the epidermal cells during photosynthesis. In all casesthe
rate of apparent photosynthesis through the stomatal bearing epidermiswas greater than through the astomatous side. In some plants
theamount
of carbon dioxide exchange through epidermal cells alone was verysignificant sometimes
being approximately equal to the amount which diffusedthrough the
stomates. In other plants of this group, particularlythose with
a thick cuticle, little or no apparent photosynthesis could bedetected
through the epidermis without stomates.4.
The data for those plants having stomates in both the upper andlower epidermis
indicate that apparent photosynthesis, in terms of CO2absorption,
may or may not show a direct correlation with stomatal frequencyand
distribution.DEPARTMENT
OF BOTANYNORTHWESTERN
UNIVERSITYEVANSTON, ILLINOIS
LITERATURE
CITED1. BLACKMAN, F. F.
Experimental researches on vegetable assimilationand respiration. II.
On the paths of gaseous exchange betweenaerial leaves
and the atmosphere. Phil. Trans. Roy. Soc. B. 186:503-562. 1895.
2. BROWN, HORACE T., and ESCOMBE,
F. Static diffusion of gases andliquids
in relation to the assimilation of carbon and translocationin
plants. Phil. Trans. Roy. Soc. B. 193: 223-294. 1900.3.
FREELAND, R. 0. Automatic electric switch for constant air pressure.Sci.
102: 231-232. 1945.4.
HEINICKE, A. J., and CHILDERS, N. F. The daily rate of photosynthesis,during
the growing season of 1935, of a young apple treeof
bearing age. Cornell Univ. Agr. Exp. Sta. Bull. 201. 1937.5-99
600
PLANT PHYSIOLOGY5. MASKELL, E. J. Experimental researches on vegetable assimilation
andrespiration. XVIII. The relation between stomatal opening and
assimilation-a critical study of assimilation rates and porometer
rates in cherry laurel.
Proc. Roy. Soc. B. 102: 488-533. 1928.6. MAXIMOV, N. A. Plant Physiology. McGraw-Hill
Book Co. 1938.7. MEYER, B. S., and ANDERSON,
D. B. Plant Physiology. D. Van NostrandCo. 1939.
8. MITCHELL, J. W. Effect of atmospheric humidity on rate of carbon
fixation
by plants. Bot. Gaz. 98: 87-104. 1936.9.
NUTMAN, F. G. Studies of the physiology of Coffea arabica. II.Stomatal
movements in relation to photosynthesis under naturalconditions. Ann. Bot.
N. S. 1: 681-694. 1937.10.
SCHNEIDER, G. W., and CHILDERS, N. F. The influence of soil moistureon
photosynthesis, respiration, and transpiration of apple leaves.Plant
Physiol. 16: 565-583. 1941.




