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Diversity of Salix reticulata L. (Salicaceae) leaf traits inEurope and its relation to geographical positionKatarzyna Marcysiak aa Department of Botany, Kazimierz Wielki University, Ossolinskich 12, Bydgoszcz, 85–093,Poland Phone: (+48) 523419015

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To cite this article: Katarzyna Marcysiak (2012): Diversity of Salix reticulata L. (Salicaceae) leaf traits in Europe and itsrelation to geographical position, Plant Biosystems - An International Journal Dealing with all Aspects of Plant Biology: OfficialJournal of the Societa Botanica Italiana, DOI:10.1080/11263504.2012.727879

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1

Salix reticulata leaf traits

Diversity of Salix reticulata L. (Salicaceae) leaf traits in Europe and its

relation to geographical position

Katarzyna Marcysiak*

Department of Botany, Kazimierz Wielki University, Ossolinskich 12, 85-093, Bydgoszcz,

Poland, tel. (+48) 523419015

*Corresponding E-mail marc@ukw.edu.pl

Abstract

Leaves of 279 individuals of Salix reticulata, collected from eight populations in

contemporary isolated parts of the species range in Europe, and for comparison, from one

population in the Rocky Mountains, were measured and analyzed statistically. The characters

describing the leaf size were more variable than the shape characters. All size characters were

statistically significantly correlated, as well as they positively correlated to the northern

latitude and eastern longitude, and negatively related to the altitude. The shape describing

characters were assumed to be independent of the environmental conditions. Relations

between samples based on multivariate analyses showed that the biogeographical structure of

the studied S. reticulata populations is not quite clear. The present results suggest early

transatlantic migrations of the species, possibly at least two migration routes to Scandinavia,

and close relations between the Tatra Mountains, the Alps and the Western Scandinavia.

Key words arctic-alpine plants, leaf shape, leaf size, morphological variability, Salix

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Introduction

Salix reticulata is a circumpolar arctic-alpine species. In Europe, it grows in the arctic

and subarctic areas and in Central European mountains (Jalas & Sominen 1973, Hedrén 2000,

Zając & Zając 2009). As an arctic plant, it played an important role during cold periods in the

Pleistocene, when the species range was much wider than today (Hedrén 2000, Birks & Willis

2008). Nowadays, it is both an element of arctic tundra and of subalpine and alpine belt in the

mountains. Together with other plants of this type of geographic range, it is possibly

endangered by the climate warming. The contemporary variability of arctic-alpine plants

species, as the consequence of their history, especially glacial and postglacial migrations,

have recently been the subject of many studies (Despres et al. 2002, Skrede et al. 2006,

Eidesen et al. 2007, Schönswetter et al. 2007, Ronikier et al. 2008, Alsos et al. 2009). Still,

Salix reticulata remains poorly investigated. It is mentioned as an element of tundra or

montane flora, sometimes as a pioneer species, in different studies (Walker et al. 2001;

Schmidtlein & Ewald 2003; Mardon 2003; Bergman et al. 2005; Nyman & Julkunen-Tiitto

2005; Tscherko et al. 2005; Gough 2006; Ling-Yu et al. 2006). The aim of the present work is

to examine the morphological diversity of Salix reticulata on the base of the leaf traits within

the species range in Europe and to compare the traits characteristics from different regions, as

well as to compare with an American sample. The question also is whether the diversity of

morphological leaf characters from the isolated populations could be related with a possible

migration history of the species.

Material and Methods

Species description

Salix reticulata is a dwarf shrub up to 10-30 cm, with a creeping stem, short twigs and

characteristic leaves: with petiole 4-23 mm long and a blade nearly orbicular to ovate or

obovate, 15-55 mm long and 7-45 mm wide, thick, with sunken veins on the adaxial side of

the blade, and prominently raised on the abaxial side. Catkins are terminal on long peduncles

(7-50 mm), appearing with the leaves or shortly after. The willow prefers moist, calcareous

soil. In the mountains, it can be found mainly in the subalpine and alpine belt, in snowbeds,

on rocky slopes, cliffs and moraines, glacials borders, sometimes on moist Dryas heath or

grasslands, rarely in forest. The willow is circumpolar except for Greenland and Iceland. In

Europe, besides the arctic and subarctic part, it grows in northern Scotland, in the

Scandinavian Mountains, the Carpathians, the Alps and the Pyreneés (Rechinger 1964,

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Pawłowski 1956, De Bolòs & Vigo J. 1990, Castroviejo et al. 1993, Hedrén 2000, Argus

2010).

Material

The analyses were based on the traits of leaves, collected from eight populations from

contemporary isolated parts of the species range in Europe and for the comparison one sample

was collected in the American part of the range (Table 1). In each examined population

approximately ten leaves from 30 individuals were sampled. The individuals growing no

closer than 3 m from each other were chosen, to avoid sampling the same genet (Max et al.

1999; Stamati et al. 2007). Leaves were put into the herbarium, and measurements were taken

on the dried material, omitting leaves damaged during transport and drying process. 2631

leaves of 279 individuals were examined altogether.

Methods

Leaves were scanned and measurements were taken from the scans with the use of

digiShape 1.9.177 (CortexNova). Typical traits used for leaf characteristic were chosen

(Elkington 1968, Santini et al. 2004, Kovačić & Nikolić 2005, Kehl et al. 2008, Viscosi et al.

2009) (Table 2).

The characters describing the leaf size, i.e. per, ll, pl, w1/2, w1/4, w3/4, were used

mainly for characteristics of the populations. On the basis of these six measured traits, further

five were calculated (per/l, l/w, p/llp, w1/4/w, w3/4/w), and together with measurements of the

leaf apex and base (aa, ba), they were treated as the shape describing features (Table 2). Only

these characters were used for multivariate analysis because they are treated as more stable

and independent on the environmental conditions (Kremer et al. 2002). The results of the

measurements were analysed statistically.

Means and standard deviations of all characters of the whole data set were calculated,

and minima and maxima were found. Afterwards, the coefficient of variation

(CV=100SD/M) for each character was calculated. Means and standard deviations calculated

for each population were presented on the boxplots.

For every sample, arithmetic means of the individuals were calculated. The matrix

constructed with them was standardized and used in further analyses.

Correlations between characters were calculated using the Pearson’s correlation

coefficient. R Spearman’s coefficient for non-parametric distributions was used to find

correlations between characters and the geographical coordinates of every sample: longitude,

latitude and altitude. This analysis was based on the means of samples.

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The analysis of variance was performed to find which characters differentiated

significantly the analyzed samples. Then Tukey’s test (RIR) was carried out in order to show

which samples differed in reference to particular features.

The multivariate relations between populations were analyzed with the use of the

discriminant analysis on the basis of the shape characters. As a result of this analysis, the

means of the populations were shown on the scatterplot of the two first discriminant variables.

The correlations between characters and the discriminant variables were also calculated to

find characters responsible for the variability observed (Sokal and Rohlf 2003).

All the calculations and analyses were conducted with the help of STATISTICA 9.0

(StatSoft, 2009).

Results

The mean length of the leaves studied in the present work was 20.0 mm and their

width was 13.5 mm, while the average petiole was 8.4 mm long (Table 2).

The mean values of the leaf characters in the examined populations varied, and the

differences between the samples means were generally greater with regard to the characters

describing the leaf size (Fig. 1) than in the case of the shape describing features (Fig. 2 A-C).

The analysis of the leaf size let distinguish the group of populations with bigger leaves: NS,

WA, T1 and T2. In these populations, with the exception of WA, the dispersion of

measurement values was also bigger. Samples: P1, WS, EC and R created the group with

smaller leaves and less dispersed values. The leaves from the population P2 were very small,

with the means evidently smaller than the others and the narrowest confidence intervals (Fig.

1).

The differences between samples means with regard to the shape characters were less

evident. The group of populations with more elongated leaves could be observed: R, WA and

P1, and the almost rounded leaves were found in population P2 (Fig. 2 A). The proportion of

the petiole length in the whole leaf length was biggest in P1, but smallest in P2 (Fig. 2B). The

leaves from R and T1 were wider in the upper part when compared to other samples, and at

the same narrower in the bottom side, with the opposite to NS, P1, T2 and EC (Fig. 2C).

Sample P2 again reached the extreme values of these characters (Fig. 2A-C).

The coefficients of variation of characters calculated for the whole data set proved that

the size characters were more variable then the shape characters (Fig. 3).

All the size describing characters were statistically significantly correlated. Among the

shape characters, both angles (aa, ba) were correlated to the ratio of the length and width of

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the leaf blade (l/w), besides, aa was significantly related to the ratio of the perimeter and the

length of the leaf (per/l), and ba to the ratio of the width measured at 1/4 of the leaf length

and the one measured at 1/2 (w1/4/w) (Table 3).

What is more, the size characters were statistically significantly correlated to the

geographical coordinates, thus their value increased from West to East and from South to

North, as these relation were positive, but diminished with the growing altitude, as that

correlation was negative. Still, the correlations were not very strong. At the same time, as far

as the shape features are regarded, the weak, but significant, correlations were found only

between ba and w1/4/w and the latitude and between l/w and longitude (Table 4).

The results of the analysis of variance showed that all characters differentiated the

samples statistically significantly (Table 5). The F statistics of the size characters were

evidently greater than in the case of the shape characters, so the former should be more

important for the analysis. At the same time, these characters were strongly correlated, as

described above, so in the multivariate analysis they were omitted.

The Tukey’s test showed that, with regard to the size characters, populations differed

from each other. However, the lack of differences was found between the Tatra populations

(T1, T2) and WA, as well as between the American sample (R) and the Eastern Carpathian

and Western Scandinavian ones (EC and WS) (Table 6). There were fewer differences

between samples with regard to the shape characters, many features differentiated only P2

from all the others, with the exception of EC. P1 also differed from some other populations,

and, interestingly, five of seven analyzed shape characters differentiated P1 from P2 (Table

7).

On the scatterplot expressing the results of the discriminant analysis, on the plain of

the two first canonical variables explaining almost 73% of the total variation, samples: T1,

T2, WA, P1 and WS create one group. P2 is a slightly more distant from the group,

especially with regard to the second variable. The populations: EC and NS are most distant

from the rest of them and also much more distant from each other with regard to the second

variable. Surprisingly, the sample collected in Rocky Mountains takes place between NS, EC

and the others (Fig. 4). The first canonical variable is influenced most by ba and per/l, and the

second by the same two characters and l/w.

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Discussion

Leaf size

The average values of S. reticulata leaves in the present study are slightly smaller than

most of the average sizes given in the Floras (Rechinger 1964, De Bolòs & Vigo J. 1990,

Castroviejo et al. 1993, Hedrén 2000). The mean leaf length observed was 20 mm and

maximal 46 mm, while European literature data indicated 15-20 mm as minimal and 50 mm

as the maximal length, and differences concerning the leaf width and the petiole length were

comparable. The average value of the leaf elongation (l/w) equaling 1.5, was similar in the

present study and in the sources.

The morphological leaf traits of S. reticulata are more variable than some other boreal

and arctic small shrubs species studied (Marcysiak & Lewandowska 2008 a, b).

The dependence of morphological characters on environmental factors is a known fact

(Huber & Wiggerman, 1997; Noda et al., 2004; Marchand et al., 2006; Baquedano et al.,

2008; Xu et al. 2009, Bruschi 2010, Fletcher et al, 2010). The findings of a present study

seem to confirm the connection of the leaf size with the environmental conditions. The

populations studied differed, sometimes considerably, with regard to the leaf size. The

interesting feature found was the relation of the leaf size to the geographical coordinates. The

positive correlation to the northern latitude and eastern longitude from one side and the

negative connections to the altitude from the other, can be treated as the same property. In

Europe, the further the North and East, the more severe environmental conditions become,

and the willow grows at the lower altitudes, down to the polar tundra in the northernmost

parts of the species range. Thus these relations may reflect the influence of the climatic

condition on the plant size. The smallest leaves were observed in the western- and

southernmost located sample Pyrenées2, although in neighbouring Pyrenées1 they were much

greater. Still, Pyrenées2 grows 2670 m asl, about 300 m higher than Pyrenées1, and actually,

slightly above the upper limit of the species in that mountain system, reported as 2600 m asl

(De Bolòs & Vigo J. 1990, Castroviejo et al. 1993), thus the growth conditions there may be

unsuitable. The sample P2 seem to be the outlier in the light of the data analyzed. Amongst

the studied European populations, very small leaves were also found in the Munti Rodnei in

the Eastern Carpathians, the easternmost location at this latitude and close to the eastern range

limit, lying 1910 m asl, which is quite high, as the highest peak of these mountains is 2305 m

high. The local adaptation to altitude, in spite of the gene flow, has been already described

(Byars et al. 2007; Gonzalo-Turpin & Hazard 2009).

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On the other hand, the small leaf size was also observed in WScandinavia, on the

position located well within the species range but exposed southward (Table 1), and thus

pretty dry, where S. reticulata grew on the shale, together with S. herbacea (Fig 5), and the

development conditions probably were again unfavourable.

It can be concluded that, although the environmental conditions were not the subject of

the study and so this data are incomplete, the results indicated the connections between the

leaf size and the climate. This may be treated as the message of the species reaction to the

climate warming, what, in the light of the present finding, would cause the diminishing of the

leaf size.

Relations between populations

As the shape characters were practically not correlated to each other (Table 3) and the

detected correlations to the geographical coordinates were very weak (Table 4), it was

assumed that these characters are independent of environmental conditions. The weaker

variability of these characters, expressed with the variation coefficients (Fig. 3), was also

considered as a sign of their independence. Thus, the shape characters may represent the

relations between populations resulting from their history, isolations and migrations routes.

It was expected that the connections between the samples based on the leaf shape could

reflect the glacial migration history of the species. The main goal was focused on the relations

within Europe, and the Rocky M. sample (R) was treated as a sort of out-group, the

population geographically being greatly distant and so probably much longer isolated from

the others, and, consequently, much different. However, this hypothesis appeared false, as

both the size and shape leaf characteristics of the Rocky M. sample well fell into the general

range of the European variability of the species (Fig. 1-2).

Besides, in the result of the discriminant analysis, the Rocky Mountains sample took the

position between the easternmost populations: NS and EC, and other populations, located

further to the west (Fig. 4). If these distances mirror the existing relationships, the conclusion

is that the glacial/postglacial migrations of the species leaded across the Atlantic Ocean. The

long distance dispersal was often proved true for the arctic-alpine species and the North

Atlantic was reported not to create a barrier during the glaciations (Gabrielsen et al. 1997,

Abbott et al. 2000, Brochmann et al. 2003, Schönswetter et al. 2007). However, the findings

based on the morphology only, could not reveal the direction of these migrations.

The P2 population, much different from the rest with regard to the all characters

analyzed (Fig. 1-2), was also distant on the graph of the discriminant analysis. This may be a

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sign of the much earlier migration and the longer isolation of these populations, as in the case

of the Pyrenéan population of Salix herbacea (Alsos et al. 2009). At the same time the low

variability of the size characters of this sample (Fig. 1) may signify decreasing variability in

the population caused by the genetic drift.

In the light of the above considerations and results of the discriminant analysis, the

origin of the EC population remains unclear, as it is closest to the American sample (Fig. 4).

This relation may point out the common source of these populations, perhaps the early

transatlantic migration.

The sample from the northern Scandinavia is the most separated from all the others in

the present results, and the source population of this part of the range might have lain

somewhere else, for instance in the Siberia, as it was suggested for another calcareous species,

Dryas octopetala (Skrede et al. 2006).

The Tatra populations (T1, T2), as well as the populations from the Alps (WA),

Western Scandinavia (WS) and the Pyrenées (P1) formed one group on the scatterplot (Fig.

4). Their origin from the same source and migrations into different directions after the

glaciations retreatment seems possible.

The distance between two analyzed Scandinavian samples: northern (eastern) and

western, should be noted. The different migration routes to Scandinavia were described for

some species (Nordal & Jonsell, 1988; Taberlet et al., 1998; Hewitt, 2000), and present results

support these findings.

The conclusion is that the biogeographical structure of the studied S. reticulata

populations is not quite clear and demands further, possibly genetic, investigations. The

present results suggest the early transatlantic migrations of the species, possibly at least two

migration routes to Scandinavia, and close relations between the Tatra Mountains, the Alps

and the Western Scandinavia, as well as the differentiated periods of isolation of populations.

Acknowledgments

I would like to thank Amelia Lewandowska for her great help in the measurement

procedures and Adam Boratyński, Krystyna Boratyńska, Anna Ronikier and Michał Ronikier

for the material collection. I am also grateful to Jason Sinicki and Alison Cowper for

proofreading.

The collection of the part of the material was possible thanks to the financial support

of Kazimierz Wielki University in Bydgoszcz, Poland.

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Fig. 1. The box-plots of the size characters of Salix reticulata samples. Populations acronyms

as in Table 1. Populations sequence shows the affinities between them. Point –

characters’ means, box – standard deviations, whiskers – 1.96 standard deviations.

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Fig. 2. The box-plots of the shape characters of Salix reticulata samples. Populations

acronyms as in Table 1. Populations sequence shows the affinities between them. Note

that sequence in A is different than in B and C. Point – characters’ means, box –

standard deviations, whiskers – 1.96 standard deviations.

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Fig. 3. Values of the coefficients of variation of Salix reticulata leaf characters, calculated for

the whole data set.

Fig. 4. Result of the discriminant analysis, the scatterplot of means of samples on the plane of

the two first discriminant variables.

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Fig. 5. Salix reticulata in Prestholtskarvet, Norway (sample WS), growing together with S.

herbacea.

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Table 1. Collection data of Salix reticulata samples.

No Location Acronym Alt. Longitude Latitude N Exposition Year of

collection

Collector

*

1 Romania, Eastern Carpathians, Muntii Rodnei EC 1910 E 24o51’52” 47o32’18” NW 2008 AR, MR

2 Poland, Tatra Mts. Kondracka Przełęcz T1 1700 E 19o57,21’ 49o15,79’ N 2005 KM

3 Poland, Tatra Mts, Czerwone Wierchy T2 1750 E 19o54,11’ 49o15,20’ E 2005 KM

4 France, Western Alps, Col du Galibier WA 2620 E 06o24’47” 45o03’56” N 2007 AB

5 Andorra, Pyrenées, Rialb P1 2330 E 01o34,12” 42o38’33” N 2007 AB

6 Andorra, Pyrenées, Casamanya P2 2670 E 01o34’08” 42o35’06” SSE 2007 AB

7 Norway, Buskerud, Geilo, Prestholtskarvet WS 1300 E 08o04,75’ 60o33,56’ S 2006 KM

8 Norway, Finnmark, Nordkapp NS 100 E 25o47’04” 71o05’36” NW 2008 KB, AB

9 USA, Montana, Carbon Country, Rocky Mountains, Beartooth Plateau

R 3000 W 109o24’32” 45o01’21” - 2008 AR, MR

* Collectors: AB-Adam Boratyński; AR-Anna Ronikier; KB-Krystyna Boratyńska, KM-Katarzyna Marcysiak; MR-Michał Ronikier

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Table 2. Statistics of analysed characters of Salix reticulata leaves.

No Character

Acr

onym

Mea

n

Min

-Max

Stan

dard

de

viat

ion

1. Perimeter of leaf blade [mm] per 54.6 14.6 - 117.0 16.342. Length of leaf blade [mm] ll 20.0 5.1 - 45.5 6.243. Length of petiole [mm] pl 8.4 0.8 - 26.2 3.754. Width of leaf blade measured at 1/2 of its

length [mm] w1/2 13.5 3.5 - 32.9 4.29

5. Width of leaf blade measured at 1/4 of its length [mm]

w1/4 11.1 2.6 - 27.7 3.57

6. Width of leaf blade measured at 3/4 of its length [mm]

w3/4 12.1 3.6 - 29.5 3.94

7. Apex angle of leaf blade [o] aa 145.8 93.6 - 171.1 12.718. Base angle of leaf blade [o] ba 130.5 59.7 - 207.8 22.199. Ratio of perimeter/length of a leaf blade

(char. 1/2) per/l 2.75 2.28 - 4.05 0.20

10. Ratio of length/width at1/2 (char. 2/4) l/w 1.50 0.82 - 3.54 0.2611. Ratio of length of petiole/sum of blade and

petiole length (char. 3/(char. 2+3)) p/llp 0.29 0.09 - 0.50 0.06

12. Ratio of width at 1/4/width at 1/2 (char. 5/4) w1/4/w 0.82 0.53 - 1.04 0.0513. Ratio of width at 3/4/width at 1/2 (char. 6/4) w3/4/w 0.89 0.73 – 1.22 0.05

Table 3. Correlation coefficients between S. reticulata leaf characteristics; correlations significant at p<0.01 are shaded.

ll 0.99 pl 0.93 0.95

w1/2 0.98 0.97 0.87 w1/4 0.97 0.96 0.86 1.00 w3/4 0.98 0.96 0.87 1.00 0.99 aa -0.61 -0.67 -0.72 -0.48 -0.46 -0.47 ba -0.70 -0.71 -0.77 -0.58 -0.53 -0.59 0.80

per/l -0.51 -0.61 -0.69 -0.46 -0.44 -0.44 0.81 0.63 l/w 0.43 0.50 0.64 0.27 0.23 0.27 -0.87 -0.82 -0.77

p/llp 0.48 0.53 0.76 0.39 0.36 0.39 -0.59 -0.63 -0.66 0.72 w1/4/w -0.48 -0.49 -0.55 -0.42 -0.34 -0.47 0.48 0.81 0.47 -0.58 -0.46 w3/4/w 0.13 0.09 0.14 0.11 0.04 0.19 0.07 -0.29 0.03 -0.13 0.14 -0.73

per ll pl w1/2 w1/4 w3/4 aa ba per/l l/w p/llp w1/4/w

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Table 4. Correlation coefficients between geographical coordinates and characters values for Salix reticulata; correlations significant at p<0.01 are shaded.

alt E N

per -0.42 0.40 0.45ll -0.38 0.36 0.43pl -0.26 0.24 0.27

w1/2 -0.48 0.50 0.52w1/4 -0.47 0.49 0.49w3/4 -0.47 0.50 0.51aa 0.07 -0.01 -0.10ba 0.14 0.02 -0.18

per/l 0.07 -0.04 -0.13l/w 0.08 -0.16 -0.03

p/llp -0.00 -0.01 -0.04w1/4/w 0.09 -0.03 -0.17w3/4/w 0.06 -0.00 0.02

Table 5. Results of the analysis of variance. SS df MS F p

per 169.23 8 21.15 52.43 0.0000ll 168.44 8 21.06 51.72 0.0000pl 126.27 8 15.78 28.02 0.0000

w1/2 154.08 8 19.26 41.89 0.0000w1/4 142.46 8 17.81 35.53 0.0000w3/4 152.79 8 19.10 41.07 0.0000aa 32.74 8 4.09 4.50 0.0000ba 76.54 8 9.57 12.90 0.0000

per/l 64.27 8 8.03 10.60 0.0000l/w 50.75 8 6.34 7.55 0.0000

p/llp 50.73 8 6.34 7.54 0.0000w1/4/w 47.96 8 6.00 7.13 0.0000w3/4/w 46.15 8 5.77 6.73 0.0000

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Table 6. Results of Tukey test for size describing characters of leaves of S.reticulata populations (acronyms as in Table 1); acronyms of characters significantly differing populations at p<0,01 are listed.

T1

per ll pl w1/2 w1/4 w3/4

T2

per ll w1/2 w1/4 w3/4

WA

per ll pl w1/2 w1/4 w3/4

pl

P1

pl

per ll w1/2 w1/4 w3/4

w1/2 w1/4 w3/4

per ll w1/2 w1/4 w3/4

P2

per ll pl w1/2 w1/4 w3/4

per ll pl w1/2 w1/4 w3/4

per ll pl w1/2 w1/4 w3/4

per ll pl w1/2 w1/4 w3/4

per ll pl w1/2 w1/4 w3/4

WS

per ll pl w1/2 w1/4 w3/4

per ll w1/2 w1/4 w3/4

per ll pl w1/2 w1/4 w3/4

pl

per ll pl w1/2 w1/4 w3/4

NS

per ll pl w1/2 w1/4 w3/4

per ll w1/2 w1/4 w3/4

per ll pl w1/2 w1/4 w3/4

per ll pl w1/2 w1/4 w3/4

R

per ll pl w1/2 w1/4 w3/4

per ll w1/2 w1/4 w3/4

per ll pl w1/2 w1/4 w3/4

per ll pl

per ll pl w3/4

per ll pl w1/2 w1/4 w3/4

EC T1 T2 WA P1 P2 WS NS

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Table 7. Results of Tukey test for shape describing characters of leaves of S.reticulata populations (acronyms as in Table 1); acronyms of characters significantly differing populations at p<0,01 are listed

T1 ba

w1/4/w

T2 w1/4/w

WA aa ba l/w

aa per/l l/w

P1 ba l/w

p/llp

w1/4/w w3/4/w

per/l p/llp

P2

ba p/llp

w1/4/w w3/4/w

ba

aa ba

per/l l/w

w1/4/w

aa ba

per/l l/w

p/llp

WS ba

w3/4/w p/llp

ba per/l

w1/4/w

NS per/l per/l w1/4/w

per/l

ba per/l

R

l/w w3/4/w ba per/l l/w

w1/4/w w3/4/w

w3/4/w

EC T1 T2 WA P1 P2 WS NS

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