Flowering and seed production of

aspen in Scotland during a heavy

seed year
R. WORRELL1, A. G. GORDON2, R. S. LEE2 AND A. MCINROY3
1 School of Forestry, Institute of Ecology and Resource Management, Edinburgh University, Mayfield Road,
Edinburgh, EH9 3JU; and Upper Park, by Aberfeldy, Perthshire, PH15 2EH, Scotland
2 Forestart, Church Farm, Hadnall, Shrewsbury, SY4 4AQ, England
3 Drumnagowan, Glen Fincastle by Pitlochry, Perthshire, Scotland

Summary
In 1996, profuse flowering of aspen (Populus tremula L.) occurred throughout Scotland. In
response, studies were initiated in Perthshire and Strathspey to investigate aspects of flowering, seed
production and seed storage of aspen. The proportions of clones flowering were high, averaging 72
per cent in Strathspey and 77 per cent in Perthshire. The sex ratios were 1.8 male clones to 1 female
clone in Strathspey and 1.3 male clones to 1 female clone in Perthshire, which are within the range
of ratios recorded for aspen on the Continent. Easy and reliable methods of sexing aspen trees in the
field during flowering are described. Seed production per catkin on four female clones in Tayside
ranged from six to 60 seeds per catkin. Seed production was clearly correlated with the number of
male clones in the vicinity of the female clones. A simple field technique for artificially pollinating
female flowers is described. Artificial pollination resulted in seed production averaging between 159
and 460 seeds per catkin, i.e. approximately a 12-fold increase over natural pollination. The viabil-
ity of seed from the two study areas was very high ranging from 94 to 98 per cent. The numbers of
seed per kilogram varied between 7 million and 10 million viable seeds kg1. Seed stored at 6 per
cent moisture content following rapid drying, and at 2C, retained levels of viability in excess of 95
per cent for at least one year. The results present excellent opportunities for increasing the supply of
planting stock of aspen from Scottish seed and fundamentally change current perceptions of the
ecological status of aspen in Scotland.

Introduction regeneration. Some ecologists had even begun to

It has been widely believed that aspen (Populus
tremula L.) neither flowered nor set seed success-
fully in Scotland, and that vegetative reproduc-
tion by suckers was the only means of natural
Institute of Chartered Foresters, 1999
speculate that aspen in Scotland might have lost
its ability to flower as a result of climate change
during the Holocene (E. Easton personal com-
munication). Most nurseries had discounted any
Forestry, Vol. 72, No. 1 , 1999
28 FORESTRY
prospect of obtaining British seed and either
propagated planting stock from rooted cuttings,
by tissue culture, or by using seed imported from
eastern Europe (Jobling, 1990; Hollingsworth
and Mason, 1991).
However there is evidence that aspen does in
fact flower and produce seed in Britain, at least
occasionally (Gray, 1949; Worrell, 1995a, b).
Grays early Forestry Commission Forest Record
The raising of aspen from seed, gave details of
seed production and propagation of aspen in
southern England. In recent years, the authors of
this paper have made several collections of viable
aspen seed from both Strathspey and Tayside
(Scotland) and Shropshire (England). The most
successful collection was from Strathspey in 1991
when Forestart seed company and Alba Trees plc
collected enough seed to raise 60 000 trees. This
raised interest in the prospect of seed production
in Scotland, and led to attempts to monitor flow-
ering more closely. This revealed that some flow-
ering of aspen in fact occurred every year in both
Strathspey and Tayside. Flowering was usually
sparse, varied radically between different clones,
but a few clones flowered relatively regularly
(Easton, 1998; R. Shearer, personal communica-
tion; I. Brodie, personal communication).
A further problem besetting those wishing to
establish seed supplies from British sources was
the conventional wisdom that stated that aspen
seed was short-lived and could not be stored
more than a few weeks at the most (e.g.
Anderson, 1950). However, there were increas-
ingly frequent reports from abroad that seed of
aspen, American aspen (Populus tremuloides
Michx.) and other Populus species could in fact
be stored for several years (Zasada and Dens-
more, 1980; Fung and Hamel, 1993; Worrell,
1995b). This led Forestart seed company to begin
seed storage trials on imported Hungarian seed
during 199295. Early trials with seed dried to
very low moisture contents either by air drying,
or in sealed containers with silica gel, and then
stored in a cold store, proved unsuccessful.
However in 1995, it was demonstrated that seed
stored at 1015 per cent moisture content, and at
2C, germinated for up to 7 months, though
with gradually reducing levels of viability.
In the spring of 1996, profuse flowering
occurred throughout Scotland and in response to
this a number of studies were initiated, at short
notice, to investigate some basic aspects of repro-
duction in aspen during a good seed year. Seed
storage trials were also carried out. The specific
aims of these studies were to:
1 record the proportion of clones flowering;
2 establish easy and reliable methods of sexing
aspen trees in the field during flowering;
3 record sex ratios;
4 evaluate seed production from a number of
contrasting female clones;
5 assess the viability of pollen and the effects of
artificial pollination on seed production;
6 assess the viability of seed;
7 investigate possible methods of long-term stor-
age of seed and assess changes in seed viability
during storage.
The overall objective of the work was to pro-
vide information that would assist seed collectors
and nurseries in obtaining viable aspen seed.
Methods
Flowering frequency and sex ratios
Aspen occurs naturally as groups of trees referred
to as clones, which have regenerated vegeta-
tively from root suckers, usually from a single
individual (Worrell, 1995a; Easton, 1998). As
aspen is dioecious, the clones are usually single
sex. A total of 52 clones in Tayside and 127 in
Strathspey/Lagganside were visited during the
period 1220 April 1996. The following informa-
tion was recorded for each clone:
whether flowering was occurring;
the sex, as determined by the morphology of
the catkins.
In addition, because catkins were frequently
beyond easy reach, features which allowed the
sex of catkins to be determined from ground level
using binoculars were recorded.
Seed production
Four profusely flowering female clones in the
Tay/Tummel catchment, Perthshire were
selected for assessing seed production. These
clones were selected to give a range in the number
of male clones known to be flowering in their
 FLOWERING AND SEED PRODUCTION OF ASPEN IN SCOTLAND
proximity; from zero to approximately 30 male
trees within 1 km of the female clones.
A sample of five catkins was collected from
each of these clones during the period 1119
June. The seed was extracted from these catkins
by drawing the hairs (pappus) and seed through a
sieve into the body of an industrial vacuum
cleaner (with bag removed). The seeds were
counted by placing them on squared paper and
viewing them through a magnifying glass.
Artificial pollination
Mature male catkins were collected from four
clones in the Tummel and Braan catchments on
18 and 19 April. These were laid out to dry on
newspaper at room temperature for 2448 h.
They were then shaken to remove the pollen onto
the paper, from where it was transferred into
plastic bags. A sample was sent for analysis of
viability using a fluorescence test at The Royal
Botanic Gardens, Kew.
On 20 April, a branch bearing female catkins
was artificially pollinated on each of the four
female clones described above. This was carried
out by placing about 25 cm3 of pollen in a large
clear plastic sack, inserting the twigs bearing the
catkins into the sack and shaking the pollen
over the catkins. Each branch typically con-
sisted of about 10 twigs, each bearing 810
catkins. The branch was marked with coloured
tape.
Random samples of five of these artificially
pollinated catkins were collected from each of the
clones during the period 1119 June. The seed
was extracted from two of these catkins using the
extraction method detailed above. Only two
catkins were assessed because of the profuse
amounts of seed, up to 500, contained in each
catkin. The catkins closest to the mean size of
catkin for each sample were assessed.
Seed viability
Two bulked samples of 80 seeds (20 from each
clone) representing the naturally and artificially
pollinated seed were sown on damp potting
compost at room temperature in a tray covered
by glass. The proportion of seed germinating
and the number of seedlings produced were
recorded.
29
Seed storage
Between 30 and 300 g of newly extracted seed
from three sources (Calvine, Perthshire; Newton-
more and Aviemore, Strathspey) were dried
quickly by placing them in open plastic contain-
ers in direct sunlight for about 30 min. The con-
tainers were agitated to prevent excess heating of
the seed. This resulted in the seed being dried to
just below 6 per cent moisture content. The lots
were placed in a cold store in sealed containers
and kept at 2 to 4C. Minute samples of these
were extracted at intervals during this period and
set to germinate. The final germination test
reported in this paper was carried out after 11
months storage (11 April 1997) when 0.1 g of
seed of each lot was tested. Germination tests
were carried out following standard procedures,
with seed grown on filter paper at 20C.
Results
Flowering
Timing Flowering was first observed on 12
April, though by this date flushing of the catkins
was reasonably well advanced. On the majority
of trees at this date, both male and female, some
catkins were fully out and others were still emerg-
ing. It should be noted that 1996 was a cold spring
and onset of flowering was about 2 weeks later
than had been observed in previous springs.
There appeared to be no obvious differences
between the timing of flushing in the male and
female clones. The peak of male flowering and
pollen production occurred during the period
1417 April. However there was considerable
variation between clones. Two of the male clones
(Kinloch Rannoch, Craignuisg) had shed most of
their pollen and the catkins had started to lose
their colour by 12 April, whereas others were still
emerging on 19 April.
Proportion of clones flowering The proportion
of clones flowering was similar in Perthshire and
Strathspey/Lagganside, at 77 per cent and
72 per cent respectively. This is somewhat in
contrast to the general pattern in observed previ-
ous years, when there was greater abundance of
trees flowering in Strathspey as opposed to
Perthshire.
30 FORESTRY
Trees tended to flower profusely or not all, and
only a small number of clones had sparse flower-
ing. Trees flowering profusely were typically
densely covered with catkins throughout the
crown; catkins were estimated as numbering sev-
eral tens of thousands in larger trees. Catkins
occurred down to ground level in open grown
clones. No obvious pattern was observed in those
flowering and those not. Some relatively young
clones (56 m tall) flowered, though not as pro-
fusely as older larger trees. Flowering was pro-
fuse in both sexes, but male trees appeared to
carry rather more catkins than female.
Sexing clones by catkin morphology
Male and female catkins are superficially similar
in aspen, and difficulties were experienced ini-
tially in determining the sex of the catkins from
ground level. This was not helped by the differing
descriptions in standard texts. For example
Mitchell (1974) describes male catkins as grey-
brown in colour, Edlin (1985) as purple and Fitter
et al. (1974) illustrate them as pale green-grey and
red!
The colour of both sexes on emergence, in
those observed in this study, was a distinct dark
red. The stigmas are red in the female and sta-
mens red or purple/red in the males. They soon
become hairy. The stamens turn yellow in the
males as the pollen is produced, while at the same
time the bracts change from green to brown and
then grey. Shortly after this they are shed. The
females lose their red after a few days and turn a
dull green.
The shape of catkins varies widely between
clones and changes as they mature. Male catkins
start by being rather stouter than female ones,
with a broad rounded tip; but as they mature and
shed their pollen they elongate, ultimately being
Table 1: Seed production of four contrasting female aspen clones
Clone Grid No. of male
ref. trees within
1 km
1. Calvine NN783669 28
2. Old Struan NN807653 15
3. Croftcat NN899524 3 17
4. Balmacneil NN973512 0 6
more slender than female catkins. Female catkins
narrow over their length to a more slender tip;
however they broaden gradually as they mature,
and eventually lose the slender tip as the individ-
ual capsules separate and become more distinct.
The line drawings in Edlin (1985) give the best
representation of catkins among the standard
texts cited above.
The most useful feature for determining sex
from a distance is that male catkins become flex-
ible soon after flushing and move in the wind,
whereas female ones are stiff and motionless at
this time in the season. Male catkins are also shed
en masse relatively quickly after flowering; so a
tree with large number of grey catkins on the
ground during AprilMay is male. Trees bearing
catkins in May or June are obviously female.
Sex ratios
The female : male ratios of the clones surveyed
were 1 : 1.8 in Strathspey/Lagganside and 1 : 1.3
in Perthshire.
Seed production
Timing Seed became mature and dispersed dur-
ing the period 1120 June. However, spring 1996
was generally cold, and flowering of many species
was delayed by approximately 714 days com-
pared with normal. In previous years, aspen seed
production in the Highlands has generally been
observed during the first week of June.
Natural pollination Seed production in natu-
rally pollinated trees varied between six and 60
per catkin (Table 1). Clones with the greater
number of flowering male trees in the vicinity had
more seeds per catkin. On the basis of the figures
in Table 1, seed collection should be concentrated
No. of seeds No. of seeds/catkin
per catkin following artificial
(natural pollination
pollination)
60 460
24 159
262
359
 FLOWERING AND SEED PRODUCTION OF ASPEN IN SCOTLAND 31

on those female clones that have several flower-
ing male trees within, say, 1 km.
It should be noted that the weather conditions
during pollination were exceptionally poor.
During the period 1220 April, the weather was
largely cloudy and damp; it rained significantly
on 4 days and was showery on a further 3 days.
Winds were between south-east and west and
were gale force on 18 April. The only frost was a
ground frost on 20 April. In years with better
pollination conditions seed production would
almost certainly be better.
Artificial pollination Seed production in artifi-
cially pollinated catkins was dramatically higher
than in naturally pollinated ones, varying from
between 160 and 460 seeds per catkin (Table 1).
The relatively low figure for clone 2 was largely a
result of seed dispersal having begun just before
the catkins were collected.
Artificial pollination increased seed production
by factors of between seven and 50 times, with an
average of 12. This demonstrates clearly that seed
production was limited by pollination during this
season. The results also suggest that artificial pol-
lination may be an effective method of enhancing
seed production in aspen.
The pollen viability was 54 per cent approxi-
mately 3 weeks after collection, which is within
the range expected for trees (R. Atkinson, per-
sonal communication).
Seed viability Seed viability was very high for
both naturally and artificially pollinated seed lots
(Table 2). Whether the catkins had been naturally
or artificially pollinated had no influence on
germination or seedling numbers. A loss of about
13 per cent of the germinated seedlings occurred.
The causes of death were not investigated.
However it was observed that some seedlings
failed to produce full sized cotyledon leaves and
may have died for this reason.
Seed storage
Seed storage was remarkably successful (see
Table 3) with the viability of the three lots
remaining above 95 per cent for the entire 11-
month period. The results show that the number
of seeds per kilogram lies in the range 710 mil-
lion seed/kg.
Discussion
Ecology
The events of 1996 and the results of this study
show conclusively that aspen in Scotland is capa-
ble of widespread flowering and seed production.
Thus the suggestion that it is capable only of veg-
etative reproduction can finally be laid to rest.
Flowering was undoubtedly triggered by the
unusually warm and dry summer of 1995, which
also resulted in heavy flowering and fruiting of
many tree species. Little can be said about the
frequency of such major flowering and seeding
years except that they are infrequent. Nothing on

Table 2: Germination of naturally and artificially pollinated aspen seed

No. of seed No. of seed No. of seedlings
sown germinating established*
No. (%) No.(%)
Naturally pollinated seed 80 76 (95) 60 (75)
Artificially pollinated seed 80 75 (94) 62 (77)
* Assessed when first true leaves opened

Table 3: Results of germination tests on 0.1 g of seed stored for 11 months from three origins of Scottish aspen
Seed origin No. of seeds No. of seeds Germinable Dead seeds Percentage
per kg tested seeds germination
Aviemore 9 240 000 924 892 32 96.5
Calvine 10 220 000 1022 986 36 96.5
Newtonmore 7 740 000 774 761 13 98.3
32 FORESTRY
the scale of 1996 has been observed previously by
the authors; at least since 1983. However a
farmer upon whose land one of the larger clones
in Strathspey is located can recall several previ-
ous years when the wood went white, suggest-
ing that major seed years have occurred in the
past (E. Easton, personal communication). The
last exceptionally warm summer similar to 1995
was 1976, and this is known to have stimulated
seed production in a wide range of tree species.
The authors have observed aspen over several
years in Perthshire and Strathspey. In Perthshire,
the typical pattern is for only a modest number of
flowers to appear on five to six clones out of the
30 observed. In Strathspey, flowering is usually
more frequent, with several clones flowering rel-
atively reliably in the majority of years.
The fact that aspen can reproduce by seed in
Scotland, albeit relatively infrequently, has con-
siderable ecological significance. First, it means
that aspen populations are not limited in their nat-
ural distribution to existing clones, but are poten-
tially capable of colonizing new sites. As the seed
of aspen is remarkably light and mobile (similar in
many characteristics to a small dandelion seed),
this colonization could take place over large dis-
tances. This goes some way to explaining how
aspen has established itself on such a wide range of
remote sites including mountain crags at high ele-
vation, sea cliffs and on all the islands (Worrell,
1995a). Second, it has also been pointed out that
failure to reproduce sexually will lead eventually
to reduced genetic diversity and poorer adaptabil-
ity of populations (Easton, 1998; Ennos et al.,
1998). The results of this study suggests that there
is some prospect of aspen maintaining levels of
genetic diversity, at least in areas where the poten-
tial for seed production is greatest.
However this study demonstrates only the
potential for regeneration, and provides no evi-
dence of successful seedling establishment. It
should be noted that successful germination of
aspen seed requires some fairly unusual site con-
ditions, such as disturbed or burnt over areas,
and that even in countries where it reproduces
more regularly, natural regeneration is only
observed relatively infrequently (Brset, 1960).
Current land use in Scotland presents some for-
midable obstacles to colonization of aspen by
natural regeneration, particularly in terms of
dense pasture swards and high grazing levels.
The female : male ratio of 1 : 1.31.8 observed
in Scotland lies between those reported by other
studies. In southern and central Italy and in the
Caucasus a ratio of 1 : 1 was reported, while in
Norway and northern Italy ratios of 1 : 2 and 1 :
2.5 respectively were reported (see Worrell,
1995a). No explanation for varying sex ratios is
given in the literature on aspen. Dioecy is a char-
acteristic of the Salicaceae to which aspen
belongs. A wide range of sex ratios is reported for
other members, with strong female bias reported
in many Salix species (Ahman, 1997; Alstrom-
Rapaport et al., 1997), ratios close to 1 : 1 in
Populus tremuloides in USA (e.g. Valentine,
1975) and strong male dominance in Populus
nigra var. beautifolia (Pursh) Torr. in Britain
(E. Cundall, personal communication). Mechan-
isms which lead to diverging sex ratios include
the operation of sex determining genes, competi-
tion between male and female determining
pollen, preferential browsing of one sex by herbi-
vores, and lower viability of male plants (Ahman,
1997).
This study highlights the critical role of the
proximity of male and female clones in relation
to seed production. Production increased
markedly with increasing numbers of male trees
in the vicinity of female clones. Even where 28
male trees were recorded within 1 km, seed pro-
duction per catkin was only 16 per cent of that
achieved by artificial pollination. However it
should be noted that pollination conditions dur-
ing the spring of 1996 were remarkably poor, and
it is likely that if weather conditions had been
better, pollination would have been far more
successful. Observations in England have shown
small amounts of seed being produced by trees
where the nearest male is well in excess of 1 km.
The problems of pollination are a graphic exam-
ple of the wide ranging ecological problems
caused by the fragmentation of the native wood-
land resource.
Seed production and collection
Seed years on the scale witnessed in 1996 present
the opportunity to collect almost limitless quan-
tities of seed. Several successful collections of
seed were made in 1996 and the nurseries have
produced many tens of thousands of aspen
plants, albeit from relatively few parent clones.
 FLOWERING AND SEED PRODUCTION OF ASPEN IN SCOTLAND
This is timely considering the increased interest
in planting aspen in native woodlands, and recent
difficulties experienced in obtaining sufficient
British stock.
The germination percentages of the seed were
remarkably high. This may be partly because
aspen seek lacks a seed coat, and so the process by
which empty seeds are produced by many other
species cannot occur in aspen.
Artificial pollination was so successful that it
presents the possibility of using this means to
secure seed supplies during years when aspen is
flowering more sparsely. A few male catkins can
produce a relatively large quantity of pollen; and
artificially pollinated female catkins can produce
200500 seeds. Thus it only takes a relatively
small number of catkins to produce several thou-
sand seeds. Producing plants by artificial pollina-
tion may be more cost-effective than by
vegetative means; especially if pollen can be
stored from year to year, as is possible with many
forest trees.
Seed storage
The results of the seed storage tests demonstrate
conclusively that aspen seed can be stored with-
out loss of viability for at least one season. It
appears to be important that the period of drying
should be as short as possible. The technique
described in this paper removes a further major
impediment to securing seed supplies in Britain.
Forest practice
This study suggests that we should pay attention
to the sex of the stock being planted, to ensure
that a mix of male and female clones is included
in schemes. Sexing could be done for vegetatively
produced material by nursery staff sexing the
clonal parents and selling lots with an equitable
number of males and female plants. Plants raised
from seed can be assumed to have an equitable
sex ratio. Clones of different sexes should ideally
be planted within, say, 1 km of each other to
ensure reasonable success in pollination. In exist-
ing woodland, a case can be made for locating
isolated female clones and establishing clonal
material from male clones in their vicinity so as to
help secure seed production in the future.
The success of seed production in aspen in 1996
33
should encourage seed collectors, nurseries and
those planting the species to rely more on indige-
nous sources. The conventional wisdom regard-
ing the inability of Scottish aspen to produce
seed, and the seed being impossible to store, has
turned out to be a myth. That these ideas should
have been so widely accepted, and so easily over-
turned, is a cautionary tale. One wonders what
other advances could be achieved by modest
targeted studies of the ecology of some of our less
well-researched native tree species. Aspen in
Scotland is not a relict species capable only of a
limited role in our forests, but a fully functional
forest tree.
Acknowledgements
The authors wish to thank the following people: Eric
Easton who carried out the survey of flowering in
Strathspey and Lagganside; Rodney Shearer of Alba
Trees provided help and encouragement and Iain
Brodie, an inveterate aspen enthusiast, who provided
some background information on recent flowering pat-
terns in Strathspey. The Scottish Forestry Trust con-
tributed to supporting this study. Many local
landowners co-operated by allowing access to aspen
clones. Finally wives and partners put up with being
abandoned for several weekends during the spring
whilst this work was carried out.
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