Journal of

Dispersal and gene flow of Populus nigra (Salicaceae)

Blackwell Publishing Ltd.

Ecology 2003
91, 447 – 456 along a dynamic river system
ERIC IMBERT* and FRANÇOIS LEFÈVRE
INRA, Unité de Recherches Forestières Méditerranéennes, Avenue A. Vivaldi, F-84000 Avignon, France

Summary
1 We used genetic markers to study gene flow of the riparian pioneer tree species Populus
nigra along the Drôme river (France). This dioecious species is supposed to have more effi-
cient dispersal mechanisms for pollen (wind) and seeds (wind and water) than other trees.
2 Seedlings belonging to the same reproduction/migration event were sampled in 22
riparian forest fragments along the river and their genetic diversity assessed through six
nuclear microsatellites.
3 We found a high level of diversity and significant differentiation among populations. The
significant isolation by distance allowed us to reject the infinite island model of migration.
4 Gene flow parameters were higher in the upper, mountainous part than in the alluvial
plain downstream. There was no accumulation of diversity downstream, indicating
migration rates were symmetrical upstream and downstream. This was confirmed by
computing individual migration parameters between adjacent populations.
5 The results are discussed with regard to the dispersal mechanisms of seeds and pollen.
The discrepancy between potential gene flow and effective gene flow is interpreted as an
effect of fragmentation, due to the alteration of the natural dynamics of the riparian
ecosystem rather than to physical barriers.
Key-words: effective gene flow, European black poplar, isolation by distance, micro-
satellite, riparian ecosystem
Journal of Ecology (2003) 91, 447–456

modifications of riparian habitats lead to an increase

Introduction
Populus nigra L. (Salicaceae) is a pioneer tree species
of the riparian forests, where it often exists as a series
of metapopulations. It regenerates through colonization
of areas of bare soil along the riverbank (Barsoum &
Hughes 1998). The major threat for this species lies
in human disturbance, particularly due to the develop-
ment of agriculture and, more recently, to urbaniza-
tion, both of which reduced the total area of
floodplain woodlands (Dynesius & Nilsson 1994). This
has caused a reduction in size of local populations of
black poplar, which are no longer continuous along
the river system. Furthermore, the natural dynamics of
the ecosystem (periodic flooding) and the lateral move-
ment of the river bed have been dramatically affected
by management so that the area available for P. nigra
regeneration is reduced and poplars and willows have
been progressively replaced by hardwood species (e.g.
Fraxinus, Acer, Ulmus, Quercus). Therefore, human
Correspondence: rançois Lefèvre (tel. +33 4 90135920, fax
+33 4 90135959, e-mail lefevre@avignon.inra.fr).
© 2003 British *Present address: I.S.E.M., CC 065, Université Montpellier 2,
Ecological Society F-34095 Montpellier, cedex 5, France.
of the geographical distance between suitable habitats
or populations, i.e. fragmentation (Jansson et al. 2000).
The effect of fragmentation and, more generally, of
landscape structure, is currently an important topic in
both evolutionary and conservation biology (Heino &
Hanski 2001; Wiens 2001). Fragmentation affects both
the exchange of individuals and gene flow (Young et al.
1996; Aldrich & Hamrick 1998; Dayanandan et al. 1999).
Habitat loss therefore leads to an important decrease in
patch occupancy, and thus in metapopulation viability
(Hanski & Ovaskainen 2000; Casagrandi & Gatto 2002).
Another consequence of population isolation is an
increase of consanguinity within populations and a
greater susceptibility to perturbations or demographic
stochasticity. Finally, fragmentation contributes to the
evolution of dispersal-related traits (Leimar & Norberg
1997; Heino & Hanski 2001). Consequently, the
implementation of conservation strategies requires the
monitoring of gene flow in relation to habitat structure.
In most riparian species, seed dispersal combines an
initial wind-mediated phase, i.e. transport of seeds from
the maternal plant to the ground or water, followed
by a secondary hydrochorous phase (Barat-Segretain

448 1996). Seeds are thought to cover several kilometres
E. Imbert & (Ridley 1930; Nilsson et al. 1991) and gene flow is
F. Lefèvre therefore supposed to be continuous and to give rise to
large population size. Furthermore, the direction of
dispersal is expected to be predominantly down-
stream. We therefore expect an accumulation of
genetic diversity in populations located in the lower
reaches of a river. Although evidence has been
found for fish (Hernandez-Martich & Smith 1997; Baer
1998) and for some plant species (Gornall et al. 1998
and Kudoh & Whigham 1997, 2001 for Potamogeton
colouratus and Hibiscus moscheutos, respectively), Ritland
(1989) and Russell et al. (1999) found no evidence for
unidirectional gene flow in Mimulus caespitosis and
Calycophyllum spruceanum, respectively.
We studied gene flow in P. nigra along an entire river
system. We analysed the structure of the genetic diver-
sity among even-aged seedlings, thus avoiding the
effect of age-structure on the genetic diversity, in order
to infer the spatial component of gene flow among the
reproductive populations. We addressed the following
questions: (i) Is gene flow limited by distance or is there
a single migrant pool along the river system? (ii) Is gene
flow similar in parts of the river showing contrasted
topography, i.e. mountains vs. large alluvial plain?
(iii) Is gene flow asymmetrical, i.e. more important in
the downstream direction?
Several genetic models exist to study gene flow (for
a review see Ouborg et al. 1999; Sork et al. 1999;
Rousset 2001). We used the model proposed by Rousset
(1997) to test whether migration rate depends on the
spatial distance between populations, i.e. isolation by
distance model. Hardy & Vekemans (1999), who for-
mulated the same model with spatial statistics for a
continuous population, showed in their simulations
that a stable genetic structure can be detected within
a few generations following colonization events (see
also Slatkin 1993). We used a separate method, pro-
posed by Beerli & Felsenstein (1999, 2001), to study the
symmetry of gene flow in both directions. This method
is based on a full matrix model, assuming a closed
system of n populations of any size that show demo-
graphic stability and exhibit no selection (these restric-
tions are discussed later). In this case, n2 parameters,
describing population sizes and number of migrants
from one population to another, are estimated indi-
vidually using the coalescence approach.
Methods
 
Populus nigra is a typical riparian pioneer tree widely
distributed across Europe and Asia, with a life-span
typically in excess of 100 years in the absence of
© 2003 British
ecological perturbations. In natural populations, the
Ecological Society, youngest reproductive individuals are reported to be 6–
Journal of Ecology, 10 years old (Zsuffa 1974). The species is dioecious,
91, 447–456 and pollination is anemophilous. Female trees produce
13652745, 2003, 3, Downloaded from https://besjournals.onlinelibrary.wiley.com/doi/10.1046/j.1365-2745.2003.00772.x by Cochrane Colombia, Wiley Online Library on [07/09/2023]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
thousands of seeds, especially when the crown is in full
light, which are dispersed first by wind, and secondarily
by water. This two-phase dispersal is possible because
the seeds are small (1450 seeds per g, Zsuffa 1974) and
carry a large hydrophobic pappus. There is no dor-
mancy (Muller & Teissier du Cros 1982) and, as seeds
remain viable for only 2 weeks, there is no soil seed
bank. Germination occurs exclusively on bare soil
(Barsoum & Hughes 1998). Hypocotyls develop within
6–8 h after moisture has reached the seed and the
pappus has degraded (Zsuffa 1974) and the seedling
dies if conditions are not favourable for further develop-
ment. The species is diploid (2n = 38) (Zsuffa 1974).
  
The Drôme is one of the important rivers of the French
Alps (about 106 km long, 44°31′ N, 5°34′ E to 44°46′ N,
4°45′ E), whose only artificial construction is a 10-km-long
dike in the downstream part, where it joins the Rhone
(Fig. 1). The water levels fluctuate under the snow-
melt in spring and increased autumn precipitation
typical of Mediterranean climates. The Drôme flows
from east to west (Fig. 1). In its upper part, where the
river is bounded by mountains (Vercors), it falls from an
altitude of 1440 –230 m (Fig. 1). The prevailing winds
are from the west and thus opposite to the water flow,
although violent winds (> 8 m s−1) are rare (source
Météo-France). In the lower part, the river flows in
an alluvial plain (230 –85 m a.s.l.). The prevailing
winds are from the North, and thus perpendicular to
the river (Fig. 1), and are more than 8 m s−1 for 4% of
the time. In the upstream part, the river is very
dynamic, with lateral movements of the riverbed
favouring the emergence of pioneer habitats in the
riparian ecosystem. In the downstream part, lateral
movements only occur locally. The riparian ecosystem
tends to be limited by the topography in the upstream
part, and by agriculture in the downstream part.
During the winter of 1999, adult trees were ran-
domly sampled in two large populations (33 and 34 in
populations 1 and 6, respectively, Fig. 1). During the
spring of 2000, sex could be determined for 53 of these
67 trees: 15 males and 11 females in population 1, 18
males and 9 females in population 6. This sex ratio was
not significantly different from 1 : 1.
Very few hybrid poplar plantations exist along the
Drôme river (one in the vicinity of population 6 and
one between populations D and E, Fig. 1), although
cultivated hybrids are more common along the Rhone
river. The fastigiate cultivar P. nigra ‘italica’ was regu-
larly observed in the landscape and fastigiate trees were
exceptionally observed within the riparian forest.
 
An inventory of all riparian forests along the Drôme
was made in 1998 (Heslouis 1999). From this inventory,
we selected 22 sampling sites (hereafter populations)

449
Dispersal in black
poplar
Fig. 1 Position of the 22 populations of Populus nigra studied along the Drôme river. Each box gives the name of a population
above its distance from the river’s source (km) and its elevation (m). Broken lines represent tributaries. The double arrow indicates
the limit between the upper part of the river (from population A to population M), and the lower part of the river (from population
5 to population 8). The two small arrows indicate the direction of prevailing winds in the upper and lower parts of the river.
based on the discontinuity of the riparian forest
(Fig. 1). In August 1999, 30 seedlings were collected per
population, except in population P where only 22 seed-
lings could be found, for a total of 652 seedlings. We
used the presence of cotyledons as a criterion to avoid
collecting vegetative offspring, and to guarantee that all
seedlings belonged to the same unique reproduction
and migration event (i.e. spring 1999). Within each
population, regeneration was spatially structured, occur-
ring in separate patches. We sampled as many different
patches as possible (11–23 per population).
We also assessed the structure of genetic diversity
among patches within two populations: population 1
in the upstream part of the river and population 6 in the
downstream part. Sixty seedlings, belonging to three
different patches, were sampled in each population. In
population 1, the distance between pairs of patches
ranged from 50 to 500 m, whereas in population 6 the
patches were separated by more than 1 km.
In the field, material was stored in plastic bags and
frozen with dry ice. In the lab, material was stored at
− 80 °C until extraction. DNA was extracted using the
protocol proposed by Fulton et al. (1995) slightly modified.
Six microsatellite markers were used. Five of them
were designed by van der Schoot et al. (2000) and Smulders
et al. (2001): WPMS09, WPMS12, WPMS16, WPMS18
and WPMS20. The sixth one (PMGC14) was obtained
from the Poplar Molecular Genetics Cooperative database
(http://poplar2.cfr.washington.edu/pmgc/). After ampli-
© 2003 British
fication, separation was made on polyacrylamide gel,
Ecological Society, and visualization was made using a silver staining protocol.
Journal of Ecology, A genetic linkage map of P. nigra recently published
91, 447–456 by Cervera et al. (2001) indicated that WPMS09 and
13652745, 2003, 3, Downloaded from https://besjournals.onlinelibrary.wiley.com/doi/10.1046/j.1365-2745.2003.00772.x by Cochrane Colombia, Wiley Online Library on [07/09/2023]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
WPMS12 belong to the same linkage group, but are
60 c distant, whereas WPMS16, WPMS20 and
PMGC14 belong to separate linkage groups. There-
fore we considered these markers as independent.
WPMS18 could not be mapped with this pedigree.
 
We first tested the genotypic linkage disequili-
brium between each pair of loci within each population,
using Fisher’s exact test available in GENEPOP 3.2a
(Raymond & Rousset 1995a). We also used GENEPOP
software to test for departure from Hardy–Weinberg
equilibrium. Several genetic diversity parameters (in-
cluding mean number of alleles per locus, unbiased
expected heterozygosity and FIS) were computed using
GENETIX 4.01 (Belkhir et al. 2000). Global and pair-wise
FST estimates (following Weir & Cockerham 1984), were
computed with GENETIX, and tests of the geno-
typic differentiation among populations were per-
formed using the exact test provided by GENEPOP
(Raymond & Rousset 1995b). For all the tests per-
formed with GENEPOP, we used the default settings.
Because many tests were repeated several times, we used
the sequential Bonferroni correction (Rice 1989).
We used the method proposed by Rousset (1997) to
test the null hypothesis of a single migrant pool over the
whole river system (island model) against isolation by
distance. Two different matrices of geographical
distances were used. The matrix in which the euclidian
distance was inferred from the geographical co-
ordinates of the centre of each population was highly
correlated with one constructed with the distance in one

450 dimension along the river system (Kudoh & Whigham
E. Imbert & 1997) (r = 0.98, P < 0.05). Results from the second
F. Lefèvre matrix are presented because this one-dimension
model fits the hypothesis we wanted to test concerning
the distribution of the riparian forests along the river.
Therefore, as data were analysed in a one-dimensional
model, we analysed the regression of FST /(1 − FST) on dis-
tance (Rousset 1997), using GENEPOP.
We analysed the spatial structure of kinship coeffi-
cients between individuals following Hardy & Vekemans
(1999). This procedure assumes a continuous popula-
tion, but it is still informative for groups of individuals.
The significance of kinship coefficients was tested
through a permutation test with 10 000 permutations
of individuals among positions. Calculations were
performed with AUTOCORG (Hardy & Vekemans 1999).
We used the program MIGRATE for n populations
(Beerli & Felsenstein 1999, 2001) to study the uniform-
ity of gene flow along the river system and to test for
symmetry of gene flow in either directions. In this pro-
gram, gene flow is estimated in a maximum-likelihood
approach based on coalescence. The output of such an
analysis can be represented by a n × n matrix. Values
on the diagonal are the product of the mutation rate
and the effective size of the population. Values below
the diagonal are 4NiMij from population i (population
i is considered as the source of the migrants) to popu-
lation j, while values above the diagonal are 4NjMji
from population j, now considered as the source, to
population i. Therefore, we can compare gene flow from
i to j, and from j to i, and thus estimate the asymmetry.
Furthermore, we can sum all the values obtained
from one given population considered as a source. This
parameter gives an idea of the number of individuals
sampled in other populations that can be assigned
to this source population.
For all analyses, the default settings of MIGRATE
were used. The time required to estimate gene flow using
this program was prohibitive and we could not therefore
obtain confidence intervals for the estimates (a data set
on another species with four polymorphic loci and 228
individuals required 2 months to obtain estimates and
confidence intervals). For the present data set, we could,
however, obtain estimates of the gene flow and effective
population size parameters (22 × 22 = 484 parameters).
We performed two independent computations using
two different workstations running at the same time.
The two sets of parameter estimates were highly corre-
lated, and their mean values were therefore used.
Results
 
The number of successfully scored seedlings ranged
from 551 for WPMS16 to 600 for WPMS18 (Table 1).
© 2003 British
Ecological Society, The number of alleles per locus ranged from 9 to 15,
Journal of Ecology, and the number of different genotypes observed per
91, 447–456 locus ranged from 27 to 68 (Table 1). For the entire data
13652745, 2003, 3, Downloaded from https://besjournals.onlinelibrary.wiley.com/doi/10.1046/j.1365-2745.2003.00772.x by Cochrane Colombia, Wiley Online Library on [07/09/2023]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
set, we obtained 473 seedlings with complete scor-
ing (i.e. amplification for each of the six loci), of which
only two had the same multilocus genotype (from the
same population). Within each population, the mean
number of alleles per locus varied from 5.5 to 7.7 (i.e.
most alleles were not present in every population). The
reduced number of single-banded patterns was indirect
evidence for the absence, or very low frequency, of null
alleles for these microsatellite loci. Finally, only one
genotypic disequilibrium (concerning WPMS09 and
WPMS12) out of 330 was significant at the 5% level
after Bonferroni correction. We therefore treated the
loci as independent.
  

The markers used in this study provided informa-
tion about potential hybridization between cultivated
hybrids and the wild black poplar in this area: we
detected no alleles specific to the exotic Populus species
currently used in cultivated hybrids such as P. deltoides
and P. trichocarpa.
Hybridization with the P. nigra cultivar ‘italica’ is
more difficult to detect as it belongs to the same species.
For each locus, we found alleles in our populations that
are shared with the cultivar. The frequency of these alleles
in our samples was highly variable: the most frequent
alleles may be considered as common alleles for the spe-
cies, whereas the rarest alleles provide an upper estimate
of possible hybridization with the cultivar. Among the
six loci, WPMS09 showed the lowest frequency of the
two alleles shared with ‘italica’ (< 0.05).
 
The expected heterozygosity for each locus in each
population was generally high but observed hetero-
zygosity was even higher (Table 1, Fig. 2). Two loci,
WPMS16 and WPMS18, showed a higher heterozygote
excess. Genotypic distribution for WPMS16 was char-
acterized by the very high frequency (33%) of one single
heterozygous genotype. For WPMS18, two hetero-
zygotes, sharing one allele, were frequent (respectively
27.3% and 18.5% of the samples).
For the multilocus estimation, 19 populations showed
a significant excess of heterozygosity, two showed a
deficit and one population was in Hardy–Weinberg equi-
librium. However, after removing the loci WPMS16
and WPMS18 from the analyses, only 14 populations
showed a significant departure from equilibrium, of
which five were in heterozygote excess while nine had a
significant deficit.
   
 
Global FST estimates ranged from 0.028 for WPMS20
to 0.068 for WPMS16 (all values were significantly
 13652745, 2003, 3, Downloaded from https://besjournals.onlinelibrary.wiley.com/doi/10.1046/j.1365-2745.2003.00772.x by Cochrane Colombia, Wiley Online Library on [07/09/2023]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
451 Table 1 Microsatellite diversity for the six primers used in this study. Data do not include the three-patch samples. (FIS:
Dispersal in black heterozygote deficit)
poplar
Number of Number of Within population Within population
Number of different seedlings with observed heterozygosity expected heterozygosity
Locus alleles genotypes amplification (mean value) (mean value) FIS

WPMS09 13 61 595 0.79 0.79 −0.01

WPMS12 15 68 579 0.78 0.80 +0.03
WPMS16 11 29 551 0.82 0.65 −0.27
WPMS18 12 29 600 0.85 0.73 −0.18
WPMS20 09 27 559 0.74 0.71 −0.04
PMGC14 13 47 563 0.71 0.72 +0.01

Fig. 2 Distribution of the observed heterozygosity (filled bars) and unbiased expected heterozygosity (open bars) per population
for each locus.

different from 0 at P < 0.0001), the overall FST estimate
being 0.047. Pairwise FST (22 populations, 231 pairs)
ranged from 0.005 to 0.117: 230 P-values were signi-
ficant at the 5% level. Therefore, even close populations
appeared to be genetically different, and the test for
isolation by distance was significant (Fig. 3). Isolation by
distance was also tested by computing auto-correlation
with the kinship coefficient. It was significant (P < 0.05)
© 2003 British
Ecological Society, for comparisons of individuals within populations
Journal of Ecology, (kinship coefficient = 0.044), as well as for the shortest
91, 447–456 distances (class 0–5 km, mean distance 3.2 km, kinship
coefficient = 0.01) and the largest distances (class
70–90 km, mean 80 km, kinship coefficient = −0.01,
Fig. 4). These patterns were not changed when exclud-
ing WPMS16 and WPMS18.
    
 
Differentiation among populations was of the same
magnitude in both parts of the river: FST = 0.046 in the
upper part and 0.041 in the lower part, both values
 13652745, 2003, 3, Downloaded from https://besjournals.onlinelibrary.wiley.com/doi/10.1046/j.1365-2745.2003.00772.x by Cochrane Colombia, Wiley Online Library on [07/09/2023]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
452
E. Imbert &
F. Lefèvre

Fig. 3 Relationship between pair-wise genetic (FST /1−FST) and

linear distances between populations along the river. The P-
values come from a Mantel test.

Fig. 5 Relationship between pair-wise genetic (FST/1−FST) and

linear distances between populations for populations in the
upper part of the river (above) and for populations in the
lower part of the river (below).

Fig. 4 Kinship coefficient and geographical distance between

individuals. For each distance class, each point represents
a single locus value. The dotted line and symbols (triangle)
represent the mean values among loci.

highly significant. In the upper part, the isolation by

distance was significant (Fig. 5). In the lower part, the
model was only marginally significant: the slope of the
regression was steeper but the number of populations
and the range of distances were reduced.
Estimations of migration rates obtained from MIG-
RATE were also used to test whether gene flow increases
downstream. We compared migration rates between adja-
Fig. 6 Diversity index (mean number of alleles, open sym-
cent populations according to the position of the source bols, and unbiased expected heterozygosity, filled symbols)
population along the river system. Spearman’s coefficient per population according to position along the river.
of correlation was not significant either when we con-
sidered gene flow from the upper population to its down-
stream neighbour population or vice versa (r = −0.34 and values), we obtained 107 positive differences and 124
r = −0.27, respectively, P > 0.2 for both values, n = 21). negatives ones (P = 0.14, sign test). Considering only
adjacent populations (21 comparisons), 13 values were
negative and eight positive (P = 0.19, sign test). Thus we
   
conclude that there was no evidence for asymmetrical
From the estimations of 4NiMij obtained from MIG- gene flow.
RATE, we computed the difference between migration Correlations between distance from the river source
from population i to population j and migration from and genetic diversity (expected heterozygosity and
j to i, considering population i to be upstream of mean number of alleles per locus) were not significant
© 2003 British
Ecological Society, population j. Therefore, if migration was predomin- (Fig. 6), and remained non-significant when WMPS16
Journal of Ecology, antly downstream, i to j minus j to i should be positive. and WPMS18 were excluded (data not shown). Out of
91, 447–456 Comparing estimates for the entire data set (231 the 73 alleles identified among the six loci, only three
 13652745, 2003, 3, Downloaded from https://besjournals.onlinelibrary.wiley.com/doi/10.1046/j.1365-2745.2003.00772.x by Cochrane Colombia, Wiley Online Library on [07/09/2023]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
453
Dispersal in black
poplar

Fig. 7 Significant clines in allelic frequencies observed along the river system (in kilometres). Alleles of (a) WPMCG14,
(b) WPMS20 and (c) WPMS18. For each allele, Spearman’s coefficient of correlation and the associated P-value are given.

Table 2 Comparison of the multilocus diversity estimates within two populations from the global samples and the seedling
patches samples. (FST: differentiation among populations, FIS: heterozygote deficit, NS: not significant, *P < 0.05)

Expected
Population Sample heterozygosity FST FIS

1 Global 0.68 −0.09

1 Patches 0.72 0.001 NS −0.05
6 Global 0.76 −0.14
6 Patches 0.74 0.016* −0.17

alleles showed a significant Spearman’s coefficient of
correlation with distance from the source after Bonfer-
roni’s correction: two alleles increased in frequency
going down the river and one allele decreased (Fig. 7).
    
   

In populations 1 and 6, the same level of genetic diver-
sity was sampled with a global sample from many
patches and from a set of three patches. In population
1, there was no genetic differentiation among patches
but in population 6 differentiation was significant due
to the higher distance between patches (Table 2).
© 2003 British
Discussion
Ecological Society,
Journal of Ecology, This genetic study of dispersal was based on a sample
91, 447–456 of contemporaneous seedlings. This sampling strategy
had a twofold advantage: (i) it was an efficient way to
avoid clonal replicates when the relative importance
of sexual vs. asexual propagation might have varied
between poplar stands (Legionnet et al. 1997; Arens
et al. 1998; Winfield et al. 1998); and (ii) it avoided the
bias of multicohort sampling given that genetic differ-
entiation between cohorts might have occurred, e.g. for
a pioneer species when the different cohorts relate to
colonizing gene pools. As there is no seed bank for this
species, the observed structure of genetic diversity was
due only to the spatial component of gene flow and not
to temporal heterogeneity of recruitment. Furthermore,
this sampling method fits the migrant pool model used
in many genetic models. However, our study was limited
by considering a single dispersal event with no replica-
tions through time (Clark et al. 1999). Nevertheless, as
we studied dispersal along the whole river-system
(> 100 km) and as no extraordinary climatic event
occurred during this year, we consider that our results
provide general information on gene flow for this species.

454 The partition of the genetic diversity among and
E. Imbert & within populations was consistent with most results
F. Lefèvre from tree species, i.e. diversity was mainly found within
populations (Adams 1992; Hamrick et al. 1992). Selec-
tion processes during the life cycle are generally
invoked to explain the significant excess of hetero-
zygosity that is often observed in tree species (Bush &
Smouse 1992; Husband & Schemske 1996). Although
we analysed young seedlings (2 months old), previous
investigations showed that for poplars, as for most pio-
neer species, competition for resources becomes import-
ant soon after germination (Braatne et al. 1996) and
thus selection might occur at a very early stage. It can
also be suggested that excess heterozygosity was due to
genetic differentiation between the male and the female
gamete pools. However, we did not find any genetic dif-
ferentiation between 33 male and 20 female adult trees
that were sampled in populations 1 and 6 (FST = 0.00).
Finally, it seemed unlikely that heterozygosity would
result from hybridization with the cultivated gene pool
as we detected no trace of introgression from exotic
species and no more than a few per cent from cultivated
P. nigra ‘italica’.
Although gene flow is thought to be important in
forest trees, in particular for anemophilous species, we
found significant genetic differentiation even between
nearby populations. Thus single migrant pool does not
exist over the whole metapopulation, i.e. the infinite
island model is rejected, and the method proposed by
Rousset (1997) leads us to conclude that there is iso-
lation by distance. This is somewhat surprising if we
consider the assumptions underlying the isolation by
distance model: several studies have shown that depar-
tures from these assumptions are common and reduce
the power of statistical test for the spatial structure of
diversity (see for example Richards et al. 1999). One
assumption of the model that appears unlikely is the
demographic stability of the populations through time.
However, the ecology and recent history of P. nigra
along the Drôme river suggest that population size
is relatively stable. The area of riparian forests has
changed noticeably during the 20th century (J. M. Faton,
personal communication), but we expect changes in
effective population size to be less important than
changes in population area. Indeed, as competition
for light determines crown development and flower
set, male and female gametes are mainly produced
by trees on the edge of the forest, or isolated, rather
than located inside the forest. Effective population
size is therefore more directly related to length of
the perimeter of the riparian forest, which suffered less
variation, than to forest area, consistent with one basic
assumption of the model.
We first estimated the parameters of the isolation by
distance model for the whole river-system, and then
© 2003 British
showed that the model still holds in each part of the
Ecological Society, river (upstream/downstream), although probably not
Journal of Ecology, with the same parameter values. Indeed, in the isola-
91, 447–456 tion by distance model, the slope of the regression
13652745, 2003, 3, Downloaded from https://besjournals.onlinelibrary.wiley.com/doi/10.1046/j.1365-2745.2003.00772.x by Cochrane Colombia, Wiley Online Library on [07/09/2023]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
between FST/1 − FST and distance is 1/4Dσ2, where D is the
density of populations, and σ2 a parameter describing
dispersal (Rousset 1997). The slope is lower in the
upper part of the river so that, if we assume identical
population density in the two parts of the river, we can
conclude that σ2 is greater in the upper than the lower
part. We expected more gene flow and less differenti-
ation among populations downstream for two reasons:
(i) there are no topographical obstacles; and (ii) the
larger area of riparian forest with adult trees favours
the production of an abundant migrant pool. These
a priori considerations refer to potential gene flow,
i.e. the number of migrants produced and the physical
constraints to migration. However, our study refers to
effective gene flow, i.e. the number of migrant seeds that
could effectively germinate and survive for at least 2
months. The discrepancy between potential and effect-
ive gene flow suggests that seedling establishment is a
limiting factor to gene flow: the lateral dynamics of the
river in the upstream part regularly produces a large
area suitable for colonization, whereas in the lower part
the river is less dynamic and the pioneer habitats are
limited. According to Cain et al. (2000) poplar would thus
be recruitment-limited rather than dispersal-limited.
Prevailing winds also contribute to the observed pat-
terns, spreading pollen among populations only in the
upper part of the river, where they follow the direction
of the valley.
The full matrix model proposed by Beerli & Felsenstein
(2001) allows for non-uniform demographic parameters
Ni, Mij, but even so we found no difference between
upstream and downstream directions. In addition
to temporal stability of demographic parameters
the model also assumes the absence of external gene
flow. This can be assumed for the upper part of this
river-system (neglecting the impact of the few small
poplar stands in the tributaries of the Drôme) but
probably does not hold for the alluvial plain down-
stream, where the influence of external populations is
more likely. Nevertheless, a symmetrical migration pat-
tern was found in both parts of the river.
The observed isolation by distance and symmetrical
gene flow lead us to conclude that gene flow among
black poplar populations occurs essentially through
pollen and that seed flow is very limited for distances
above 1–3 km. This was also supported by the signific-
ant differentiation among the well-separated seedling
patches in population 6. Thus, for black poplar, wind
seems to be the main dispersal factor. Primary wind
dispersal may prevail even for seeds as their viability is
reduced and they need to find suitable conditions for
germination as soon as hydration occurs (van Splunder
et al. 1995; Barsoum & Hughes 1998). The importance
of dispersal of pollen compared with that of seeds has
been demonstrated for several tree species using differ-
ent methods (e.g. Latta et al. 1998; Oddou-Muratorio
et al. 2001). We observed no accumulation of genetic
diversity in the downstream part of the river, in con-
trast with studies on some other riparian species (see

455 Introduction). Clinal variation, in both directions, was
Dispersal in black observed for three alleles from three independent loci:
poplar these probably reflect a diffusion process currently in
progress.
Casagrandi & Gatto (2002) showed that the effects
of fragmentation on metapopulations depend on dis-
persal rates: species with high dispersal are more
affected by habitat destruction than by reduction of the
habitat’s carrying capacity. Their model is not, how-
ever, spatially explicit (island model) and does include
a spatial description of dispersal. Although we do not
have precise estimates of the demographic parameters
used in our model, we can speculate on some conse-
quences for P. nigra conservation in our river-system.
We found clear differences between P. nigra popu-
lations in the upstream and the downstream parts.
Upstream, population size is generally smaller due to
topography and land uses but, as the river is still
dynamic, suitable sites are available for seedling estab-
lishment and gene flow is more important. Priority
here should therefore be to avoid the loss of suitable
habitats for P. nigra and to prevent discontinuities
emerging in the riparian forest. Conversely, in the lower
part, although large populations are still found (several
hundreds of reproductive trees), suitable areas for seed-
ling establishment are scarce and gene flow is more lim-
ited. Attention might therefore focus on the main-
tenance of large riparian forests that do have a capacity
for local regeneration.
The current structure of genetic diversity might also
reflect the recent history of the species along the river
(Petit et al. 1997), but still suggests a similar restriction
in effective gene flow as Hardy & Vekemans (1999)
showed that spatial structure of diversity rapidly
reaches quasi-equilibrium.
A general framework for in situ conservation activi-
ties of this pioneer species has been developed by the
Populus nigra EUFORGEN Network (Lefèvre et al.
2001). Monitoring gene flow in European black poplar
for in situ conservation purposes would require both
a demographic and ecological management strategy.
The development of mechanistic models of recruitment
would be very useful (Mahoney & Rood 1998).
Acknowledgements
Many thanks to Jeffrey H. Braatne, Bruno Fady,
François Rousset and three anonymous reviewers for
their comments on an earlier version of this paper. We
are also particularly indebted to Joop Ouborg for help-
ing us discover MIGRATE, to Peter Beerli for his help
with installing MIGRATE on workstations, to Benard
Orth for providing some light about the UNIX system,
and to Bruno Jouaud for technical assistance in the lab-
oratory. This study has been carried out with financial
© 2003 British
support from the Commission of the European Com-
Ecological Society, munities, Agriculture and Fisheries (FAIR) specific
Journal of Ecology, RTD programme, PL97-3386 ‘Genetic diversity in river
91, 447–456 populations of European Black Poplar for evaluation
13652745, 2003, 3, Downloaded from https://besjournals.onlinelibrary.wiley.com/doi/10.1046/j.1365-2745.2003.00772.x by Cochrane Colombia, Wiley Online Library on [07/09/2023]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
of biodiversity, conservation strategies, nature develop-
ment and genetic improvement’. It does not necessarily
reflect its views and in no way anticipates the Commis-
sion’s future policy in this area. This publication is no.
2003–008 of the Institut des Sciences de l’Evolution.
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