UV-B-Mediated Changes on Below-Ground Communities Associated with the Roots of

Acer saccharum
Author(s): J. N. Klironomos and M. F. Allen
Source: Functional Ecology , Dec., 1995, Vol. 9, No. 6 (Dec., 1995), pp. 923-930
Published by: British Ecological Society

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Functional
Ecology 1995
UV-B-mediated changes on below-ground communities
9, 923-930
associated with the roots of Acer saccharum

J. N. KLIRONOMOS and M. F. ALLEN

Biology Department and Soil Ecology and Restoration Group, San Diego State University, San Diego, CA
92182-0057, USA

Summary

1. Little is known about how exposure to UV-B radiation affects rhizosphere

microbes. Rhizosphere organisms are fed primarily by root-derived substrates and
fulfil functions such as mineralization, immobilization, decomposition, pathogeneity
and improvement of plant nutrition; they form the base of the below-ground food web.
2. In this study, we exposed Sugar Maple (Acer saccharum) seedlings to UV-B
radiation in order to determine if UV-B influences the activities of mycorrhizal and
non-mycorrhizal fungi, bacteria and microbe-feeding arthropods in the rhizosphere.
3. Below-ground organisms are greatly affected by UV-B radiation. Overall, carbon-
flow in the plant soil system was shifted from a mutualistic-closed, mycorrhizal-
dominated system to an opportunist-open, saprobe/pathogen-dominated one.

Key-words: Acari, arbuscular mycorrhiza, bacteria, Collembola, global change, ultraviolet-B

Functional Ecology (1995) 9, 923-930

Introduction ties and a significant proportion of available carbon is

Global change models predict continual reductions in
the earth's protective ozone layer (Gurney, Foster &
Parkinson 1993). Such reductions are of concern
because this layer is the primary attenuator of solar
ultraviolet-B radiation (UV-B: 280-320 nm). Even
though UV-B represents a small proportion of the total
solar electromagnetic spectrum, it exerts profound
effects on living organisms because it can be absorbed
by macromolecules such as proteins and nucleic acids.
Biological effects of UV-B radiation on higher
plants have been studied extensively and many of the
deleterious effects have been documented (Caldwell,
Teramura & Tevini 1989). Many plants have proper-
ties that help them tolerate higher UV-B fluxes
(Sullivan, Teramura & Ziska 1992), especially those
found in high latitudes and altitudes. However, plant
species found in lower altitudes and latitudes tend to
be more vulnerable to UV-B irradiance. This is of
concern because significant decreases in total ozone
have been reported throughout the globe over the past
decade (Gurney et al. 1993).
A large body of literature is available on UV-B
effects on above-ground organs, tissues and cells at
the physiological, structural and biochemical levels
(Caldwell et al. 1989). However, little attention has
been paid to the effects of UV-B irradiance on below-
ground organisms and their functioning, even though
it is recognized that these effects may be important
?C 1995 British (Moorhead & Callaghan, 1994; Gehrke et al. 1995).
Ecological Society Roots are a major source of energy for soil communi-
allocated below ground (Agren et al. 1980), and ulti-
mately fuels the soil ecosystem (Coleman 1985).
We chose to work with Sugar Maple (Acer saccha-
rum Marsh. L.) because it is an economically important
tree species in eastern North America and has been
declining over the past decade (Hendershot & Jones
1989). Also, Sugar Maple is a shade-tolerant tree
species. Seedlings and saplings can grow slowly in the
sub-canopy layers for many years before being exposed
to direct solar radiation during gap formation, when
young seedlings will require high rates of nutrient sup-
plies to compete with neighbours for the upper canopy.
During preliminary observations we did not detect
any obvious visual stress symptoms of Sugar Maple to
UV-B exposure. In this study, we exposed Sugar
Maple seedlings, which were growing in intact maple-
forest soil, to UV-B radiation in order to determine if
UV-B influences the activity of mycorrhizal and non-
mycorrhizal fungi, bacterial community composition
and microarthropods below ground.
Materials and methods
On 16 April 1992, 120 pre-germinated Sugar Maple
seeds were planted at random within a 100m2 area
(Klironomos & Kendrick 1995a) of a Sugar Maple for-
est near Waterloo, Ontario, Canada (Klironomos et al.
1993; Klironomos & Kendrick 1995a). The dominant
tree species throughout the site is Sugar Maple and the
surface soil is a sandy loam with a low fertility and
water holding capacity.

923

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924 Sixteen months later, on 4 August 1993, 80 of the
J. N. Klironomos seedlings, each with c. 1 litre of intact soil, were
& M. F. Allen removed by driving a 10 cm diameter, 15 cm deep steel
cylinder into the ground with a 12 kg weight that slides
along a steel rod (Brundrett, Melville & Peterson 1994).
Each of these cores contained one 16 month old Sugar
Maple seedling. Less than 5% of the root biomass was
lost as a result of soil coring. The soil and plant were
ejected from the steel ring into plastic lined 1 litre pots
and brought back to the laboratory. Twenty plants
were harvested immediately to record baseline data.
The remaining 60 plants were placed in growth cham-
bers set at 22 'C, with a 16 h photoperiod. Visible light
(400-700 nm, 100 gmol m-2 s-1) at the leaf surface was
generated with white fluorescent lamps, suspended
above and perpendicular to the plants, and filtered
through clear 008 mm-thick polyester film (320 nm
cut-off, Transil Wrap, Inc., Toronto, Ont.) to screen
out extraneous UV-B and UV-C radiation. The three
light treatments began 2 weeks later. Field capacity (%
water) levels were determined for samples of each soil
and used to calculate the weight of each core when it
was watered to field capacity. No additional nutrients
were added throughout the experiment.
UV TREATMENT
Sixty pots, each containing a 1 year old Sugar Maple
seedling, were subjected to one of three light treat-
ments, 20 pots per treatment. Light treatments were
either (a) visible light, (b) visible light plus UV-A
radiation or (c) visible light plus UV-B radiation,
given in 16h photoperiods. Three growth chambers
were used for the experiment, so plants and treatments
were randomly rotated at weekly intervals to avoid
chamber effects. Visible light was generated at
100 rmol m-2 s-1, as described above. UV-B and UV-in a blender with 250 ml of water at high
agitated
A were generated with photoreactor lamps RPR-3000
and RPR-3500, respectively (Southern New England
Ultraviolet Co., Hamden, CT). The UV-B was filtered
through clear, 0 08 mm, cellulose acetate film to elim-
inate extraneous UV-C (<290 nm). UV-A radiation
was filtered through clear, 0-08 mm, polyester film to
remove both UV-C and UV-B radiation (<320 nm).
The spectral outputs of these sources are shown in
Fig. 1. The fluence rate of either UV-A or UV-B radi-
ation applied at the plant leaf surface was 6 gmol m-2
s-1 and was controlled by changing the number oftwo 50 ml centrifuge tubes, floated on top of a 60%
lamps. The plants were exposed to these treatments
for 8 weeks and then harvested.
From each pot, a 4 cm diameter core for arbuscular-
mycorrhizal (AM) fungal spore extractions and bacte-
rial analyses, and a 4 cm diameter core for hyphal
extractions, were taken. For the bacterial analyses,
only soil still attached to roots (rhizosphere) after
shaking individual root pieces was used. Shoots were
?) 1995 British
Ecological Society,
then cut off and roots were carefully separated from
Functional Ecology, the soil. A subsample of the roots was cut into 1 cm
9, 923-930 fragments and 20 feeder root pieces (recognized by
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- '. 1 l UV-A
o 0 o75
Fig. 1. Sectr of V-A nd U-B lght ourcs us0.. d UV-
0
*6 0.5-
0.9
Ca
ai: 0.25-
0~
0 0 0 0 LO 0 0O 0 0
10 0 "i t 0 i 0O L 0 10 0 - f
Wavelength (nm)
Fig. 1. Spectra of UV-A and UV-B lig
study.
their characteristic beaded appearance) (Kessler
1966) were removed and stored in formalin-acetic
acid (FAA) for subsequent assessment of root colo-
nization. The remaining roots were used to count the
number of young AM fungal spores still attached to
them. The remaining potting media were placed in
polythene bags for arthropod extractions. Shoots and
roots were oven-dried at 80 'C for 24 h to determine
dry mass.
AM FUNGAL SPORE EXTRACTION FROM SOIL
Spore abundance was calculated directly by extracting
AM spores from soil using a modified wet-sieving
technique (Klironomos et al. 1993). Each sample was
speed for 1 min. The resulting suspension was passed
through two sieves (mesh 1 0 mm and 500 gim) using a
high-pressure water hose, collected in a large plastic
tub and then passed through another two sieves (mesh
250 and 45 gim). Material retained by the 45 gm sieve
was resuspended in 1 litre of water, allowed to stand
and passed through another 45 gm sieve. This process
was repeated three times. The resulting material was
collected in 40-60 ml of water.
The spore suspension was equally divided between
sucrose solution, and then centrifuged at 688 xg for
20 min. Material in the water and at the water-sucrose
interface was collected with a Pasteur pipette and
passed through a 1.2gm millipore filter. The spores
were collected on gridded (3 x 3 mm) nitrocellulose fil-
ter paper and washed with distilled water, under a fil-
tration vacuum, to spread spores evenly over the grid.
Populations of spores were estimated by counting
all spores present on 10 randomly chosen squares
from the entire grid. A subsample of spores from the
grid was mounted in a polyvinyl-alcohol mounting
925 medium, examined under differential interference-
Below-ground contrast illumination and identified using the compila-
responses to tion of AM fungi by Berch (1988).
UV-B
AM FUNGAL SPORES ATTACHED TO ROOTS
Roots were soaked in a 0.5% solution of sodium hexa-
metaphosphate (as described by Moutoglis et al. 1995)
and gently agitated to remove most of the adhering soil.
They were then rinsed in tap water until the root system
was free of all or most debris, leaving the attached extra-
matrical AM hyphae and spores. The number of spores
on each plant was counted and samples of spores were
transferred to FAA (formalin, alcohol, acetic acid) for
later quantification and identification.
HYPHAL LENGTH
Total hyphal lengths were estimated by extracting
hyphae from the soil (Miller, Reinhardt & Jastrow
1995) and measuring lengths by a gridline-intersect
method. Two 5 g portions of soil were removed
from each sample, then suspended in 250 ml dis-
tilled water. To break up aggregates, 30ml of 3.6%
w/v sodium hexametaphosphate solution was added
and left for 16-18 h. Samples were then stirred to
break up any remaining aggregates. The soil suspen-
sion was agitated at high speed in a blender for
2 min. The suspension was then stirred with an elec-
tronic stir-bar at such a speed that the vortex was
about half way between the top of the suspension
and the bottom of the beaker. One 6 ml aliquot per
sample was removed from half way between the
beaker edge and the vortex and transferred to
another beaker. To this, 250 ml distilled water were
added along with 30ml 3.6% w/v sodium hexa-
metaphosphate solution. The diluted suspension
was slowly stirred again to resuspend hyphae, then
10ml aliquots were taken and transferred to 50ml
centrifuge tubes.
Samples were centrifuged at 1000 xg for 8 min and
the supernatant was discarded. After five repetitions
of the extraction protocol, no additional hyphal frag-
ments were extracted. The efficiency of the first
extraction was calculated to be 61%. To the remain-
ing pellet, 10 ml 50% glycerol was added and the pel-
let was resuspended with a vortex mixer and then described in Washington (1981). Bacterial isolates
centrifuged at 75 xg for 30 s. The supernatant was fil-were stab-inoculated and grown at 22 'C for 2 weeks.
tered onto a 1.2 gm nitrocellulose filter paper. Filters Bacteria capable of protein hydrolysis were those that
were cut in half, placed on glass slides and dried at
35?C for 15min. For microscopy, the filters were
made transparent by mounting in low-viscosity
immersion oil.
Slides were viewed at x 100 magnification through a
lOx 10 grid reticule placed in the eyepiece. Using the
gridline intersect method, intersections were counted
(C 1995 British
for six horizontal and six vertical alternating lines.
Ecological Society,
Functional Ecology, This was done for 70 fields of view and hyphal length
9, 923-930 g-' dry soil was calculated as in Newman (1966).
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ROOT INFECTION
Feeder roots were stored in FAA for at least 24 h.
Roots were then cleared by autoclaving for 15 min in
10% potassium hydroxide, bleached in 35% hydrogen
peroxide for 30 min, acidified in FAA for 5min and
stained using Chlorazol Black E (Brundrett, Piche &
Peterson 1984). Fungal infection was quantified using
the magnified intersections method (McGonigle et al.
1990) by inspecting intersections between the micro-
scope eyepiece cross-hair and roots at x 200 magnifi-
cation. The proportions of root length containing
arbuscules, vesicles, hyphal coils and hyphae were
determined. The hyphae of non-mycorrhizal fungi
were distinguished from those of AM by careful
observation of characters normally missing in the lat-
ter, such as melanization, clamp connections or regu-
larly septate hyphae.
BACTERIAL ANALYSES
Total bacterial populations were determined by the
dilution plate method using a soil extract agar
medium (James 1958) and total actinomycete popula-
tions using a Kuster' s agar medium (Kuster &
Williams 1964). After incubation and enumeration
the colonies of bacteria and actinomycetes were
picked from either the entire area or a representative
sector of agar plates, purified and maintained to study
the morphological and physiological characters. A
total of 50 bacteria and 30 actinomycetes were
obtained from each pot.
The bacterial isolates from the root zon were stud-
ied for their gram reaction and spore for tion. They
were also studied for certain physiological character-
istics: starch hydrolysis, urea hydrolysis and protein
hydrolysis. One per cent starch agar was prepared as
described in Washington (1981). Amylolytic activity
was detected after flooding the plates with an iodine
solution. A yellow zone around the colony indicated
starch hydrolysis; otherwise the starch reacts with the
iodine to turn the medium blue. Two per cent urea
agar was prepared as described in Washington (1981).
Bacterial isolates were streaked on the agar slants and
slants which changed from light orange to magenta
colour were defined positive for hydrolysis of urea.
Twelve per cent gelatin agar was prepared as
liquified gelatin after 30 min at 4 'C.
MICROARTHROPOD EXTRACTION
Microarthropods were extracted using a high ef
canister-type soil-arthropod extractor (Lussenhop
1971). Microarthropods were sorted, counted and then
stored in 70% ethanol. Specimens requiring further
examination were mounted on slides using Hoyer' s
medium.
926 STATISTICAL ANALYSES To help determine which individual variables were
J. N. Klironomos significantly affected by the light treatments, univari-
The data were analysed with a multivariate analysis of
& M. F. Allen ate analyses were used. Total root colonization by AM
variance (ANOVA). The independent variable was light
fungi did not change with light treatment; however,
treatment and the dependent variables were all mea-
drastic changes were observed with type of infection
surements made after harvest (shoot and root biomass,
(Fig. 3). Under UV-A and control, the frequencies
arbuscular, vesicular, coil and hyphal colonization,
of arbuscules and hyphal coils were relatively high,
melanized, non-melanized and clamped hyphal
whereas the frequency of vesicles was low. Under
length, spore abundance, collembolans, oribatid mites,
UV-B, the number of hyphal coils remained high but
non-oribatid mites, predatory mites, bacteria and acti-
shifts were seen with arbuscular and vesicular infection.
nomycetes). Results of evaluation of assumptions
of normality, homogeneity of variance covariance
Arbuscular colonization was decreased (F2,57 = 7 05,
P = 0.002) by 69%, whereas vesicular infection was
matrices, linearity and multicollinearity were satisfac-
tory. All variables were also analysed further using increased (F2,57 = 20.91, P = 0 .0001) by 343%. Infection
of the young maple roots by non-mycorrhizal fungi
univariate ANOVA to help determine which variables
contribute to any significant differences observed in also increased significantly (F2,57= 585, P=0.005),
more than doubling with UV-B radiation.
the multivariate analysis. The Tukey post-hoc test was
The density and species composition of AM fungal
used to test for differences among means. All statisti-
spores extracted from the soil did not differ with light
cal analyses were performed using the SYSTAT soft-
treatment (data not shown). The species detected were
ware (Wilkinson 1990).
identified as Acaulospora foveata, Glomus geospo-
rum, Glomus hoi and Glomus macrocarpum. More
Results details on the occurrence of AM fungal propagules in
this field site are provided in Klironomos & Kendrick
At harvest, the Sugar Maple seedlings showed no
(I 995a). However, the average number of young spores
apparent direct visual effects in response to the various
found attached to the roots was three-fold higher
irradiance treatments. Among all treatments the plants
(F2,57= 19 34, P = 0.0001) with UV-B radiation (Fig. 4).
seemed equally healthy, and shoot (F2,57 = 0 32,
All attached spores belonged to the genus Glomus but
P > 0.05) and root (F2,57 = 1-56, P > 0*05) biomass did
were too young to be identified to species level.
not change as a result of UV-A or UV-B radiation
Extraradical hyphal lengths were also highest under
(Fig. 2). However, below-ground organisms associated
the UV-B treatment. Hyphal fragments were catego-
with the roots of the young Sugar Maple seedlings
rized into three groups: i.e. with melanized thalli, non-
were greatly affected. With the use of the multivariate
melanized, and with clamp connections, as seen in
Wilks' criterion, the combined dependent variables
Fig. 5, and all three showed increases (P < 0 .05). AM
(shoot and root biomass, bacterial and actinomycete
fungal hyphae would fall into the non-melanized
colonies, arbuscular, vesicular, coil and hyphal colo-
grouping. In this soil, however, saprobes belonging to
nization, melanized, non-melanized and clamped
the Zygomycota with similar hyphal morphologies
hyphal length, spore numbers and animal abundances)
have been isolated (Klironomos 1994) so we do not
were significantly affected by light treatment
know whether the observed increase was a result of
(F32,84=4.102, P=0.0001). The results reflect a
increasing AM fungal extraradical growth or growth
strong association between light treatment and the
of other saprobes.
combined dependent variables, p2=0.76.
The populations of total bacteria and actinomycetes
in the root zones are presented in Table 1. The UV-B
800 treated seedlings showed significant increases in the
a Shoot biomass
total bacterial (F2,57= 8.36, P= 0.001) and actino-
U Root biomass
mycete (F2,57= 3.77, P = 0.029) populations compared
600 - with control. Patterns of morphological and physio-

E T T logical groups of bacteria occurring in the rhizosphere

are summarized in Table 2. The control and UV-A
treated plants supported high relative amounts of
Gram-positive and Gram-negative bacteria. Under
UV-B treatment, the difference between Gram types
200 T-
was much greater (F2 57= 22.34, P = 0.000 1), as there
were many more Gram-negative types. Judging from
the increase in the total number of bacteria, it was the

Baseline Visible Visible + Visible + Gram-negatives which thrived in the UV-B treatments.
UV-A UV-B The relative incidence of spore forming bacteria
?C 1995 British
Ecological Society,
was much less with UV-B radiation (F2,57=18.01,
Fig. 2. Effects of ultraviolet radiation on shoot and root
Functional Ecology, biomass of Acer saccharum seedlings. Values are the mean P =0.0001) and this is not surprising because the rela-
9, 923-930 ?SE. tive amounts of Gram-negatives to Gram-positives

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 927 E Arbuscules
Below-ground 80 - Vesicles
responses to l2 Hyphal coils T
UV-B
U Non-AMF hyphae
60-
0 I
20 T T T
T the two symbionts (Bonfante-Fasolo 1986), but arbus-
0 1994). Hyphal coils are longer-lived structures
Baseline Visible Visible + Visible +
only UV-A UV-B
Fig. 3. Effects of ultraviolet radiation on the arbuscular
mycorrhizal and non-arbuscular mycorrhizal fungal (AMF)
infection of Acer saccharum feeder roots. Values are the
mean ? SE.
increased under that treatment. The rhizosphere of
Sugar Maple harboured urea- and protein-hydrolysers
but both types decreased with UV-B treatment
(P <0.05). The relative abundance of starch hydroly-
sers was different among all radiation treatments
(P < 0.05), the highest found with UV-B irradiance.
Microarthropods' responses to UV treatment are
shown in Fig. 6. We categorized the arthropods into
four groups, the collembolans, oribatid mites, non-ori-
batid mites and predatory mites. Only collembolans
(F2,57=6*91, P=0.003) and non-oribatid mites seedlings and saplings that would require higher
amounts of inorganic nutrients to compete with other
(F2,57 = 4-01, P = 0.018) showed significantly higher
populations with UV-B treatment. No response was
detected to UV-A.
Discussion
The data from this study clearly show that below-
ground microbes are greatly affected by UV-B radia-
tion. A significant proportion of the available carbon
in a plant is directed to roots and much of that is used
to support rhizosphere micro-organisms (Finlay &
S6derstr6m 1992). It is therefore not surprising that
microbial populations associated with the rhizosphere
would be affected, when one considers that UV-B can
have adverse effects on the process of photosynthesis
(Caldwell & Flint 1994).
Mycorrhizal fungi associated with the roots of most
terrestrial plants, including Sugar Maple, help the
acquisition of inorganic nutrients from soil, in
exchange for organic compounds derived from photo-
synthesis (Harley & Smith 1983). In our initial
hypothesis, these fungi were expected to respond to
UV radiation. We did not detect a significant change
in total AM fungal infection of Sugar Maple roots;
?C 1995 British
Ecological Society, however, the morphology of the mycorrhizal associa-
Functional Ecology, tion changed drastically, largely switching from the
9, 923-930 production of arbuscules to that of vesicles and hyphal
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coils. There are a number of reasons why these changes
can be associated with stress. First, vesicles are rest-
ing structures that are capable of enduring adverse
conditions (Brundrett 1991). An increase in these
structures implies that there is a decrease of carbon
from the plant that is available to the fungus. Second,
it has been previously shown that, in Sugar Maple,
arbuscules develop from hyphal coils (Cooke,
Widden & O'Halloran 1992), both arbuscules and
coils can serve as sites of nutrient exchange between
cules are hypothesized to be more efficient and more
expensive to maintain (Duckmanton & Widden
(Cooke et al. 1992), less metabolically expensive and
have been shown to predominate under other unrelated
stressful conditions (Duckmanton & Widden 1994).
A healthy mycorrhiza, with plenty of arbuscules, is
essential for adequate nutrient uptake by the plant
(Harley & Smith 1983) and so a shift in the develop-
ment of AM fungi in roots of Sugar Maple may be a
prelude to stress that will be observed later on the tree.
Overall, it seems that instead of spending more energy
on the important function of nutrient-exchange
(arbuscules) the fungus allocated more energy to stor-
age and long-term survival (spores and vesicles). Both
vesicles and spores are considered to be resting organs
which are capable of reinitiating infections once con-
ditions become favourable. Also, as UV-B induced
stress would occur during the formation of gaps in the
canopy, this may be most harmful to the younger
plants for establishment in the upper canopy.
In the rhizosphere, bacteria and non-mycorrhizal
fungi increased significantly. It must be remembered,
however, that only a small proportion of bacteria can
be isolated using the dilution plate method (Torsvik,
Goks0yr & Daae 1990), so changes are not absolute,
but rather relative. Regardless, the data strongly indi-
150 -
)C T
aL 100 -
U)
4 -
0
U)
o 50-
6
z
TT
0
Baseline Visible Visible + Visible +
only UV-A UV-B
Fig. 4. Effects of ultraviolet radiation on spore production
by arbuscular mycorrhizal fungi associated with feeder roots
of Acer saccha rum. Values are the mean ? SE.
928 ^ 1500- rE Non-melanized if the plant requires greater rates of N-uptake from the
0
J. N. Klironomos (0 E Melanized soil to maintain this function, the altered mycorrhizal
E
& M. F. Allen XClamped T structures (see above) may not be able to support the
plant's needs (Azcon-Aguilar & Barea 1992).
w 1000 Infection of the feeder-roots by non-mycorrhizal
fungi, characterized by narrow, septate, often melanized
hyphae, or those with clamp connections, also increased
significantly, more than doubling with UV-B radia-
500- tion. Such an increase was coupled with the increased

0~~~~ T~~~~~ extraradical rhizosphere mycota and it seems that with

(0
? Baseline Visible Visible + Visible +
UV-A UV-B
contact
Fig. 5. Effects of ultraviolet radiation onwith
theroot tissues and/or
growth
extraradical fungal hyphae. Values are the mean ? SE.
cate that UV-B caused an increase in rhizodeposition,
either as root exudates or as cell sloughing, or as both.
Soil microbes are carbon limited (Zak et al. 1994) and
so are stimulated by root deposits. Also, rhizodeposits
were probably qualitatively altered, as was demon-
strated by shifts in the proportions of urea-, protein-
and starch-hydrolysing bacteria. The decreased occur-
rence of bacteria that could hydrolyse urea and protein
indicates that the plant deposited proportionally fewer
compounds which were nitrogen-rich. This is consis-
tent with the results of other studies which show
plants altering their secondary metabolism in response
to UV-B, in effect utilizing more N for protective
functions, particularly in leaves, i.e. the synthesis of
greater quantities of flavonoids (see Hahlbrock &
Grisebach 1979). Consequently, if this is the case, and
Table 1. Effects of ultraviolet radiation on total bacterial
and actinomycete densities in the rhizosphere of Acer sac-
charum (cfu g-1 oven-dry rhizosphere soil)
UV-B radiation, Sugar Maple roots may have become
more susceptible to infection by fungal saprobes,
pathogens and parasites.
Microbes associated with plant roots are likely to be
the first to respond to UV-B because they are in direct
ofrhizodeposits; how-
ever, other organisms, such as soil animals, which are
not directly associated with roots can also be affected.
The majority of collembolan and mite species are
microbial feeders, so the higher microbe levels asso-
ciated with UV-B radiation also supported higher
microarthropod densities. The consequences of this
are difficult to predict. Grazing by soil animals may
influence the mobilization of nutrients in soil
(Seastedt & Crossley 1984). For example, fungal
mycelium can contain a major proportion of the N, K
and other cations in the soil pool (Dowding 1981).
This may provide a positive feedback loop to the plant
by increasing the pool of available nutrients for root
uptake, especially in soils where supplies of nutrients
such as N and P are limited (Huhta, Setala & Haimi
1988; Setala et al. 1990). On the other hand, negative
consequences are also possible. Collembolans and
mites prefer to feed on melanized non-mycorrhizal
fungi (Klironomos & Kendrick 1995b), but high
microarthropod densities in the rhizosphere can lead
to increased disruption of the delicate mycorrhizal
fungal network (Finlay 1985; Harris & Boerner 1990)
which is very important for proper mycorrhizal func-
tioning (Friese & Allen, 1991). Whatever the conse-

Irradiance Bacteria (x 107) Actinomycetes (x 104) quences, it is evident from the present data that UV-B
may alter patterns of interactions in soil food webs.
Baseline 1.98 2.09 In conclusion, we have shown that microbes and
Visible 1.71 2.33 invertebrates associated with the roots of Acer saccha-
Visible + UV-A 1.42 2.76
rum do respond to UV-B radiation. Overall, it appears
Visible+UV-B 3.10 3.88
that carbon flow in the plant-soil system was shunted
from a mutualistic-closed, mycorrhizal dominated

Table 2. Influence of ultraviolet radiation on morphological and physiological groups of bacteria in the
rhizosphere of Acer saccharum (% of total tested)

Gram reaction
Spore Urea Starch Protein
Irradiance + - formers hydrolysers hydrolysers hydrolysers

Baseline 41.3 58.7 22.5 29 6 39.1 27.0

?) 1995 British
Visible 38.6 61.4 28.7 17.9 20.9 33-5
Ecological Society,
Visible+UV-A 44 1 55.9 24.3 22 6 34.8 31 5
Functional Ecology, Visible + UV-B 217 78.3 12.3 6.7 59.0 1112
9, 923-930

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929 EJ Collembola
Below-ground E Oribatid mites
responses to
1250 E: Non-Oribatid mites
UV-B
a Predatory mites
1000
co
v' 750 T
Baseline
UV-A UV-B
Fig. 6. Effects of ultraviolet radiation on Ecological
soil microarthropod
Interactions in Soil (eds A. H. Fitter, D.
densities. Values are the mean ? SE.
system to an opportunist-open, saprobe/pathogen dom-
Finlay, R. and S6derstrom, B. (1992) Mycorrhiza and carbon
inated one. Effects could also be noticed as our focus
shifted away from the root and into soil food webs. ThisPlant-Fungal Process (ed. M. F. Allen), pp. 134-160.
indicates that UV-B global change may lead to far lessChapman
predictable consequences than previously thought.
Acknowledgement
The authors wish to thank Anastasia Harizanos for
technical assistance.
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