Plant Physiol.
(1992) 100, 2106-2108
0032-0889/92/100/21 06/03/$01 .00/0
Communication
Effect of Atmospheric Pressure on Maize Root Growth and
Ethylene Production'
1. Sarquis2, Wayne R. Jordan, and Page W. Morgan*
Jorge
Department of Soil & Crop Sciences, Texas A&M University, College Station, Texas 77843-2474
ABSTRACT
Maize (Zea mays) seedlings were exposed to elevated atmos-
pheric pressures while growing in moist sand in open plastic
envelopes to evaluate the effects of directly applied atmospheric
pressure on ethylene production and root growth. Effects were
evaluated after 24 h. The threshold pressures necessary to promote
ethylene production and decrease root elongation were about 600
and 400 kPa, respectively. Direct atmospheric pressure, at levels
up to 300 kPa, mimicked the control decrease in root diameter and
increased diameter only slightly at 500 to 1200 kPa. In contrast, in
previous work it was shown that physical impedance resulting from
compression of the growth medium by external application of 100
kPa increased the ethylene production rate 4-fold and the root
diameter 7-fold while reducing elongation 75% in 10 h. The relative
insensitivity of roots to direct atmospheric pressure suggests that
they perceive physical impedance, achieved experimentally by
compressing the growth medium, via a surface mechanism rather
than via a pressure-sensing mechanism.
The involvement of ethylene in the inhibition of elongation
and the promotion of radial enlargement (swelling) in maize
(Zea mays) roots exposed to mechanical impedance has been
demonstrated (8). In this experimental system, pressure was
applied externally to the growth medium ('soil') surrounding
the growing tissues, without altering the atmospheric pres-
sure within the growth medium. Subsequent work showed
that 1-aminocyclopropane-1-carboxylic acid oxidase, 1-ami-
nocyclopropane-1-carboxylic acid synthase, and N-malonyl-
transferase respond rapidly to the impedance stress (9); how-
ever, the question of how the mechanical stimulus is
perceived and transduced into increased ethylene production
remains unanswered. We now report results of an indirect
approach to address the question of the perception of physical
impedance. We have determined the effect of pressurizing
the whole system in contrast with applying an extemal
pressure to the growth medium (8). This approach allowed
us to compare the direct effect of atmospheric pressure on
plant growth with the effect of increasing the mechanical
resistance of the growth medium on elongation of the grow-
'Texas Agricultural Experiment Station manuscript TA-30667.
2Present address: Departamento de Bioquimica, Facultad de
Quimica, Universidad Nacional Aut6noma de Mexico, Ciudad Univ-
ersitaria 04510, Mexico D.F., Mexico.
2106
Received for publication June 24, 1992
Accepted August 14, 1992
ing tissues. Differences in partial pressures of gases and water
relations were ignored in this comparison.
MATERIALS AND METHODS
Maize seeds (Zea mays L. cv Tx 5855) were surface sterilized
in 1% NaOCl for 10 min, rinsed, and soaked under running
water overnight before planting in vermiculite wet to satu-
ration with i0- M CaCl2 solution. After 48 h in the dark,
seedlings selected for uniform root length (30 ± 2 mm) were
transferred to plexiglass boxes (20 x 30 cm) the covers of
which were marked with a 1-mm grid to allow monitoring
of the root elongation rate for 10 h. After this period, seed-
lings were selected for uniformity in root elongation rate (2.0
± 0.2 mm/h). Individual seedlings were placed in sand
(particle size <2 mm) in tissue culture bags provided with
five pockets. Three such bags (i.e. 15 seedlings) were used
for each treatment in each experiment. The bags were placed
on test tube racks for vertical support, and the racks were
placed inside pressure plates (7). The pressure plate appara-
tus, which served as a chamber, was sealed and pressurized
with breathing air (20.9% 02) for 24 h. Each pressure treat-
ment was applied to a separate pressure plate. Seedlings
treated identically but with no elevation of pressure above
atmospheric represented the controls. After 24 h, the pressure
was released and seedlings were used for either growth
measurement or ethylene production measurement.
Root diameter was measured 5 mm behind the root tip
under a microscope equipped with an ocular micrometer.
Root length was measured on a millimeter grid under a
stereoscopic microscope. Ethylene was determined by incu-
bating five seedlings in a 20-mL test tube for 3 h in the dark
and then transferring 1 mL by syringe from the gas phase
into a gas chromatograph (Beckman 72-5) equipped with an
activated alumina column and a flame ionization detector.
All seedling handling operations were performed under a
dim green light (maximum transmission at 525 nm and
negligible transmission at <475 or >575 nm, maximum irra-
diance 3.5 x 10-3 j.m-2.s). Data are means of at least three
experiments.
RESULTS
The ethylene production rate in control seedlings was on
the order of 40 pmol seedling-' h-' (Fig. 1). Pressures up to
400 kPa had little or no effect on this rate; production
increased first at 800 kPa. Higher pressures increased the
 ATMOSPHERIC PRESSURE AS A STRESS ON ROOT GROWTH 2107

Juu -

250- '-4

4) 4)
4)
4.) 200 - El
04a
84) 150- 10co
0
2
1-- 100- .9
on
i) Ca

500v
U
,
60 2 4 6 8 10 12 14

Applied pressure (kPal102) Applied pressure (kPa 10-2)

Figure 1. Effect of elevated atmospheric pressure for 24 h on the
rate of ethylene production during the next 3 h at atmospheric
pressure. Data ± SD are averages for five seedlings per sample from
three separate experiments. The curve is a regression line calculated
from the data.
ethylene production rate, which peaked at 1300 kPa and
declined rapidly at higher pressures. Root elongation de-
creased quite linearly over the range of pressures tested (Fig.
2). However, a significant reduction in root elongation was
observed only at 400 kPa or higher pressures. Roots from
control seedlings typically became thinner after 24 h (Fig. 3).
Application of pressures up to 300 kPa did not alter this
trend; however, higher pressures, from 500 to 1200 kPa,
resulted in a very slight increase in diameter above control
roots.
Seedlings were examined after pressure treatments, and no
physical injury such as surface wounds, discoloration of
tissue, or leaking of fluid was evident. Seedlings left in the
sand in the envelopes for several days survived and devel-
oped normally. Thus, although the highest pressure treat-
ments were severe, they apparently did not cause extensive
cellular collapse or death.
5 ('soil') around seedlings, thereby increasing resistance to
E growth. In contrast, the present study showed that the
4io
4)
Figure 3. Effect of elevated atmospheric pressure for 24 h on the
change in root diameter from the beginning to the end of that
period. Negative values indicate that root diameter 5 mm behind
the primary root tip decreased during the 24-h treatment period.
Data ± SD are averages for 15 seedlings per sample from three
separate experiments. The curve is a regression line calculated from
the data.
DISCUSSION
The pressure plate chambers were several liters in volume
and contained only 15 seedlings in sand, which remained
moist throughout the experiment. Thus, although effects of
gas partial pressure and water potential differences cannot
be totally eliminated, the major variable clearly was the large
differences in pressure applied. The difference between sub-
jecting an entire plant to elevated pressure versus compress-
ing its growing medium with extemally applied pressure is
the focus of this report. Both approaches employed intact,
germinating seedlings. For reference, 100 kPa equals 1 bar
and 1000 kPa equals 1 MPa.
Pressures of 25 to 50 kPa will induce substantial ethylene
production (8) and/or modification of plant tissue growth (1,
4, 6), provided the plant tissue itself is kept at atmospheric
pressure and only the growth medium is pressurized. In such
studies, elevated pressure compacted the growth medium
amount of pressure required to promote substantial ethylene
production and modify the primary root-growth pattem was

s 3 much higher when the whole system (plant and soil) was
under elevated atmospheric pressure, which did not compress
4)

.0

0
2

10
!0501 X,,<
r
~~~~~~~~~~~~~~~~~~~~~~~~
the medium and which, therefore, did not increase the re-
sistance to growth. Significant enhancement of ethylene pro-
duction did not occur until the pressure reached 800 kPa,
and it peaked at 1300 kPa (Fig. 1). The exact threshold for
OI * * .
elevation of ethylene production may be between 400 and
0 2 4 6 8 10 12 14 800 kPa. Significant reduction of root elongation rate oc-
Applied pressure (kPa-10-2) curred only at 400 kPa, and 800 kPa were necessary to
achieve a 50% reduction in elongation after 24 h (Fig. 2). At
Figure 2. Effect of elevated atmospheric pressure for 24 h on root low atmospheric pressures (0-300 kPa), the normal small
elongation during that period. Data ± SD are averages for 15 reduction in root diameter occurred, whereas a slight increase
seedlings per sample from three separate experiments. The curve in diameter occurred at pressures from 500 to 1500 kPa (Fig.
is a regression line calculated from the data. 3). In contrast, when the tissue was kept at atmospheric
2108 SARQUIS ET AL.
pressure and only the growth medium was pressurized ex-
temally, 100 kPa was enough to induce a 4-fold increase in
ethylene production rate, a 7-fold increase in root diameter,
and a 75% reduction in root elongation over 10 h (8).
If the mechanism by which primary roots and shoots sense
normal contact pressures, such as barriers or impedances to
growth, involves processes sensitive to atmospheric pressure,
we would expect to see sensitive responses to atmospheric
pressures in the present experiments. However, the atmos-
pheric pressures necessary to duplicate the effects of physical
pressure or impedance at the surface of the growing tissues
are at least 30 times higher. Thus, it is likely that the physical
interaction between the plant root and shoot surfaces and
the growing medium is part of the sequence of events that
leads to a modified growth pattern under mechanical imped-
ance. Likewise, it is unlikely that this mechanism is also
sensitive to atmospheric pressure per se. It appears that this
physical interaction, possibly localized in or near exterior
cells, is the initial event involved in the perception of the
mechanical stimulus.
Another way to evaluate the data presented here is in
relation to the fact that plants and other organisms are
naturally exposed directly to elevated hydrostatic pressures
in aqueous environments such as ocean depths (2). Homeo-
viscous adaptation, involving the alteration of membrane
lipid composition, occurs in response to increasing pressure
(5). The site of high hydrostatic pressure injury to cells is the
plasma membrane (3). If the lower levels of pressure em-
ployed here (Figs. 1-3) impact the cell membranes, then the
peak in ethylene production at 1300 kPa may represent a
membrane property change, presumably throughout the or-
ganism. In contrast, in the physical impedance experiments
the stress should occur initially or primarily at the organ
surface (8, 9). Thus, the pressure differences between the two
kinds of experiments are not directly comparable; however,
the magnitudes involved are interesting. We found that a
direct pressure of 1300 kPa promoted ethylene production
Plant Physiol. Vol. 100, 1992
and inhibited root elongation (Figs. 1 and 2); tissue culture
cells of Bromus inermis survived pressure from 25 to 100 MPa
if treated with ABA (10), and 17.2 to 69 MPa induced
membrane lipid compositional changes in a marine bacterium
(5). Because compressing the growth medium with 25 kPa of
pressure has significant effects on production of ethylene and
elongation of roots (8), such treatment is clearly distinguished
from those that require 10-fold (Figs. 1 and 2) to 1000-fold
(5, 10) higher levels of pressure applied directly.
LITERATURE CITED
1. Barley KP (1963) Influence of soil strength on growth of roots.
Soil Sci 96: 175-180
2. Bernhardt G, Jaenicke R, Ludemann H-D, Konig H, Stetter
KO (1988) High pressure enhances the growth rate of ther-
mophilic Archebacterium Methanococcus thermolithotrophicus
without extending its temperature range. Appl Environ Micro-
biol 54: 1258-1261
3. Chong PL, Cossins AR (1983) A differential polarized phase
fluorometric study of the effects of high hydrostatic pressure
upon the fluidity of cellular membranes. Biochemistry 22:
409-415
4. Collis-George N, Yoganathan P (1985) The effect of soil
strength on germination and emergence of wheat (Triticum
aestivum L.). Aust J Soil Res 23: 577-587
5. DeLong EF, Yayanos AA (1985) Adaptation of the membrane
lipids of a deep-sea bacterium to changes in hydrostatic pres-
sure. Science 228: 1101-1102
6. Goss MJ (1977) Effect of mechanical impedance on root growth
in barley (Hordeum vulgare). I. Effects on elongation and
branching of seminal root axis. J Exp Bot 28: 96-1 11
7. Reginato RC, Van Bavel CHM (1962) Pressure cells for soil
cores. Soil Sci Soc Am Proc 21: 1-3
8. Sarquis JI, Jordan WR, Morgan PW (1991) Ethylene evolution
from maize (Zea mays L.) seedling roots and shoots in response
to mechanical impedance. Plant Physiol 96: 1171-1177
9. Sarquis JI, Morgan PW, Jordan WR (1992) Metabolism of 1-
amino-cyclopropane 1-carboxylic acid in etiolated maize seed-
lings grown under mechanical impedance. Plant Physiol 98:
1342-1348
10. Tanino KK, Chen THH, Fuchigami LH, Weiser CJ (1992)
Abscisic acid increases terrestrial plant cell resistance to hy-
drostatic pressure. Plant Physiol 98: 745-748