PHYSIOLOGIA PLANTARUM 109: 244–251.
2000
Printed in Ireland —all rights reser!ed
Effects of ultraviolet-B radiation on leaf elongation, production and
phenylpropanoid concentrations of Deschampsia antarctica and
Colobanthus quitensis in Antarctica
Christopher T. Ruhland* and Thomas A. Day
Department of Plant Biology and The Photosynthesis Center, Life Sciences E-Wing, Room 218, Box 871601, Arizona State Uni!ersity,
Tempe, AZ 85287 -1601, USA
*Corresponding author, e-mail: ruhland@imap2.asu.edu
Received 15 September 1999; revised 1 March 2000
Stratospheric ozone depletion by anthropogenic chlorofluoro-
carbons has lead to increases in ultraviolet-B radiation (UV-B;
280–320 nm) along the Antarctic Peninsula during the austral
spring. We manipulated UV-B levels around plants of Antarc-
tic hair grass (Deschampsia antarctica; Poaceae) and Antarc-
tic pearlwort (Colobanthus quitensis; Caryophyllaceae) for
one field season near Palmer Station along the west coast of
the Antarctic Peninsula. Treatments involved placing frames
over naturally growing plants that either (1) held filters that
absorbed most biologically effective radiation (UV-BBE; ‘re-
duced UV-B’, 22% of ambient UV-BBE levels), (2) held filters
that transmitted most UV-BBE (‘near-ambient UV-B’, 87% of
ambient UV-BBE levels), or (3) lacked filters (‘ambient UV-
B’). Leaves on D. antarctica exposed to near-ambient and
ambient UV-B were 16–17% shorter than those exposed to
reduced UV-B, and this was associated with shorter epidermal
cells at the leaf base and tip. Leaves on C. quitensis exposed
to near-ambient and ambient UV-B tended to be shorter
(P =0.18) and epidermal cells at the leaf base tended to be
smaller than those under reduced UV-B (P !0.10). In order
to further explain reductions in leaf length, we examined leaf
concentrations of insoluble (cell-wall bound) phenyl-
propanoids, since it has been proposed that wall-bound phenyl-
Introduction
Stratospheric ozone depletion by anthropogenic chlor-
ofluorocarbons has led to increases in the flux of ultravio-
let-B radiation (UV-B; 280–320 nm) reaching the earth’s
surface (Madronich et al. 1995), and these increases have
been particularly severe during spring in Antarctica. For
example, the average November irradiance of 298-nm UV-
B at Palmer Station (64°47" S; 64°04" W) along the
west coast of the Antarctic Peninsula has doubled from
244
Copyright © Physiologia Plantarum 2000
ISSN 0031-9317
propanoids such as ferulic acid may constrain cell expansion
and leaf elongation. In both species, HPLC analysis revealed
that ferulic and p-coumaric acid were major components of
both insoluble and soluble phenylpropanoids. Although there
were no significant differences in concentrations between UV-
B treatments, concentrations of insoluble ferulic acid in D.
antarctica tended to be higher under ambient and near-ambi-
ent UV-B than under reduced UV-B (P =0.17). We also
examined bulk-leaf concentrations of soluble (methanol ex-
tractable) UV-B-absorbing compounds and found that concen-
trations were higher in plants exposed to near-ambient and
ambient UV-B than in plants exposed to reduced UV-B. We
also assessed the UV-B-screening effectiveness of leaves that
had developed on plants at the field site with a fiber-optic
microprobe. Leaf epidermal transmittance of 300-nm UV-B
was 4.0 and 0.6% for D. antarctica and C. quitensis, respec-
tively, which is low compared to grasses and herbaceous
dicotyledonous plants found in more temperate climates.
While the leaves of Antarctic vascular plants are relatively
effective at screening UV-B, levels of UV-B in Antarctica are
sufficient to reduce leaf epidermal cell size and leaf elongation
in these species, although the mechanisms for these reductions
remain unclear.
1988 to 1996, and this increase is negatively correlated
with atmospheric ozone concentrations measured with the
Nimbus 7 Total Ozone Mapping Satellite (Booth et al.
1998). Although concentrations of some chlorofluorocar-
bons in the atmosphere have declined (Butler et al. 1992,
Montzka et al. 1996), enhanced levels of UV-B are expected
to continue well into the 21st century (Madronich et al.
1995).
Physiol. Plant. 109, 2000
 Antarctic hair grass (Deschampsia antarctica Desv;
Poaceae) and Antarctic pearlwort (Colobanthus quitensis
(Kunth) Bartl.; Caryophyllaceae) are the only vascular
plants native to Antarctica, where they occur along the west
coast of the peninsula. Only one study has examined the
effect of UV-B on these two species. Day et al. (1999) found
that ambient and near-ambient levels of UV-B reduced
growth of both D. antarctica and C. quitensis. In other
species of plants, enhanced levels of UV-B reduce growth, as
manifested by reductions in leaf elongation (Liu et al. 1995,
Sullivan et al. 1996, Nogués et al. 1998) and leaf area
(Sullivan et al. 1994), although the mechanisms behind these
reductions are poorly understood and not necessarily related
to reductions in photosynthesis (Allen et al. 1998). One
possible explanation for reductions in leaf elongation is
UV-B-induced production of UV-B-absorbing phenyl-
propanoids such as ferulic acid in the cell-wall matrix of the
epidermis. These phenylpropanoids may form crosslinks
between pectin and/or extensin molecules in the epidermis
(Fry 1986, Fry and Miller 1989) and could conceivably
constrain epidermal cell elongation and subsequent leaf
expansion (Liu et al. 1995).
We report the results from an experiment in which we
manipulated ambient UV-B levels around naturally growing
D. antarctica and C. quitensis plants near Palmer Station
along the west coast of the Antarctic Peninsula. We placed
filters that either absorbed or transmitted most of the ambi-
ent UV-B over plants and assessed what effect this had on
leaf length and production. In order to provide possible
explanations for changes in leaf length, we also assessed
epidermal cell size and concentrations of insoluble p-cou-
maric and ferulic acid, since these wall-bound compounds
have been suspected of constraining epidermal cell expan-
sion. In addition to these field experiments, we also exam-
ined the UV-screening effectiveness of leaves which had
developed on plants at our field site using a fiber-optic
microprobe.
Materials and methods
Study site
The field experiment was conducted on the easternmost
island of Stepping Stones (64°47" S; 64°00" W), a group of 3
islands, approximately 3 km SE of Palmer Station, along the
west coast of the Antarctic Peninsula. The island is approx-
imately 1.75 ha in size. About 60% of it’s surface area is
covered by terrestrial vegetation consisting of several species
of mosses, algae and lichens along with among the largest
populations of D. antarctica and C. quitensis in Antarctica
(Komárková et al. 1985). The climate is maritime Antarctic
with annual air temperatures and precipitation averaging
−2.3°C and 38 cm annually (Smith et al. 1996). A more
detailed description of the field site is provided by Day et al.
(1999).
Field manipulations
Fifteen cylindrical frames (21 cm height ×26 cm diameter)
were placed over a mixed community of D. antarctica and
Physiol. Plant. 109, 2000
C. quitensis on the NE-facing slope of the island in early
November 1997. The frames were constructed of galvanized
steel fencing (10-cm square). Frames were arranged in
groups or blocks, based on proximity, with each block
containing one frame of the 3 different UV treatments:
ambient, near-ambient and reduced UV-B. The ambient
treatment consisted of unfiltered frames. Near-ambient and
reduced UV-B treatment frames were covered with 0.2-mm
thick Aclar (Type 22A, ProPlastics, Linden, NJ, USA),
which transmits most UV-B (sharp transmission cut-off
below 270 nm) or Mylar (Cadillac Plastic & Chemical,
Phoenix, AZ, USA), which absorbs most UV-B (sharp
cut-off below 325 nm). In order to reduce passive warming
under the filter treatments, the filters that we wrapped
around the near-ambient and reduced UV-B treatment
frames did not extend completely around each frame. On
the southern side of the frame the filters were open for 260°
around the top of the frame and slanted to an opening of
90° around the bottom of the frame. This design reduced
passive warming while filtering direct UV-B at all solar
angles. Filter tops placed over these frames had 9 2.5-cm
diameter holes that further reduced passive warming and
allowed precipitation to enter the frames. Over the growing
season (November through February), mean diurnal air
temperatures (2 cm height) in the near-ambient and reduced
UV-B filter frames were within 0.2°C of the open frames
(Day et al. 1999). Biologically effective UV-B irradiance
(UV-BBE ; Caldwell 1971, normalized to 300 nm) was mea-
sured with a broadband UV-B dosimeter (SKU 430, Skye
Powys, UK) calibrated against an SUV-100 scanning spec-
troradiometer (Biospherical Instruments, San Diego, CA,
USA) following Day et al. (1999). Diurnal UV-BBE levels
under the near-ambient and reduced UV-B treatments aver-
aged 87 and 22%, respectively, of those under open frames
over the growing season. Diurnal levels of PAR, measured
with a quantum sensor (LI-190SA, Li-Cor, Lincoln, NE,
USA), under the near-ambient and reduced UV-B treat-
ments averaged 96 and 89% of those under open frames
(Day et al. 1999).
Leaf length and production
We marked 7 tillers of D. antarctica and 7 shoots of C.
quitensis under each frame in mid-November 1997 and made
monthly measurements until mid-March 1998. Each tiller
was on a different plant. During the initial census for D.
antarctica, each tiller was marked by wrapping a colored
wire loop (0.5-mm diameter) around its base. A small
black-ink dot was placed at the tip of the second to
youngest leaf on each tiller using a water-proof marker. The
lengths of the marked leaf and longest leaf were measured
with calipers. In addition, the total number of leaves (green
and dead) and the number of tillers produced since the
previous census were recorded.
During the initial census of C. quitensis, each shoot was
marked with a small colored pin (20 mm height ×1 mm
diameter) placed "0.5 cm from its base. A small black-ink
dot was placed on the tip of the uppermost pair of green
leaves. On subsequent monthly censuses, the length of each
leaf above the marked pair of leaves was measured with
245
calipers. In addition, the total number of leaves (green and
dead) and branches produced since the previous census were
recorded.
At the end of the field season in mid-March 1998, all 7
tillers of D. antarctica and shoots of C. quitensis under each
frame were harvested. One tiller and shoot were oven dried
at 60°C for 48 h and weighed to determine specific leaf mass
(SLM). Epidermal cell size was estimated in another tiller
and shoot. These were rehydrated in water for 24 h and cell
size was estimated with microscopy (see below). The leaf
areas of the remaining 5 tillers and shoots under each frame
were measured with a leaf-area meter (CI-202, CID, Van-
couver, WA, USA), oven dried and transported to Arizona
State University for HPLC analysis of phenylpropanoid
compounds (see below).
Epidermal cell size
We assessed epidermal cell size by measuring epidermal cell
length (D. antarctica) or epidermal cell area (C. quitensis)
on one tiller and shoot harvested from each frame. Since
epidermal cell number and cell size dictate leaf size (Dale
1988, 1992), we measured epidermal cell length (D. antarc-
tica) and cell size (C. quitensis) to see if differences in leaf
length appeared attributable to smaller epidermal cells. In
D. antarctica, the length of 10 randomly selected adaxial
epidermal cells were measured at the base, the mid-length
and the tip of the longest leaf of each tiller following
Kubı́nová (1994). Measurements of epidermal cell lengths
were facilitated by observing UV-induced blue–green
fluorescence emission from epidermal cell walls using an
epifluorescence microscope (16WL, Carl Zeiss Inc., Thorn-
wood, NY, USA) equipped with a UV-excitation filter (365
nm) and an ocular micrometer under 400 ×magnification.
Epidermal cells of some dicotyledonous plants such as C.
quitensis have undulating anticlinal cell walls, which make
epidermal cell lengths vary, depending on where across the
cell this is measured. Hence, we estimated the mean adaxial
epidermal cell area on the longest leaf of each shoot using
the unbiased two-dimensional sampling rule following Ku-
bı́nová (1994). A 62 500-!m2 sampling frame was used with
the epifluorescence microscope by using an ocular grid
under 400×magnification. Average epidermal cell area was
measured in one frame at the base, the mid-length and the
tip of each leaf. In both species we also measured leaf and
epidermal thickness on fresh cross-sections at mid-length of
the leaves, halfway between the leaf margins in D. antarctica
and halfway between the mid-vein and margin in C.
quitensis.
Bulk soluble UV-B-absorbing compound concentrations
We measured bulk concentrations of UV-B-absorbing com-
pounds in fully expanded leaves on one tiller of D. antarc-
tica and one shoot of C. quitensis that we collected in
February 1998 from plants adjacent to those with the
marked tillers and shoots under each frame. For each
measurement, a leaf area of 0.30–0.36 cm2 was placed in a
20-ml Erlenmeyer flask containing 5 ml of acidified
246
methanol (MeOH:HCl:H2O; 90:1:1). Solutions were heated
(60°C) and stirred for 10 min, cooled at room temperature
for 15 min and filtered through 90-!m screens. Concentra-
tions of soluble UV-B-absorbing compounds were estimated
by measuring absorbance at 300 nm with a UV/visible
spectrophotometer (Lambda 4, PerkinElmer, Norwalk CT,
USA) following Day et al. (1994).
Soluble and insoluble p-coumaric and ferulic acid
concentrations
We modified the HPLC technique of Liu et al. (1995) for
isolation and quantification of insoluble and soluble p-cou-
maric and ferulic acids. Dried leaf samples were ground to a
fine powder in liquid N2 with a mortar and pestle and
weighed with an analytical balance. Soluble phenyl-
propanoids were extracted from the powder with acidified
MeOH for 48 h and centrifuged at 1200 g for 10 min. The
supernatant and pellet were considered the source of the
soluble and insoluble phenylpropanoids, respectively. p-
Coumaric and ferulic acid esters in both insoluble and
soluble samples were converted to free p-coumaric and
ferulic acid by alkaline hydrolysis with 2 M NaOH under N2
gas for 1 h. The insoluble and soluble samples were brought
to a pH of #1.0 with HCl and extracted twice with diethyl
ether, once with ethyl acetate and dried under N2 gas.
Samples were then resuspended in 0.5 ml of MeOH. p-Cou-
maric and ferulic acid were separated and identified on a
Merck Lichrocart reverse-phase column (125×4 mm; EM
Science, Hawthorne, NY, USA) at a flow rate of 1 ml
min − 1. Two solvents were used: 1% (v/v) H3PO4 in HPLC-
grade H2O, and pure acetonitrile (MeCN). The elution was
started at 16% MeCN for 0.5 min and increased to 60% over
25 min. Detection was at 314 and 320 nm using a multi-
wavelength detector (model 1040A, Hewlett Packard, Palo
Alto, CA, USA). Determination of concentrations of p-cou-
maric and ferulic acid were compared against commercial
standards (Sigma-Aldrich Chemical, St Louis, MO, USA).
Epidermal transmittance of UV
We measured the UV epidermal transmittance of leaves that
had developed on plants growing under ambient conditions
at our field site during the 1995 growing season. Whole
plants were collected and transported in insulated boxes
(4°C) to Arizona State University. The plants were kept in
growth chambers at 12°C with a 12-h photoperiod of 500
!mol m − 2 s − 1 PAR for 30 days prior to measurements.
Plants received no UV in the growth chambers. A fiber-optic
microprobe (Vogelmann et al. 1991) was used to measure
epidermal transmittance of 280–380 nm UV in an older leaf
of 10 plants of each species following Day et al. (1994). The
microprobe consisted of a fused-silica optical fiber (125-!m
OD), which tapered to a 15-!m diameter radiation-collect-
ing tip. Leaves were mounted in a leaf holder and irradiated
on the adaxial surface with broadband radiation from a
150-W xenon-arc lamp (model 6602, Oriel Corp., Stratford,
CT, USA). The microprobe was positioned at the abaxial
surface of the leaf with a micromanipulator and advanced
through the leaf at an orientation of 0° (microprobe facing
Physiol. Plant. 109, 2000
radiation source) to the adaxial epidermis/mesophyll inter-
face. The location of this interface was determined on fresh
cross-sections of adjacent leaf segments by measuring epi-
dermal thickness with a light microscope (BX50, Olympus,
Lake Success, NY, USA) equipped with an ocular microme-
ter. During measurements, the other end of the optical fiber
was attached to a UV/visible spectroradiometer (model 742,
Optronic Laboratories, Orlando, FL, USA). Epidermal
transmittance of UV was assessed by scanning from 280–
380 nm in 1-nm increments with the spectroradiometer.
In addition, UV-absorption spectra of bulk soluble UV-
absorbing compounds were measured in 10 leaves that were
adjacent to the leaves used for measurements of epidermal
transmittance. Compounds were extracted in acidified
methanol as previously described. Absorbance of the aci-
dified methanol extracts was measured every 1 nm from
280–380 nm using the spectrophotometer and concentra-
tions were corrected for leaf area.

Statistical analysis
The general linear model procedure was used with two-way
analysis of variance (ANOVA; Statistix 1996) to examine
UV-B and block effects. The least significant difference test
(LSD; Statistix 1996) was used to compare treatment means. Fig. 1. Effect of UV-B reduction on (A) length of the longest leaf
and epidermal cell length in (B) the leaf tip, (C) middle of the leaf,
and (D) the leaf base of D. antarctica. Treatments: ambient UV-B
(unfiltered frames), near-ambient UV-B (Aclar filters) and reduced
UV-B (Mylar filters).Values are means (n =5 plants)$ SE.
Results
Leaf length and production UV-B (LSD, P !0.05). Epidermal cells at the leaf base
averaged 1257 and 1678 !m2 in size under near-ambient and
UV-B had a significant effect on the length of the longest
ambient UV-B, respectively, compared to 1785 !m2 under
leaf in D. antarctica at the end of the growing season
(ANOVA, P !0.05). Leaves under near-ambient and ambi-
ent UV-B were 16 and 17% shorter, respectively, than those
under reduced UV-B (LSD, P !0.05; Fig. 1). There were no
significant UV-B effects on tiller or leaf production in D.
antarctica (data not shown). UV-B tended to have an effect
on leaf length in C. quitensis (ANOVA, P =0.18); leaves
under near-ambient UV-B were 10% shorter than those
under reduced UV-B (LSD, P !0.10; Fig. 2). There were no
significant UV-B effects on leaf or shoot production in C.
quitensis or on SLM in either species (data not shown).

Epidermal cell size

In D. antarctica, epidermal cells were significantly shorter at
two leaf locations (leaf base and tip) in plants growing
under near-ambient and ambient UV-B compared to those
under reduced UV-B (LSD, P !0.05). Epidermal cells at the
leaf base, mid-leaf and leaf tip averaged 131, 123 and 73 !m
long under near-ambient UV-B and 135, 148 and 75 !m
long under ambient UV-B compared to 165, 149 and 90 !m
long under reduced UV-B (Fig. 1). While epidermal cells of
D. antarctica were shorter under near-ambient and ambient
UV-B, the adaxial epidermis was thicker under near-ambi-
ent (16.8 !m) and ambient UV-B (14.8 !m) than under
reduced UV-B (12.7 !m; LSD, P !0.05; data not shown). Fig. 2. Effect of UV-B reduction on (A) leaf length, and epidermal
cell area in (B) the leaf tip, (C) middle of the leaf, and (D) the leaf
In C. quitensis, epidermal cells were smaller at the base of base of C. quitensis (treatments in Fig. 1).Values are means (n = 5
the leaf under near-ambient UV-B than under reduced plants) $ SE.

Physiol. Plant. 109, 2000 247


Fig. 3. Effect of UV-B reduction on concentrations of bulk soluble
UV-B-absorbing compounds (assessed by absorbance at 300 nm of
whole-leaf extracts) on a leaf-area basis in (A) D. antarctica and (B)
C. quitensis (treatments in Fig. 1). Values are means (n =5
plants)$ SE.
reduced UV-B. There were no significant UV-B effects on
epidermal cell area at the middle or the tip of the leaf (Fig.
2), or epidermal thickness in C. quitensis (data not shown).
Bulk soluble UV-B-absorbing compound concentrations
There were no significant UV-B effects on concentrations of
soluble UV-B-absorbing compounds in leaves of D. antarc-
tica (Fig. 3). However, in C. quitensis, concentrations of
soluble UV-B-absorbing compounds in leaves were signifi-
cantly higher under near-ambient and ambient UV-B than
under reduced UV-B (LSD, P !0.05; Fig. 3).
Soluble and insoluble p-coumaric and ferulic acid
concentrations
Both p-coumaric and ferulic acid were major components of
insoluble and soluble UV-B-absorbing compounds in leaves
(Fig. 4). There were no significant UV-B effects on insoluble
or soluble concentrations of these compounds in either of
the species (Figs. 5 and 6). However, concentrations of
insoluble ferulic acid tended to be higher in D. antarctica
leaves under near-ambient and ambient UV-B than reduced
UV-B (LSD, P =0.17).
Epidermal transmittance of UV
In both species, epidermal UV transmittance was low in
leaves that had developed on plants at our field site and
subsequently placed in growth chambers. Leaf epidermal
248
transmittance of 300-nm UV-B averaged 4.0 and 0.6% in D.
antarctica and C. quitensis, respectively (Fig. 7). Leaf con-
centrations of bulk soluble UV-B-absorbing compounds
averaged 0.43 and 0.35 A300 cm − 2 in D. antarctica and C.
quitensis, respectively (Fig. 7). In general, troughs in epider-
mal transmittance mirrored peaks in absorbance and vice
versa across the 280–380 nm waveband (Fig. 7).
Discussion
Ambient and near-ambient levels of UV-B reduced leaf
elongation in D. antarctica (Fig. 1) and there was a tendency
for this in C. quitensis (Fig. 2). These findings are consistent
with Day et al. (1999) who found that the most consistent
UV-B effect on several growth parameters measured in D.
antarctica was a reduction in leaf length. We found that
leaves of D. antarctica exposed to ambient UV-B were 17%
shorter than leaves exposed to reduced UV-B, which is
similar to the 20% reduction in leaf length Day et al. (1999)
observed. Possible explanations for these reductions in leaf
length include: (1) UV-B-induced reductions in photosynthe-
sis (Greenberg et al. 1989, Melis et al. 1992, Jansen et al.
1993, 1996), (2) stimulation of a UV-B photoreceptor, possi-
bly a flavin (Ballaré et al. 1995), that reduces cellular
division (Dickson and Caldwell 1978, Wellman 1983,
Nogués et al. 1998), (3) stimulation of insoluble UV-B-ab-
sorbing phenylpropanoids that are bound in the cell-wall
matrix of epidermal cells and may constrain epidermal cell
elongation and leaf expansion (Liu et al. 1995).
Although possible, it seems unlikely that these reductions
in leaf elongation in D. antarctica were associated with
photosynthetic targets or other photoreceptors in the meso-
phyll. Leaf epidermal transmittance of UV-B in plants
brought back from the field site was low in both D. antarc-
tica (4.0%) and C. quitensis (0.6%; Fig. 7). Although other
Fig. 4. Representative elution profiles from HPLC analyses of
insoluble (cell-wall bound) and soluble phenylpropanoids isolated
by alkaline hydrolysis from leaves of D. antarctica (A and B) and C.
quitensis (C and D) growing under ambient UV-B. Peak identifica-
tion: 1 = p-coumaric acid (Rf # 5.7 min); 2 =ferulic acid (Rf # 6.1
min). Detection was at 314 nm.
Physiol. Plant. 109, 2000
 near-ambient and ambient UV-B (P =0.17), which could
conceivably explain reductions in leaf length.
Concentrations of wall-bound ferulic and p-coumaric acid
in D. antarctica and C. quitensis appear to be relatively high
compared to related species examined to date. High concen-
trations of ferulic and p-coumaric acid are relatively uncom-
mon in plants, occurring in conifers (Strack et al. 1988,
Sánchez et al. 1996, Hoque and Remus 1999), certain mono-
cotyledonous plants including grasses (Harris and Hartley
1980, Lichtenthaler and Schweiger 1998), and to a lesser
extent among dicotyledonous plants in the Caryophyllaceae
and other families in the order Caryophyllales (Hartley and
Harris 1981). Lichtenthaler and Schweiger (1998) found that
the grasses Zea mays, Triticum aesti!um, and A!ena sati!a
contained much higher concentrations of insoluble p-cou-
maric (1–5 mg g − 1) and ferulic acid (2–6 mg g − 1) than
plants in the order Caryophyllales. The latter (Amaranthus
caudatus, Spinacia oleracea and Portulaca oleracea) had no
detectable levels of p-coumaric and !1 mg g − 1 of ferulic
acid. In contrast, we found higher concentrations of p-cou-
maric acid in the grass D. antarctica (5–8 mg g − 1; Fig. 5)
and much higher concentrations of both wall-bound com-
pounds in C. quitensis. Concentrations of p-coumaric and
ferulic acid in the latter ranged from 5–8 mg g − 1 and 7 – 10
mg g − 1 of p-coumaric and ferulic acid, respectively (Fig. 6).
Leaf epidermal transmittance of 300-nm UV-B was sur-
prisingly low in both species, averaging 4.0 and 0.6% in D.
Fig. 5. Effect of UV-B reduction on concentrations of insoluble (A)
p-coumaric and (B) ferulic acid and soluble (C) p-coumaric and (D)
ferulic acid (assessed by HPLC at 314-nm detection) on a dry-mass
basis in leaves of D. antarctica (treatments in Fig. 1). Values are
means (n = 5 plants)$ SE.

studies have found that shorter leaves under UV-B are

associated with reductions in the number of cells (Dickson
and Caldwell 1978, Logemann et al. 1995, Nogués et al.
1998), these reductions can also be accompanied by smaller
epidermal cells (Nogués et al. 1998). We found that near-
ambient and ambient UV-B reduced adaxial epidermal cell
size by 21 and 18% at the leaf base and 19 and 17% at the
leaf tip, respectively, in D. antarctica (Fig. 1). Near-ambient
UV-B reduced adaxial epidermal cell size by 30% at the leaf
base in C. quitensis (Fig. 2). Recent studies have suggested
that UV-B-induced increases in cell-wall bound ferulic acid
may play a role in reducing leaf expansion (Liu et al. 1995).
Transcription of key enzymes in the phenylpropanoid path-
way, most notably phenylalanine ammonia lyase (PAL), can
be stimulated by UV-B (Li et al. 1993, Liu and McClure
1995) and at least one study has shown that concentrations
of wall-bound ferulic acid increase under enhanced levels of
UV-B (Liu et al. 1995). Although the role of these phenyl-
propanoids in the cell-wall matrix is not completely under-
stood (Lu and Ralph 1998), they form side-chains on
polysaccharides and can crosslink cellulose/hemicellulose
microfibrils in the cell-wall matrix. It has been proposed that
these crosslinks may limit cell-wall expansion (Fry 1986, Fry
and Miller 1989). Increased concentrations of wall-bound or Fig. 6. Effect of UV-B reduction on concentrations of insoluble (A)
insoluble ferulic acid may indicate more crosslinks in the p-coumaric and (B) ferulic acid and soluble (C) p-coumaric and (D)
ferulic acid (assessed by HPLC at 314-nm detection) on a dry-mass
epidermal cell walls. Concentrations of insoluble ferulic acid basis in leaves of C. quitensis (treatments in Fig. 1). Values are
tended to be higher in D. antarctica plants grown under means (n =5 plants)$ SE.

Physiol. Plant. 109, 2000 249


Fig. 7. Leaf epidermal UV transmittance (assessed with a fiber-op-
tic microprobe) and soluble UV-absorption spectra (assessed from
whole-leaf acidified methanol extracts) in (A) D. antarctica and (B)
C. quitensis (bottom). Leaves for measurements developed under
ambient conditions at the field site and then were placed in a
growth chamber without UV 30 days prior to measurements. Val-
ues are means (n= 10 plants).
antarctica and C. quitensis, respectively. These values are
lower than those found in other grasses and herbaceous
dicotyledonous plants from more temperate climates. For
example, in a survey of naturally growing plants in West
Virginia, USA, Day (1993) found that epidermal transmit-
tance of 300-nm UV-B ranged from 18–58% in grasses and
7–58% in herbaceous dicotyledonous plants. While leaves of
D. antarctica and C. quitensis were relatively effective at
screening UV-B, bulk concentrations of soluble UV-B-ab-
sorbing compounds in leaves of the plants we made these
measurements on were relatively low, averaging 0.43 and
0.35 A300 cm − 2 for D. antarctica and C. quitensis, respec-
tively. One explanation for the surprisingly effective UV-B
screening in these species in view of their low concentrations
of soluble screening compounds, is their high concentrations
of wall-bound phenylpropanoids. A major pathway for UV-
B transmittance through the epidermis in leaves which lack
substantial concentrations of UV-B screening compounds, is
through anticlinal cell walls of the epidermis (Day et al.
1993). The relatively high concentrations of p-coumaric and
ferulic acid in cell walls of D. antarctica and C. quitensis
would provide a much more spatially uniform filter and
explain their low UV-B epidermal transmittance.
Our findings show that ambient and near-ambient levels
of UV-B in Antarctica reduce leaf length in D. antarctica
and C. quitensis. These reductions are associated with
smaller epidermal cells in both species, although the mecha-
nisms behind these reductions are unclear. Both of these
species have relatively high concentrations of wall-bound
250
p-coumaric and ferulic acid, which have been implicated in
constraining epidermal cell elongation and overall leaf ex-
pansion, although we were unable to detect significant in-
creases in concentrations of these compounds under ambient
levels of UV-B.
Acknowledgements – This work was supported by NSF grant
OPP-9615268. Logistical support provided by personnel of Antarc-
tic Support Associates at Palmer Station is greatly appreciated.
Fusheng Xiong and Erin C. Mueller provided field assistance and
W. Dennis Clark and Christopher Byrd assisted with HPLC mea-
surements. This is publication number 422 from The Photosynthesis
Center at Arizona State University.
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