Exudate #Avonoid Aglycones in the Alpine Species of Achillea Sect
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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 includ- ing 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 avail- able 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