P1. Syst. Evol.

1,15, 193--201 (1984)

Plnnt V.stemntits und Eonlution

by Springer-Verlag 1984

Seed Morphology in Some European Aconites (Aconitum,

Ran unculaceae)
By Elsa M. Cappelletti and Livio Poldini

(Received July 18, 1983)

Key Words: Angiosperms, Ranunculaceae, Aconitum.--Seed morphology, seed coat SEM microcharacteristics, taxonomic importance, adaptative ecological significance. Abstract: The seed coat morphology, investigated in taxa representative of the main European groups of Aconitum, are in good agreement with the current taxonomy of the genus. The seed coat microcharacteristics (warty epidermal cells) are very constant. There is a trend for the reduction of longitudinal wings on the edges concomitant with the development of ridges and transverse wings on the faces. Another morphological progression leads from smooth to rugulose and eventually to transverse wing-bearing seed faces. A working hypothesis suggests an ecological adaptative significance to these changes.

Owing to their great constancy (stressed b y DAvis & HEYWOOD 1963), features of seed morphology potentially have great taxonomic value (STEBBINS 1974). This also applies to Aconitum and its infrageneric classification (GAYER 1909, TUTIN & MERXMULLER 1964, SEITZ 1969). We have further investigated the seed morphological features of the genus, taking into account seed coat microcharacteristics (by SEM), in an a t t e m p t to evaluate their taxonomic significance at the infrageneric as well as at the infraspecific level.

Materials and Methods Nomenclature within Aeonitum napellus L. s. 1. is according to SmTz (1969), and for the remaining groups according to TUTI~T& MERXMULLER(1964). Ripe seeds from specimens representative of all the main groups described by T~ZTIN& MEI~XMULLER(1964) within the genus Aconitum for Europe have been studied: 13"

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1. Aconitum anthora L.: wild specimens from the Conservatoire et Jardin Botaniques de la Ville de Gen~ve (G), Switzerland; 2. Aconitum napellus group: 2.1. A. napellus L. subsp, tauricum (WuLs.) GAYEa: wild plants from the Carnie Alps and Mr. Generoso (Lombard Pre-Alps), Italy; 2.2. A. napellus L. subsp, hians (RmcHE~m) GAYS~: cultivated plants from the Zaktlad Farmakologii, Pracownia Agrotechniki, Krakdw (Poland); 2.3. A. napellus L. subsp, neomontanum (WuLF.) GAYE~:wild plants from Val Sassina, Western Alps, Italy; 3. Aconitum variegatum group: 3.1. A. variegatum L.: wild plants from Mt. Nanos, Slovenia, Yugoslavia; 3.2. A. paniculatum LAM.: wild plants from the Carnic Alps, Italy; 3.3. A. angustifolium BEaNH.: wild plants from the Julian Alps and Pre-Alps, Italy. 4. Aconitum vulparia group: 4.1. A. vulparia REICgENB.:wild plants from the Conservatoire et Jardin Botaniques de la Ville de Gen~ve (G), Switzerland; 4.2. A. lamarckii Rmo~E~m: wild plants from the Carnic Alps, Italy. Voucher specimens--as far as available--have been deposited in the herbarium of the Institute of Botany, University of Trieste (TSB). The seeds were coated with gold in an Edwards S 150 A Sputter Coa.ter and observed under a Scanning Electron Microscope Cambridge Stereoscan 250 at the accelerating voltage of 25kV at the Centro Universitario Grandi Apparecchiature Scientifiche (CUGAS) of the University of Padova.
Results

The seeds of all the investigated A c o n i t u m species and subspecies are triangular and p y r a m i d a l . A base, at the center of which the region of the hilum is found, a n d three faces can be distinguished. On the three edges, longitudinal wings m a y be present. As regards the longitudinal wings, the following basic seed types were found: - presence of three longitudinal wings of c o m p a r a b l e size; -- presence of three longitudinal wings, one of which is distinctly more developed t h a n the other two; - presence of only one longitudinal wing; -longitudinal wings absent. Figs. 1-12.--Figs. 1 4 . Aconitum anthora. -- Fig. 1. seed with longitudinal wings of comparable size on the edges, x 15.--Figs. 2, 3. epidermal cells with roundtopped tubercles. Fig. 2. x 50; Fig. 3. x 150.--Figs. 4, 5. aligned tubercles on adjacent cells. Fig. 4. 50; Fig. 5. x 150.--Fig. 6. warty cell ornamentation, x 250. --Fig. 7. smooth cells on the top of longitudinal wings, x 150.--Fig. 8. smooth cells of the hilum region, 100. --Fig. 9. A. napellus subsp, hians: one longitudinal wing (right) more developed than the other two, x 20.--Figs. 101 l. A. napellus subsp, tauricum. -- Fig. 10. ridges on the faces, x 70.--Fig. 11. small transverse expansion, x 250.--Fig. 12. A. napellus subsp, hians: transverse grooves on the less developed longitudinal wings, x 20

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Figs. 1-12

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CAPPELLETTI & L. POLDI~:

The seed faces are either smooth (Fig. 1) or rugulose (Fig. 10) owing to the presence of ridges which sometimes extend into membranous expansions (transverse wings, Figs. 17-21). In the following a brief description of the seed morphology of all taxa studied is presented.
1. Aconitum anthora: the seeds are characterized by the occurrence of three longitudinal wings of comparable size on the edges (Fig. 1 ). The seed faces are smooth with only one round-topped tubercle occurring on every single rectangular-shaped epidermal cell (Figs. 2-3). Sometimes the tubercles occurring on adjacent cells are aligned (Fig. 4) forming a transversal pattern (Fig. 5). Generally, epidermal cells exhibit warty ornamentations (Fig. 6), which are missing only on the cells situated on the top of the longitudinal wings running along the edges (Fig. 7) and on the polygonal-outlined cells of the hilum region (Fig. 8). Quite similar epidermal cell microcharacteristics have been observed in all the other Aconitum species and subspecies studied. 2. Aconitum napellus group: seeds with three longitudinal wings, one of which is distinctly much more developed than the other two (Fig. 9). 2.1. A, napeIlus subsp, tauricum: seed faces with ridges more or less developed (depending on the specimen) (Fig. 10) and sometimes scarcely identifiable transverse expansions (Fig. 11). 2.2. A. napelIus subsp, hians: ridges on the seed faces expanded into more or less developed membranous transverse wings (Fig. 9). The two less developed longitudinal wings somewhat w a v y (Fig. 9), sometimes touched by the ridges, and with transverse grooves in that case (Fig. 12). 2.3. A. napeIIus subsp, neornontanum: numerous well marked ridges on the seed faces which extend to the less developed longitudinal wings and give rise to transverse grooves (Figs. 13-14).

Figs. 13-24.--Figs. 13-14. Aconitum napellus subsp, neomontanum: marked ridges on the faces and on the less developed longitudinal wings. --Fig. 13. 20; Fig. 14. 40.--Figs. 15 17. A. variegatum.--Fig. 15. seed with only one longitudinal wing (left), 15.--Fig. 16. large transverse wings on the face opposite to the longitudinal wing, 12. --Fig. 17. transverse wings, 40. --Fig. 18. A. paniculatum: seed with only one longitudinal wing (right) and transverse wings on the opposite face, 15.--Fig. 19. A. anguat~folium: very numerous transverse wings on the face opposite to the longitudinal wing, x 30. --Figs. 2021. A. vuIparia. -- Fig. 20. wavy appearance of the longitudinal wing, 20.--Fig. 21. transverse membranous wings, x 20.--Fig. 22. A. napellu8 subsp, tauricum: transverse expansion through "pinching" of adjacent epidermal cells, x 300.--Figs. 23-24. transverse expansions through outwards projection of adjacent epidermal cells. --Fig. 23. A. paniculatum, 150. --Fig. 24. A. vulparia, x 150

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Figs. 13 24

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CAPPELLETTI ~5 L POLDINI:

3. Aconitum variegaturn group: 3.1. A. variegatum: only one longitudinal wing present (Fig. 15). A few (3-4) ridges expanding into large membranous wings (Figs. 16-17). The transverse wings more developed on the face opposite to the edge bearing the longitudinal wing than on the other two faces (Fig. 15). 3.2. A. panieulatum: only one longitudinal wing. The transverse membranous wings more numerous (5-6) but less expanded than in A. variegatum (Fig. 18). 3.3. A. angustifolium: only one longitudinal wing. Many transverse ridges that expand into membranous wings (Fig. 19). 4. Aconitum vulparia group: 4.1. A. vulparia: very often the longitudinal wing not apparent; when present, the longitudinal wing with a marked wavy appearance (Fig. 20). On all faces the transverse ridges expand into membranous wings (Fig. 21). 4.2. A. lamarckii: wing pattern quite comparable to that described for A. vulparia.
Discussion

Great constancy of seed microcharacteristics has been observed within the genus. All the Aconitum species and subspecies studied exhibit a warty ornamentation of the integument epidermal cells except on the top of longitudinal wings (when present) and on the hilum region. This kind of ornamentation is not an unusual pattern of epidermal cell surface, and frequently occurs for instances on trichomes, nor can it be considered an exclusive feature of Aconitum seeds, since it has also been found on seeds of another Ranunculaceous species: Cimicifuga europaea SCHIPCZ. (CAPPELLETTI,unpubl.). As regards to seed morphology, marked differences have been observed within the genus Aconitum. A seed morphological trend has been pointed out, characterized by the reduction of longitudinal wings on the edges with a concomitant development of ridges on the seed faces expanding into transverse membranous wings. The seed provided with three equally developed longitudinal wings along the edges and with faces devoid of ridges (as found in A. anthora), may be regarded as the simplest type. Another seed type with reduction of two longitudinal wings and appearance of ridges on the faces is apparent in all the subspecies of A. napellus studied. The infolding of the less developed longitudinal wings, never found in the subsp, tauricum, is already present in the subsp, hians and even more apparent in the seeds of the subsp, neomontanum. In a third type, the complete disappearance of two longitudinal wings is coupled with considerable development of ridges

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expanding into membranous wings (A. variegatum, A. paniculatum, A. angustifolium). The disappearance even of the remaining longitudinal wing along with ridge development on all the faces and edges, gives rise to the seed type of the A. vulparia group (for instance A. vulparia and A. lamarckii). These seed morphological patterns clearly indicate an inverse correlation between the occurrence of longitudinal wings on the edges and the development of transverse wings and their expansion onto the faces. The overall seed coat morphological pattern of the genus Aconitum is in good agreement both at the infragenerie and infraspeeifie levels with the current taxonomic scheme (WARNCI(E1964, GOTZ1967, SmTZ 1969), thus confirming DAWS & ItEYWOOD'S (1963) statement about the great potential taxonomic value of seed characteristics. The peculiar seed morphology of A. anthora (the only species with three equally developed longitudinal wings, smooth faces and cells provided with tubercles) seems to stress its isolated taxonomic position (HEeI 1912). Great significance for understanding the possible origin of the transverse membranous wings has to be attached to the tubercles found on the seed integument cells of A. anthora. In fact we can recognize in the alignment of the tubercles of a row of contiguous cells (Figs. 4-5) the beginnings of the transverse ridges and wings. It is possible to explain this expansion process through a "pinching" of the middle portion of a series of adjacent epidermal cells (Fig. 22). In such an instance the transverse expansions (recognizable only with SEM) can be considered as an accentuation of a series of aligned tubercles as seen in A. anthora. A second possibility by which the transverse expansions may originate is through an outward projection of a series of adjacent epidermal cells (Figs. 23-24), giving origin to structures easily recognizable even at low magnification. In the A. napellus group the first possibility of transverse expansion is the one most common, while in the other Aconitum groups outward projections of a row of cells can be observed. Transverse expansions according to the first possibility, however, also occur in A. variegatum, A. ioaniculatum and A. angustifoliurn, but only in close proximity of the edge still bearing the longitudinal wing. CytologieM studies provide evidence that A. angustifolium is an allopolyploid between a species of the A. variegatum group and A. napellus (most probably subsp, tauricum) (SmTZ 1969). Chemical data seem to confirm this, since small amounts of aeonitine--the alkaloid characterizing all the members of the A. napellu8 group (KaTz & STAE~IELIN 1978)--have been found in A. angustifolium along with mesaeonitine and/or hypaconitine as main components (KATz & STAEHELIN1979). On the contrary, the available chemical evidence fails to confirm affinities with the A. variegatum group (KATz & STAEHELIN

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1979, 1980). Seed morphology of A. angustifolium is not reminiscent of the A. napellus group, but exhibits great resemblance to the A. variegatum group. In A. angustifolium even further morphological progression with respect to both A. variegatum and A. paniculatum can be observed as regards the n u m b e r and development of the m e m b r a n o u s transverse wings, which even tends to reach the remaining longitudinal wing. The simple seed morphology of A. anthora, a species regarded as a xerothermic relict (I~EGI 1912), suggests t h a t ecological a d a p t a t i v e significance m a y be a t t r i b u t e d to the more elaborate seed coat features occurring in the other aconites. This hypothesis is supported b y the fact t h a t within the A. variegatum and A. vulpa~"ia groups we find m a r k e d l y mesophilous-hygrophilous nemoricole species, while the aconites of the napellus group grow in open habitats, often on rocky soils, indicative of ecological requirements intermediate between A. anthora and the species of the A. variegatum and the A. vulparia groups. Ecological adaptations in connection with improved floating ability was suggested for the extent of seed transverse wing expansion in Aconitum (DAMBOLDT & ZrSISfEI~5'~ANN1974) and of nutlet tuberculation in Limnanthes (HAuPTL~ & al. 1978). We also feel t h a t the transverse m e m b r a n o u s seed wings of Aconitum m a y affect the seed dispersal efficiency (floating ability, hygroscopic m o v e m e n t s distancing one seed from the other) and m a y perhaps assure o p t i m u m moisture condition for germination. Obviously, such a working hypothesis needs to be experimentally verified.

R e f e r e n c e s

DAvis, P. H., HEYWOOD,V. H., 1963: Principles of Angiosperm Taxonomy. -Edinburgh: Oliver. DAMBOLDT, J., ZIMMERMARrN, W., 1974: Aconitum. -- In HEal, G.: Illustrierte Flora yon Mitteleuropa 3 (3), 152--177 (3ra Ed.). -- Berlin, Hamburg: P. Parey. GOTZ,E., 1967: Die Aconitum variegatum-Gruppe und ihre Bastarde in Europa. -- Feddes Repert. 76 (1-2), 1--62. HAUPTLI,H., WEBSTER,B. D., JalN, S., 1978: Variation in nutlet morphology in Limnanthes. -- Amer. J. Bot. 65 (6), 615--624. HEGI, G., 1912: Aconitum. -- In HEGI, G., Illustrierte Flora yon Mitteleuropa 3 (4), 492--507 (F t Ed.). -- Miinehen: Lehmann. KATz, A., STAEHELI~, E., 1978: Diinnsehichtchromatographiseher Vergleieh einiger europ//iseher Aeonitarten und -Handelsmuster. -- Planta Medica 33 (3), 287--288. -- 1979: DC-Untersuchung der europ/iisehen Aconitum napellus-Gruppe. -Pharm. Acta HeN. 54 (9/10), 253--265. -- 1980: Further investigations pertaining to European Aconites. -- Planta Mediea 39 (3), 209--210. SEITZ, W., 1969: Die Taxonomic der Aconitum napellus-Gruppe in Europa. -Feddes Repert. 86 (1), 1-76.
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STEBBINS~G. L., 1974: Flowering Plants: Evolution Above the Species Level. -Cambridge: Belknap Press. TUTIN, T. G., MERXlVI~LI~ER,H., 1964: Aconitum L. -- I n TUTI~ & al.: Flora Europaea l, 211--213. -- Cambridge: University Press. WARNCKE,K., 1964: Die europgischen Sippen der Aconitum lycoctonum-Gruppe. Diss. d. Ludwig-Max.-Univ. Miinchen, 1--66.
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Addresses of the authors: Prof. E. M. CAPPELLETTI, Istituto di Botanica e Fisiologia vegetale, Via Orto Botanico 15~ 1-35100 Padova, Italy. -- Prof. L. PoLm~TI, Istituto Botanieo, Via A. Valerio 30, L34100 Trieste, Italy.