Chemotaxonomical investigations of theSymphytum officinale polyploid complex andS. asperum (Boraginaceae): The pyrrolizidine alkaloids

By means of thin layer chromatography in conjunction with mass spectrometry the pyrrolizidine alkaloid patterns derived fromSymphytum asperum, several cytotypes ofS. officinale agg. and the artificial hybrids of the former taxa, were compared. The obtained patterns were not essentially affected by variation in cytotype, harvesting times and -location of plants. Lycopsamine, acetyl-lycopsamine and symphytine or their isomers were generally found in theS. officinale cytotypes, echimidine and symphytine inS. asperum. The interspecific hybrids contained all alkaloids mentioned. The definite lack of echimidine in the 2 n=40 cytotype proves that it is conspecific withS. officinale and does not belong to a hybrid-swarmS. asperum × S. officinale with 2 n=48. Part I of this series of contributions.     

Chemo- and karyotaxonomic studies on some rhizomatous species of the genusSymphytum (Boraginaceae)

InS. tuberosum subspp.tuberosum andnodosum, S. grandiflorum andS. ibericum the presence of the pyrrolizidine alkaloids lycopsamine, echimidine and symphytine could be demonstrated. The taxonS. tuberosum contains an unknown compound that seems to be specific for this taxon. This compound is not the pyrrolizidine alkaloid anadoline which has previously been reported for this species. It is possibly represented by a peak on GC/MS with a molecular ion peak at m/z 623 (as TMS derivative) and can be used as a chemotaxonomic marker for the speciesS. tuberosum. The pyrrolizidine alkaloid pattern of the two subspecies ofS. tuberosum reinforces the close relationship. Fresh material ofS. tuberosum contained the triterpene isobauerenol, but in herbarium material isobauerenol was lacking. InS. grandiflorum, neither fresh nor dried material contains isobauerenol. In herbarium material ofS. ibericum also no isobauerenol could be found. More extensive chemotaxonomical research is necessary to support the view thatS. abchasicum is more closely related toS. ibericum than toS. grandiflorum.     

Heteromerikarpie und Fruchtpolymorphismus beiMicroparacaryum,gen. nov., (Boraginaceae)

Microparacaryum (M. Pop. exH. Riedl)Hilger & Podlech is described as a new genus of theBoraginaceae-Cynoglosseae. It comprises the annual species hitherto included inParacaryum (DC.)Boiss. andMattiastrum (Boiss.)Brand. Distribution maps are given for all 3 genera.Microparacaryum consists of two species,M. salsum (Boiss.)Hilger & Podlech (M. s.) andM. intermedium (Fresen.)Hilger & Podlech (M. i.). ParticularlyM. i. is a very variable species, and most of the species formerly recognized belong here. Scattered all over the range of the genus, plants occur with nutlets exhibiting flat or incurved marginal wings, often in mixed populations. This fruit polymorphism is taxonomically treated by recognizing formae. In addition, the following new infraspecific taxa and combinations are described:M. i. var.intermedium formaparacaryoides Hilger & Podlech,M. i. var.stellatum (H. Riedl)Hilger & Podlech,M. i. var.stellatum formamattiastroides Hilger & Podlech,M. s. formamattiastroides Hilger & Podlech.

     

Seed coat structure ofCrocus vernus agg. (Iridaceae)

Cytotypes of theCrocus vernus aggregate differ slightly in their seed surface patterns.Crocus albiflorus may be grouped together withCrocus vernus subsp.vernus, both are relatively easily distinguishable by their seed surface patterns from theCrocus scepusiensis—Crocus heuffelianus group.     

Chromosomal differentiation withinCrocus vernus agg. (Iridaceae) in the Carpathian Mts.

Western and eastern Carpathian populations ofCrocus heuffelianus s. lat. (incl.C. scepusiensis) have 2n = 18 but differ in karyotype. While western populations are chromosomally monomorphic, eastern populations exhibit geographical karyotypic differentiation.     

Rapid cytofluorimetric determination of leaf nuclear DNA content in the polyploid seriesRanunculus marsicus (R. auricomus agg.,Ranunculaceae)

A cytofluorimetric method for DNA amount determination on nuclei released from fixed leaves has been successfully tested on samples belonging to the polyploid series ofRanunculus marsicus Guss & Ten. (sect.Auricomus). The procedure was devised to screen rapidly for different ploidy levels within topodemes and progenies.     

Wachstum und Ausbildungsformen des Gynoeceums vonRochelia (Boraginaceae)

The ontogeny of the gynoecium ofRochelia disperma has been investigated by LM and SEM. From the floral apex only one carpel primordium arises abaxially and eventually shifts into a subterminal position. Neither an initial stadium of a second carpel nor an adaxial vascular strand in gynobase and style could be observed. InR. stylaris two vascular strands run through the style and two undifferentiated lobes in adaxial position may be regarded as rudimentary mericarps. Only from comparison with related taxa the conclusion can be drawn thatRochelia is really pseudomonomerous, more so inR. disperma than inR. stylaris. The primary gynoecial bulge splits up into three parts inR. disperma: style/stigma, nutlets with gynobase, and disc. While the mericarps originate in mostBoraginoideae from the symplicate region and the ascidiate one is restricted to the very basal parts, inR. disperma the ascidiate part extends and forms the nutlets. The hood-shaped mouth of the carpel (the plicate zone) is closed to a triangular slit in lateral position, the stigma.The nutlets are triangular with broad base and do not surround the adaxial part of the gynobase inR. disperma, R. persica, R. bungei, R. stylaris, andR. macrocalyx. In contrast,R. peduncularis, R. cardiosepala, andR. cancellata have nutlets clasping the gynobase; they may be more closely related than was assumed up to now. The glochids ofRochelia are fascicled unicellular hairs (with different shapes) and not emergences as in theCynoglosseae. There is an evolutionary trend towards fruit formation with only one mericarp, especially inR. disperma.

     

Labellum micromorphology in theOphrys bertolonii agg. and some related taxa (Orchidaceae)

Micromorphology and distribution of trichomes in six taxa of theOphrys bertolonii aggregate (O. bertolonii, O. bertoloniiformis, O. benacensis, O. aurelia, O. drumana, O. catalaunica) are described. A fairly good homogeneity between all the taxa, with a moderate degree of variability even within the same taxon, has been observed. Comparison of these characters with those ofO. promontorii andO. garganica corroborates the inclusion of the former species into theO. sphegodes aggregate. Trichome analysis of a hybrid is exemplified byO. ×chiesesica (O. drumana ×O. fuciflora).     

A contribution to the karyology ofCrocus vernus agg. (Iridaceae) from the Iserian Mts. (Sudety Mts., Poland)

The Iserian Mts. form ofCrocus vernus agg. has 2n = 16 and a karyotype clearly deviating fromC. heuffelianus s. lat.     

Embryology ofCordia sebestena (Boraginaceae)

Sporogeneses and gametogeneses inCordia sebestena are described. Anther wall develops according to the Dicotyledonous type. Pollen grains are three-celled at the time of dehiscence. The occurrence of enlarged, precociously developing, densely cytoplasmic cells toward the proximal end of the anther and between adjacent pollen sacs is a unique feature in this species. The possible functions of these cells are discussed. A placental obturator develops simultaneously with the ovules. The dual origin of parietal tissue (from nucellar epidermis and the archesporial initial) is an unusual feature in this species. Megagametophyte development is monosporic or (occasionally) bisporic. Megaspore tetrads are linear or T-shaped. Double dyads and tetrads are frequently observed.—Maintainance of subfamily status forCordioideae is supported.     

Flower and fruit development in the Macaronesian endemicCeballosia fruticosa (syn.Messerschmidia fruticosa,Boraginaceae, Heliotropioideae)

The ontogeny of the flower and the fruit of the Macaronesian endemicCeballosia were investigated morphologically and anatomically by SEM and LM. The fruit does not break into four mericarps, but splits into two two-seeded carpids. Exo- and mesocarp wither after fruit-ripening and the endocarp constitutes the remaining outer wall. Within the stony endocarp tubular parenchymatic isles develop which are linked with the mesocarp. Subsequent disintegration pretends additional locules in the mature fruit. Similar pericarp formations are also found in someHeliotropium species but result from a different ontogeny. Therefore, although a close relationship ofCeballosia toHeliotropium is obvious, the taxon should be treated as a separate genus.Carpological investigations in theHeliotropioideae (Boraginaceae) 2. For part 1 seeHilger (1987).     

Embryology ofHeliotropium scabrum andH. strigosum (Boraginaceae)

The embryology ofHeliotropium scabrum andH. strigosum has been studied. The development of the anthers follows the dicotyledonous type, the tapetal cells become binucleate. The pollen grains are shed at the two-celled stage. Megaspore tetrads are linear and the development of the megagametophyte corresponds with the Polygonum type. The endosperm is cellular. The embryo development follows the Onagrad Type, i.e. the Capsella variation inH. scabrum and the Nicotiana variation of the Solanad Type inH. strigosum. The pericarp is differentiated into a one-layered epicarp with bulbous-based, unicellular hair, a 5–6-layered chlorenchymatous mesocarp and a 6–7-layered endocarp. The seed coat consists only of the thickened portions of the epidermis.     

Cynoglossopsis somaliensis,sp. nov., und die GattungCynoglossopsis (Boraginaceae)

Cynoglossopsis somaliensis H. Riedl, sp. nov., is described as a new species of the hitherto monotypic genusCynoglossopsis Brand which is closest related toCynoglossum L. but has to be included inBoraginoideae-Eritrichieae from the way the nutlets are attached to the gynobasis.

     

Chemical constituents from Stenosolenium saxatile (Pall.) Turcz. (Boraginaceae)

Chemical investigation on the whole plants of Stenosolenium saxatile led to the isolation of three naphthoquinones (1–3), five pyrrolizidine alkaloids (PAs) (4–8), and six phenolic acids (9–14). The structure of the isolated compounds was elucidated by different spectrometric methods (¹H NMR, ¹³C NMR and ESI-MS). These results are the first chemical data on constituents of this genus. The chemotaxonomic relationships between the genera Stenosolenium, Onosma, Arnebia, Echium and Lithospermum are also discussed.     

Relationships betweenParacaryum intermedium andP. boissieri on Sinai,and the generic separation ofParacaryum andMattiastrum (Boraginaceae)

Paracaryum intermedium andP. boissieri differ only in their inflexed or flattened fruit wings, and often occur in mixed populations on Mt. Katherina, Sinai. This and cultivation experiments suggest that they may represent forms of a single species only. The fruit characters mentioned therefore are hardly useful for a generic separation ofParacaryum andMattiastrum.     

ThePulmonaria dacica group: Its affinities with central and south-east European allies and with the genusParaskevia (Boraginaceae)

Comparative morphological, karyological and chorological studies prove the distinctness of the SE European and Asiatic populations ofPulmonaria mollis s.l., to be treated asP. dacica (Figs. 4, 5).P. mollis s.str. is interpreted as a relatively young taxon, which probably has evolved from an older stock ofP. dacica ancestors during the Pleistocene (Figs. 6, 8) and might have migrated, perhaps with oak woodland communities, from SE to C. Europe. A similar evolution can be postulated for S—SE European dysploidP. rubra-carnica-stiriaca-vallarsae group; this may have involved still unknown tetraploids (P. carnica?, Fig. 7). This group is linked via the hypertriploid speciesP. vallarsae with theP. saccharata group and at least the eastern species of theP. australis group. The monotypic genusParaskevia apparently marks the earliest divergence from the common Tertiary ancestral stock (Fig. 8). It exhibits a tetraploid chromosome number (2n = 28) but has preserved the most primitive characters. — Some comments on the systematics and nomenclature ofP. australis andParaskevia are added.     

Anatomy and evolution in MacaronesianEchium (Boraginaceae)

The endemic Macaronesian members of the genusEchium (Boraginaceae) were examined anatomically to elucidate their relationships. Vessel length to breadth ratio was used as the initial guide to the relative primitive or derived nature of each taxon. Together with other features (e.g. trichome structure) it was possible to establish the phylogenetic pathways within this recently evolved relict group (Fig. 1).     

Alkaloids of theSedum acre-group (Crassulaceae)

The 16 species of theSedum acre-group were investigated for the presence of alkaloids. They areS. acre ofS. ser.Acria, S. alpestre, S. annuum, S. apoleipon, S. borissovae, S. euxinum, S. grisebachii, S. laconicum, S. multiceps, S. sexangulare, S. tuberiferum, S. tuberosum, S. ursi, andS. urvillei ofS. ser.Alpestria, S. samium ofS. ser.Samia, andS. litoreum ofS. ser.Litorea. S. acre differs significantly from the other species. It contains sedamine, hydroxy sedamine, and a number of 2,6-disubstituted piperidine alkaloids. The leafy parts of the species ofS. ser.Alpestria, S. ser.Samia, andS. ser.Litorea contain 4 piperidine alkaloids which also occur inS. acre, and in addition 4 pyrrolidine alkaloids not present inS. acre. The composition of the alkaloid fraction agrees with the infrageneric classification (series) based on the hybridization patterns of the species (comparia).     

Seed protein polymorphism inVicia sativa agg. (Fabaceae)

TheVicia sativa aggregate contains plants with different chromosome numbers karyotypes; they are still cross-compatible and have partially fertile F₁-hybrids. There is also wide adaptive radiation and geographic distribution. The seed protein profiles ofV. sativa agg. are remarkably variable but no band or bands could be related to a specific karyotype, chromosome number or taxon, and variation within and between karyotypes are of the same magnitude. It is suggested that variation in the seed proteins ofV. sativa agg. might be an important agent for acquiring ecological flexibility, and that it is probably enhanced by interkaryotypic hybridization.     

Pollen morphology in theBoragineae (Boraginaceae) in relation to the taxonomy of the tribe

The results of a comparative survey on pollen morphology in theBoragineae by means of light, scanning and transmission electron microscopy are presented and discussed in relation to the taxonomy of the tribe. Multivariate analyses lead to the recognition of 15 pollen morphotypes for which a discriminant key is proposed. The discriminatory characters concern mainly the stereostructural and ultrastructural features of the grains, such as tectum sculpture and aperture morphology, while the number of apertures appears variable within genera and even single species. Seven out of the 12 investigated genera, as currently circumscribed, are matched by palynological data:Anchusella, Borago, Brunnera, Elizaldia, Lithodora, Symphytum andTrachystemon. On account of aperture shape,Lithodora is however closer to theLithospermeae than to theBoragineae. Other genera, and in particularNonea, show a wide variation in tectum ornamentation, shape of grains and number of apertures. Palynological data do not support a broad concept of the genusAnchusa, and point to the autonomy of the satellite generaHormuzakia, Gastrocotyle, Phyllocara, Pentaglottis andCynoglottis.