Plant Syst. Evol. 248: 143–169 (2004) DOI 10.1007/s00606-004-0171-x A re-evaluation of morphological characters in European Gentianella section Gentianella (Gentianaceae) J. Greimler1, B. Hermanowski2, and C.-G. Jang1 1 Department of Systematics and Evolution of Higher Plants, Institute of Botany, University of Vienna, Vienna, Austria 2 Department of Ultrastructure Research and Palynology, Institute of Botany, University of Vienna, Vienna, Austria Received September 8, 2003; accepted March 9, 2004 Published online: July 20, 2004
Springer-Verlag 2004 Abstract. Morphological traits were investigated in Gentianella section Gentianella by morphometrics and Scanning Electronic Microscopy (SEM). Variation in vegetative measures and in calyx, corolla, ovary, and gynophor was analyzed in populational samples and herbarium material. Special emphasis was given to micro-morphology and variation of papillae on the calyx using SEM. Three types of papillae on the calyx lobes were found: (A) short conical (G. amarella-group, G. insubrica, G. germanica); (B) long conical often curved (G. campestris, G. anisodonta, G. engadinsesis, and G. liburnica; (C) long cylindrical (G. aspera and G. pilosa). G. austriaca, G. caucasea, G. crispata, G. fatrae, and G. ramosa usually lack any papillae. Plants without and with short conical papillae were found in G. bulgarica and G. lutescens as well as in the intermediate taxa G. bohemica and G. stiriaca. The different types of papillae together with other calyx characters (sinus, shape and margin of lobe) are of high systematic importance and provide more stable characters than morphometric flower measures. Principal component and correlation analysis revealed a strong response of nearly all morphometric traits to the environmental variable altitude. Adaptive and historical causes of morphological variation as well as taxonomical consequences are discussed and a determination key is provided. Key words: Gentianella section Gentianella, calyx, papillae, morphology, SEM, morphometrics, determination key. The genus Gentianella Moench comprising ca. 260 species (von Hagen and Kadereit 2001) is a member of the large subtribe Swertiinae of the primarily temperate-alpine Gentianeae (Struwe et al. 2002). Two taxa, segregated from Gentianella on the basis of morphological characters, Gentianopsis (Ma 1951) and Comastoma (Toyokuni 1961), are clearly confirmed by molecular data (Yuan and Küpfer 1995, von Hagen and Kadereit 2001) as genera that do not belong to the monophyletic uninectariate and biennial (occasionally annual) Gentianella s. str. With one exception (G. aurea) all European taxa of Gentianella s. str. are members of the fimbriate group which corresponds to Gentiana sect. Endotricha (Wettstein 1896) or Gentianella section Gentianella (Pritchard and Tutin 1972). Section Gentianella comprises 16 species plus five uncertain taxa (because they are weakly differentiated narrow endemics or intermediates or recently not observed) in 144 J. Greimler et al.: Morphology in European Gentianella Europe (Appendix 1) according to taxonomy applied in Pritchard and Tutin (1972) which is based essentially on the framework given by Kerner and Kerner (1882), and Wettstein (1892, 1896). Geographical variants raised to specific rank: G. fatrae (Holub 1983) and G. stiriaca (Maurer 1998) we treat as separate entities as well as G. germanica samples from the Alps (= G. rhaetica, Kerner and Kerner 1882) and outside. Except three taxa all with unique calyx morphology, G. caucasea (falcate calyx lobes), G. crispata (crispate calyx lobes) and G. columnae (4 calyx lobes, larger not enclosing smaller ones), all taxa of the section can be assigned to one of the three groups of G. amarella, G. campestris and G. germanica. High character differentation and species diversity within the G. germanica group (large 5merous flowers, mostly stalked ovary, with 9 species plus 4 intermediate or uncertain taxa) is contrasted by little differentiation in the groups of G. amarella (small 5-merous flowers, sessile ovary, with 3 species) and G. campestris (large 4-merous flowers, two larger enclosing smaller ones, with 2 species plus 1 uncertain taxon). However, taxonomy in these two groups has been questioned in recent studies (Renobales Scheifler 2003, Winfield et al. 2003). High morphological variation essentially in vegetative characters within and among species in section Gentianella is complicated by seasonal dimorphism (Wettstein 1896) or ecotypic polymorphism (Zopfi 1991). The high variation in these characters being more or less correlated with flowering time and ‘‘the intersection of these morphological gradients with other more definite characters’’ (Pritchard and Tutin 1972, p. 64) is a major reason for the confusing taxonomy in European floras. As can be judged from the investigated herbaria populations of the early flowering variants (aestival forms) probably always have been rare in Middle Europe at least in the Alps and close to them. Certainly these populations have declined rapidly in the last decades (Zopfi 1991, Rosenbauer 1996, Skalicky pers. com., Kirschner pers. com.) apparently due to impact from human activities. Complications from seasonal dimorphism or ecotypic polymorphism in vegetative characters are usually critical in delimitation of intraspecific taxa. High polymorphism, however, is also found in floral characters. Problems of specific delimitation can be summarized as follows: (1) The exact distribution of many species in Europe is yet unclear as characters used for taxonomic limitation show high intraspecific variation and little differentiation among taxa (Holub 1983, Dostal 1989, Zopfi 1991, Petanidou et al. 1998). (2) Some of the endemics (including ‘‘seasonal vicariant taxa’’) that have been described (Samuelsson 1922, Kunz 1940, Ritter-Studnicka 1955, Mayer 1969, Skalicky 1969, Holub 1983, Renobales et al. 2002) exhibit only minor differences from their closest relatives. (3) There is confusion about specific characters namely, ciliae and papillae on the calyx (e.g. Hess et al. 1972, Pritchard and Tutin 1972, Pignatti 1983). In this study we investigate variation and differentation of morphological features in European Gentianella section Gentianella. Applying a nested sampling with two datasets (populations and taxa) we focus on: (1) variation in specific characters of the flower: calyx, corolla, gynophor; (2) micro-morphology and variation of papillae located on the calyx; (3) relationships among morphological traits and environmental/geographical variables; and (4) the systematic relevance of these characters. Materials and methods Plant material. Population samples of Gentianella section Gentianella were taken during field excursions between 1996 and 2003. Details on the 75 samples (including accessions provided by other collectors) and sites are given in Appendix 2. Specimens are kept in WU. Variation of critical characters was also studied in specimens from the herbaria GZU, LI, M, NMGW, PRC, RUEB, TSB, UDM, W, and WU to complete results from our population samples and to overcome bias caused by the preponderance of central European populations in our sampling. Nomenclature follows Pritchard and Tutin (1972) except for taxa raised to J. Greimler et al.: Morphology in European Gentianella specific rank since then (Holub 1983, Maurer 1998) and the two taxa within G. germanica (Kerner and Kerner 1882). Preparation. Collections made on the field trips were immediately prepared for morphological analysis. Flat preparations of calyx and corolla were dried with the vouchers. In the lab these preparations and vouchers were photographed using the digital equipment Olympus E-10. Photographs have been modified in Adobe-Photoshop 6.0 and then displayed directly or used as templates for drawings of morphological details. Scannning Electron Microscopy (SEM). Freshly collected flowers were preserved directly in FAA (formaldehyde : acetic acid : distilled water: 70% ethanol; 2 : 1 : 7 : 10) or in 70 % ethanol which was later replaced by FAA. Calices were prepared for analysis of epidermal structures with SEM: (A) Calyx lobes of flowers fixed in FAA were cut off, transferred to fresh FAA and cleaned in the Ultrasonic Cleaner B-32 (Branson) for max. 45 minutes. (B) The cleaned lobes were dehydrated in formaldehyde dimethyl acetal (FDA) and then critical-point-dried in the CPD 030 Critical Point Dryer (Balzers). The lobes of calices fixed in 70% ethanol were transferred to fresh 70% ethanol and cleaned in an ultrasonic cleaner. Then they were dehydrated in acetone for 24 hours and criticalpoint-dried. The dried lobes were sputtered with gold for 5 minutes. Adaxial (C) and abaxial (D) surfaces of lobes were examined with the Jeol JSM35CF Scanning Microscope and digitally documented using the Digital Image Processing System Version 2.4.5.8. (Point electronic GmbH, Halle/ Saale, Germany). Digital images were reworked in Adobe Photoshop 6.0. Morphometry, statistics. Morphometric data (counts, lengths) were gathered from 75 population samples by measuring plant size, number of internodes (above the rosette-like agglomerated leaves at the stem base), number of flowers, pedicel, total calyx (base of tube to apex longest lobe), calyx tube, total corolla, corolla tube, gynophor (= carpophor, the stalk of the ovary), and ovary. Measures of pedicel and floral traits in full anthesis were taken either from terminal flowers of the main stem or from the subterminal nodes (which are most similar in all dimensions) when the former was missing or not accessible (occasionally in attached vouchers). Between four and 14 individuals per population were measured (single charac- 145 ters occasionally could be measured in only two or three individuals). From these data the individual lengths of calyx lobes and corolla lobes and the ratio lobe/tube for calyx and corolla, corolla/calyx, and pedicel/corolla were calculated. Data distribution and total character variation were investigated using the software SPSS 10: histograms (normal distribution fit) and box-plots whereby a box includes 50% of cases and median (horizontal line), bar ranges from smallest to largest observed value, that is not an outlier (more than 1.5 box lengths from 25th or 75th percentile) or extreme (more than three box lengths from 25th or 75th percentile). Median, quartiles, minimum and maximum, were calculated for each of the 75 population samples for data set 1. Mean, standard deviation, minimum, maximum were calculated from all samples per taxon for data set 2, i. e. the 75 population samples plus additional data (2–3 individuals/voucher) from herbaria collected for the above mentioned characters except number of flowers, ratio corolla lobe/tube and ovary length, which turned out negligible after analysis of data set 1. Four qualitative characters were scored on an ordinal scale: (i) Calyx papillae: 1 absent, 2 shortconical, 3 long-conical, 4 long-cylindrical (scored with a 10–16 fold lense); (ii) Calyx sinus between two lobes: 1 obtuse, 2 acute; (iii) Shape of calyx lobes: 1 linear, 2 lanceolate; (iv) Margin of calyx lobes: 1 flat, 2 slightly revolute, 3 strongly revolute (so that the margins, [nearly] touch each other [Fig. 6d]). Measuring the proper lobes is important in samples with very unequal ones as there is a size hierarchy among them as can bee seen also in ± equal (= subequal) ones ( Fig. 1a, b). Calyx sinus and shape was scored between/on the smaller (inner) lobes: sinus between lobe 3 and 5 (not always feasable in attached herbarium specimens) and shape on lobe 3, 4 or 5; margins on the larger (outer) lobes 1 or 2. Means calculated from the individual assignments were used as population scores in subsequent multivariate analyses (dataset 1). For the taxon samples (dataset 2) the percentage of occurrence of each character (character state) was calculated. One way ANOVA (analysis of variance) was applied on population samples to calculate variance components within and among populations in each of the larger samples (N = 5 or more populations) i. e. G. anisodonta, G. aspera, G. austriaca, 146 J. Greimler et al.: Morphology in European Gentianella Fig. 1. Calyx and corolla (flat preparations). Basic calyx types: a G. campestris, b G. germanica. c Corolla: f = fimbriae, g = gynophor; bar = 1 cm G. germanica (Alps), and G. stiriaca. Principal components analyses (PCA with correlation matrix, unrotated solution and Varimax rotation) were performed on the medians and scores of the populations (labeled with taxon acronyms) using (i) all samples with complete data (72 populations of data set 1) and (ii) a subset (46 populations) comprising the Alps and the adjacent north eastern lowlands. Correlations among the medians were investigated calculating the Pearson productmoment correlation coefficient. Number of flowers and plant size were log-transformed prior to these analyses. Altitude as a surrogate of environmental variables and geographical longitude, latitude (potential indicators of biogeographic differentiation) were included in these analyses as well as in a final PCA of the subsample with the characters as OTUs. For statistical analyses the software Splus 6.1 and SPSS 10 was used. Results Vegetative characters. Plant size, number of internodes and number of flowers showed higher variation among populations than within in all taxa sampled across a wide range of habitats and elevations. ANOVAs were significant (F = 3.9 to 64.7; p < 0.005) in all cases. This causes high variation within species (Table 1). For length of pedicel we found significantly higher variation among populations (F = 3.5; p < 0.005) only in G. germanica (Alps). Number of flowers ranged from 1 to 175 (n = 591) with an extreme left-sided distribution [median 8 (quartiles 5/16)] caused by a similar pattern in nearly all taxa. Investigating patterns of variation among taxa revealed (Table 1): (1) Small plant sizes combined with low variation in G. bulgarica and G. liburnica. (2) Lower numbers of internodes in G. uliginosa than in G. amarella. (3a) Shortest pedicels in G. liburnica, however, to a high proportion (ca. 50%) within the lower range of G. anisodonta and G. engadinensis. (3b) An overlapping range of variation in length of pedicel for G. bulgarica and G. ramosa. Calyx. Two basic calyx types are shown in Fig. 1a,b. The tetramerous type A with flat lobes is restricted to G. campestris: two ovoidlanceolate larger sepals of equal size at least partly cover two smaller narrowly lanceolate or nearly linear ones (Fig. 1a) and G. columnae: two larger triangular-lanceolate subequal sepals usually not covering the smaller narrowly lanceolate or nearly linear ones. The reduced number of sepals, although rarely, can be found in single flowers of most usually fivemerous taxa. Within the pentamerous type, which is typical for the G. germanica group J. Greimler et al.: Morphology in European Gentianella 147 Table 1. Metric data (min.-max., mean, standard deviation, number of individuals) for length of calyx, pedicel, gynophore (mm), stem (cm); ratios of pedicel/corolla and calyx/corolla; number of internodes Taxa Calyx Pedicel R Pe/Co R Ca/Co Gynophor Stem N Internodes amare 7–15 10.3±1.8 35 8–20 13.6±3.0 35 13–27 19.9±3.4 34 10–36 19.5±4.4 77 11–20 15.9±2.4 26 11–23 16.3±3.1 40 10–25 16.4±3.6 29 11–21 15.4±2.7 12 9–19 13.4±2.5 24 8–15 11.0±2.0 28 12–26 18.4±3.8 14 11–24 15.9±2.8 75 6–21 12.7±2.7 54 12–27 18.4±4.8 22 6–10 8.0±1.3 7 5–32 12.6±5.4 22 2–25 10.0±5.5 25 13–43 23.4±8.2 26 5–25 16.0±4.8 35 6–23 14.2±5.4 17 3–23 10.0±6.2 22 6–30 17.2±6.8 23 5–40 19.1±9.1 13 5–25 16.3±7.7 9 3–15 6.7±3.0 17 3–23 12.3±6.1 12 6–25 14.1±5.6 48 7–35 16.4±7.5 27 7–29 17.5±7.0 12 1–5 3.0±1.4 7 0.3–2.0 0.7±0.3 22 0.2–1.0 0.4±0.2 24 0.4–1.2 0.7±0.2 26 0.2–0.8 0.5±0.2 35 0.2–1.1 0.5±0.2 17 0.1–1.0 0.4±0.3 20 0.3–1.1 0.6±0.2 23 0.2–1.7 0.8±0.4 10 0.3–1.1 0.7±0.3 9 0.2–0.8 0.4±0.2 17 0.1–0.8 0.4±0.2 11 0.2–1.0 0.5±0.2 48 0.2–0.9 0.6±0.2 23 0.3–1.2 0.7±0.3 11 0.1–0.3 0.2±0.1 7 1.2–1.9 1.7±0.2 22 1.3–2.3 1.8±0.3 24 1.5–2.3 1.8±0.2 26 1.2–2.6 1.7±0.3 38 1.3–2.1 1.8±0.2 17 0.9–1.8 1.5±0.2 28 1.3–2.2 1.7±0.2 28 1.3–2.0 1.7±0.3 10 1.3–1.7 1.4±0.1 10 1.3–2.5 1.6±0.4 18 1.4–2.3 1.8±0.3 12 1.1–2.3 1.7±0.2 48 1.6–3.5 2.2±0.4 44 1.0–1.9 1.5±0.3 20 1.6–2.2 1.9±0.2 7 9–43 25.3±10.3 21 2–23 7.6±4.5 67 8–53 22.8±10.8 48 8–40 19.3±8.2 78 9–51 20.3±9.0 22 3–10 5.7±2.0 30 5–25 13.5±4.5 34 9–22 16.2±3.8 15 3–14 8.6±4.1 16 2–18 5.6±3.6 48 7–27 14.9±4.9 15 4–41 13.0±7.2 88 4–44 17.7±10.8 37 8–22 14.6±3.7 18 3–7 3.6±1.5 7 6–13 9.1±2.4 21 2–8 4.4±1.4 67 3–15 8.0±3.1 48 4–14 8.3±2.3 57 5–12 8.7±2.2 19 3–7 4.5±1.0 30 3–7 4.8±0.9 34 3–7 5.1±1.2 14 2–6 4.2±1.3 15 2–7 3.5±1.3 42 5–9 6.5±1.0 16 3–12 5.9±1.6 82 3–13 8.5±2.5 38 4–8 5.8±1.0 14 2–6 3.4±1.3 7 Min.–Max. Mean±Std N aniso Min.–Max. Mean±Std N asper Min.–Max. Mean±Std N austri Min.–Max. Mean±Std N bohem Min.–Max. Mean±Std N bulga Min.–Max. Mean±Std N camp Min.–Max. Mean±Std N caucas Min.–Max. Mean±Std N colum Min.–Max. Mean±Std N engad Min.–Max. Mean±Std N fatrae Min.–Max. Mean±Std N germ_A Min.–Max. Mean±Std N germ_O Min.–Max. Mean±Std N insub Min.–Max. Mean±Std N liburn Min.–Max. Mean±Std N 0–2 0.5±0.6 20 0–4 1.7±1.2 18 0–7 2.9±2.4 19 1–10 4.3±2.3 52 1–6 3.9±1.4 11 0–10 3.7±2.9 21 1–4 2.7±1.1 12 2–6 3.9±1.1 11 0–2 0.6±0.9 5 0–3 1.4±0.9 14 1–5 3.1±1.6 13 0–6 2.2±1.1 49 2–7 4.5±1.5 22 0–4 2.1±1.1 12 1–2 1.3±0.5 4 148 J. Greimler et al.: Morphology in European Gentianella Table 1. (continued) Taxa lutesc pilosa ramos styria uligin Total Min.–Max. Mean±Std N Min.–Max. Mean±Std N Min.–Max. Mean±Std N Min.–Max. Mean±Std N Min.–Max. Mean±Std N Min.–Max. Mean±Std N Calyx Pedicel R Pe/Co R Ca/Co Gynophor Stem N Internodes 9–31 16.8±6.2 25 12–24 18.2±3.4 28 9–17 12.8±2.4 25 13–23 18.2±2.7 51 10–18 14.0±2.3 16 6–36 15.8±4.4 657 5–22 13.0±5.7 14 5–19 11.5±4.3 20 2–15 5.8±3.2 24 5–30 17.0±5.6 22 13–70 34.5±20.6 15 1–70 14.6±9.0 410 0.2–1.0 0.5±0.2 14 0.2–0.7 0.4±0.1 20 0.1–0.7 0.3±0.1 24 0.2–0.9 0.5±0.1 21 0.7–4.6 2.2±1.3 15 0.1–4.6 0.6±0.5 397 1.2–2.2 1.6±0.3 14 1.3–2.1 1.7±0.2 21 1.3–1.8 1.5±0.2 22 1.5–2.5 1.9±0.3 21 0.9–1.8 1.2±0.3 16 0.9–3.5 1.7±0.3 446 7–35 21.9±7.7 24 6–39 19.4±9.1 29 4–16 9.6±3.1 29 7–36 16.5±6.6 43 3–16 9.6±3.7 16 2–53 14.4±9.0 685 4–10 6.9±1.5 23 4–11 7.4±2.2 28 4–7 5.3±1.1 29 4–10 6.7±1.4 46 2–4 2.7±0.7 15 2–15 6.2±2.5 645 and G. amarella group (Fig. 1b: G. germanica, Alps) there is a great variation concerning proportions, margins, sinuses between adjacent lobes (Table 2), and the presence and shape of papillae on the margin and midrib of the lobes (Table 3). As a rule the sepals decrease in size according to a 2/5 spiral. The larger (outer) sepals occasionally have ‘‘wings’’ in their connate part forming the calyx tube. Within the basic type B two calyx types are unique: (1) crispate calyx lobes with often blackish margins in Gentianella crispata; (2) falcate and very delicate, narrowly linear lobes and the tube often split down on one side in G. caucasea. In the remaining taxa belonging to the G. germanica group and G. amarella group three major calyx types are found (Fig. 2): (1) Lobes triangular-lanceolate to ovoidlanceolate, partly enfolding the inner ones, extremly unequal in size, margins often conspicuously recurved (= lobe revolute), sinus acute: G. anisodonta (Figs. 2a, 6d), G. engadinensis, and G. liburnica. (2) Lobes (often narrowly) lanceolate, ± equal in size, margins not or slightly recurved, sinus acute: G. aspera (Fig. 2b), G. pilosa 0–4 2.7±1.5 15 2–5 3.5±1.0 11 1–5 3.1±1.1 15 1–8 4.5±1.8 32 0–1 0.5±0.6 4 0–10 3.0±2.0 360 (Fig. 2c), G. germanica, and G. ramosa (Fig. 2d). (3) Lobes linear, ± equal in size, margins not or slightly recurved, sinus obtuse: G. austriaca (Fig. 2e), G. bulgarica, G. fatrae (Fig. 2f), G. lutescens (Fig. 2g), and often in G. amarella (Fig. 2h). However, in single individuals from populations of all taxa we found intermediate calyx types due to occasionally high variation in the characters sinus, lobe, margin. Such intermediate calyx types were often found in G. bohemica, G. insubrica, and G. stiriaca but also in G. amarella (Table 2). We observed high metric variation of calyx features: The ratio lobe/tube revealed significantly higher variation among populations than within in all five largest taxon samples (ANOVA, F = 2.6 to 7.8; p < 0.01). There is, however, also high differentiation among taxa (Fig. 3). Variation in total calyx length was significantly higher among populations (F = 3.2 to 8.2; p < 0.005) in G. aspera, G. austriaca, and G. germanica (Alps). Micromorphology of papillae on the calyx. In taxa with papillae, these occur on the 735 24 40 70 30 61 34 5 38 42 22 100 71 29 18 82 11 16 85 46 37 48 15 27 43 57 68 32 29 36 18 100 97 3 100 46 78 31 17 57 26 23 52 48 92 8 72 28 49 51 53 36 11 89 18 100 10 90 29 91 9 23 52 48 31 86 14 78 6 94 47 100 63 81 19 Margin 1 2 3 N N 69 31 42 Lobes 1 2 42 16 81 19 50 50 28 37 63 27 15 85 46 12 88 90 100 100 28 16 36 N 83 17 751 100 11 86 14 42 27 73 22 45 55 38 43 58 40 42 58 24 751 17 83 24 20 80 40 13 87 38 100 22 81 19 42 100 11 11 89 27 9 91 46 3 97 90 100 100 28 100 18 100 100 29 61 39 23 35 65 31 86 14 78 100 47 100 63 74 26 42 1 2 Sinus St. amare aniso asper austri bohem bulga camp caucas colum engad fatrae germ_A germ_O insub liburn lutesc pilosa ramos styria uligin Tot. Char. Table 2. Discrete calyx character states (percent of observations given): Sinus between lobes: 1 obtuse, 2 acute; shape of lobes: 1 linear, 2 lanceolate; margin of lobes: 1 flat, 2 slightly revolute, 3 strongly revolute = margins (nearly) touch each other J. Greimler et al.: Morphology in European Gentianella 149 margin of the leaves, gradually becoming more pronounced towards the inflorescence. The papillae are most conspicuous on the margins of the calyx lobes from which they often run down the tube along the fusion lines of the sepals. They also occur (often decreased in size) at least in part of the adaxial surface and (in two taxa) also on their abaxial midrib or the whole abaxial surface of the tube. Table 3 gives a quantiative summary about papillar structures as shown in detail by the schematic drawing (Fig. 4) and the SEM photographs (Figs. 5–8): We found three types of papillae: (A) Short-conical (in herbario: short-triangular) papillae, about as long as broad (basal diameter), sharply pointed, ± asymmetrical and pointing slightly towards apex of sepal were found in the G. amarella-group (Fig. 5ac), G. germanica (Fig. 5d-f), G. insubrica. As the following types of papillae often become shorter towards the apex, papillae approaching type A could be found in all taxa near the very apex. (B) Long-conical (long-triangular), about twice as long as broad, occasionally curved papillae with sharp or rounded tip, ± pointing towards apex were found typically in G. campestris (Fig. 6a–c), G. anisodonta (Fig. 6d–f), G. columnae, G. engadinensis, and G. liburnica. These taxa, however, show some variation in often having additionally (occasionally only) short type A papillae or also longer papillae approaching type C but with a broader base. (C) These are long-cylindrical (long-linear) with blunt or rounded tip, usually much more than twice as long as broad, typically found in G. pilosa (Fig. 7a–c) and G. aspera (Fig. 7d–f), where they also occur (often shortened in G. pilosa) on the abaxial midrib. However, in both G. aspera occasionally and more often in G. pilosa we found the midrib smooth and the margins only short-papillose (approaching types B and A) or nearly smooth, rarely perfectly smooth. Non-papillose taxa with smooth margins of the calyx lobes (G. austriaca, Fig. 8a,b; 150 J. Greimler et al.: Morphology in European Gentianella Table 3. Overview of occurrence and types of calyx papillae in European Gentianella sect. Gentianella. Descriptions are based on freshly fixed flowers (see SEM-photographs) and herbarium observations by a 10–16 fold lense (in parentheses). N: number of individuals (1209 total) and percent of observed papillae type per taxon (main type bold) Taxon N Papillae absent (%) Papillae present (%) (A) shortconical (= shorttriangular) G. G. G. G. G. G. G. G. G. G. G. G. G. G. G. G. G. G. G. G. G. amarella anisodonta aspera austriaca bohemica bulgarica campestris caucasea columnae crispata engadinensis fatrae germanica Alps germanica Out insubrica liburnica lutescens pilosa ramosa styriaca uliginosa 108 66 195 98 46 39 51 15 24 30 38 16 109 107 20 11 25 36 88 44 29 11 98 37 36 89 9 15 2 63 64 27 Location (B) longconical (= longtriangular) (C) longcylindrical (=longlinear) 76 11 15 74 73 margin lobe (%) 89 100 100 2 63 64 100 midrib lobe (%) 84 100 87 88 11 3 60 3 98 18 20 25 13 8 12 89 95 100 18 40 19 2 82 80 G. fatrae, Fig. 8c; G. ramosa, Fig. 8d,e; G. caucasea; G. crispata) occasionally have somewhat undulate-uneven (rarely very short papillose) margins towards the apex [the crispate margin of G. crispata is a feature of the total lobe and not of the epidermal cells]. High variation between the smooth and short-papillose (type A) was found in G. bohemica, G. bulgarica, G. lutescens, G. stiriaca (Fig. 8f), and G. uliginosa . Corolla and gynophor. The obconical or nearly cylindrical corolla with four or five lobes, which is fimbriate in the throat (Fig. 1c), offers few characters: Corolla size, color (could not be scored reliably from herbaria) and the ratio lobe/tube (often difficult to see in old specimens). In corolla size 75 87 5 2 82 42 36 100 13 100 12 89 97 100 100 40 97 2 82 80 56 (Fig. 9) we found a common size range of more than 75 % for the pairs G. engadinensis/ G. anisodonta and G. austriaca/G. lutescens. This is due to high within species variation, resulting from high differentiation among populations (ANOVA for corolla size, F = 8.1 to 13.8, p < 0.001) in G. anisodonta, G. austriaca and G. germanica (Alps). The ratio corolla lobe/tube calculated for the population samples exhibited some geographical correlation (see below) due to high values in G. stiriaca and clinal variation in G. germanica towards the East and G. austriaca towards the West, respectively. Comparing taxa, where this might be a useful additional character, e.g. in G. aspera and G. anisodonta we found a widely common range for this ratio J. Greimler et al.: Morphology in European Gentianella 151 Fig. 2. Drawings after flat preparations of the calyx of (a) G. anisodonta (note the very unequal lobes), (b) G. aspera, (c) G. pilosa, (d) G. ramosa – all with acute sinuses; (e) G. austriaca, (f) G. fatrae, (g) G. lutescens, (h) G. amarella – all with obtuse sinuses; bar = 1 cm 5 Calyx, ratio lobe/tube 4 3 2 1 austri bohem bulga camp caucas colum 14 67 41 19 23 26 22 47 16 uligin asper 27 styria 24 ramos 12 pilosa 23 lutesc 30 insub 21 germ_O 77 germ_A 29 fatrae 29 engad 28 aniso N= amare 0 Taxa Fig. 3. Variation in the ratio calyx lobe/tube within and among taxa (abbreviated epitheta) of Gentianella section Gentianella. Horizontal line = median, circle = outlier, asterisk = extreme. germ_O: populations of G. germanica outside the Alps; germ_A: in the Alps 152 J. Greimler et al.: Morphology in European Gentianella Fig. 4. Schematic drawings of papillae in the middle of the calyx lobes (abaxial view) with approximate size relations among the three types: (a) short-conical, about as long as broad, sharply pointed, ± asymmetrical and pointing slightly towards apex of sepal; (b) long-conical, about twice as long as broad, often curved with sharp or rounded tip, ± pointing towards apex; (c) long-cylindrical (long-linear) with blunted or rounded tip, usually more than twice as long as broad. For exact size measures compare Figs. 5–7 (> 50%) in a boxplot view (not shown) despite a statistically significant difference (t-test, p < 0.001) in the means. High variation in length of gynophor was found in nearly all taxa. Again ANOVA revealed significantly higher varition among than within populations (F = 2.1 to 31.1; p < 0.05) in the large samples except in G. stiriaca. Sessile ovaries are not confined to the G. amarella-group and G. engadinensis. We found them in flowers of seven other taxa (Table 1). On the other hand, gynophor length in rare cases reaches up to 2 mm in G. amarella and occasionally up to 3 mm in G. engadinensis. Correlations, PCA. Correlation analyses performed on a subsample of dataset 1 (46 populations of the Alps and adjacent areas) revealed the high importance of the environmental variable altitude which explained 57% of total variation in number of internodes (r = )0.758, p < 0.001), 36% in plant size (r = )0.602, p < 0.001), and 17% in number of flowers (r = )0.407, p = 0.005). Altitude also explained high proportions of variation in floral characters: 20% in calyx length (r = )0.445, p = 0.002), 38% in corolla length (r = )0.614, p < 0.001), 36% in ovary length (r = )0.598, p < 0.001), and 29% in gynophor length (r = )0.482, p = 0.001). Both geographical variables longitude and latitude (which were inversely related to altitude due to the west-northeast sampling along the Alps) showed high positive correlations with the above variables. For the ratio corolla lobe/tube, which was not significantly associated with altitude we also found high correlations with both, longitude (r = 0.559, p < 0.001) and latitude (r = 0.515, p < 0.001). Vegetative characters were highly correlated with floral characters: number of internodes with corolla length (r = 0.543, p < 0.001), ovary length (r = 0.543, p < 0.001), and gynophor length (r = 0.434, p = 0.003); plant size with corolla length (r = 0.588, p < 0.001), ovary length (r = 0.549, p < 0.001), and gynophor length (r = 0.413, p = 0.004). A principle components analysis on all populations of data set 1 did not reveal any groups (not shown). Similar results were obtained with the subsample as used for correlation analysis (Varimax rotated solution shown in Fig. 10a). Therefore the characters were ordinated in another PCA together with altitude of the sampling site and geographical coordinates. This unrotated PCA underlined the high importance of altitude (Fig.10b) in J. Greimler et al.: Morphology in European Gentianella 153 Fig. 5. Short-conical (type A) papillae (SEM photographs of calyx lobes, abaxial): G. amarella: (a) surface, (b) margin, (c) midrib; G. germanica: (d) surface, (e) margin, (f) midrib. Bar: 100 lm constituting the first component, which explained 40% of the total variance. The correlations reported above were essentially displayed on this first axis. Together with the second (14%) and third component (11%) 65% of the total variance was explained. Patterns, however, along the other axes did not provide additional insights except demonstrating the high association of the ratio corolla lobe/tube with geographical variables in all dimensions. As neither PCA with all characters (Fig. 10a) nor ordination of selected metric characters (Fig. 10c) did reveal groups in the data we tested the ordinal characters alone. The population scores of these characters (character states see Table 2 and 3) did reveal groups in the data. The best resolution was obtained by including (1) papillae type, (2) shape of lobe, and (3) margin. (Fig. 10d). Discussion Character variation. Focusing on micromorphology of epidermal structures on the calyx revealed that there are three different types of papillae and only few taxa with no papillae (Figs. 4–8). The calyx lobes where they occur have been discribed in the past by so many subtle terms, such as ‘‘smooth, glabrous, (somewhat) scabrid, (strongly) ciliate, shortly hirsute, papillose, papillose-hirsute’’ (Pritchard 154 J. Greimler et al.: Morphology in European Gentianella Fig. 6. Long-conical (type B) papillae (SEM photographs of calyx lobes, abaxial): G. campestris: (a) surface, (b) margin, (c) midrib; G. anisodonta: (d) surface, (e) margin, (f) midrib. Bar: a, d 1 mm; b, c, e, f 100 lm and Tutin 1972 p. 63 ff) or ‘‘glatt, schwachrauh, knotig-rauh, kurz-rauhhaarig, rauh-gewimpert’’ (Jäger and Werner 2002 pp. 515–516). We propose a more straightforward terminology by distinguishing between smooth and papillose and defining the three types of papillae and their occurence (Fig. 4, Table 3). By this we hope to provide a more efficient tool for identification and help reduce confusion about these characters in many European determination keys. Calyx features (number of sepals, shapes, sinuses between the lobes, ratio lobe/tube, presence/absence of papillae) have always been central diagnostic characters in section Gentianella since Kerner and Kerner (1882) and Wettstein (1892, 1896). A biogeographical pattern in these characters as suggested by Wettstein (1896) and Skalicky (1969) is obvious in the G. germanica group (including G. caucasea) although G. bulgarica and G. ramosa do not fit well into this: (1) Smooth or only shortly papillose often linear lobes and obtuse calyx sinuses are typical for taxa of eastern and south-eastern Europe (incl. the Caucasus region). The very delicate calyx tubes often split down on one side in G. caucasea are occasionally found also in G. austriaca, G. fatrae, and G. lutescens. (2) Long conical papillae, combined with acute sinuses and lanceolate lobes are found in the southern Alps and Dinarids (G. anisodonta, G. engadin- J. Greimler et al.: Morphology in European Gentianella 155 Fig. 7. Long-cylindrical (type C) papillae (SEM photographs of calyx lobes, abaxial): G. pilosa (a) surface, (b) margin, (c) midrib. G. aspera: (d) surface, (e) margin, (f) midrib; Bar: d 1 mm; a, b, c, e, f 100 lm ensis, G. liburnica). (3) Long cylindrical papillae combined with lanceolate lobes and acute sinuses are found only in a small area of the southern eastern (G. pilosa) and along the northern Alps extending to adjacent central Europe (G. aspera). (4) Short–conical papillae are typical for G. germanica of central and north-western Europe and the southern Alps narrow endemic G. insubrica. These patterns of calyx variation, however, are not always correlated with taxonomic entities. Taxa with intermediate characters such as G. bohemica (Skalicky 1969) and G. stiriaca (Wettstein 1892, 1896; Hayek 1911–14; Maurer 1998) have been considered transgressive. However, clinal variation can be found in many Gentianella taxa. Such variation might be due to inter-/postglacial introgression events or even intergradation in refugia during glaciation as supposed for populations of Pritzelago alpina (Kropf et al. 2003). Molecular markers revealed high genetic similarities among populations across Gentianella taxa of the Eastern Alps (Jang and Greimler, unpublished). Characters of the corolla (color, size) and gynophor length are of limited diagnostic value although often used (Samuelsson 1922, Mayer 1969, Pritchard and Tutin 1972). In some cases unusual characters might point to hybridization as e.g. the long gynophor found in an Austrian population of G. amarella 156 J. Greimler et al.: Morphology in European Gentianella Fig. 8. Smooth margins (SEM photographs of calyx lobes, abaxial): G. austriaca: (a) surface, (b) margin; G fatrae: (c) margin; G. ramosa: (d) surface, (e) margin; G. styriaca: (f) margin. Bar: 100 lm (in this region sympatric with G. germanica). Hybridization of G. amarella with taxa of the G. germanica group is not uncommon (Pritchard 1961, Moravec and Vollrath 1967). We have no explanation for the higher ratios lobe/ tube in the Nort-Eastern Alps in G. stiriaca and populations of G. austriaca, G. germanica and G. aspera occuring there. Floral similarities across taxa often reflect local adaptations to attract similar pollinators as found in orchids (Borba et al. 2002). We found evidence that high proportions of metric variation especially of vegetative traits in Gentianella sect. Gentianella can be explained by altitude used as a complex environmental variable. In a detailed investi- gation of G. austriaca (Greimler and Dobeš 2000) we also found high intraspecific variation in vegetative characters connected with different habitats and different population histories. This agrees with Zopfi (1991) who favoured a model of ecotypic polymorphism in contrast to Wettsteins (1896) simple seasonal dimorphism model (aestival plants with fewer and longer internodes compared to the autumnal plants). There are, however, clear patterns of seasonal differentiation in populations outside the central European mountain ranges in the G. amarella group and in G. campestris (Lennartsson 1997, Rich et al. 1997). In the central European mountain taxa of Gentianella, there are very few extant populations J. Greimler et al.: Morphology in European Gentianella 157 60 50 Corolla length, mm 40 30 20 10 austri bohem bulga camp cauca colum crispa 13 96 102 21 8 21 23 23 47 23 uligin asper 34 styria 16 ramos 21 pilosa 25 lutesc 30 liburn 26 insub 39 germ_O 71 germ_A 67 fatrae 42 engad 111 aniso N= amare 0 Taxa Fig. 9. Variation in corolla length within and among taxa (abbreviated epitheta) of Gentianella section Gentianella. Horizontal line = median, circle = outlier, asterisk = extreme. germ_O: populations of G. germanica outside the Alps; germ_A: in the Alps (Skalicky 1969 and pers. comm., Zopfi 1991, Wagner and Mitterhofer 1998) showing seasonally and morphologically seperated cohorts. Calyx features (sinus, shape, proportions) as used traditionally are central for understanding systematic relationships in the section and for distinguishing taxa looking beyond the concept of Pritchard and Tutin (1972). Our more detailed investigation of epidermal structures on the calyx, i.e. the presence/absence and different shapes of papillae, provided useful critical characters that so far have been underestimated. As morphometric characters of calyx, corolla and gynophor as well as vegetative characters display a great withinspecies variation partly due to environmental modification, such additional characters help in systematic delimitation. However, all diagnostic characters can become less definitive towards the edge of distribution of many central European taxa. Morphologically this has been interpreted as reflecting introgression or perhaps ancient hybridization (Wettstein 1892; 1896; Hayek 1911–14; Skalicky 1969). Conclusions towards a modified taxonomy. We here summarize various investigations and our observations on European Gentianella section Gentianella with respect to the determination key proposed below. Short 158 J. Greimler et al.: Morphology in European Gentianella comments on taxa for which we have no original data are given in Appendix 1. We partly resolve the G. amarella group in distinguishing between G. amarella and G. uliginosa. However, due to some taxonomic confusion and many missidentifications in the G. amarella group we excluded many herbaria samples of G. uliginosa which by far did not agree with the pre-selective criteria as given in Pritchard and Tutin (1972) and Holyoak (1999). These characters have also been found too variable for clearly identifying Dutch populations of G. amarella and G. uliginosa (Petanidou et al. 1998). Also we did not observe the more outcurved calyx lobes (as in British samples: Rich 1997, Holyoak 1999) in Middle and Northern European samples of G. uliginosa. Taking the characters given in Pritchard and Tutin (1972) it is very difficult to distinguish between G. uliginosa and G. anglica due to the high variation of these characters (upper leaves, calyx lobes) in what we consider G. uliginosa. From the data given in Rich et al. (1997), G. anglica well fits into the range of G. uliginosa and therefore we include it in the latter taxon. Considering the poor genetic differentiation within the G. amarella-group (data in Winfield et al. 2003) this rather reflects ecotypic variation of one species occuring with various fully compatible variants in various habitats than differentiation on the species level (with subsequent hybridizations) as suggested by Rich et al. (1997) and Holyoak (1999). Additionally there is some geographical differentiation. Because of high proportions of tetramerous flowers, often smooth calyx margins and high variation in the gynophor length Spanish populations of G. ama- rella were considered a different species (G. hispanica) by Renobales et al. (2002). However, recently (Renobales Scheifler 2003) this taxon was included again in G. amarella. G. engadinensis is a variant of G. anisodonta with smaller and more reddish flowers and nearly sessile ovaries as found by Samuelsson (1922). However, there is some variation in color within G. anisodonta. Moreover variants with very small flowers and nearly sessile ovaries we also found in other regions of the large distribution area. Such small variants we cannot distinguish from G. engadinensis, at least not in the herbaria. G. liburnica, another variant with small and dirty-violet flowers was distinguished from G. engadinensis by more narrow and acute leaves, smaller flowers and a longer gynophor. Summarizing our observations and considering the diagnosis of G. liburnica (Mayer 1969), none of these characters holds. We cannot distinguish clearly between all these variants of G. anisodonta and therefore treat it as a group here. Apart from problems of clearly characterizing all the variants, this group requires further investigation focusing on patterns of differentiation in context with the many refugial areas (Tribsch and Schönswetter 2003) in the Southern Alps and Illyric mountain ranges and possible hybridization zones. G. aspera shows some clinal variation. In the Bavarian Prealps the taxon often lacks long papillae in contrast to the densely longpapillose populations of the Alps. Towards the western and northern border of its distribution area this might also indicate introgression by G. germanica. Samples of such probably intermediate variants were not included in the statistics and key. c Fig. 10. (a) PCA 1 (varimax rotated) ordinating populations (labeled with first two characters of epitheta) does not reveal groups. (b) PCA 2 shows (i) high negative relationships between the environmental variable altitude (alt) and metric variables: lengths of calyx (cx), corolla (co), gynophor (gy), ovary (ov), plant size (sh); number of internodes (ni) and flowers (nf); the ratios lobe/tube of calyx (rcx) and corolla (rco); and high association between rco and the geographical variables longitude (lon) and latitude (lat). (c) Ordinating populations by selected metric characters (gynophor, corolla, ratio lobe/tube of calyx) does not reveal structure. (d) Ordinating populations by ordinal characters shows core groups in accordance with taxonomy despite high variation also in these characters J. Greimler et al.: Morphology in European Gentianella 159 160 J. Greimler et al.: Morphology in European Gentianella G. austriaca includes G. fatrae as we cannot distinguish the two entities by the longer and narrower calyx lobes of the latter (as given by Holub 1983) or by any other character we have investigated. For the same reason we have to include also G. lutescens. This taxon, however, shows high variation in all characters across its wide distribution area and requires further investigation. For now we include it into an informal G. austriacagroup. In G. campestris there is no differentiation of plants in the Pyrenees justifying a separate species G. hypericifolia. Spanish authors have included this taxon in G. campestris (Morales et al. 1996, Renobales et al. 2002). According to Renobales Scheifler (2003) the sepals in these populations are mostly elliptical, ovate, not obovate as given in Wettstein (1896: plate 4, fig. 2) and therefore not different from the sepals in G. campestris. We here propose to follow Kerner and Kerner (1882) in assigning the alpine populations of G. germanica to a separate taxon G. rhaetica although most differential characters given by the authors (relative leaf size and shape, corolla size) are too variable to separate the populations from the Alps and the regions of northern Middle and Western Europe. Another character given by the authors, the length of gynophor in relation to calyx tube is difficult to observe in herbaria. This character, however, shows significant differentiation due to the more delicate calyx and longer gynophor in G. germanica s.str. as well as the total relation of corolla to calyx (t-tests, all p < 0.001) which we scored additionally (Fig. 11). Other characters as given in Lauber and Wagner (2001): calyx lobes outcurved (some regional differentiation within G. germanica s.str.) and calyx winged do not show differentiation of taxonomic significance. As we have evidence for high genetic differentiation between these two taxa (Jang et al., unpublished) differentiation of all characters will be tested again including further samples of other regions of widespread G. germanica s.str. Due to high intraspecific polymorphism, however, morphometry often fails to provide clues for taxonomy despite genetic differentiation (Woo et al. 2002). Finally, we have to deal with two obvious intermediate taxa and one taxon combining characters of other taxa. G. bohemica is a morphological and geographical intermediate between G. austriaca and G. germanica (Skalicky 1969). Intermediate populations between G. austriaca, G. aspera, and G. rhaetica in the Eastern Alps have been assigned to G. stiriaca on various taxonomic ranks (Wettstein 1892, 1896; Hayek 1911–14). Recently this taxon has been raised to specific rank again (Maurer Fig. 11. Variation in calyx length (white), ratio corolla/calyx (light grey), gynophor length (grey), and ratio gynophor/calyx tube (bricks) within and between G. germanica (Alps) (= G. rhaetica) and G. germanica s.str. (Out); all lengths in cm J. Greimler et al.: Morphology in European Gentianella 1998: sub Gentiana). The case of G. insubrica is less clear. Although Kunz (1940) empasized that this is a distinct taxon to us it seems likely that it is a southern isolated variant of G. germanica or G. rhaetica with smaller flowers. However, in G. insubrica the calyx lobes are often very unequal (as in G. anisodonta) and elongated (as in G. ramosa). We do not agree with the inclusion of this taxon (Pignatti 1983) in G. anisodonta. Genetic evidence has to be considered also in deciding wether these taxa should be included elsewhere. Determination key. This is a multiple access key based on the investigated material. Terms describing frequency refer to statistical counts. Exclusive characters (>95% of all observations) are given without adverb. In other cases terms discribing frequencies are used: usually (>70% of all observations), often (>50%) frequently (>30%), occasionally (>5%), rarely (<5%). Measures include minimum and maximum (in parentheses) and the range covered by the standard deviation (all rounded to next integer). The papillae type can be judged with a ten-fold lense. Data from Rich et al. (1997) were including for the G. amarella group. Caveat: green or hyaline epidermal cells alternating with darkpurplish cells can produce the impression of short papillae which are actually not present. 1 Calyx lobes with strongly crispate margin; margin usually strongly blackish and without papillae G. crispata * Calyx lobes with flat or recurved, but not crispate, margin 2 2 Flowers 4- (rarely 5-) merous; margin of calyx lobes usually flat 3 * Flowers 5- (rarely 4-) merous; margin of calyx lobes flat or recurved 5 3 Margin of calyx lobes usually with longconical, occasionally with short-conical papillae; calyx lobes usually very unequal 4 * Margin of calyx lobes smooth or with short-conical papillae; calyx lobes subequal. 4-merous variants (incl. G. hispanica) of the G. amarella-group 13 161 4 2 large ovate-lanceolate calyx lobes (partly) enclosing the 2 smaller linear-lanceolate ones G. campestris (incl. G. hypericifolia) * 2 large triangular-lanceolate calyx lobes not enclosing the 2 smaller linear-lanceolate ones G. columnae 5 Calyx lobes falcate, linear; calyx tube often split down one side G. caucasea * Calyx lobes not falcate 6 6 Margin of calyx lobes usually with longconical or long-cylindrical papillae 7 * Margin of calyx lobes smooth or with short-conical papillae 9 7 Midrib of calyx lobes without papillae; calyx lobes usually strongly revolute and very unequal, margin usually with long-conical, occasionally with short-conical or long-cylindrical papillae G. anisodonta-group * Midrib of calyx lobes often with papillae; calyx lobes revolute or not, usually subequal, margin with long-cylindrical or long-conical papillae, occasionally with short-conical papillae 8 8 Calyx lobes usually broadly-lanceolate, (1.1-)1.3-2.0(-2.6) · as long as tube; margin of calyx lobes usually with long-cylindrical, occasionally with long-conical or short-conical papillae G. aspera * Calyx lobes narrowly-lanceolate, (1.4-)1.9-2.7(-3.0) · as long as tube; margin of calyx lobes with long-conical or longcylindrical, occasionally with short-conical papillae or rarely smooth G. pilosa 9 Calyx sinus usually obtuse; calyx lobes usually linear or ovate-lanceolate-widest towards apex 10 * Calyx sinus usually acute; calyx lobes (triangular-) lanceolate 12 10 Calyx lobes usually strongly revolute, often ovate-lanceolate-widest towards apex G. bulgarica * Calyx lobes flat or rarely slightly revolute, usually linear 11 11 Corolla (10-)14-19(-22) mm; gynophor 0-1(-2) mm; margin of calyx lobes usually with short-conical papillae G. amarella 15 * Corolla (21-)22-40(-51) mm; gynophor (0-)1-7(-10) mm; margin of calyx 162 J. Greimler et al.: Morphology in European Gentianella lobes without (Central Europe) or frequently with short-conical papillae (Eastern Europe) G. austriaca-group 12 Calyx lobes (2.0-)2.1-3.8(-4.5) · as long as tube; corolla usually much shorter than 25 mm 13 * Calyx lobes (0.8-) 1.1-1.6(-2.4) · as long as tube 16 13 Tall plants, usually > 15 cm, > 7 internodes (autumnal variant); stems simple or branched from above the base or smaller plants with < 4 internodes and long ascending branches and terminal internode plus pedicel longer than leaves G. amarella-group 15 Note: Rare(?) aestival variants of G. amarella with < 7 internodes can be distinguished from the following taxa by at least 3 internodes much longer than the leaves, the longest internode 3-7 · as long as the leaves below. * Small pants, < 15 cm, usually 4–7 internodes; stems usually densely branched from the base; plants often of caespitose cushion-like habit; terminal internode plus pedicel shorter than leaves 14 14 Calyx lobes usually lanceolate, margin flat or slightly revolute, smooth; leaves in middle of stem often ovate and (nearly) blunt G. ramosa * Calyx lobes linear or lanceolate, margin occasionally strongly revolute, often with short-conical papillae; leaves in middle of stem ovate and acute or lanceolate G. bulgarica 15 Usually > 4 internodes; tall plants (5-)15-80(-109) cm; terminal pedicel and uppermost internode together forming 1030(-40)% heigth of plant; calyx often delicate G. amarella * Usually < 4 internodes; small plants (4-)6-20(-55) cm; terminal pedicel and uppermost internode together forming (30-)50-90 % heigth of plant; calyx often enlarged G. uliginosa (incl. G. anglica) 16 Corolla (10-)14-19(-22) mm; gynophor 0-1(-2) mm G. amarella-group 15 * Corolla (15-)22-33(-50) mm; gynophor (0-)1-7 mm G. germanica s.l. 17 17 Gynophor (0-)0.1-0.5(-0.8) · as long as calyx tube; Corolla (1.1-) 1.5-2.0(-2.3) · as long as calyx G. rhaetica * Gynophor (0.4-)0.6-1.1(-1.4) · as long as calyx tube; Corolla (1.6-) 1.9-2.6(-3.5) · as long as calyx G. germanica s.str. Additional key for taxa with intermediate characters Plants without long-conical or longcylindrical papillae; the calyx lobes are often narrowly-lanceolate, approaching linear; sinuses somewhat intermediate, more acute than obtuse; often within one individual both lanceolate and linear lobes as well as acute and obtuse sinuses were found. 1 1 or 2 calyx lobes usually much larger than the others, (1.1-)1.5-3.3(-3.5) · as long as tube, with short-conical papillae G. insubrica * Calyx lobes subequal, (1.0-)1.3-2.0(-2.4) · as long as tube, with short conical papillae or smooth 2 2 Calyx lobes usually linear or narrowlylanceolate; corolla (21-)25-32(-35) mm G. bohemica * Calyx lobes often broadly-lanceolate; corolla (22-)29-37(-47) mm G. stiriaca This research was supported by the Austrian Science Fund (FWF, P15346-B03). We further want to thank: A. Alegro, N. Boscaiu, C. Dobeš, F. Gruber, M. Fischer, M.A. Fischer, H. Niklfeld, M. Onete, F. Prochazka, G. M. Schneeweiss, P. Schönswetter, L. Schratt-Ehrendorfer, E. Sinn, A. Tribsch, and B. Wallnöfer for various help in collecting; the herbaria GZU, LI, M, NMGW, PRC, RUEB, TSB, UDM, W, and WU for supplying vouchers via loan or during visits; T.C.G. Rich for comments; H. Rainer for technical support; A. Kästner for the calyx drawings; T.F. Stuessy for reading through an earlier version of the manuscript. J. Greimler et al.: Morphology in European Gentianella 163 Appendix 1. Synopsis of European taxa of Gentianella section Gentianella with short distribution notes and comments on doubtful taxa. Bold letters indicate taxa that have been accepted or given for Europe (G. caucasea see note) in Pritchard and Tutin (1972); numbers indicate taxa or informal groups, that can be distinguished according to our observations 1 2 3 4 5 6 G. caucasea (Loddiges ex Sims) J. Holub G. columnae (Ten.) J. Holub G. crispata (Vis.) J. Holub G. amarella group G. amarella (L.) Börner incl. G. hispanica López Udias, Fabregat & Renob. G. uliginosa (Willd.) Börner incl. G. anglica (Pugsley) E.F. Warburg G. campestris group G. campestris (L.) Börner incl. G. hypericifolia (Murb.) Pritchard G. laevicalyx (Rohlena) Rohlena G. germanica group G. albanica (Jav.) J. Holub 7 8 9 10 11 12 13 14 15 16 17 G. anisodonta group incl. G. anisodonta (Borbas) A. & D. Löve incl. G. liburnica E. Mayer & H. Kunz incl. G. engadinensis (Wettst.) J. Holub G. aspera (Hegetschw. & Heer) Dostal ex Skalicky, Chrtek & Gill G. austriaca group incl. G. austriaca (A. & J. Kerner) J. Holub incl. G. fatrae (Borb) J. Holub incl. G. lutescens (Velen.) J. Holub G. bohemica Skalicky G. bulgarica (Velen.) J. Holub G. germanica (Willd.) E.F. Warburg G. insubrica (H. Kunz) J. Holub G. pilosa (Wettst.) J. Holub G. ramosa (Hegetschw.) J. Holub G. rhaetica (= Gentiana rhaetica A. & J. Kerner, not combined in Gentianella) G. stiriaca (= Gentiana stiriaca Wettst., not combined in Gentianella) Caucasus, ? (presence in European part of Turkey questioned) C. Appennini Balkans, S. Italy N., W. and C. Europe extending to S.E. Europe C. Spain N., N.C. and W. Europe S. England N. W. and C. Europe, Pyrenees Pyrenees ?, no voucher seen, diagnosis not seen; former Jugoslavia ?, only one voucher (two small plants) seen, diagnosis not seen; Albania S.C. and S.E. Alps, N.W. Dinarids S.E. Alps, N.W. Dinarids (E.)C. Alps N. and E.C. Alps, C. Europe N.E. Alps, E.C. Europe W.Carpathians E. Europe, N. Balkans C. Europe C. Balkans, S. Carpathians N.C. and W. Europe S.C. Alps S.E. Alps C. and S.W. Alps E. Alps N.E. Alps 164 J. Greimler et al.: Morphology in European Gentianella Appendix 2. Vouchers of population samples with collection number (deposited in WU), location, altitude, coordinates East/North, and collector. Additional vouchers (in italics) studied from public herbaria are given with data base accession number (as numbers on vouchers are missing in many cases), collector, and herbarium G. albanica Greece: 42, W. Greuter, M. G. amarella Austria: 347, A. Polatschek, W; 348, R. Seipka, W; 349, R. Seipka, W; 350, A. Polatschek, W; 351, A. Polatschek, W; 352, A. Polatschek, W. Czech Republic: 28a, Southern Bohemia, Sudslavice N Vimperk, 600 m, 1347’, 4905’, J. Greimler; 86, Southern Bohemia, Sudslavice N Vimperk, 600 m, 1347’, 4905’, J. Greimler; 91, Southern Bohemia, Jaroskov, 750 m, 1340’, 4907’, J. Greimler. 356, F. Kovar, W; 408, F. Svestka, WU. Danmark: 342, N. Jakobsen & J. Svensden, W. Germany: 43, F.G. Dunkel, M; 44, O. Angerer, M; 45, C. Correns, M; 46, P. Junge, M. Hungary: 407, V. Borbas, WU. Lettland: 354, K. Starcs, W; 398, K. Starcs, WU; 403, K. Starcs, WU. Norway: 341, B. Federley, W; 344, P.Wendelbo & A. Roesvik, W. Russia: 393, T.A. Teploukhoff, WU. Slovac Republic: 45, Mala Fatra, Chata Vratna S Terchova, 760 m, 1902’, 4912’, J. Greimler & G.M. Schneeweiss. 353, F. Svestka, W; 357, J. Chrtek & B. Deylova, W. Sweden: 345, K.H. Rechinger, W; 346, V. Samuelsson, W; 395, Enander, WU; 399, F. Vierhapper, WU. United Kingdome: 47, S.L. Jury, M; 48, M. Weigend, M; 197, R.&M. Gulliver, NMGW; 200, W.A. Shoolbred, NMGW; 201, W.A. Shoolbred, NMGW; 202, W.A. Shoolbred, NMGW; 203, F. Rose, NMGW; 206, E. Vachell, NMGW; 207, A.E. Wade, NMGW; 209, A.H. Trow, NMGW; 211, H.J. Dawson, NMGW; 212, R. Lewis, NMGW; 231, R.A. Boniface, NMGW; 232, C.E. Hubbard, NMGW; 233, T.C.G. Rich, A. McVeigh & J. Carey, NMGW; 234, T.C.G. Rich, A. McVeigh & J. Carey, NMGW; 235, T.C.G. Rich, A. McVeigh, NMGW; 355, E.K. Horwood, W; 400, F. Vierhapper, WU. G. anisodonta Austria: 53, Salzburg, Schaldminger Tauern, W Steirischer Kalkspitze, 2280 m, 1337’, 4717’, J. Greimler; 55, Salzburg, Radstädter Tauern, Zauchensee, 1500 m, 1327’, 4716’, J. Greimler; 81, Salzburg, Hohe Tauern, Goldberggruppe, Frauenkogel, 2200 m, 1310’, 4710, G.M. Schneeweiß; 84, Salzburg, Hohe Tauern, Goldberggruppe, Heukareck, 2100 m, 1310’, 4718’, G.M. Schneeweiß. Croatia: 59, Gorski-Kotar mountains, Snjeznik, 1450 m, 1435’, 4526’, A. Alegro. Italy: 42, Sondrio, Alpi Orobie, Passo San MarcoMonte Azzarini, 2200, 0940’, 4603’, L. Schratt-Ehrendorfer. 459, H. Becker, LI; 468, L. Poldini, TSB; 469, L. Poldini, TSB; 470, L. Poldini, TSB. G. aspera Austria: 14, Oberösterreich, Totes Gebirge, Hinterstoder, 650 m, 1407’, 4742’, J. Greimler; 68, Steiermark, Totes Gebirge, Bad Aussee, Loser, 1550 m, 1347’, 4740’, J. Greimler; 82, Salzburg, Hohe Tauern, Goldberggruppe, Plattenberg, 1650 m, 1257’, 4709’, G.M. Schneeweiß; 83 Salzburg, Hohe Tauern, Glockner-Gruppe, Fusch, 1900 m, 1247’, 4713’, G.M. Schneeweiß. 50, Arnold, M; 51, F. Schuhwerk, M; 53, J. Sellmair, M; 54, J. Sellmair, M; 55, W. Freiberg, M; 56, E. Dörr, M; 57, J. Koch, M; 58, J. Koch, M. Czech Republic: 17, Southern Bohemia, Kocelovice, 450 m, 1350’, 4928’, J. Greimler. Germany: 59, Arnold, M; 60, F. Vollmann, M; 61, Arnold, M; 62, Arnold, M; 63, Sendtner, M; 64, C.J. Mayer, M; 65, E. Hepp, M; 66, F. Vollmann, M; 67, G. Hegi, M; 68, Diehm, M; 69, H. Paul, M; 70, E. Dörr, M; 71, P. Eggensberger, M; 72, P. Eggensberger, M; 73, R. Urban, M; 74, H. Merxmüller, M; 75, W. Lippert, M; 76, R. Urban, M; 77, D. Lemp, M; 78, C. Niederbichler, M; 79, H. Roessler, M; 80, Eschelmüller & Dörr, M; 81, E. Dörr, M; 82, F. Schuhwerk, M; 83, F. Eberlein, M; 84, F. Eberlein, M; 85, F. Eberlein, M; 86, A. Lang, M; 87, F. Eberlein, M; 88, F. Eberlein, M; 89, F. Eberlein, M; 90, Dr. Wolley, M; 91, H. Löffelmann, M; 92, W. Kortenhaus, M; 93, W. Kortenhaus, M; 94, W. Kortenhaus, M; 95, A. Mayer, M; 96, G. Weisenbeck, M; 97, W. Lippert, M; 98, R. Grützmann, M; 99, E. Dörr, M; 100, C. Niederbichler, M; 101, J. Mayer, M; 102, R. Binsfeld, M; 103, F. Schuhwerk, M; 104, H.J. Tillich, M; 105, O. Angerer, M; 106, O. Mergenthaler, M; 107, G. Hegi, M; 108, F. Vollmann, M; 109, F. Vollmann, M; 110, Ernst, M; 111, Arnold, M; 112, H. Wild, M; 113, R. Urban, M; 114, W. Kortenhaus, M; 115, N. Müller, M; 116, E. Dörr, M; 117, E. Dörr, M; 118, F. Schuhwerk, M; 119, J. Mayer, M; 120, Dihm, M. J. Greimler et al.: Morphology in European Gentianella 165 Appendix 2 (continued) G. austriaca Austria: 3, Niederösterreich, Raxplateau, 1610 m, 1545’, 4744’, J. Greimler; 4, Niederösterreich, Schneeberg, 1100 m, 1550’, 4744’, J. Greimler; 5, Niederösterreich, Gutensteiner Alpen, Pernitz, 470 m, 1556’, 4753’, J. Greimler; 6, Niederösterreich, Wienerwald, Lindabrunn, 360 m, 1609’, 4755’, J. Greimler; 7, Niederösterreich, Wienerwald, Gaaden, 350 m, 1613’, 4803’, J. Greimler; 8, Steiermark, Prein/ Rax, above Preiner Gscheid, 1350 m, 1543’, 4741’, J. Greimler; 9, Burgenland, Seewinkel, Weiden am See, 120 m, 1651’, 4754’, J. Greimler; 44, Niederösterreich, Wiener Becken, Moosbrunn, 180 m, 1626’, 4800’, J. Greimler. Hungary: 58, Günser Gebirge, Bozsok SE Köszeg, 380 m, 1629’, 4720’, J. Greimler. G. bohemica Austria: 461, J. Kerner, GZU; 462, J. Kerner, GZU. Czech Republic: 15, Southern Bohemia, Onsovice, 650 m, 1346’, 4907’, J. Greimler; 16, Southern Bohemia, Javornik NW Vimperk, 900 m, 1339’, 4908’, J. Greimler. 306, L. Krajcr, PRC; 307, M. Protiva, PRC; 308, R. Schreiber, PRC; 309, J. Simak, PRC; 310, A. Matatko, PRC; 311, J. Diener, PRC; 312, B. Vopravil, PRC; 313, P. Hora, PRC; 314, Sourek-Sadova, PRC; 315, M. Jerist, PRC; 316, S. Hejny, PRC; 318, J. Obdrzalek, PRC; 319, Rohlena, PRC; 320, J. Rohlena, PRC; 321, J. Rohlena, PRC; 322, ?, PRC. G. bulgarica Albania: 421, Dimonie, WU; 423, I. Dörfler, WU. Bulgaria: 420, I.K. Urumoff, WU; 422, J. Wagner, WU; 424, J. Wagner, WU; 425, St. Gheorghieff, WU; 426, Velenovsky, WU. Romania: 76-79, Eastern Carpathians, Muntii Bucegi, Sinaia, 1950-2210 m, 2528’, 4524’, J. Greimler & C-G. Jang. G. campestris Austria: 127, F. Schuhwerk, M. France: 155, L. Giraudias, M. Spain: 128, P. Montserrat, M; 129, A. Meebold, M; 130, P. Garin, M; 131, P. Montserrat, M. Switzerland: 25, Graubünden, Maloja SW St. Moritz, 2400 m, 0940’, 4625’, J. Greimler; 30, Graubünden, Pass dal Fuorn (Ofenpaß), 2200 m, 1018’, 4639’, J. Greimler; 89, Luzern, Alpnachstad-Pilatus, 2050 m, 0816’, 4657’, J. Greimler. 126, H. Hertel, M. G. caucasea Georgia: 60, Chewi, Cauacasus, Kasbegi, 2040, 4446’, 4234’, C.-G. Jang; 62, Chewi, Cauacasus, Mt. Kasbek, 2430, 4437’, 4240’, C.-G. Jang; 99, Kartli, Trialetis kedi, Didgori, 1700 m, 4427’, 4145’, P. Schönswetter & A. Tribsch; 100, Chewi, Cauacasus, Kasbegi, 1900 m, 4440’, 4239’, P. Schönswetter & A. Tribsch. G. columnae Italy: 47, Abruzzo, ĹAquila-Gran Sasso, 1760 m, 1340’, 4225’, J. Greimler & B. Wallnöfer; 96, Umbria, Perugia, Monte Sibillini, NE Casteluccio, 1250 m, 1315’, 4251’, J. Greimler & B. Wallnöfer; A3, Abruzzo, Gran Sasso, Mte. Tremoggia, 2100 m, 1342’, 4226’, G.M. Schneeweiß & P. Schönswetter. G. crispata Bosnia: 446, E. Brandis, WU; 447, F. Vierhapper, WU; 448, K. Maly, WU; 449, H. Raap, WU; 450, F. Fiala, WU; 451, T. Pichler, WU; 452, F. Fiala & G. Beck, WU; 453, F. Fiala, WU; 454, H. Handel-Mazzetti, WU; 455, S. Murbeck, WU; 456, H. Handel-Mazzetti & E. Janchen, WU; 457, H. Handel-Mazzetti & E. Janchen, WU. G. engadinensis Italy: 29, Alto Adige, Ortler-Alpen, Sulden, 1950 m, 1035’, 4631’, J. Greimler; 51, Alto Adige, 1900 m, Ortler-Alpen, Trafoi, 1032’, 4632’, J. Greimler; 97, Sondrio, Alpi Retiche, Lago di S. Giacomo, 1950 m, 1015’, 4633’, L. Schratt-Ehrendorfer. Switzerland: 98, Graubünden, Pass dal Fuorn (Ofenpaß), 2100 m, 1018’, 4640’, P. Schönswetter. G. fatrae Slovac Republic: 43, Mala Fatra, Mala Fatra, Velky Krivan, 1670 m, 1902’, 4911’, J. Greimler & G.M. Schneeweiß; 38, Velka Fatra, Velka Fatra, Tlsta E Blatnica, 1350 m, 1858’, 4856’, J. Greimler & G.M. Schneeweiß. 166 J. Greimler et al.: Morphology in European Gentianella Appendix 2 (continued) G. germanica (Alps) Austria: 22, Tirol, Igls-Patscherkofel, 2000 m, 1127’, 4713’, J. Greimler & C. Dobes; 40, Tweng, 1250 m, 1335’, 4712’, J. Greimler; 49, Steiermark, Rottenmanner Tauern, Scheibel-Alm, 1650 m, 1426’, 4726’, J. Greimler; 50, Steiermark, Rottenmanner Tauern, Großer Bösenstein, 1950 m, 1425’, 4726’, J. Greimler; 54, Salzburg, Schladminger Tauern, Obersee, 1880 m, 1336’, 4717’, J. Greimler; 56, Steiermark, Wölzer Tauern, Niederer Zinken - Kleiner Zinken, 2150 m, 1421’, 4716’, P. Schönswetter; 69, Salzburg, Schladminger Tauern, Preber, 1600 m, 1352’, 4711’, C.-G. Jang; 70, Kärnten, Hohe Tauern, Goldberggruppe, Fragant, 1850 m, 1301’, 4657’, C.-G. Jang; 71, Tirol, Ötztaler Alpen, Vent, 2000 m, 1054’, 4651’, J. Greimler; 72, Tirol, Ötztaler Alpen, Rofen, 2450 m, 1053’, 4652’, J. Greimler; 121, Salzburg, Hohe Tauern, Sportgastein, Nassfeld, 1600 m, 1303’, 4703’, F. Gruber. 458, H. Wittmann & P. Pilsl, LI; 471, A. Kerner, WU; 472, A. Pernhoffer, WU. Italy: 23, Alto Adige, Ötztaler Alpen, Plawenn, 2050 m, 1035’, 4646’, J. Greimler & C. Dobes. Switzerland: 24, Graubünden, Fuldera, 1620 m, 1022’, 4637’, J. Greimler & C. Dobes. G. germanica (outside Alps) Belgium: 325, M. Crepin, PRC; 326, A. Hardy, PRC; 327, E. Peaubert, PRC; 409, A. Hardy, WU. Czech Republic: 323, J. v. Sterneck, PRC; 324, K. Hosic, PRC. France: 258, F. Rose, NMGW; 328, A. Schmidely, PRC; 384, H. Caron, W; 389, A. Huguenin, W; 391, Abbe F. Riguet, W; 392, C. Billot, W. Germany: A27, Sachsen-Anhalt, Trebitz near Kloschwitz, 100 m, 1145’, 5132’, J. Greimler. 135, Dihm, M; 136, F. Schuhwerk, M; 137, F. Schuhwerk, M; 138, E. Dörr, M; 141, W. Freiberg, M; 142, W. Freiberg, M; 143, W. Freiberg, M; 144, W. Freiberg, M; 145, Ruppert, M; 146, W. Freiberg, M; 147, W. Freiberg, M; 148, W. Freiberg, M; 149, J. Koch, M; 150, J. Koch, M; 151, W. Freiberg, M; 152, W. Freiberg, M; 153, W. Freiberg, M; 366, A.Vocke, W; 367, J. Bornmüller, W; 368, J. Bornmüller, W; 369, Hausknecht, W; 370, T. Vogel, W; 371, T. Vogel, W; 372, Westram, W; 373, W. Freiberg, W; 374, G. Sennholz, W; 375, Girth, W; 376, C.J. Mayer, W; 377, Girth, W; 378, H. Neumann, W; 380, G. Sennholz, W; 381, H. Schulz, W; 410, F. Wirtgen, WU; 411, J. Bornmüller, WU; 412, A. Hayek, WU; 413, Sagorski, WU. Switzerland: 87, Jura mountains, Dittingen S Basel, 350 m, 0731’, 4726’, J. Greimler; 88, Jura mountains, Diegten SE Basel, 550 m, 0749’, 4725’, J. Greimler. United Kingdome: 276, R.A. Boniface, NMGW; 277, W.H. Griffin, NMGW; 280, E. Vachell, E. Knowling & J. Davy, NMGW; 281, J. E. Lousley, NMGW; 282, A. McVeigh & J. Carey, NMGW; 283, T.C.G. Rich & A. McVeigh, NMGW; 292, A. McVeigh & J. Carey, NMGW; 294, A. McVeigh & J. Carey, NMGW; 295, A. McVeigh & J. Carey, NMGW; 296, A. McVeigh & J. Carey, NMGW. G. insubrica Italy: 27, Lombardia, Monte Generoso 1680 m, 0903’, 4556’, J. Greimler & C. Dobes. Switzerland: 463, W. Koch, RUEB; 464, W. Lüdi, RUEB; 465, Jaquet, RUEB; 466, J. Coaz, RUEB; 467, W. Koch, RUEB. G. liburnica Slovenia: 337, G. Sauli, TSB. G. lutescens Poland: 32, Krakow, Vysokie Tatry, Tatry Zachodnie, 1400 m, 1959’, 4915’, P. Schönswetter & A. Tribsch; 34, Krakow, Vysokie Tatry, Tatry Zachodnie, 1950 m, 1954’, 4914’, P. Schönswetter & A. Tribsch. Romania: 73, Eastern Carpathians, Piatra Craiului, Zarnesti, 800 m, 2517’, 4534’, J. Greimler & C-G. Jang; 75, Eastern Carpathians, Piatra Craiului mica, 1710 m, 2516’, 4533’, J. Greimler & C-G. Jang. 475, J. Römer, WU; 476, Porcius, WU; 477, A. Rehmann, WU; 478, A. Scherfel, WU. G. pilosa Italy: 18, Friuli-Venezia Giulia, Alpi Giulie, Cave del Predil, 980 m, 1334’, 4626’, J. Greimler & C-G. Jang; A4, Veneto, Prealpi Carniche, Mte. Tiarfin, 1235’, 4628’, G.M. Schneeweiß & P. Schönswetter. 479, I. Dörfler, WU; 480, O. Krebs, WU; 481, M. Statzer, WU; 482, Petter, WU; 483, Ressmann, WU; 484, I. Dörfler, WU; 485, M.& I. Dörfler, WU; 486, S. Rizzardini, UDM; 487, E. Tomasi, UDM; 488, F. Martini, UDM. J. Greimler et al.: Morphology in European Gentianella 167 Appendix 2 (continued) G. ramosa Italy: 427, H. Handel-Mazzetti, WU; 431, G.B. Traverso, WU;436, Muret, WU; 439, R. Wettstein, WU; 440, R. Wettstein, WU. Switzerland: 90, Gotthardgebiet, Oberalp, 2040 m, 0846’, 4640’, J. Greimler; 26, Graubünden, Maloja SW St. Moritz, 2100 m, 0941’, 4624’, J. Greimler. 428, H. Handel-Mazzetti, WU; 429, Wolf, WU; 430, H. Handel-Mazzetti, WU; 432, F.O. Wolf, WU; 433, R. Wettstein, WU; 434, K. Ronniger, WU; 435, Favrat, WU; 437, F. Vierhapper, WU; 438, Favrat, WU; 441, H. Handel-Mazzetti, WU; 442, H. Handel-Mazzetti, WU; 443, H. Handel-Mazzetti, WU; 444, H. Handel-Mazzetti, WU; 445, R. Wettstein, WU. G. stiriaca Austria, all Steiermark: 12, Hochschwab, Seetal, 1050 m, 1513’, 4737’, J. Greimler; 13, Ennstaler Alpen, Lugauer, 1530 m, 1442’, 4733’, J. Greimler; 48, Hochschwab, Aflenzer Staritzen, 1350 m, 1516’, 4738’, J. Greimler; 52, Schneealpe, Neuberg, 1080 m, 1536’, 4740’, M.A. Fischer; 57, Ennstaler Alpen, Johnsbach, 700 m 1435’, 4733’, J. Greimler. 460, A. Lonsing, LI; 473, J. Nevole, WU; 474, A. Klammerth, WU. G. uliginosa Finnland: 414, H. Lindberg, WU; 415, H. 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Barbara Hermanowski, Department 169 of Ultrastructure Research and Palynology. All: Institute of Botany, University of Vienna, Rennweg 14, A-1030 Vienna, Austria.