a multivariate approach to assess the taxonomic utility of morphometric characters indoronicum...

Folia Geobotanica 36: 423-444, 2001

A MULTIVARIATE APPROACH TO ASSESS THE TAXONOMIC UTILITY OF MORPHOMETRIC CHARACTERS IN DORONICUM ( ASTERA CEAE, SENEClONEAE)

In6s Alvarez Fernfindez & Gonzalo Nieto Feliner

Real Jardln BoMnico. CSIC, Plaza de Murillo 2, E-28014 Madrid, Spain/ fax +34 914200157, e-mail [email protected]

Keywords: Discriminant analysis, Ordination techniques, Principal components analysis, Taxonomy

Abstract: A multivariate morphometric study based on 22 characters was carried out on samples of the 26 accepted species of the Eurasian and north-African genus Doronicum, to assess whether quantitative, mainly continuous, characters were of use in suggesting, delimiting and structuring natural groups. The study concentrated on three-medium sized to small groups: Mediterranean (D. plantagineum group), European (1). grandiflorum group), and central Asian. Two questions were addressed for each of these groups: species delimitation within them and the inclusion/exclusion of species of uncertain, but putative close affinities. Principal components analysis and discriminant analysis (canonical variates analysis), gave moderately satisfactory results. The addition of doubtfully-related species to presumably natural groups increased cohesiveness of the groups because the between-species differences were minimized when placed in a broader more diverse context. The relative status of the uncertain species, either as outliers or true members, was appropriately depicted by the scatterplots, but more convincingly solved when the species turned out to be outliers. Discrimination between species, which belong to cohesive groups was effective in some cases at least when using discriminant analysis, and the exceptions involved particularly problematic taxa. It was concluded that morphometric characters are a useful complementary source of information in groups where qualitative (in particular, shared) characters are scarce as is the case in Doronicum.

INTRODUCTION

The genus Doronicum (Asteraceae, Senecioneae) is represented by perennial rhizomatose herbs with alternate, simple leaves, beating one to several radiate yellow-flowered capitula. Phyllaries are all uniform, herbaceous, and arranged in 2-3 rows. Cypselae are broadly elliptic-oblong and have 10 longitudinal fibs. A pappus o f bristles is present in all cypselae, except for the ligulate florets o f the heterocarpic species, where it is lacking. The representatives o f this genus are distributed in Asia, Europe, and northern Africa, and occur on open or forested habitats from sea level up to 5,000 m o f elevation.

Although several taxonomic studies o f the genus Doronicum circumscribed to particular regions are available (WIDDER 1925, EDMONDSON 1973, 1975, 1978, AVETISJAN 1980, CHACON 1987, Pt~REZ MORALES & PENAS 1990, DUVIGNEAUO 1992, Pf/REZ MORALES et al. 1994), the only comprehensive revision o f this genus is that by CAVmLIER (1907, 1911). He recognized 34 species arranged in 3 sections: Doronicastrum CAVILL., Soulieastrum CAVILL., and Hookerastrum CAVILL. The latter section included one species nowadays placed in another genus within a different tribe, Nannogtottis hookeri (C.B. CLARKE ex HOOK. f.) KITAM. While sect. Soulieastrum is monospecifie (D. stenoglossum MAXIM.), Cavillier subdivided his main

424 I. ~,lvarez Fernhndez & G. Nieto Feliner

Table 1. Outline of the taxonomy of the genus Doronicum proposed in/~LVAREZ FERNANDEZ (1999). Informal

~ roups, mainly based on morphology and geography, are considered instead of formal infrageneric categories. - in ALVAREZ FERNANDEZ & NIETO FELINER (2000); 2 _ in ALVAREZ FERNANDEZ (2001).

Group Taxa

1). caviUieri ,~LVAREZ FERNANDEZ et NIETO FEL. 1 1), kamaonense (DC.) ,~LVAREZ FERNANDEZ 2 (D. roylei DC.) D. oblongifolium DC. D. pardalianches L. D. stenoglossum MAXIM.

D. austriacum group D, austriacura JACQ, D. carpetanura BOISS. et REUT. ex WILLK. subsp, carpetanum D. carpetanum subsp, diazii (Pl~REZ MORALES et PENAS) ,~LVAREZ FERNANDEZ 2 D. carpetanura subsp, kuepferi (R. CHACON) ,~LVAREZ FERNANDEZ 2 D. carpetanum subsp, pubescens (P~REZ MORALES, PENAS, LLAMAS et ACEDO) AIZPURU D. cataractarura WIDDER D. corsicura (LOISEL.) POIR.

D. plantagineum group D. carpaticum (GRISEB. et SCHENK) NYMAN D. coluranae TEN. D. hungaricum RCHB. f. D. orientale HOFFM. D plantagineum L, (D. atlanticum CHABERT)

SW Asian group D. cacaliifolium BOISS. et HELDR. D, dolichotrichum CAVILL. D. haussknechtii CAVILL. D. macrophyllum FISCH. subsp, macrophyllura (19. macrolepis FREYN et SINT.) D macrophyllura subsp, sparsipilosum (J.R. EDM.)/~LVAREZ FERNANDEZ 2 D. maximum BOISS. et A. HUET D. reticulatum BOISS.

Central Asian group D. altaicum PALL. D. briquetii CAVILL. D. falconeri C.B. CLARKE (D. turkestanicura CAVlLL,)

D. grandiflorum group D grandiflorum LAM. D. clusii (ALL.) TAUSCH D. glaciale (WULFEN) NYMAN

section, Doronicastrum, into 7 subsections. This treatment needed a thorough revision since the subsections are not defined on the basis of exclusive (synapomorphic) characters, and since the variability of those characters used to define subsections actually extends far beyond the limits of his subsections (e.g., leaf size and shape, plant size, number of capitula). As a consequence, the placement of several species within Cavillier's treatment is rather arbitrary as is the circumscription of most of his subsections.

A taxonomic revision of the genus based on ca. 4,500 herbarium specimens from 47 herbaria has resulted in a new taxonomic treatment (ALVAREZ FERNANDEZ 1999, 2001, ALVAREZ FERNANDEZ & NIETO FELINER 1999, 2000), which recognizes 26 species and 4 subspecies.

Morphometric characters in Doronicum 425

Taxonomy is problematic, in part due to the scarcity of putative synapomorphies (primary homologies sensu DE PINNA 1991) within the genus. From a raw matrix of 44 characters (not shown), only ten could be selected as potentially informative for a phylogenetic analysis (JkLVAREZ FERNANDEZ et al. 2001). This reduction resulted from the removing of quantitative continuous characters, autapomorphies, intra-taxon polymorphic characters, and those which, despite superficial similarity, were presumably non-homologous. Even after this previous selection, only three of the remaining ten characters contained no homoplasy in a simultaneous (total evidence) parsimony analysis based on three different data sets: morphology, nuclear ribosomal ITS DNA sequences, and chloroplast trnL-trnF sequences (/kLVAREZ FERNANDEZ et al. 2001). These were the ciliate phyllaries, the scapose stem, and the triplinerved leaf venation. These three characters support the recognition of the D. plantagineum group (Tab. 1). The simultaneous analysis of the three data sets constitutes a homology test for each character against the rest in the data set (PATTERSON 1988). The remaining seven characters contained different amounts of homoplasy, and thus failed a test of homology by congruence with other characters: homocarpy, the presence of hairs and buds in the rhizome, the shape and arrangement of nodes and internodes in the rhizome, the pinnate-reticulate leaf venation, the plant architecture displayed by the SW Asian species (a few scattered very large leaves and several capitula), and the glandular cypselae. As a consequence of the low reliability of morphological characters at the above-species level, infrageneric taxa are not recognized, but several informal groups of species are named and are referred to below and in Tab. 1.

To a lesser extent, problems also exist at lower taxonomic levels. Species delimitation is, in a few cases, based on autapomorphic morphological characters. However, most of the species are defined by combinations of non-exclusive characters. The fact that some of these are continuous and their ranges of variation are wide can make species recognition equivocal.

Given the paucity of qualitative characters in Doronicum, and their questionable reliability for delimiting natural groups, we decided to assess whether quantitative, primarily continuous, characters were useful for taxonomic purposes. The objective of the present paper is thus to determine if morphometric multivariate patterns of variation can help in suggesting, delimiting and structuring natural groups. For operability, we have chosen three smaller groups of species that are presumably natural and involve unresolved taxonomic problems. In particular, two different questions have been addressed based on those groups: species delimitation within them and the inclusion/exclusion of species of uncertain affinities in these selected groups.

MATERIAL AND METHODS

Plant material and data sets

A total of 291 specimens were scored from the following herbaria: B, BM, BRNM, E, G, GAZI, GH, GZU, K, LE, MA, MO, NY, and W. The origin of the material is given in Appendix 1. These specimens are representatives of the 26 species and 4 subspecies recognized in our taxonomic treatment (,h, LVAREZ FERNANDEZ 1999). An outline of such treatment as well as authorship of the taxa are given in Tab. 1. However, to allow visualization and discussion of their morphometric behavior, specimens representing two previously recognized species that we consider synonyms have been labeled separately. These are D. atlanticum and D. turkestanicum, synonyms ofD. plantagineum and D. falconeri, respectively. For most of

426 I. Alvarez Fernandez & G. Nieto Feliner

the taxa, ten specimens spanning its geographic distribution were selected, sometimes including all the available material.

A global matrix including all the samples was used in a preliminary analysis. Even though it is likely to provide only a coarse picture of morphometric variation for taxonomic purposes, this matrix was analyzed to determine if there is a general pattern of variation that is consistent with any of the infrageneric arrangements proposed. To facilitate examination of the scatterplot, only five species were labeled independently (D. stenoglossum, D. oblongifolium, D. cavillieri, D. pardalianches, D. kamaonense), D. kamaonense being the correct name for D. roylei (ALVAREZ FERNANDEZ 2001). These five species are either morphologically very distinct or cannot be included in other groups with certainty. The rest of the species were gathered for labeling under five different groups based on morphological characters recorded during the taxonomic revision (Tab. 1). The species included in those groups share morphological, geographical and often ecological features, although they do not necessarily represent natural groups.

These groups are: (1) D. austriacum group: European tall herbs with leafy stem and several capitula. It includes D. austriacum, D. carpetanum (subsp. carpetanum, subsp, diazii, subsp. kuepferi, and subsp, pubescens), D. cataractarum, and D. corsicum. (2) D. plantagineum group, including European taxa with ciliate phyllaries, scapose stem, and triplinerved leaf venation. It is a seemingly natural group constituted of D. hungaricum, D. orientale, D. plantagineum, D. carpaticum, and D. columnae. Because D. carpaticum and D. columnae are not core members, we refer to the remaining three species as D. plantagineum s. str. group. (3) SW Asian group: species from Asia Minor, Caucasus, and Iran, integrated by tall herbs with a few very large cauline leaves, and several to many capitula. It includes D. cacaliifolium, D. dolichotrichum, D. haussknechtii, D. macrophyllum (subsp. macrophyllum and subsp. sparsipilosum), D. maximum, and D. reticulatum. (4) Central Asian group, integrated by species with one capitulurn (very rarely up to four), leaves scattered along the stem, pinnate-reticulate venation and numerous ligulate florets (30). It includes D. altaicum, D. briquetii and D. falconeri. (5) D. grandiflorum group, integrated by homocarpic European short herbs with one capitulum and several leaves scattered along the stem, occurring in open mountain areas. It includes D. grandiflorum, D. clusii, and 1). glaciale.

Three of these groups were selected for further analyses based on three criteria: they were presumably monophyletic, they stand as one of the putative closest relatives to species whose placement was doubtful, and/or they involved internal taxonomic problems that could be addressed with multivariate techniques. These were the D. plantagineum group (no. 2), the D. grandiflorum group (no. 5), and the central Asian group (no. 4). Two species of uncertain affinities (D. carpaticum and D. columnae) were analyzed with the first two groups, while another three (D. cavillieri, D. kamaonense and D. stenoglossum) were included in the third.

Characters

Since floral and fruit characters are not shown simultaneously in the same specimen, we were forced to exclude all characters relative to the cypselae except for the occurrence of heterocarpy, which can be easily visualized by the absence of a pappus in the ligulate florets. Most of the measurements were made with a Brown & Sharpe Plus digital calliper (model 599-571-3).


Table 2. Characters scored for the multivariate analyses (DA and PCA). All distance measures are in mm except for LT. Number in parentheses denote coding of discrete characters.

1. ABE: phyllary maximum width 2. AMP: receptacle base width 3. CAL: ratio of medium eauline leaf width to medium cauline leaf length 4. CALBE: ratio of phyllary maximum width to phyllary length 5. CALL: ratio of ligule maximum width to total ligulate floret length 6. CBL: ratio ofphyUary length to ligulate floret length 7. CHB: ratio of basal leaf maximum width to basal leaf length 8. CLMLB: ratio of phyllary length (from the base to the maximum width) to phyllary total length 9. CLPLT: ratio of peduncle length (uppermost leafless portion of the stem) to stem length

10. CPL: ratio of basal leaf petiole length to basal leaf blade length 11. DIAM: diameter of the eapitulum (phyllaries and ligulate florets excluded) 12. HB: basal leaves absent (0) vs. present (1) at flowering 13. HET: homocarpic eapitulum (0), heterocarpic capitulum (I) 14. LA: anther length 15. LF: disk floret length 16. LHCM: medium cauline leaf length 17. LL: ligulate floret length 18. NC: number of eapitula 19. NDA: number of teeth at the ligule apex 20. NEN: number of internodes in the stem 21. NL: number of ligulate florets per eapitulum 22. LT: stem length (categorized into three states < 0.6 m, 0.6-1 m, > 1 m)

On the whole, twenty-two characters were scored (Tab. 2). However, depending on the group analyzed, some characters were not considered due to a lack o f variation or the inapplicability for the matrix in question. For instance, when analyzing the global matrix eight characters that could not be measured in all the 30 taxa were eliminated. The characters removed from each analysis due to the above reasons are given under the results section. As a consequence, the number o f characters scored varied from 14 to 20. Eight o f them are quantitative continuous, another eight are ratios, four are quantitative discrete and two are binary (homocarpy vs. heterocarpy, presence o f basal leaves at flowering). One o f the quantitative continuous variables (stem length, LT) could not be measured exactly in incomplete large herbarium specimens and was therefore categorized into three different states (< 0.6 m, 0.6-1 m, > 1 m). The values in the data matrices are averages o f the three measurements in each specimen except for CHB, CLPLT, CPL, DIAM, FIB, HET, NC, NDA, NEN, NL, LT.

Analyses

Principal components analyses (PCA), based on the correlation matrix (SNEATH • SOKAL 1973, ABBOTT et al. 1985, MARCUS 1990), were performed in order to see whether data reduction obtained through the new set o f variables (PCs) revealed a pattern o f variation that is consistent with the taxonomy. Although these analyses are not designed for grouping, clusters can be produced when the largest components o f the overall variance are contributed by differences among groups.

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Discriminant analysis (DA) was used mainly as an ordination procedure, i.e., what is sometimes called canonical variates analysis (ABBOTT et al. 1985, MARCUS 1990). It was used to check if specimens from each species could be discriminated efficiently from congeners on the basis of morphometric characters when plotted against the canonical discriminant functions that maximize between-group variation relative to within-group variation. A stepwise procedure was followed to select the combination of variables that maximize the discrimination of predetermined groups. The low number of specimens per group could cause problems of degrees of freedom when using ca. 20 variables, although any number of variables below the total number of specimens minus the number of groups is acceptable for other authors (KLECKA 1980). Since the number of variables selected by the stepwise procedure was in all the analyses three to four, we avoided technical problems of this type. The selected variables in each of the analyses are indicated under the results section. Besides the ordination, we have also used the classificatory approach of the DA. This uses a different set of discriminant functions as compared to the canonical ones with which the scatterplots are drawn. The percent of "correctly" classified specimens was obtained by the stringent procedure of cross-validation. To refine the classification results, probabilities of pertaining to each group for every specimen were also examined although they are not given in this paper. All the analyses were carried out using SPSS 10.0.5 for Windows.

RESULTS

Global matrix PCA

Characters excluded from this analysis are CAL, CHB, CLPLT, CPL, HB, LA, LHCM, and NEN (Tab. 1). The first three PCs explain 56% of the variance (25%, 20%, and 11%, respectively). Nine PCs are needed to account for 90% of the overall variance. The highest loadings on the I st PC correspond to characters CALBE, CLMLB, and ABE. This component


is related with phyllaries shape. The variables with the highest loadings on the 2nd PC are DIAM, AMP, and LL. These variables are related with the appearance and size of the capitula.

In the scatterplot of the 291 specimens against the first two PCs, no discrete clusters are apparent (Fig. 1). Nonetheless, an examination of the placement of each species and group reveals a clear trend determined by the variables contributing most to the first two PCs. Specimens with narrow acuminate phyllaries are on the left side of the plot, corresponding to negative values for the PC1 (D. plantagineum group, D. stenoglossum, D. pardalianches, and most of the central Asian group). Species presenting wide phyllaries are placed on the other extreme of the plot (most of the D. austriacum group, most of the SW Asian group, and D. oblongifolium). The remaining species (D. kamaonense and the D. grandiflorum group) appear on the central portion of the plot.

Doronicum plantagineum group

Characters excluded from the analyses of this group are CAL, CHB, CPL, HB, LA, and LHCM (Tab. 1). The first three PCs explain 54% of the variance (23%, 17%, and 14%, respectively). Ten PCs are needed in order to explain 90% of the overall variance. The variables with highest loadings on the 1st PC are LT, DIAM, and AME This component is related to the size of the plant and the capitula. The variables with the highest influence on the 2rid PC are CLPLT, NEN, and LL. These mostly depict plant architecture.

In the scatterplot against the first two PCs, no clear-cut clusters are apparent except maybe for the samples of D. orientale located towards the negative values for PC2 (Fig. 2A). The specimens belonging to D. carpaticum and D. columnae occur on the upper left quadrant, partially overlapping with the remaining three species (D. atlanticum, D. hungaricum, and D. plantagineum), which tend to cluster at the upper right quadrant.

In the discriminant analysis, four variables were selected using the stepwise procedure (NEN, LT, LF, LL). The first two canonical discriminant functions account for 94% of the variance, the first of them being highly (negatively) correlated with NEN and the second with LT. Classification results yielded 43 cases out of 59 (73%) assigned to the correct group when six groups are considered. When confusions within D. plantagineum (i.e., between specimens labeled as such and as "D. atlanticum") are excluded, the number of correctly-classified specimens increases to 48. Four of the l l misclassifications involve confusions between D. carpaticum and D. columnae. The remaining 7 involve confusions between D. carpaticum or D. columnae and one of the three species in the D. plantagineum group. In the scatterplot against the first two canonical discriminant functions, specimens belonging to D. columnae and D. carpaticum are largely overlapped (Fig. 2B), as are those previously recognized as D. atlanticum with respect to the remaining D. plantagineum. The core members of the D. plantagineum group are not clearly separated from D. columnae and D. carpaticum.

Doronicum plantagineum s. str. group

Characters excluded from the analyses of this group are CAL, HB, HET, LA, and LHCM (Tab. 1). The variance explained by the first three PCs is 59% (26%, 18%, and 15%, respectively). Eight PCs are needed in order to explain 90% of the overall variance. The highest loadings on the 1st PC correspond to CLPLT, NL, CHB and NEN. Therefore, the first component is related mainly with plant architecture and the shape of the basal leaves. The

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variables with the highest loadings on the 2nd PC are mainly related to the capitula appearance (ABE and LF).

In the scatterplot against the first two PCs, specimens are arranged in very loose and slightly overlapped clusters (Fig. 3A). The most distinct is formed exclusively by D. hungaricum on the lower right corner of the plot. The specimens belonging to D. orientale although rather dispersed and adjacent to the remaining two species, are confined to the space with negative values for the PC1. The samples of D. plantagineum show wide overlapping with their conspecifics labeled as "D. atlanticum".


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Fig. 3. Morphometric multivariate analysis of the Doronicum plantagineum s. str. group. A - principal components analysis: scatterplot of the specimens against the first two principal components axes. B - discriminant analysis: scatterplot of the specimens against the first two canonical discriminant functions.

The variables remaining in the discriminant analysis after the completion of the stepwise procedure were the same (NEN, LT, LF, LL) as for the group enlarged to include also D. columnae and D. carpaticum (D. plantagineum group). The first two canonical discriminant functions account for 96% of the variance, the first of them being highly correlated with NEN and the second with LT, as in the D. plantagineum group. Classification results yielded 35 cases (90%) assigned to the correct group. The four misclassifications involved confusions between "D. atlanticum'" and D. plantagineurn. Therefore, 100% of the specimens were classified into the predetermined species when our taxonomy (Tab. 1) is

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followed. The scatterplot arranges the spec imens into three loose groups corresponding to D. hungaricum, D. orientale, and D. plantagineum (Fig. 3B).

Doronicum grandiflorum group plus D. carpaticum and D. columnae

Characters excluded from the analyses of this group are CHB, CPL, LA, and LHCM (Tab. 1). The first three PCs explain only 49% of the variance (22%, 14%, and 13%, respectively). Eleven PCs are needed to explain 90% of the overall variance. The highest


loadings on the 1 st PC correspond to characters DIAM, AMP, LF, and ABE. This component is related to the size and the appearance of the capitula. Variables CALL, LL, and LT contribute the most to the 2nd PC. This component is thus related to the size and the shape of ligulate florets and to plant size.

In the scatterplot against the first two PCs, discrete clusters are lacking (Fig. 4A). However, most of the samples ofD. carpaticum and D. columnae are placed on the left half of the space defined by the first two PCs, corresponding to negative values for PC1. The species of the D. grandiflorum group overlap on the central and right areas.

Four variables remained in the discriminant analysis after the completion of the stepwise procedure (LF, CAL, ABE, LT). The first two canonical discriminant functions account for 96% of the variance, the first of them being highly correlated with LF and the second with CAL. Only 29 of the 50 specimens (58%) were correctly classified, and 20 of the 21 misclassifications involved confusions between D. carpaticum and D. columnae or between D. grandiflorum, D. clusii and D. glaciale, that is, only one crosses the borders of the two clear subgroups recognizable in this analysis (D. carpaticum + D. columnae, on one side, the D. grandiflorum group on the other). Furthermore, the second most likely group for all the 50 specimens did not cross the borders of the two subgroups either. The seatterplot against the first two canonical discriminant functions provides a good synthesis of this result, i.e. D. carpaticum and D. columnae widely overlapped and separated from the three species of the D. grandiflorum group, which are also overlapped among themselves (Fig. 4B).

Central Asian group plus D. cavillieri, D. kamaonense and D. stenoglossum

Characters excluded from the analyses of this group are CHB and CPL (Tab. 1). The first three PCs explain 58% of the variance (27, 20, and 11%, respectively). Ten PCs are needed to account for 90% of the overall variance. The highest loadings on the 1st PC are those of characters LL, LF, and NC. This component is related with flower size and number ofcapitula. CALL, CLMLB and CALBE contribute most to the 2nd PC. These variables are related with shape of ligulate florets and phyllaries.

In the scatterplot of the specimens against the first two PCs, the three added species form reasonably distinct clusters, although D. cavillieri only partially so (Fig. 5). The three members of the central Asian group (D. altaicum, D. briquetii and D. falconeri (incl. D. turkestanicum)) form a single cluster. Therefore, ordination obtained with PCA already suggests that the three doubtful species are outliers. As expected, the discriminant analysis reinforces such outcome (results not shown).

Central Asian group

Characters excluded from the analyses of this group are CHB and CPL (Tab. 1). The first three PCs explain 50% of the variance (24%, 15%, and 11%, respectively). Eleven PCs are needed in order to explain 90% of the overall variance. The highest loadings on the 1 st PC are those of characters LL, LF, and DIAM. This component is related to the size of the flowers and the capitula. Variables contributing the most to the 2nd PC are CLMLB, ABE, and NDA. This component depicts the shape of the phyllaries and ligules.

In the scatterplot against the first two PCs, no discrete clusters are shown, but the position of D. altaicum is marginal and only slightly overlapped with other species (Fig. 6A).

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PC 1 Fig. 5. Principal components analysis of the central Asian taxa in Doronicum (central Asian group + D. stenoglossum, D. kamaonense, D. cavillieri). Scatterplot of the specimens against the first two principal components axes.

Four variables remained in the discriminant analysis when the stepwise procedure was followed (LF, LHCM, CAL and CLPLT). The first two canonical discriminant functions account for 93% of the variance, the first one being correlated with LF and LHCM while the second was with CAL. Classification results yielded 31 cases out of 40 (77%) assigned to the correct group. Three of the 9 misclassifications involved confusions between "D. turkestanicum" and D.falconeri. Therefore, 85% of the specimens were classified into the predetermined species when our taxonomy (Tab. 1) is considered. The scatterplot does not provide a clear discrimination for the three species (Fig. 6B). Clear-cut clusters are not present, and the most distinct is that of D. briquetii. Specimens of D. falconeri labeled as "D. turkestanicum" are adjacent, but not much overlapped with the remaining conspecific samples. However, the structuring of the dots in the plot is consistent with the taxonomy of the group.

DISCUSSION

Global matrix

A matrix containing data from 30 taxa is not likely to yield discrete clear-cut clusters corresponding to taxonomic groups when analyzed with PCA. This is due to various reasons. First, because PCA is not a clustering technique, but one that is designed to reduce data by examining relationships among characters and producing a set ofuncorrelated new variables. Second, when 30 different biological entities are involved, it is expected that a significant number of components of variation may occur in the data. Only a few of these components will be represented in the scatterplots generated from the first PC axes. Characters that contribute poorly to the main components of variation will thus be under-represented in the first PC axes. Groups that are diagnosed on the basis of such characters will not be discriminated by scatterplots of the first PCs. This situation applies particularly when the first three PC axes


O.

-2 -3

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r~

o

o 0

o

o

o o

oo o 0

0 0

+

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+

O

.,~.+ +

-2 -1 0 1

-IF

-I$

+

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PC 1

A

o D. a l fa icum

<> D. briquet#

D. falconeri

+ D. falconer/ " D . turkesfanicura"

(N LI. 0

3

2

1

0

-1

-2

-3

-4 -4

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0

O o o

0 0 oo o

4- 0

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~4- o

B

o D. alfalcum

0 D. br/que#i

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+ D. falconer/ "D. turkestanicum"

6 -2 0 2 4

DF 1 Fig. 6. Morphometric multivariate analysis of the central Asian group in Doronicum. A - principal components analysis: scatterplot of the specimens against the first two principal components axes. B - discriminant analysis: scatterplot of the specimens against the first two canonical diseriminant functions.

together only explain half o f the overall variance. Additionally, the characters that contribute the most to the first PC axes may be highly variable within species, a situation that also produces a lack o f clear-cut clusters. Part o f the difficulties in obtaining clear-cut groups, and thus in testing the taxonomic usefulness of morphometric characters, can be overcome by reducing the number o f taxa to be analyzed. As we remove species from the analysis, the number o f components o f the overall variance decreases, and thus the taxonomically-relevant characters, even if only for a given pair of species, are likely to move their influence onto the first PC axes. For this reason, reduced groups within Doronicum have been analyzed.

436 I. Alvarez Ferndndez & G. Nieto Feliner

However, the fact that the first PC axis is primarily influenced by characters related to the shape of the phyllaries - and not by, e.g., the size of vegetative organs - is in principle a positive symptom about the ordination produced. Phyllaries are taxonomically-important characters not only in Doronicum but also in other Asteraceae (BREMER 1994). Therefore, the trends detected in the scatterplot of the 291 specimens against the first two PCs are not taxonomically meaningless. The fact that the samples belonging to the same group (e.g. D. plantagineum group, central Asian group) and species (e.g.D. stenoglossum) are plotted on the same area of the multivariate space indicates that the taxonomic treatment is not completely arbitrary. Not much more about taxonomic ordination, however, can be said on the basis of such a large and complex data set.

Placement of species with uncertain affinities

The first purpose in analyzing the three reduced groups in this study was to examine the position of several species of uncertain affinities with respect to the core members of well-defined groups. In particular, we anticipated some clarification about the relative affinities of D. columnae and D. carpaticum by analyzing them together with the core representatives of the two putative closest groups (D. plantagineum and D. grandiflorum groups). Under the same expectations, we examined the relative position of the Asian species D. caviltieri, D. kamaonense and D. stenoglossum, but compared them to a single group (central Asian) because no other putative close groups were envisaged.

The scatterplot of the specimens of the D. plantagineum group against the first two PCs does not provide a conclusive discrimination for D. columnae and D. carpaticum (Fig. 2A). Although confined in the upper left quadrant of the plot, the specimens from both species appear intermingled with the rest. According to this result, these specimens cannot be considered clear outliers with respect to the D. plantagineum s. str. group. It should be noted that PCA is characterized by faithful representation of the distances between major groups of clusters while distances between close neighbors may be distorted (SNEATH & SOKAL 1973). This is one of the reasons why ordination should be further checked with DA. Quite surprisingly, the scatterplot of the first two canonical discriminant functions also fails to separate D. columnae and D. carpaticum from the D. plantagineum s. str. group (Fig. 2B). This is consistent with the fact that 7 of the 11 misclassified specimens involved confusions between the D. plantagineum s. str. group and D. columnae or D. carpaticum. If patterns of variation within the D. plantagineum s. str. group were markedly different to those in the two species, we would expect the scatterplots against the first two canonical discriminant functions to show a clear gap between the two species and the remaining members of the group. The fact that no such gap is found, even though DA is a powerful technique for discrimination, suggests that the two species are actually part or close relatives of the D. plantagineum s. str. group. A comparison, therefore, is needed with their behavior in an analysis of the other closest putative group.

The inclusion of D. columnae and D. carpaticum in the analyses of the D. grandiflorum group does not provide a conclusive answer to their taxonomic placement either. However, when the results of this analysis are compared to those of the D. plantagineum group, morphometric data suggest a closer affinity to the latter. Despite the fact that the PCA of the D. grandiflorum group plus D. columnae and D. carpaticum does not reveal discrete clusters (Fig. 4A), the two species (D. columnae in particular) are confined to the area with negative


values for the PC1. Unlike the situation in the other group, the discriminant analysis does effectively separate these two species from D. grandiflorum, D. glaciale and D. elusii (Fig. 4B). The percentage of misclassified specimens (42%) is higher than in the analysis of the D. plantagineum group. However, most confusions (20 out of 21) are due to problems of identification either within the D. grandiflorum group (D. grandiflorum, D. glaciale and D. clusii ) or within the D. columnae + D. carpaticum pair. The fact that the second most likely classification for all the 50 specimens also remained within the same limits is a strong argument for considering that D. eolumnae and D. carpaticurn are outliers to the D. grandiflorum group.

The closest relationship of the two species with the D. plantagineum s. str. group is consistent with molecular data from the ITS sequences, but further evidence is needed since the available sequence of a chloroplast marker (trnL-trnF) for D. coluranae suggests otherwise (~kLVAREZ FERNANDEZ et al. 2001). On morphological grounds, oneof the synapomorphies of the D. plantagineum s. str. group (the ciliate phyllaries) does not consistently appear in all populations of D. columnae and D. carpaticum.

Besides the close relationships between D. columnae plus D. carpaticum and the D. plantagineum s. str. group, a conclusion can be drawn from the analyses of the latter group. This is the taxonomic affinities between D. columnae and D. carpaticum suggested on morphological grounds (fikLVAREZ FERNANDEZ 1999). These two species largely or completely overlap in the plots (Figs. 2A, 2B) and 4 of the l l misclassifications involve confusion between them. Further, the second most-likely classification for 17 of the 20 specimens of both species was the other one (i.e., D. carpaticum instead olD. columnae or vice versa).

The same strategy of adding taxonomically-uncertain species to reasonably-delimited groups also gave positive results in the case of the three Asian species. The fact that the specimens of D. kamaonense, D. stenoglossum and D. cavillieri form discrete clusters with respect to the core of the central Asian group in the PCA scatterplot suggests that the three species are outliers (Fig. 5). The situation ofD. cavillieri is less clear than the situation of the other two in this respect. The single cluster in which D. altaicum, D. briquetii and D. falconeri are integrated indicates that the group they form (central Asian) has morphometric support. Thus, the ordination obtained with the PCA is consistent with the taxonomic arrangement mainly based on qualitative morphological characters and on molecular data (,/kLVAREZ FERNANDEZ 1999).

Species delimitation within cohesive groups

The second purpose in analyzing reduced groups was to explore whether morphometric characters could discriminate species forming seemingly natural assemblages within Doronicum. Multivariate methods have been recently applied to similar problems in other groups of Senecioneae (HODALOVA & MARHOLD 1996, 1998). As a whole, in our study the results depended on the kind of analysis and to a lesser degree on the group. The PCA of the D. plantagineum s. str. group does not produce clear-cut clusters, but the marginal position of D. hungaricum and D. orientale with respect to D. plantagineum is apparent, as is the coincident behavior of specimens labeled "D. atlanticum" as compared to the remaining D. plantagineum (Fig. 3A). The discriminant analysis emphasizes the same pattern (Fig. 3B). Two conclusions follow. First, there is no clear morphometric support for the taxonomic recognition of N African populations of D. plantagineum ("D. atlanticum") against the

438 I. Alvarez Fern6ndez & G. Nieto Feliner

European populations. Still, the possibility of introgression from D. orientale in N African populations of D. plantagineum exists (~,LVAREZ FEW'qANDEZ et al. 2001). The fact that the second most likely classification for two of the ten N African specimens ofD. plantagineum is D. orientale leaves some room for this hypothesis.

Unlike the morphometric analysis of the D. plantagineura s. str. group, the results of the central Asian group (D. altaicum, D. briquetii, D. falconeri) do not provide a consistent discrimination among species. However, even in the absence of clear-cut clusters, the arrangement of the dots in the scatterplot from the DA seems to be a reflection of the taxonomy (Fig. 6A, 6B). The placement of D. falconeri with respect to those conspecific populations previously known as D. turkestanicura (adjacent but only partially overlapped) is consistent, but not conclusive in supporting our taxonomic criterion that they are synonyms. The characters supporting a non-overlapping position of both species are primarily the length of the disk and of the ray florets. We had recorded such partial differences, but consider that their taxonomic relevance is at most doubtful. Inclusion of the binary variable HET improves discrimination between the heterocarpic D. falconeri (incl. D. turkestanicum) and the homocarpic D. altaicum and D. briquetii. Multivariate techniques are quite robust to small departures from normality (MARCUS 1990, LEGENDRE & LEGENDRE 1998). Therefore, including one or two qualitative characters in the framework of an exploratory analysis is acceptable. However, since we are primarily interested in the potential utility of continuous variables, it is preferable not to alter the selection conducted by the stepwise procedure in the DA, which excluded HET from the analysis.

Between-species discrimination based on morphometric characters in the third group considered in this study (D. grandiflorum) was rather poor (results not shown). Neither PCA nor DAproduced discrete clusters corresponding to D. grandiflorum, D. glaciale and D. clusii. This outcome is consistent with our taxonomic study based on morphological characters, under a univariate approach, which failed to detect any differences between these species other than the type of indumentum (CAVILLIER 1911, ALVA~Z FERNANDEZ 1999). The inability of morphometric techniques to separate the species of the D. grandiflorum group is also consistent with molecular data, and not attributable to the properties of the multivariate analyses.

In conclusion, the application of morphometric multivariate techniques to taxonomy in Doronicum gave moderately satisfactory results. These are restricted to low taxonomic levels and to groups containing a reasonably low number of species. On the whole, morphometric characters are a complementary source of information in groups where qualitative (in particular, shared) characters are scarce, as is the case in Doronicum. This does not imply their use in phylogenetic analysis, which poses serious problems (CrtAPeILI. 1989, STEVENS 1991, RAE 1998, THIELE 1993). However, in the context of an exploratory analysis (MARCUS 1990) they can be very useful, by facilitating a scrutiny of patterns of variation and co-variation of characters, and thus helping to make decisions regarding species recognition, taxonomic status, as well as group limits or composition.

Acknowledgements: The authors are grateful to Javier Fuertes, Karol Marhold, Pablo Vargas as well as to two anonymous reviewers for their constructive comments, which have substantially improved the manuscript, and to Laura Barrios for assistance with statistical matters. This work has been supported by grant DGES PB96-0849 of the Spanish Direccirn General de Enseflanza Superior e Investigaci6n Cientifica.


REFERENCES

ABBOTT L.A., BISBY F.A., & ROGERS D.J. (1985): Taxonomic analysis in biology. Computers, models and databases. Columbia University Press, New York.

ALVAREZ FERN/kNDEZ I. (1999): Sistem&tica del g~nero Doronieum L. (Asteraceae, Senecioneae)y anddisis filogen~tico basado en fuentes morfol6gicas y moleculares. Unpubl. Phi) dissertation. Universidad Complutense, Madrid.

/kLVAREZ FERNANDEZ I. (2001): Four new combinations in Eurasian Doronicum L. (Asteraceae, Senecioneae). Novon 11: 294-295.

• h, LVAREZ FERNANDEZ I. & NIETO FELINER G. (1999): Leetotypification of 16 species names in Doronicum (Asteraceae, Senecioneae). Taxon 48: 801-806.

• h, LVAREZ FERNANDEZ I. & NIETO FELINER G. (2000): A new species ofDoronicum (Asteraceae, Senecioneae) from central China. Ann. Bot. Fenn. 37: 249-253.

ALVAREZ FERNANDEZ I., FUERTES AGUILAR J., PANERO J.L. & NIETO FELINER G. (2001): A phylogenetic analysis of Doronicum (Asteraceae, Senecioneae) based on morphological, nuclear ribosomal (ITS) and chloroplast (trnL-F) evidence. Molec. Phy•. EvoL 20: 41---64.

AVETISJAN V.E. (1980): Rod Doronicum L. v Armenii (The genus Doronicum in Armenia). Biol. Zhurn. Armenii 33: 532-534.

BREMER K. (1994): Asteraceae. Cladistics & classification. Timber Press, Portland, Oregon. CAVILLIER F. (1907): l~tude sur les Doronicum a fruits homomorphes. Annuaire Conserv. ,lard. Bot. Gen~ve

10: 177-251. CAVILLIER F. (1911): Nouvelles 6tudes sur le genre Doronicum. Annuaire Conserv. Jard. Bot. Gendve 13-14:

195-368. CHACON R. (1987): Contribuci6n al estudio taxon6mico del g6nero Doronicum L. (Compositae) en la

Peninsula Ib6rica. Anates =lard. Bot. Madrid 43: 253-270. CHAPPILL J.A. (1989): Quantitative characters in phylogenetic analysis. Cladistics 5: 217-234. DE PINNA M.C.C. (1991): Concepts and tests of homology in the cladistic paradigm. Cladistics 7: 367-394. DUVIGNEAUD J. (1992): Le genre Doronicum L. en Belgique et dans les r6gions voisines. Nat. Mosana 45:

81-92. EDMONDSON J.R. (1973): Notes on Doronicum L. in SW Asia. Notes Roy. Bot. Gard. Edinburgh 32: 255-258. EDMONDSON J.R. (1975): Doronicum L. In: DAVIS P. H. (ed.), Flora of Turkey and the East Aegean Islands 5,

Edinburgh University Press, Edinburgh, pp. 137-145. EDMONDSON J.R. (1978): The genus Doronicum L. in Iran. Notes Roy. Bot. Gard. Edinburgh 37: 67-73. HOD/kLOV./~ I. & MARHOLD K. (1996): Sympatric populations ofSenecio ovatus subsp, ovatas, S. germanicus

subsp, germanicus (Compositae) and their hybrid in the Carpatians and the adjacent part of Pannonia. Flora 191: 283-290.

HOD/kLOVA I. & MAR/-IOLD K. (1998): Morphometrie comparison ofSenecio germanicus and S. nemorensis (Compositae) with a new species from Romania. Bot. J. Linn. Soc. 128: 277-290.

HOLLIS S. & BRUMMITT R.K. (1992): World geographical scheme for recording plant distributions. Carnegie Mellon University, Pittsburgh.

KLECKA W.R. (1980). Discriminant analysis. Quantitative applications in the social sciences. Sage University paper no. 19. Newbury Park.

LEGENDRE P. & LEGENDRE L. (1998): Numerical ecology. Elsevier, Amsterdam. MARCUS L.F. (1990): Traditional morphometrics. In: ROHLF F.J. & BOOKSTEIN F.L. (eds.), Proceedings of the

Michigan Morphometrics Workshop, University of Michigan, Ann Arbor, pp. 77-122. PATTERSON C. (1988): Homology in classical and molecular biology. Molec. Biol. Evol. 5: 603-625. PI~REZ MORALES C. & PENAS A. (1990): Sobre algunos Doronicum ib6ricos. Lagascalia 15: 151-160. PI~REZ MORALES C., PENAS A., LLAMAS F. & ACEDO C. (1994): Doronicumpubescens sp. nov. Lazaroa 14:

5-12. RAE T.C. (1998): The logical basis for the use of continuous characters in phylogenetic systematics. Cladistics

14: 221-228. SNEATH P.H.A. & SOKAL R.R. (1973): Numerical taxonomy. The principles and practice of numerical

classificaaon. W. H. Freeman & Co., San Francisco.


STEVENS P.F. (1991): Character states, morphological variation and phylogenetic analysis: a review. Syst. Bot. 16: 553-583.

THIELE K. (1993): The Holy grail of the perfect character: the cladistic treatment of morphometric data. Cladistics 9: 275-304.

WIDDER F.J. (1925): Eine neue Pflanze der Ostalpen - Doronicum (Subsectio Macrophylla) cataractarum - und ihre Verwandten. Repert. Spec. Nov. Regni Veg. 22:113-184.

Received 10 July 2000, revision received and accepted 16 March 2001 Encl. Appendix pp. 441 4~4


A P P E N D I X 1

List of the specimens used in the multivariate study. Main geographic subdivisions follow HOLLIS & BRUMMITT (1992).

Doronicum cavillieri ALVAREZ FERNANDEZ @t NIETO FEL. China (south-central): T'ao river basin (VI. 1925 J.F. ROCK BM); ibidem (VI. 1925 J.F. ROCK 12389 NY);

ibidem (VI. 1925 J.F. ROCK E); ibidem (VI.1925 J.F. ROCK 12192 NY); Tebbu country (1925 J.F. ROCK GH); ibidem (1925 J.F. ROCK K); ibidem (1925 J.F. ROCK NY); ibidem (1925 J.F. ROCK W).

Ooronicum kamaonense (DC.) ALVAREZ FERNANDEZ Nepal: Balangra pass (28.VII. 1952 O. POLUNIN et al. E); Ghurchi Lagua (22.VI. 1952 O. POLUNIN et al, (3).

Jammu-Kashmir: Gurais (6.IX.1892 J.F. DUTHtE BM); Khelanmarg (2.VIII.1956 O. POLUNIN B); ibidem (2.VIII.1956 O. POLUNIN BM); Rampur (5.VII.1940 F. LUDLOW & G. SHERRIFF BM); Sinthan pass (10.VIII.1943 F. LUDLOW & G. SHERRIFF E). Pakistan: Hazara district (VI.1955 G.L. WEBSTER 86 J. SACK G).

Ooronicum oblongifolium DC. Turkey: Artvin, Valniz~am Silsilesi (10.VII.1967 ALBURY et al. K); Coruh, Kordevan da~ (28.VI.1957

PH. DAVIS & I.G. HEDGE K); ibidem (28.VI.1957 P.H. DAVIS & I.G. HEDGE W). North Caucasus: Daghestan (without date G.F.R. RADDE (3); Zagedan (7.VII.1989 A. DOLMATOVA et al. LE). Transeaueasus: Aragac (l 3 .VII. 1975 E. GABRIELIAN E); ibidem (13.VII. 1975 E. GABRIELIAN G; 4.VI. 1988 K. TAMANJAN G); Armenia (without collector and date E); Carthalinia (VI. 1881 A.H. BROTHERUS & V.F. BROTHERUS 501 G).

Doronicum pardalianches L. Belgium: Esneux (28.V. 1976 J. DUVIGNEAUD B). France: Loire (20.V.1894 H.E. JEANPERT B); Macou

(without date and collector B). Germany: Brimav (21 .V. 1894 FERGES B); Mark Branderburg (19.V. 1974 H. SHOLZ B). Great Britain: Stainton-in-Peak (18.VI.1862 J. BALL MA 171821). Italy: San Valmazzo (14.V. 1934 BASCHANT B). Spain: Barcelona, Valvidriera (without date and collector MA 129791); Huesca, sierra de Guara (24.VI. 1995 I. ALVAREZ et al. MA); L6rida, sierra del Cadi (29.VI. 1996 I. ALVAREZ et al. MA).

Doronicum stenoglossum MAXIM. China (north-central): Ganzu, T'ao river basin (VII. 1925 J.F. ROCK GH); Tebbu country (VIII. 1926 J.F.

ROCK E); ibidem (VIII. 1926 J.F. ROCK NY). China (Tibet-Qinghai): Dari-Xian (14.VIII. 1993 T.N. HO et al. MO); Reting (28.VII.1942 F. LUDLOW & G. SHERRIFF BM). China (south-central): Lichiang range (VIII. 1906 G. FOP, REST BM); Likiang, Tungshan (V. 1923 J.F. ROCK GH); west Yunnan (1930 G. FORREST E); Yunnan (without date T.T. YO GH); Yunnan, Mull (21.X.1937 T.T. YO E).

Central Asian group

Doronicum altaicum PALL. Russia (Siberia): Altay (VII. 1947 A. KUMINOVA & PAVLOVA GH); ibidem (without date PALLAS BM);

ibidem (14.VIII.1923 V. SAPOSHNIKOV W); ibidem (without date and collector E); Sayan (1.VII.1964 E. PENKOVSKAYA & I. KRASNOBOROV LE); Tomsk, Kara-Kokshi (21.VII.1915 P.N. KRYLOV LE); Tuva (12.VII.1970 I. KRASNOBOROV LE); ibidem (2.VIII.1975 M. LOMONOSOVA & D. SHAULO LE); ibidem (3 .VII. 1971 S.N. VIDRINA LE); Siberia (30.VII. 1902 V.I. KOMAROV LE),

Ooronicum briquetit CAVILL. China (south-central): Dokerla (VIII.1935 C.W. WANG GH); Hung-po 0/I.1932 J.F. ROCK GH); ibidem

(VI. 1932 J.F. ROCK K); Konka mount (VIII. 1928 J.F. ROCK GH); Mekong on Kaakerpo (1932 J.F. ROCK GH); Peimashan (VII,1923 J.F. ROCK K); Wuli-la (1932 J.F. ROCK GH). China (Tibet-Qinghai): Kongbo (21.VI.1947 F. LUDLOW et al. BM); ibidem (21.VI.1947 F. LUDLOW et al. (3); ibidem (16.VIII.1947 F. LUDLOW et al. BM).

Doronicum falconeri C.B. CLARKE India: Dhaola Dhar (7.VII.1939 G. SHERRIFF BM); ibidem (7.V11.1939 G. SHERRIFF 7405, sheet 2 BM);

Rotang (ll.VIl.1941 N.L. BORE); Shonthar Gall (2.VIII.1935 F. LUDLOW & G. SHERRIFF E). Jammu-Kashmir: Baltistan (22.VIII. 1936 W. KOELZ GH); ibidem (3.VIII. 1940 R.R. STEWART NY); Bringhi


(23.VIII. 1943 F. LUDLOW & G. SHERRIFF BM); Gittidas (15.VII. i 953 F. SCHM1D G); Haramukh (29.VII. 1940 F. LUDLOW & G. SHERRIFF BM); Kero Lugma glacier (24.VII. 1939 RUSSELL BM). China (Xinjiang): Bagadushung (14.VI.1879 A. REGEL E*); Turkestania, Bogdo-ola (VIII.1908 G. MERZBACHER W*). Kazakhstan: Alatau, Lepsa (1841 G.S. KARELIN & I.P. KIRILOV BM*); Katon-Karagaj (10.VIII.1930 P.A. SMIRNOV W*); Ketmenj (1.VII.1964 I. ROLDUGIN G*); ibidem (1.VII.1964 I. ROLDUGIN MA 358613*). Kirgizistan: Kungei Alatau (17.VII. 1896 V.F. BROTHERUS G*); Terskei Alatau (without date and collector GH*); Vemenskij (without date N.D. SOKALSKU E*). Russia (Siberia): Altay, Kurkure (21 .VII.1977 MARIN LE*).

* Labeled as 1). turkestanicum CAVILL. in the analyses (see Material and methods)

Doronicum austr iacum group

Doronicurn austriacurn JACQ. Andorra: Pifineos (VII.1949 LOSA & MONTSERRAT MA 155589). Austria: Styfia inferior (1842 R.C.

ALEXANDER E). Bulgaria: Sitovo (VIII.1896 V. STi~iBRNY B). Italy: monte Nevon (30.VI.1935 V. ENGELHARDT B). France: Cerdagne Carlit (3 I.VII. 1916 F. SENNEN MA 129784). Germany: Heilbrunn (VI.1890 J. JAh"N B). Greece: Kastoria (1965 O. POLLrNIN E); Kerkini, mt. Belles (4.VII.1981 A. STRIDet al. B); Pella, Kaimaktsalan (13.VIII. 1977 W. GREUTER G). Yugoslavia: Bitola (22.VII. 1970 J.R. EDMONDSON E).

Doronicum carpetanum BOISS. et REUT. ex WILLK. subsp, carpetanum Spain:/kvila, Hoyos de1 Espino (28.VI. 1987 GOMEZ MANZANEQUE et al. MA); B urgos, pico Mancillas

(14.VI1.1984 BENEDI[ et al. MA 334336); Logrofio, Canalcs de la Sierra (6.VII.1995 I. ALVAREZ et al. MA); Ezcaray (20.VII.1991. PI~REZ et al. MA 560311); Lugo, Trcs Obispos (27.VII.1986 G. NIETO FELINER & J. PEDROL MA 502993); Madrid, Peitalara (VIII.1911 F. BELTRAN MA 129727); ibidem (7.VII.1986 M. LUCEIqO & P. VARGAS MA 448157); Salamanca sierra de Candelario (21.VI.1986 LANSAC & G. NIETO FELINER MA 421399); Sofia, Santa In6s (27.VII. 1972 A. SEGURA ZUBIZARRETA MA 360095).

Doronicum carpetanum subsp, dlazii (PI~.REZ MORALES et PENAS) ALVAREZ FERNANDEZ Spain: Le6n, Abeigas (27.VII.1988 E. PUENTE & C. PI~REZ MORALES MA 489306); ibidem (27.VII. 1988

E. PUENTE & C. PI~REZ MORALES MA 502699); Le6n, Cumin (without date and collector E); Soria, sierra de Urbi6n (I0.VII.1935 L. CEBALLOS & C. VICIOSO MA 129726); ibidem (10.VII.1935 L. CEBALLOS & C. VICIOSO MA 129739); ibidem (11.VI.1978 P. HARROLD & R.J.D. MCBEATH E); ibidem (VII.1958 N.Y. SAhTDWITrt K); ibidem (VII. 1958 N.Y. SANDWlTH, sheet 2 K).

Doronicum carpetanum subsp, kuepferi (R. CHACON) ALVAREZ FERNANDEZ Spain:/kvila, sierra de Gredos (without date and collector MA 445748); C~ceres, sierra de Majarreina

(7.VIII. 1946 S. RIVAS GODAY MA 155587); ibidem (7.VIII. 1946 S. RIVAS GODAY MA 348763); Salamanca, Calvitero (26.VII. 1989 S. RIVAS MARTgNEZ et al. MA 538496); ibidem (30.VI. 1985 R. VOGT B); Salamanca, Candetario, (18.VII. 1980 E. VALDI~S BERME/O et al. MA 445798); ibidem (18.VII. 1980 E. VALDI~S BERME.IO et al. MA 445798a); ibidem (18.VII.1980 E. VALDI~S BERMEJO et al. MA 445798b); Salamanca, E1 Trampal (10.VII. 1989 G. NIETO FELINER et al. MA 472502); Salamanca, sierra de B6jar (28.VII. 1982 F. NAVARRO & C.J. VALLE MA 286039).

Doronlcum carpetanum subsp, pubescens (P(=REZ MORALES, PENAS, LLAMAS et ACEDO) AIZPURU

Portugal: serra da Estrela (19.VII.1997. I. ,~LVAREZ et al. MA). Spain: Asturias, lagos de Saliencia (26.VII. 1988 M. LUCEI~O & P. VARGAS MA 449967); Cantabria, monte Cmlatrapa, (2.VII. 1983 C. AEDO MA); Cantabfia (27.V.1990 S. PATINO et al. MA 537973); Coruila, Caaveiro, (25.IV.1981 F. AMICH et al. MA 311144); Corur'fa, Puente Carreiras (29.V. 1953 F. BELLOT MA 504155); Le6n, circo CeboUero (16.VII. 1974 B. CASASECA & F.J. FERNANDEZ DIEZ MA 197037); ibidem (16.VII.1974 B. CASASECA & F.J. FERNANDEZ DIEZ MA 345401); Le6n, Posada de Valde6n, (14.VII. 1978 A. CHAP, PIN B); Palencia, pto. de Piedrasluengas (14.VII. 1995 I./kLVAREZ et al. 919/4 MA); ibidem (14.VII. 1995 I./~LVAREZ et al. 919/10 MA).

Ooronicum cataractarum WIDDER Austria: Koralpe (3.VIII. 1935 B. FEST B); ibidem (without date LEMPERG K); ibidem (16.VII. 1947 F.J.

WIDDER GZU); ibidem (9.IX.1925 F.J. WIDDER B); ibidem (I.VIII.1926 F.J. WIDDER K); Seebach (28.VIII. 1939. F.J. WIDDER B); ibidem (24.VIII. 1936 F.J. WlDDER G).


Doronicum corsicum (LOISEL.) POIR. France (Corsica): A'itone (1885 E, REVERCHON B); ibidem (1885 E. REVERCHON E); ibidem (1885 E.

REVERCHON, sheet 2 E); ibidem (1885 E. REVERCHON NY); Fiumorbo (28.VII. 1849 KRALIK E); Guagno (VIII. 1847 REQUIEZ B); Incudine (2,VIII. 1986 J. LAMBINON MA 367224); Mangamullo (18.VII. 1906 H. GYSPERGER B); Tavignano (13.VIII.1849 KRALIK E); Vizzabona (without date and collector B).

D. grandiflorum group

Doronicum clusii (ALL.) TAUSCH Austria: Ferwallgruppe (2.VIII.1942 W. FREIBERG MA 358526); Stilfser Joch (10.VIII.191I M.

SCHREIBER MA 129736). Slovalda: Vysok6 Tatry (6.VIII. 1933 J. DOSTAL MA 411781); ibidem (24.VII. 1893 L. GIRAUDIAS MA 129718); ibidem (26.VI.1915 J.B. KOMMERLE & G. TIMK0 B); ibidem (8.VII.1918 K. RONNIGER G). Italy: Bormio, passo deUo Stelvio (4.VIII. 1997 I. ,~LVAREZ et al. MA); Trento, coU de1 Cuc (3.VIII.1997 I./~LVAREZ et al. MA). Poland: Zakopane (20.VI.1961 W. HEMPEL B). Switzerland: Bemina (18.VII.1990 S. CASTROVlEJO et al. MA 484983).

Doronicurn glaciale (WULFEN) NYMAN Austria: Hochechwabgebirge (5.VIII. 1887 H. STEININGER B); Hochplateau (VI. 1906 J. BORNMOLLER B);

Maushen (18.VII.1907 V. ENGELHARDT B); Menspitze (19.VII.1988 F. JACQUEMOUD G); Raxalpe (27 .VI. 1906 J. BORNMULLER B); Weissenbach (30.VI. 1900 G. TREFFER G); ibidem (30,VI. 1900 G. TREFFER, sheet 2 (3). Germany: Hasenthal Prettan (VIII. 1882 G. TREFFER B). Italy: Trento, coil Rodella (2.VIII. 1997 I. ALVAREZ et al. MA); Gr6den (18.VII. 1907 J. BORNMOLLER B).

Doronicum grandiflorum LAM. Austria: Bondone (8.VII.1899 S. CIMAROLLI G). France: Ari6ge (27.VII.1982 A. CHARPIN & M.

DITTRICH G); Galibier (3.VIII. 1858 C. OZANON G); Pyr6n6es, vall6e d'Ossoue (2.VIIL 1996 A. HERRERO MA). Italy: Belluno ( t 1 .VII. 1970 F. DUSA & G. MORTIN MA 360098); Cuneo (4.VIII. 1982 A. CHARPrN & R. SALAMAN G); Tuscone di Lumpelli (5.VIII. 1912 P. CHENEVARD G). Spain: Huesca, pico Aneto (26.VII. 1997 T. ALMARAZ MA); Le6n, Pefia Prieta (14.VII. 1990 M.L. GIL Z0"~IIGA & J.A. ALEJANDRE MA 532701); Le6n, Pefia Ubifia (8.VII. 1990 C. AEDO MA).

D. plantagineum group

Doronicum carpaticum (GRISEB. et SCHENK) NYMAN Hungary: without locality (VII.1917 ADAMOVIC K). Romania: Bulea (23.VII.1914 J. TUZSON B);

Ceahlau (11.VII.1922 I. GRINTESCU G); Transsilvania (without date SCHLrR B); Vertop (20.IV.1873 L.P. StMKOVICS B); Vertop et Vertopul (without date L. SIMO~,AI B); ibidem (without date L. SIMONKA[ G); Zemest (30.VI.1900 WALZ MA 129725). Ukraine: Bliznitza mts. (28.VI.1950 K.N. IGOSHrNA MO); Chomogora (16.VII.1938 G. KOZIJ MO).

Doronicum colurnnae TEN. Austria: Praxersee (without date A. KERNER E). Bulgaria: Salzica (16.VI.1974 M. MARKOVA et al. E).

Greece: Parnassi (24.VII.1854 T.G, ORPHANIDES G); Sterea Ellas (27.VI.1982 I. HAGEMANN et al. B); Thessalischer Olymp. (VII. 1960 F. SORGER B); Trapeza (11.VI. 1937 E.K. BALLS & W.B. GOURLAY E). Italy: Comiolo (28.V.1976 F.L. CHIAPELLA & L. POLDINI MA) 358528; Gr6den (18.VII.1907 L BORNMULLER B). Romania: Schuler (30.V. 1886 ROMER B). Yugoslavia: Frebovic (V. 1888 BECK & FIALA G).

Doronicum hungaricum RCHB. f. Austria: Wolfstha] (10.V.1889 W. STEINITZ E). Hungary: Budam (without date J. KOVATS E);

Farkasv61gy (without date FILARSZKY & SCHILBERSZKY BM); ibidem (FILARSZKY & SCHILBERSZKY E); Solymos (11.V.1867 M. VRABI~LYI E). Romania: Ia~i (7.V1.1967 M. TOMA BM); Vlasca (V.1934 P. CRETZOIU B); Yugoslavia: Kis (without date JLII~ B); Tapeider (1888 J. BORNMOLLER B).

Doronicum orientale HOFFM. Austria: Thcresienthale (l 0.VI. 1895 E. KHEK B), Greece: Mitilinis (27.IV. 1987 A. STRID et al. G). Italy:

Potenza (12.V.1933 O. GAVIOLI MA 129723); Vico del Gargano (16.IV.1976 M. NYDEGGER G). Turkey: Antalya (28.IV. 1959 E. HENNIPMAN et al. B); Byzantii in silva Belgradensi (7.V. 1899 J. BORNMULLER B);

444 I. Alvarez Fernfindez & G. Nieto Feliner

Istanbul, Baheek6y (8.1V.1959 E. HENNIPMAN et al. B); Z6k6ri6keny (12.IV.1904 G.V. AZNAVOUR B). Yugoslavia: Hudowa (10.1V.1918 J. BORNMOLLER B); Rakovica (IV.1887 S. PETROVIC G).

Doronicum plantagineum L. France: Haute-Marne, Louze (6.V. 1991 B. RETZ MA 532469); Verfi~ res-le-Buisson (12.V. 1973 B. RETZ

MA 358614). Great Britain: Great Saling, Essex (20.V. 1886 H.E. FOX B). Portugal: Estremadura, Malveira (29.IV.1976 B. RAINHA MA 129715). Spain: C~idiz, Grazalema (21.IV.1982 G. LOPEZ & R. MORALES MA 443484); Cuenca, Talayuelas (V. 1980 G. MATEO MA 442886); Le6n, Soto de Sajambre (4.V. 1967 H. SCHOLZ & P. HIEPKO B); Madrid, sierra de Guadarrama (V.1912 C. VIcIOSO MA 129704); Salamanca, Linares de Riofrio (17.V.1968 B. CASASECA MA 191689); Zaragoza, sierra de Vicort (22.IV.1908 C. VIC[OSO MA 129694). Algeria: Kabylie (VI.1898 E. REVERCHON BM**); ibidem (VI.1898 E. REVERCHON G**); Tala Guilef (19.VI.1975 P.H. DAVIS BM**); ibidem (19.VI.1975 P.H. DAVIS E**); Teniet El Had (2.V.1937 A.H.G. ALSTON & N.D. SIMPSON BM**); ibidem (31.V.1975 P.H. DAVIS E**). Morocco: Ai'n Leuh (21.V. 1924 E, JAHANDIEZ MA 129782**); Dai" et Achlet (4.VI. 1923 E. JAHANDIEZ BM**); Ifrane (6.VI. 1971 G. BOCQUET BM**).

** Labeled as D. atlanticum CHABERT in the analyses (see Material and methods).

South-west Asian group

Doronicum cacallifolium Bolss. et HELDR. Turkey: Isauria, Geyik da[~ (28.VIII. 1947 P.H. DAVIS E); ibidem (28.VIII. 1947 P.H. DAVIS G); ibidem

(28.VIII. 1947 P.H. DAVIS, sheet 1 K); ibidem (28.VIII. 1947 P.H. DAVIS, sheet 2 K); ibidem (28.VIII. 1947 P.H. DAVIS W).

Doronicum doiichotrichum CAVILL. Iran: Dimelo (4.VI. 1956 F. SCHMID G); Maku (22.VI. 1974 SIAMI W); Azerbaydzhan, Kalibar (26.V1.1978

TERMEH et al. G). Iraq: Helgord range (3.IX.1957 A. RAWI & I. SEKHANG g). Turkey: Artvin, Yalniz~am-Gebirge (4.VIII. 1981 RAUS B); (~oruh, Kordevan daj~ (29.VI. 1957 P.H. DAVIS & I.G. HEDGE E); Kars, Yat~nurlu da~ (13.VI.1957 P.H. DAVIS & I.G. HEDGE BM). Transeaucasus: Ossetia, Didi Liachva (22.VII.1881 A,H. BROTHERUS & V.F. BROTHERUS K); Ermani (4.VI.1956 F. SCHMID W).

Doronicum hauasknechtii CAVILL. Turkey: Berytdagg (5.VIII.1865 H.K. HAUSSKNECHT W); Giresun, Karag61 (5.VIII.1965 C. TOBEY E);

Kayseri, lsikda~i (21 .VII. 1992 H. DUMAN & Z. AYTAR GAZI); Maras, Binbo~a da~ (15.VII. 1952 P.H. DAVIS et al. E).

Doronicum macrophyllum FISCH. subsp, macrophyllum Irun: Qareh Da~a (20.VII.1971 J. LAMOND & F. TERMEH K). North Caucasus: Checheno-Ingushiya

(9.VII. 1988 D. GELTMAN ¢t al. LE); Kavardino-Balkariya (I 8.VII. 1990 MENITSKIJ et al. LE). Transcaucasus: Abkhaziya (20.VIII. 1990 DOLMATOVA et al. LE); Armeniya (without date and collector W); Azerbaydzhan, Kuba (8.VII.1935 KAILIAGIN NY); Georgia (1838 R.F. HOHENACKER K); Allahverdi (without collector and date BRNM); Tiflis (without date and collector MO). Turkey: Rize, Izikdere (21.VII. 1984 A. GONER & M. VURAL GAZI); Trabzon, Bayburt (11.VII.1960 J.D.A. STAINTON & HENDERSON K).

Doronicum macrophyllum aubsp, sparsipilosum (J.R. EDM.) ALVAREZ FERNANDEZ Turkey: Bolu, Ala da~ (12 .VII. 1962 P.H. DAVIS & COODE E); ibidem (12. VII. 1962 P.H. DAVIS & COODE

K); Kastamonu, Ilgaz Da[~lari (27.VI. 1971 LR. EDMONDSON E); ibidem (27.VI. 1971 JR. EDMONDSON, sheet 2 E); ibidem (28.VII.1962 P.H. DAVIS et al. E); ibidem (5.VII.1984 M. NYDEGGER G).

Doronicum maximum BOISS. et A. HUET Turkey: Erzurum, Karliova-(~at (21.VII.1982 M. NYDEGGER G); Giresun, Balabandaglari (7.VIII.1952

P.H. DAVIS et al. BM); ibidem (7.VIII.1952 P.H. DAVIS et al. E); ibidem (7.VIII.1952 P.H. DAVIS et al. K).

Ooronicum reticulatum BOISS. Turkey: Bolu, Ala da~ (17.VI1.1967 ALPAY E); ibidem (12.VII.1962 P.H. DAVIS & COODE E); ibidem

(12.VII. 1962 P.H. DAVIS & COODE, sheet 2 E); ibidem (12.VII. 1962 P.H. DAVIS & COODE K); Bursa, Uluda~, (24.VII.1993 Z. AYTAq & M. EKICI GAZI); ibidem (9.VII.1978 BERNARDI G); Phrygia, Wilajet Konia (9.VII.1899 J. BORNMOLLER B).

a multivariate approach to assess the taxonomic utility of morphometric characters indoronicum...

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