Cytotoxic triterpenoid saponins from the fruits of Aesculus pavia L

Continued chemical investigation on the fruits of North American Aesculus pavia L. resulted in the isolation and identification of 13 polyhydroxyoleanene pentacyclic triterpenoid saponins, named aesculiosides IIe-IIk (1-7), and IIIa-IIIf (8-13), together with 18 known compounds: aesculiosides Ia-Ie (14-18), IIa-IId (19-22), IVa-IVc (23-25), 3-O-[beta-D-galactopyranosyl(1-->2)]-alpha-L-arabinofuranosyl(1-->3)-beta-D-glucuronopyranosyl-21,22-O-diangeloyl-3beta,15 alpha,16 alpha,21 beta,22 alpha,28-hexahydroxyolean-12-ene (26), 3-O-[beta-D-glucopyranosyl(1-->2)]-alpha-L-arabinofuranosyl(1-->3)-beta-D-glucuronopyranosyl-21,22-O-diangeloyl-3beta,16 alpha,21 beta,22 alpha,24 beta,28-hexahydroxyolean-12-ene (27), 3-O-[beta-D-galactopyranosyl(1-->2)]-alpha-L-arabinofuranosyl(1-->3)-beta-D-glucuronopyranosyl-21,22-O-diangeloyl-3beta,16 alpha,21 beta,22 alpha,28-pentahydroxyolean-12-ene (28), R(1)-barrigenol (29), scopolin (30), and 5-methoxyscopolin (31). The structures of these compounds were elucidated by spectroscopic and chemical analyses. Compounds 14-22 and 26-28 were tested in vitro for their activity against 59 cell lines from nine different human cancers including leukemia, non-small cell lung, colon, CNS, melanoma, ovarian, renal, prostate, and breast. It was found that compounds with two-acyl groups at C-21 and C-22 had cytotoxic activity for all cell lines tested with GI(50) 0.175-8.71 microM, while compounds without acyl groups at C-21 and C-22 had weak or no cytotoxic activity. These results suggest that the acyl groups at C-21 and C-22 are essential for their activity.     

Triterpenoid saponins from the fruits of Aesculus pavia

The compounds, named aesculiosides Ia-Ie, IIa-IId, and IVa-IVc, were isolated from an ethanol extract of the fruits of North American Aesculus pavia, along with two known compounds. Their structures were characterized as polyhydroxyoleanene pentacyclic triterpenoid saponins by spectroscopic and chemical analyses. These saponins were divided into three elution zones by chromatography according to the polarity because of the acyl substitution at C-21 and C-22 of the aglycone saponins moiety. These are structurally different from those isolated from Eurasian Aesculus hippocastanum and Aesculus chinensis in their oligosaccharide moieties.     

Survival and long-term maintenance of tertiary trees in the Iberian Peninsula during the Pleistocene: first record of Aesculus L. (Hippocastanaceae) in Spain

The Italian and Balkan peninsulas have been places traditionally highlighted as Pleistocene glacial refuges. The Iberian Peninsula, however, has been a focus of controversy between geobotanists and palaeobotanists as a result of its exclusion from this category on different occasions. In the current paper, we synthesise geological, molecular, palaeobotanical and geobotanical data that show the importance of the Iberian Peninsula in the Western Mediterranean as a refugium area. The presence of Aesculus aff. hippocastanum L. at the Iberian site at Cal Guardiola (Tarrasa, Barcelona, NE Spain) in the Lower–Middle Pleistocene transition helps to consolidate the remarkable role of the Iberian Peninsula in the survival of tertiary species during the Pleistocene. The palaeodistribution of the genus in Europe highlights a model of area abandonment for a widely-distributed species in the Miocene and Pliocene, leading to a diminished and fragmentary presence in the Pleistocene and Holocene on the southern Mediterranean peninsulas. Aesculus fossils are not uncommon within the series of Tertiary taxa. Many appear in the Pliocene and suffer a radical impoverishment in the Lower–Middle Pleistocene transition. Nonetheless some of these tertiary taxa persisted throughout the Pleistocene and Holocene up to the present in the Iberian Peninsula. Locating these refuge areas on the Peninsula is not an easy task, although areas characterised by a sustained level of humidity must have played an predominant role.     

Genetic diversity among populations and size classes of buckeyes (Aesculus: Hippocastanaceae) examined with multilocus VNTR probes

 Little is known about genetic variation in members of the genus Aesculus (Hippocastanaceae), in particular A. flava (yellow buckeye) and A. glabra (Ohio buckeye). Here, three synthetic DNA probes (composed of tandemly repeated, core sequences) that reveal alleles at multiple variable-number tandem-repeat (VNTR) loci in these two species were used to investigate: 1) levels of genetic variation in one stand of A. flava and three isolated stands of A. glabra; 2) whether the stands of A. glabra are genetically differentiated from one another; 3) whether there has been selection for more heterozygous individuals through time in one stand each of A. flava and A. glabra; and 4) whether a possible genetic bottleneck had occurred during the formation of either species of Aesculus. First, variation of VNTR genetic markers within and among three populations of A. glabra separated by 60–180 km was examined. In each one hectare (ha) population, 22 individuals were randomly sampled. Among the three populations, the mean number of bands scored per individual was 80.35 and the average number of estimated loci surveyed was 54.17. Mean similarity and estimated heterozygosity within populations ranged from 0.634 to 0.743 and from 0.342 to 0.486, respectively. The mean similarity across populations was 0.657, while the mean estimated heterozygosity across populations was 0.484 for A. glabra. The most isolated site was the most genetically differentiated as indicated by differences in levels of similarity, heterozygosity, and Fst value comparisons. In a separate experiment, genetic variation in 22 large (reproductively mature; dbh > 8 cm) individuals was compared with that in 22 small (not yet reproductive; dbh < 1 cm) individuals collected within one ha stands for both A. flava and A. glabra. Mean similarity values among large versus small individuals of A. flava were 0.665 versus 0.662, while for A. glabra the corresponding values were 0.686 versus 0.691, respectively. Permutation tests of these similarity data detected no evidence for size class genetic differentiation in either species (both p-values > 0.050). Further, permutation tests for the number of bands per individual (average band number should be higher in more heterozygous individuals) detected no significant differences between size classes for either species. Thus, evidence of pronounced inbreeding and/or selection altering population genetics within small relative to large individuals was not detected. In addition, comparable similarity and heterozygosity values between these two closely related species (which still maintain an active zone of hybridization) suggests that either: 1) no extreme genetic bottleneck has accompanied the formation of these species from a common ancestor; or 2) signs of such a bottleneck have largely been eliminated. These studies demonstrate the utility of multilocus VNTR DNA probes for investigating genetic variation within and among plant populations, between size classes within a population, and between closely related species. Received May 15, 1998 Accepted September 11, 2001     

Variation and taxonomy of Pyxidanthera (Diapensiaceae)

Brittonia - Morphological and ecological studies of seven populations ofPyxidanthera barbulata Michx. in North and South Carolina indicate that the variation among populations is clinal and...     

Variation and taxonomy ofFritillaria graeca (Liliaceae) in Greece

The geographic variation inFritillaria graeca, an endemic of Greece, is explored, using multivariate techniques. Eleven morphological features were measured in each of 630 individual plants from 38 localities throughout the range of the species.Scheffe's a posteriori test revealed an almost continuous geographic variation and did not identify groups clearly. Discriminant analysis of the groups derived from an F significance test ofMahalanobis distance resulted in five major groups on the first two discriminant axes representing the subspeciesgraeca, ionica, guicciardii, thessala and its var.othria. A diagnostic key to theF. graeca subspecies is provided.This paper is a part of a doctoral dissertation dealing withFritillaria in Greece, to be submitted to the department of Biology of the Aristotelian University of Thessaloniki.     

Variation and taxonomy in Hordeum depressum and in the H. brachyantherum complex (Poaceae)

The variation pattern in the perennial Hordeum brachyantherum complex and in the annual H. depressum are described. The diploid form of H. brachyantherum s. lat., endemic to California in USA, previously recognized as a separate species is here treated as a subspecies ( H. brachyantherum ssp. californicum ). Despite its restricted distribution it shows a considerable variation and overlap in morphology with the tetraploid ssp. brachyantherum , and no unambiguous determination based on morphology between the two tax a is possible. The tetraploid cytotype has a large distribution area in western North America and easternmost Asia and a very wide morphological variation. It has also a small disjunct distribution area in easternmost Canada. A single hexaploid population from California is referred to ssp. brachyantherum .     

The effects of desiccation on seeds of Acer saccharinum and Aesculus pavia: recalcitrance in temperate tree seeds

This study was undertaken to determine how the results from lipid, moisture, and differential scanning calorimetry analyses conducted on silver maple (Aceraceae: Acer saccharinum L.) and red buckeye (Hippocastanaceae: Aesculus pavia L.) compared with those obtained from previous studies on white and water oaks (Fagaceae: Quercus alba and Q. nigra), and the tropical zone species American muskwood (Meliaceae: Guarea guidonia) and carapa (Meliaceae: Carapa guianensis). Seeds were air-dried at room temperature for 9-11 days. At intervals, germination was tested, moisture determined, and lipids extracted. It was found that, like the other recalcitrant seeds, (1) viability was greatly reduced or lost after 11 days of drying, (2) percentage changes in individual fatty acids were not related to seed viability, and (3) results from the differential scanning calorimetry studies revealed a strong relationship between enthalpy/onset data from the embryo and cotyledon tissues and loss of viability. Also, silver maple seeds experienced a 50% reduction in viability by day 5 of drying and retained an axis moisture content over 25% throughout the experiment. However, unlike the other recalcitrant seeds surveyed, both silver maple and red buckeye had a significant reduction in the total amount (mg/g) of cotyledon lipids as the experiment progressed. However, no decrease in the unsaturated/saturated fatty acid ratio was found, so we conclude that in these species lipid peroxidation is not a marker of declining seed viability. Also, red buckeye seeds did not lose 50% viability until after day 8 of the experiment, and axis moisture content fell well below 20% as the seeds dried.     

Cytogenetics and taxonomy of the genus Globba L. (Zingiberaceae) in Malaya

Hexaploid chromosomal races of Globba cernua Bak. and G. patens Miq. show overlap in geographical distribution. Hybrid swarms in three separate localities on the Main Range in Malaya are analysed both morphologically and cytologically. Evidence indicates that introgressive hybridization of G. cernua Bak. by G. patens Miq. has taken place.     

Microtubules in Interphase Nuclei of Aesculus hippocastanum L.

Interphase nuclei of cambial cells and their derivatives indormant and active, juvenile and mature stems of Aesculus hippocastanumL. contain bundles of microtubules. Each bundle comprises betweenten and 50 microtubules arranged parallel to one another andoften in a paracrystalline hexagonal array. One end of the bundleis associated with the nucleolus and merges into it. No distinctterminal structures have been found, although in some casesthe end distal from the nucleolus lies close to the nuclearmembrane. No definite role can be suggested for these structuresas yet, but their presence in interphase nuclei, coupled withtheir association with the nucleolus, is indicative of involvementin the normal activity of the nucleolus. Aesculus hippocastanum L, microtubules, nucleolus, nucleus     

Cryopreservation of horse-chestnut (Aesculus hippocastanum L. ) somatic embryos using three different freezing methods

Cryopreservation of somatic embryos of Aesculus hippocastanum L. cultured on nutritive media containing abscisic acid (ABA) at concentrations of 0.75 μM, 7.5 μM and 75.0 μM was evaluated for three cooling methods: (i) slow freezing with cryoprotectants, (ii) fast freezing with cryoprotectants, and (iii) fast freezing with desiccation techniques. The ‘cryoprotectant’ freezing techniques included the embryo pretreatment on ABA containing medium for 4 days, followed by cryoprotective treatment in liquid medium containing 0.5 M dimethylsulfoxide, 0.5 M glycerol, 1.0 M sucrose, and cooled at slow, and rapid rates. Embryos pretreated on a medium containing 0.75 μM ABA, and cooled to −35 °C at 1°C /min, held for 30 min at this transfer temperature and then immersed in liquid nitrogen (LN) had the best embryo recovery (43%). The ‘desiccation’ method involved an air drying step of similar ABA-pretreated, non-cryoprotected embryos followed by rapid cooling. Embryos precultured on 0.75 μM ABA, then subjected to a 4 h period of air desiccation (water content reduction to 13%) showed about the same level of survival (46%) as found with the ‘cryoprotectant’ slow freezing technique. The air-dry ‘desiccation’ method is easier to apply than the more complicated ‘cryoprotectant’ method. This revised version was published online in June 2006 with corrections to the Cover Date.     

Leaf micromorphology and taxonomy of North African species of Sideritis L. (Lamiaceae)

REJDALI M., 1991. Leaf micromorphology and taxonomy of North African species of Sideritis L. (Lamiaceae). This study reports on the structure of trichomes, epidermal cells, stomata and venation patterns. The trichomes are either glandular or eglandular. The latter are unicellular or multicellular, soft or rigid pointed hairs. The glandular hairs are short or long stalked, the latter are generally flattened and may be sessile. The epidermal cells are variously shaped with sinuous or straight walls usually randomly orientated. Stomata are either anomocytic or diacytic and evenly distributed throughout the epidermis without any definite pattern of orientation. Venation is of the semicraspedodermous type.     

Variation and Evolution in the Karyotype of Lycoris (Amaryllidaceae) V. Chromosomal Variation in L. sanguinea Maxim.

Abstract In 554 bulbs from 38 populations of Lycoris sanguinea , several chromosomal variations have been discovered. Most of the bulbs have a common karyotype consisting of 22 acrocentric ( A ) chromosomes. Though their frequencies are low, some rearranged chromosomes which are aberrant have been found. The aberrants are as follows: 1. Subtelocentrics ( ST ); 2. Telocentrics ( T' ); 3. Metacentrics ( M' ); 4. Very small acrocentrics (a); 5. Very small metacentrics (m); 6. Acentric fragments ( Ac ); and 7. Dicentrics ( Di ) chromosome. All can be easily suspected to be derived from A s. Some aberrations of the satellite chromosomes have been observed also. In addition, a new karyotype composed of 2n=32=31 A + 1 M' chromosomes has been found.     

A seasonal cycle of cell wall structure is accompanied by a cyclical rearrangement of cortical microtubules in fusiform cambial cells within taproots of Aesculus hippocastanum (Hippocastanaceae)

Aspects of the structure and ultrastructure of the fusiform cambial cells of the taproot of Aesculus hippocastanum L. (horse chestnut) are described in relation to the seasonal cycle of cambial activity and dormancy. Particular attention is directed at cell walls and the microtubule and microfilament components of the cytoskeleton, using a range of cytochemical and immunolocalization techniques at the optical and electron-microscopical levels. During the dormant phase, cambial cell walls are thick and multi-layered, the cells possess a helical array of cortical microtubules, and microfilament bundles are oriented axially. In the early stages of reactivation, vesicle-like profiles are associated with the cell walls, whereas arrangement of the cytoskeletal elements remains unchanged. In the succeeding active phase, the cell walls are thin, and cortical microtubules form a random array, although microfilament bundles maintain a near-axial orientation. The observations are discussed in relation to the seasonal cycle of wall structure and cortical microtubule rearrangement within the vascular cambium of hardwood trees. It is suggested that the cell-wall thickening at the onset of cambial dormancy, which is associated with the presence of a helical cortical microtubule array, should be considered to be secondary wall thickening, and that selective lysis of this secondary wall layer during cambial reactivation restores the thinner, primary wall found around active cambial cells.     

Seed morphology and taxonomy of the North African species of Sideritis L. (Lamiaceae)

REJDALI, M., 1990. Seed morphology and taxonomy of the North African species of Sideritis L. (Lamiaceae). Seed morphology data are shown to support data from general morphology and palynology; they can be used for taxonomy at the sectional and specific levels. At times clear differences are apparent at the infraspecific level. Seed sculpturing was found to be of great value for separating taxa at all levels of the hierarchy.     

Comparative developmental morphology of two populations of Ceratophyllum L. (Ceratophyllaceae) and their taxonomy

Developmental morphology of two natural populations of Ceratophyllum inhabiting the waters of Delhi has been investigated under natural conditions and in aseptic culture. Both the populations show an extreme degree of adaptability to hydrophily. The embryo is unusually highly developed and contains 12–14 whorls of leaves and a few lateral buds. The two populations differ significantly in colour and dichotomy of leaf; structure and position of male flowers; number, position and colour of horns on the fruit; stylar length and distribution of sexes. The latter two parameters are probably responsible for the reproductive isolation of the populations, despite their sympatric association. On account of these differences, the two populations are assigned to two distinct species C. demersum L. and C. echinatum Gray, respectively and their taxonomy is discussed; C. echinatum is a New World species not previously recorded from India.     

Variation in expression of trimorphic incompatibility in Pontederia cordata L. (Pontederiaceae)

Summary Pontederia cordata L. (Pontederiaceae), a perennial diploid, possesses the rare genetic polymorphism tristyly. A controlled pollination programme was conducted over a three year period, under glasshouse conditions, on 36 clones of P. cordata var. cordata to examine the nature of the self-incompatibility system. The three major findings of the pollination study were: (1) the three floral morphs display different levels of self-incompatibility, (2) pollen from the two anther levels within a flower exhibits different compatibility behaviour in self-pollinations, (3) considerable individual genetic variation in the expression of self-incompatibility is evident among clones within floral morphs. Similar results were also obtained from a smaller study on 15 clones of P. cordata var. lancifolia conducted over a 6 month period. In common with other Pontederia species the mid-styled morph (M) of P. cordata produces large amounts of seed when self-pollinated with pollen from long-level anthers. A developmental model is proposed to explain the high level of self-compatibility of the M morph in Pontederia species. Self-pollination of segregating progenies from M and S morphs of known incompatibility status demonstrated that the expression of incompatibility is closely associated with style length. It is suggested that overall differences in incompatibility behaviour among the floral morphs may be due to the pleiotropic effects of major genes controlling sub-characters of the tristylous syndrome, rather than linked modifier genes. However, the variable expression of trimorphic incompatibility within floral morphs suggests that this variation may be polygenic in origin.