Exudate #Avonoid Aglycones in the Alpine Species of Achillea Sect

Biochemical Systematics and Ecology 29 (2001) 149}159

Exudate #avonoid aglycones in the alpine species of Achillea sect. Ptarmica: Chemosystematics of A. moschata and related species (Compositae}Anthemideae) Karin M. Valant-Vetschera! *, Eckhard Wollenweber"

!Institut fu( r Botanik der Universita( t Wien, Rennweg 14, A-1030 Wien, Austria "Institut fu( r Botanik der TU Darmstadt, Schnittspahnstrasse 3, D-64287 Darmstadt, Germany Received 23 November 1999; received in revised form 15 February 2000; accepted 14 March 2000

Abstract

In completion of our studies on Achillea exudate #avonoids, 11 alpine species of Achillea sect. Ptarmica were analyzed for their aglycone pro"les. The study focuses on species commonly associated with A. moschata. The major #avonoid constituents found in exudates of most taxa were 6-hydroxy#avonol 3,6,4-trimethyl ethers, except in A. ageratifolia and its subspecies, which are characterized by the accumulation of the 3,6,7-trimethoxy and 6-hydroxy#avones. Infraspeci"c variation was particularly high in A. abrotanoides and A. moschata. Results are discussed in relation to published data for related species and within the context of evolutionary aspects in the genus Achillea. The 3,6,4-trimethoxy substitution is regarded as a basic chemical trend within the genus Achillea. Geographical and ecological aspects are brie#y addressed, and a summary on known exudate aglycone composition of species from all sections of Achillea is included. ( 2001 Elsevier Science Ltd. All rights reserved.

Keywords: Achillea sect. Ptarmica; Alpine taxa; Compositae; Exudate #avonoid aglycones; Flavone- and #avonol methyl ethers; Chemosystematics

* Corresponding author. Tel.: #43-1-4277-54102; fax: #43-1-4277-9541. E-mail addresses: [email protected] (K.M. Valant-Vetschera), [email protected] mstadt.de (E. Wollenweber).

0305-1978/01/$- see front matter ( 2001 Elsevier Science Ltd. All rights reserved. PII: S 0 3 0 5 - 1 9 7 8 ( 0 0 ) 0 0 0 3 3 - 8 150 K.M. Valant-Vetschera, E. Wollenweber / Biochemical Systematics and Ecology 29 (2001) 149}159

1. Introduction

Achillea sect. Ptarmica (Mill.) W.D.J. Koch comprises about 30 species, depending on delimitation. The major feature of this section is the tendency to form large capitula with ligulae numbering 8}20 or more, and the general lack of transversely oriented primary lea#ets. Basically, the section may be divided into two larger parts: the species related to A. ptarmica L., and a larger group of alpine species. Both groups are morphologically and ecologically well de"ned (see Heimerl, 1884). Apart fromHeimerl (1884), no further attempts have been made towards infrasectional classi"cation including all species of this section. Considerable variation in morphological characters may be found within the alpine taxa (Heimerl, 1884; Valant-Vetschera, 1981), leading sometimes to di!erent concepts of species delimitation (Richardson, 1976; Pignatti, 1982). In earlier studies, several groups within the alpine taxa were distinguished on the basis of #avonoid glycoside pro"les and morphologial characters (Valant, 1978; Valant-Vetschera, 1981). Thus, species were grouped around the following taxa: A. atrata L., A. moschata Wulf. including A. clavennae and A. ageratifolia (Sibth. and Smith) Boiss., and the small groups such as A. fraasii Schultz-Bip., and A. lingulata Waldst. & Kit. These groups are in most parts in accordance with later concepts (FranzeH n, 1986, 1988a, 1991; Pignatti, 1982), but they do not represent taxonomically recognized units. Other phytochemical investigations mostly relate to polyacetylenes and terpenoids. Roots of A. ageratifolia subsp. serbica have been intensively studied, to yield new isobutylamides (Greger et al., 1983) and pyrrole amides (Greger et al., 1987a), while A. chamaemelifolia Poir. of the A. erba-rotta group produced simple alkamides (Greger et al., 1987b). Achillea abrotanoides (Vis.) Vis. of the same group was analyzed for its volatile constituents with a view to its replacing A. moschata Wulf. and other species used in production of herb liqueur (Bicchi et al., 1988; Hanlidou et al., 1992). A survey reports on essential oils and chromosome numbers of A. erba-rotta, A. moschata (of the same group) and A. nana L. (of the A. atrata-group; Ma!ei et al., 1989). Sesquiterpene lactones were identi"ed from A. abrotanoides of the A. moschata group (Stefanovic et al., 1989). Volatiles and other chemical constituents of A. moschata are listed by Duke (1992). The presence of chamazulene in this species may be responsible for its various uses in ethnopharmacology. Detailed #avonoid-distribution studies at the populational level, comprising both aglycones and glycosides have been carried out on taxa associated with A. ageratifolia and A. clavennae, respectively (FranzeH n, 1988b). Earlier studies revealed the presence of free aglycones in the exudates of A. abrotanoides, A. moschata, A. ageratifolia (Valant-Vetschera and Wollenweber, 1985), and of A. umbellata Sibth. and Smith (Wollenweber et al., 1987). The aglycone pro"les of the species related to A. clusiana Tausch ("A. atrata group) were reported recently (Valant-Vetschera and Wollen- weber, 1996a). In completion of our systematic survey on Achillea #avonoid aglycones (Valant-Vetschera and Wollenweber, 1996b, 1999), we wish to report on the pro"les of the remaining alpine species of sect. Ptarmica. Infraspeci"c variation of aglycone

The infrasectional concept of Heimerl (1884) does not follow the acknowledged nomenclatural rules and is therefore nomenclaturally invalid. Hence, its taxonomic application would be incorrect. K.M. Valant-Vetschera, E. Wollenweber / Biochemical Systematics and Ecology 29 (2001) 149}159 151 composition and the signi"cance of accumulation trends for infrageneric relationships form the focus of this publication.

2. Material and methods

Plant material originates from various collections, including plants from natural sites and from cultivation. Voucher specimens have been deposited in the Herbarium of the Institute of Botany, University of Vienna (WU) and as indicated.

List of collections A. ageratifolia Sibth. & Smith (1) Greece, Olympus (Sorger, 1960, WU) A. ageratifolia subsp. aizoon (Griseb.) Heimerl (2) Greece, Mt. Siniatsikon, Franzen 227, 1980, WU (3) Greece, Olymp, Zbuzek 1987, WU A. ageratifolia subsp. serbica (Nyman) Heimerl (4) Bosnia, near Visegrad, K#H Vetschera, 1982, WU (5) Cult. Bot. Garden, Univ. Vienna A. umbellata Sibth. & Smith (6) Greece, Evvoia, Ep. Halkidas, Mt. Dirkis, Franzen and Baden No. 34/1980, WU (7) Bulk material of same collection A. pindicola Hausskn. (8) Greece, Evritania, Mt. Timfristos, Franzen and Baden no. 94/1980, WU A. chamaemelifolia Poir. (9) Cult. Bot. Garden, Univ. Vienna (10) France, PyreH neH es Orientales: Casteill. Neyrout, 2.7.1911, W A. abrotanoides (Vis.) Vis. (11) Bosnia, Mt. Maglic near Tientiste, K. & H. Vetschera 1982, WU (12) Bosnien, Treskavica; Ginzberger 1929 (13) Yugoslavia, E-Montenegro, Pivljan, E Kolasin, Janchen 1916, WU (14) Greece, Ionninon, Mt. Tim", Franzen and Akeroyd no. 149/1980 (15) Cult. Bot. Garden, Univ. Vienna (16) N-Albania, Distr.Luna, DoK r#er No. 922, 1918, WU (17) Cult. from 13 A. ambrosiaca (Boiss. and Heldr.) Boiss. (18) Cult. Bot. Garden, Univ. Vienna A. fraasii Schultz-Bip. (19) Greece, Ioanninon, Mt. Tim", Franzen and Akeroyd no. 194/1980, WU A. erba-rotta All. (20) Italy, Alpi di Facetoo Juglio, Rostan 1880 (21) France, Savoie, Sillot 1893 A. moschata Wulf. (22) Austria, Tyrol, OG tztal, Vitek WU (23) Italy, S-Tyrol, Suldental, Valant 1976, WU (24) Austria, Styria. Rottenmanner Tauern, Statzer 1894, WU 152 K.M. Valant-Vetschera, E. Wollenweber / Biochemical Systematics and Ecology 29 (2001) 149}159

(25) Switzerland, Furkapa{,K.#H. Vetschera 1982, WU A. moschata var. calcarea Huter (26) Italy, Calabria, Castrovillari, Huter s.d., WU (27) Italy, Lucania. Abriola, Serra di Monteforte. Gavioli No. 2968, 1925 (28) Italy, Valle d'Aosta, Coumayeur, Ferrari 1904, WU (29) Italy, Calabria, Castrovillari, Huter, Porta and Rigo No 379, 1877 WU A. rupestris Huter (30) Italy, Calabria, Mte Pollino, Rigo No. 417, 1898, WU (31) Italy, Calabria, Mte Pollino, Huter, Porta and Rigo, No 467/1877 WU A. clavennae L. (32) Austria, Lower Austria, Schneealpe, K#H Vetschera, 1983, WU (33) Austria, Lower Austria, Reichenau, Schefzik 1961, GJO (34) Austria, Lower Austria, Schneeberg, Wettstein 1891, WU (35) Croatia, Plesevica near Plitvice, Ginzberger 1909, WU

Air-dried aerial parts excluding in#orescences were brie#y rinsed with acetone at room temperature to dissolve the exudate material.After evaporation of the solvent, the residue was chromatographed over Sephadex LH-20, eluted with methanol, to separate the #avonoids from the dominating terpenoids. Individual #avonoids were identi"ed in relevant fractions by co-chromatography with authentic samples available in E.W.'s lab. Fractions were monitored on TLC and comparisons with markers were performed on silica with toluene/ MeCOEt (9 : 1), or with toluene/MeCOEt/ MeOH (12 : 5 : 3) and with toluene/ dioxane/ glacial acetic acid (18 : 5 : 1), and on polyamide with toluene/petrol 100}140/ MeCOEt/ MeOH (12 : 6 : 2 : 1) and toluene/ dioxane/ MeOH (8 : 1 : 1). Chromatograms were viewed under UV before and after spraying with `Natursto!reagenz Aa (NA). For additional structural con"rma- tion, UV-spectra and mass spectra were recorded when necessary.

3. Results and discussion

Within sect. Ptarmica, the production of free #avonoid aglycones in leaf exudates was observed mainly in the alpine taxa, whereas the species grouped around A. ptarmica rarely accumulated such compounds (Valant-Vetschera and Wollenweber, 1999). However, a few alpine taxa did not yield exudate aglycones, as far as could be judged from chromatographic analysis. This concerns particularly the odorless species of the A. atrata group, i.e. A. atrata (Valant-Vetschera and Wollenweber, 1996a), and the Italian endemics A. mucronulata Bertol. and A. barrelieri Ten.. Similar results may be expected from A. oxyloba (DC.) Schultz-Bip. and A. schurii Schultz-Bip. of the same group. As had earlier been observed, there is a strong correlation between production of free aglycones and terpenoids (Wollenweber and Valant-Vetschera, 1996; Valant-Vetschera and Wollenweber, 1996a). However, bulk material might yield traces of free aglycones in all these odorless taxa. A survey of #avonoid aglycone variation within 11 species associated with A. moschata is given in Table 1. In contrast to earlier arrangements (Valant, 1978; K.M. Valant-Vetschera, E. Wollenweber / Biochemical Systematics and Ecology 29 (2001) 149}159 153

Valant-Vetschera, 1981), A. fraasii was also included in this group (see also FranzeH n, 1988b). In Table 1, species are arranged according to their aglycone pro"le with consideration of infraspeci"c concepts. Material of A. barbeyana Heldr. and Heimerl associated with this part of sect. Ptarmica was not available for analysis. Similarly, known hybrid taxa have not been included in this study, due to lack of material. Infraspeci"c variation of aglycone pro"les is often notable (Table 1), and in accordance with earlier studies on alpine species from Greece (A. ageratifolia, A. clavennae, A. umbellata, A. pindicola, A. ambrosiaca and A. fraasii; FranzeH n, 1988a). We therefore refrained from investigations on a larger scale in these taxa. Within populations of A. ageratifolia, a remarkable lack of correlation between geographical distribution and #avonoid variation has been observed (FranzeH n, 1988a). Similar results were obtained from cultivated and natural popluations of A. chamaemelifolia, with the pro"le of a cultivated collection (no. 9) being less diversi"ed than that of the much older, unrelated herbarium collection (no. 10 in Table 1). Similarly, quantitative and qualitative variation was apparent in exudates of A. abrotanoides (nos. 12}17 in Table 1). Material from natural sites (no. 11 in Table 1) did not correlate with cultures grown from its seeds (no. 17 in Table 1), and the overall degree of variation was quite high. There may be some correlation with observed variations in essential oil composition (Hanlidou et al., 1991). A similar degree of variation was observed in populations of A. moschata, with exception of the endemic var. calcarea (nos. 25}28). Also, the degree of variation was quite low in samples of A. clavennae (nos. 32}35) from Central Europe and Croatia, although FranzeH n (1988b) had observed inconsistencies in populations from Greece. Morphological variation as expressed by di!erent concepts of species delimitation (Richardson, 1976; FranzeH n, 1991; Valant-Vetschera, 1981) is apparently unrelated to aglycone variation. Generally, the pro"les appear to be rather simple in terms of substitution patterns and with regard to the predominance of #avonols. 6-Hydroxy#avonol 3,6,4-trimethyl ethers occurred throughout the group, except in A. ageratifolia and its subtaxa. By comparison, the A. ageratifolia assemblage accumulated 3,6,7-trimethoxy isomers and simple #avone derivatives which distinguish these taxa from the rest of the group (Table 1). Only quercetagetin 3,6,7-trimethyl ether occurs in traces in one population each of A. ambrosiaca, A. moschata and A. clavennae. Thus, this substitution trend is of limited distribution in this group. More highly substituted #avonols were observed in exudates of A. abrotanoides populations as well as in A. ambrosiaca and in one population of A. moschata (see Table 1). These data are in accordance with earlier reports on other populations of these taxa (Valant-Vetschera and Wollenweber, 1985; Wollenweber et al., 1987; FranzeH n, 1988b). Despite the infraspeci"c aglycone variation observed, some accumulation trends can be used to distinguish taxa and to interprete relationships. In particular, A. ageratifolia and subspecies (Greece, S-Yugoslavia) are well separated from the remaining taxa, both by aglycone chemistry and morphological features. Achillea umbellata, A. pindicola (both from Greece) and A. chamaemelifolia (Pyrenean Mountains) occupy an intermediate position, sharing 6-hydroxykaempferol 3,6 dimethyl ether with A. ageratifolia and 6-hydroxy#avonol 3,6,4-trimethyl ethers with the remaining taxa (Table 1). Relatively complex aglycone patterns are found in both A. abrotanoides 154 ..Vln-eshr,E Wollenweber E. Valant-Vetschera, K.M.

Table 1 Distribution of exudate aglycones in species of Achillea sect. Ptarmica!

Achillea Scu-6 6Lu-6 6K- 6K- 6K- 6K- 6Q- 6Q- 6Q- 6Q- 6Q- K 3,6 3,6,7 3,6,4 3,6,7,4 3,6 3,6,7 3,6,4 3,6,7,4 3,6,7,34 /

1. ageratifolia ᭜᭜ 149 (2001) 29 Ecology and Systematics Biochemical subsp. ageratifolia 2. ageratifolia subsp. aizoon ᮀᮀ ᭜ 3. ageratifolia subsp. aizoon ᭜᭜ 4. ageratifolia subsp. serbica ᭜ # ᭜ 5. ageratifolia subsp. serbica ᭜᭜ 6. umbellata ᭜᭜ 7. umbellata # ᭜᭜# ᮀ 8. pindicola ᭜ 9. chamaemelifolia ᭜᭜ 10. chamaemelifolia # ᭜ ## 11. abrotanoides ᭜ 12. abrotanoides ᭜᭜ᮀ 13. abrotanoides ᭜᭜ 14. abrotanoides ᭜ # ᭜ # ᭜ 15. abrotanoides ᭜᭜᭜ 16. abrotanoides ᮀ ᮀᮀᮀ 17. abrotanoides ᮀᮀ }

18. ambrosiaca ᮀᮀ # ᮀᮀ 159 19. fraasii # ᭜᭜ 20. erba-rotta ᭜ᮀ 21. erba-rotta ᭜᭜ᮀ 22. moschata ᮀ᭜ ᭜ Wollenweber E. Valant-Vetschera, K.M. 23. moschata # ᭜᭜ 24. moschata ᭜᭜ ## 25. moschata ᮀ 26. moschata var. calcarea ᭜ 27. moschata var. calcarea ᭜ 28. moschata var. calcarea # ᭜ 29. moschata var. calcarea # ᭜ 30. rupestris ᮀ 31. rupestris ᮀ᭜ 32. clavennae ᭜ # ᭜ 33. clavennae ᭜᭜ 34. clavennae # ᭜

35. clavennae # ᭜ / iceia ytmtc n clg 9(01 149 (2001) 29 Ecology and Systematics Biochemical

!Scu-6"scutellarein 6-methyl ether (hispidulin), 6Lu-6"6-hydroxyluteolin 6-methyl ether (nepetin), 6K-3,6"6-hydroxykaempferol 3,6-dimethyl ether, 6K-3,6,7"6-hydroxykaempferol 3,6,7-trimethyl ether (penduletin), 6K-3,6,4"6-hyroxykaempferol 3,6,4-trimethyl ether, (3-methylbetuletol) 6K-3,6,7,4"6- hyroxykaempferol 3,6,7,4-tetramethyl ether, 6Q-3,6"quercetagetin 3,6-dimethyl ether (axillarin), 6Q-3,6,7"quercetagetin 3,6,7-trimethyl ether (chrysosplenol-D), 6Q-3,6,4"quercetagetin 3,6,4-trimethyl ether (centaureidin), 6Q-3,6,7,4"quercetagetin 3,6,7,4-tetramethyl ether (casticin), 6Q-3,6,7,34" quercetagetin 3,6,7,3,4-pentamethyl ether (artemetin), K"kaempferol 3,4-dimethyl ether. (᭜)major#avonoids, (ᮀ)minor#avonoids, (#)traces. } 159 155 156 K.M. Valant-Vetschera, E. Wollenweber / Biochemical Systematics and Ecology 29 (2001) 149}159

(Greece, S-Yugoslavia) and A. ambrosiaca (Greece). FranzeH n in his studies of the group (1986, 1988b) considered A. umbellata to be positioned within the A. clavennae group (Central Europe, Balkans), together with A. ambrosiaca, A. pindicola amd A. fraasii (S-Yugoslavia, Greece, Turkey). With exception of A. fraasii, the remaining taxa also share common accumulation tendencies of #avonoid glycosides (Valant-Vetschera, 1981). Taxa associated with A. erba-rotta (W-Alps) like A. moschata (W-Alps, Central Europe, Italy) incl. var. calcarea (Italian endemic) and A. rupestris (Italian endemic; Richardson, 1976) exhibit a more or less uniform #avonoid composition, with 6- hydroxy#avonol 3,6,4-trimethyl ethers dominating (Table 1). In this respect, they relate to the studied collection of A. clavennae from Central Europe. Similar results were obtained from populations of A. clavennae from the Balkans (FranzeH n, 1988a). The aglycone pro"le of A. fraasii suggests a$nities both to the A. erba-rotta and A. clavennae assemblage, although this species uniquely accumulates C-glycosyl#avones, which do not occur in the other taxa (Valant-Vetschera, 1981). Some species studied here are narrow endemics (e.g. A. moschata var. calcarea, A. rupestris, Table 1). Earlier, it was assumed that #avonoid pro"les of narrow endemics consist mainly of methylated aglycones (see also FranzeH n, 1988b). In the present study, endemism could not be correlated with #avonoid aglycone expression (compare the more widespread A. moschata in Table 1). Similarly, preferences for a certain geology (e.g. limestone versus silicate rocks) appears to have no in#uence on aglycone composition, as in A. moschata (silicate rocks) and its var. calcarea (limestone rocks; compare also Valant-Vetschera and Wollenweber, 1986). Di!erences in aglycone composition distinguish the A. moschata group from the species of the A. atrata- group, with exception of A. multixda (DC.) Boiss. (Valant- Vetschera and Wollenweber, 1996a). Whereas A. clusiana and A. nana L. accumulate 6-hydroxy#avonol 3,6,7-trimethyl ethers, A. multixda produces the 3,6,4-trimethoxy isomers instead. In this respect, A. multixda resembles most taxa of the presently studied group, such as A. clavennae, A. moschata, and A. abrotanoides. Morphologi- cally, there is no doubt as to the position of A. multixda within the A. atrata-group, which was indicated by their common accumulation trends of #avonoid glycosides (Valant-Vetschera, 1981). Finally, the pro"les of all alpine taxa studied so far in A. sect. Ptarmica,di!er markedly from that known from one species of the A. ptarmica group (A. sibirica; Valant-Vetschera and Wollenweber, 1999). This diversi"cation is in line with accumulation trends of #avonoid glycosides (Valant-Vetschera, 1985), indicating two well separated units within A. sect. Ptarmica which would merit taxonomic recognition.

4. Signi5cance of aglycone accumulation trends within Achillea

Some 57 species of the 100 or more species aligned to Achillea have been studied regarding infrataxon variability of aglycone pro"les. The earlier published results are summarized in Table 2 (incl. references), by grouping species according to botanical features in the context of the current sectional concept. The groups most probably represent closer related species; their taxonomic status, however, still needs to be Table 2 Wollenweber E. Valant-Vetschera, K.M. Summary on aglycone accumulation trends in studied groups of Achillea!

Flavonols Flavones Literature"

K Q K3Q3Q3K3Q3Q3K3Q3Q3Ap6LuAp6Lu6Ap6Lu6Ap6Lu6Lu6 36 36 67 67 63 64 64 6767676736774 3 4 74 74 73 3 4 4 4 4

Achillea sect. Ptarmica ptarmica gr. ᮀᮀ᭜ ᮀ 1 atrata gr. ᮀ᭜# ᮀᮀ ## # 2 moschata gr. ᮀ ### ᭜᭜# ᮀᮀ### This publication Achillea sect. Millefolium subsect. Millefoliatae millefolium gr. #### ᭜᭜ ## ᮀ #### ᭜ ## # 3 nobilis gr. ᮀᮀᮀᮀ# ᮀᮀ ᮀᮀᮀ#### ## 4 / iceia ytmtc n clg 9(01 149 (2001) 29 Ecology and Systematics Biochemical Achillea sect. Millefolium subsect. Filipendulinae bieberst.gr. ᭜᭜ᮀᮀ᭜ ## 5 gerberi gr. ## ᮀ ## ## 5 ageratum gr. ᮀ # ᭜ # ᭜ ####5 clypeolata gr. # ᮀ # ᭜ ## ᭜ ## 5 ochroleuca gr. ## ᭜ᮀ ## ᮀ # ᮀ # ᭜ ## ᭜ 5 Achillea sect. Santolinoidea ##ᮀ᭜ # ᭜ ## ᭜᭜᭜6 Achillea sect. Babounya ᭜ ###᭜ ###6 Achillea sect. Arthrolepis 2 Analysed species: no aglycones detected 7

!Favonols: K 3 6"6-hydroxykaempferol 3,6-dimethy lether; Q 3 6" 6-hydroxyquercetin ("quercetagetin) 3,6-dimethyl ether (axillarin); K 3 6 7"6-hydroxykaempferol 3,6,7-trimethyl ether (penduletin); Q 3 6 7"quercetagetin 3,6,7-trimethyl ether (chrysosplenol-D); Q 3 6 3" quercetagetin 3,6,3-trimethyl ether (jaceidin); K 3 6 4"6- hydroxykaempferol 3,6,4-trimethyl ether (3-methylbetuletol); Q 3 6 4"quercetagetin 3,6,4-trimethyl ether (centaureidin); K 3 6 7 4"6-hydroxykaempferol 3,6,7,4-tetramethyl ether;Q3673"quercetagetin 3,6,7,3-tetramethylether(chrysosplenetin);Q3674" quercetagetin 3,6,7,4-tetramethylether(casticin);Q3673 4"quercetagetin 3,6,7,3,4-pentamethyl ether (artemetin). Flavones: Ap6"6-hydroxyapigenin (" scutellarein) 6-methyl ether (hispidulin); Lu6"6-hydroxyluteolin 6-methyl ether (nepetin); Ap6 7"scutellarein 6,7-dimethyl ether (cirsimaritin); Lu 6 7"6-hydroxyluteolin 6,7-dimethyl ether (cirsiliol); Ap 6 4"scutellarein 6,4-dimethyl ether (pectolinarigenin); Lu 6 3

4"6-hydroxyluteolin 6,3,4 trimethyl ether (eupatilin); Ap 6 7 4"scutellarein 6,7,4-trimethyl ether (salvigenin); Lu 6 7 4"6-hydroxyluteolin 6,7,4-trimethyl ether (eupatorin); Lu } 159 673 4"6- hydroxyluteolin 6,7,34-tetramethyl ether. gr."group. Relative amounts: (᭜)majortrend;(ᮀ)minortrend;(#)traces. "Literature: (1) Valant-Vetschera and Wollenweber (1999); (2) Valant-Vetschera and Wollenweber (1996a); (3) Valant-Vetschera and Wollenweber (1988a); (4) Valant-Vetschera and 157 Wollenweber (1988b); (5) Valant-Vetschera and Wollenweber (1996b); (6) Valant-Vetschera and Wollenweber (1994); (7) Valant-Vetschera and Wollenweber, (unpublished). 158 K.M. Valant-Vetschera, E. Wollenweber / Biochemical Systematics and Ecology 29 (2001) 149}159 formally assessed. Similarly, the sectional arrangement warrants taxonomic reconsid- eration. Since not all taxa of Achillea accumulate exudate aglycones in detectable quantities, the mere presence of such aglycones may be perceived as a chemical character. This is true, e.g. for xerophytic species of sect. Arthrolepis (Table 2) or some species of the alpine A. atrata group (Valant-Vetschera and Wollenweber, 1996a). It should be assumed that species growing under similar ecological conditions (e.g. increased UV-raditation in alpine and xeric habitats) would react in a similar way to cope with environmental stress factors. Apart from these quantitative di!erences, qualitative di!erences between pro"les of alpine taxa (#avonol-dominated) and xerophytes (#avone dominated) were observed (Table 2). Both the systematic and ecological signi"cances of this observation are open to speculation. Apart from the strong tendency towards 6-methoxylation of #avonols and #avones, additional substitution patterns and their combinations characterized the groups listed in Table 2. Basic trends are represented by

E Predominant formation of 3,6,4-trimethyl ethers of 6-hydroxy#avonols: A. sect. Ptarmica (alpine species), A. sect. Millefolium subsect. Millefolium and subsect. Filipendulinae (p.p.). E Predominant formation of (polymethoxy)#avonols with 3-methylation: A. atrata group, A. biebersteinii group. E Predominant formation of (polymethoxy)#avones: A. sect. Santolinoidea; A. sect. Babounya. E Predominant formation of highly substituted #avonols and #avones (the latter partly higher substituted): species of the A. millefolium group and of the A. nobilis group; A. ochroleuca group.

Further aspects of #avonoid aglycone diversi"cation have been discussed in a preced- ing study on A. sect. Millefolium subsect. Filipendulinae (Valant-Vetschera and Wol- lenweber, 1996b). In particular, the predominance of 6-hydroxy#avonol 3,6,4- trimethyl ethers in many taxa of the Balkans and SE-Europe versus the predominance of 6-hydroxy#avone methyl ethers in those of Turkey indicates a correlation with the geographic evolution of Achillea. It is assumed that the production of mainly #avonols is a basic evolutionary trend in this genus, whereas the more derived taxa are characterized by pro"les rich in #avones. Thus, the origin of the genus Achillea in the East-Mediterranean region, where the genus clearly has its center of diversity (Heimerl, 1884; see also Valant-Vetschera, 1981), appears to be con"rmed by aglycone diversi"cation.

References

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