The phylogeography of five flowering plant species (Cakile maritima, Eryngium maritimum, Salsola kali, Halimione portulacoides, Crithmum maritimum) widespread along the European coasts was investigated across their entire European range using AFLP evidence. Both similarities and dissimilarities were found. All species contain a distinct Black Sea/Aegean Sea cluster, and all except E. maritimum contain a distinct Adriatic Sea cluster or group of genetically very similar clusters. All species except Cr. maritimum contain a distinct Atlantic Ocean/North Sea/Baltic Sea cluster clearly separate from the Mediterranean material. In Ca. maritima a distinct Baltic Sea subcluster was found. In the western Mediterranean, two species groups can be recognized. Whereas in S. kali and E. maritimum material from this area falls into only one cluster or a group of genetically very similar clusters, it falls into two clusters or groups of clusters of either more Atlantic or more central Mediterranean similarity in Ca. maritima, H. portulacoides and Cr. maritimum. Similarities and dissimilarities in patterns found are discussed in terms of a combination of historical and extant abiotic and biotic factors. Thus, the distribution range of all species in the eastern Mediterranean area was not affected by Quaternary temperature changes, resulting in phylogeographic congruency here. The existence of distinct Black Sea/Aegean Sea and Adriatic Sea clusters or groups of clusters is the result of sea currents isolating these regions from each other. In the western Mediterranean basin the more cold‐sensitive species (H. portulacoides, Cr. maritimum) but not the less cold‐sensitive species (S. kali, E. maritimum) had to retreat from northern coasts. Re‐colonization of these areas from two different directions is implied by their phylogeographic pattern. The existence of a distinct Gibraltar gap is explained in terms of extant sea currents. Comparison of phylogeographic patterns found with those observed in either marine or terrestrial organisms leads to the conclusion that marine dispersal is of overriding importance in these coastal plants.

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Iris haynei and I. atrofusca are two closely related narrow endemics distributed vicariously along an ecogeographical north-south gradient in Israel and the West Bank. To obtain baseline information of the taxonomic status, conservation and population history of these taxa, we investigated patterns of phenotypic variation and the partitioning of genetic variation within and among populations using dominant random amplified polymorphic DNA (RAPD) markers. Multivariate (principal components analysis) and taxonomic distance analyses based on morphometric traits from eight populations revealed no unambiguous separation into two distinct groups. Results of genetic analyses for nine populations differed only slightly when either allele- or marker-based approaches were employed. Mean within-population diversity was high (0.258 for Nei's expected heterozygosity), but there was no significant relationship between genetic diversity and either population size or latitude. Although the range-wide estimate of GST ( approximately 0.20) revealed relatively high differentiation among populations this value was inflated because of a small, but significant, component of molecular variance among regions viz. taxa ( approximately 5%). Limited long-distance dispersal capabilities in conjunction with a linearized habitat distribution are proposed to contribute to the approximate isolation by distance pattern observed. It also appears that extant populations are currently deviating from equilibrium conditions because of primary divergence of a formerly more widespread ancestral population. Given the absence of deep genetic and phenotypic subdivision among northern (I. haynei) vs. central/southern (I. atrofusca) populations, we argue for a revision of their species status. Nonetheless, we recommend conservation attention to these geographically differentiated segments as separate management units, which can be seen as an instructive example of incipient species formation.

Long-distance seed dispersal, clone longevity and lack of phylogeographical structure in the European distributional range of the coastal Calystegia soldanella (L.) R. Br. (Convolvulaceae)The investigation of the geographical distribution of intraspecific genetic variation and intraspecific phylogenetic or genetic lineages in a large number of plant and animal taxa from different parts of the world has resulted in the detection of often clear phylogeographical patterns (reviewed in, e.g. Soltis et al. ABSTRACTAim To explore the relative effects of Quaternary climatic history vs. speciesspecific biological properties (high seed dispersability, high seed longevity, clonal growth) on phylogeographical structure in European Mediterranean and Atlantic Ocean and Black Sea material of the coastal dune plant Calystegia soldanella (L.) R. Br. Location Black Sea and European Mediterranean and Atlantic Ocean coasts.Methods Variation in amplified fragment length polymorphism was analysed at two different sampling levels. First, an entire-range sample from the Black Sea to the North Sea, including single individuals from sites evenly spread along this entire coast was analysed. Second, in a population-level sample, seven populations from the European Mediterranean and Atlantic Ocean coasts were analysed.Results Neither the entire-range nor the population-level sampling resulted in clear phylogeographical patterns. Instead, individuals from geographically distant areas were often genetically more similar to each other than individuals from the same area. Non-significant isolation-by-distance was found for both sampling approaches, and comparatively low levels of intrapopulational genetic variation were observed. Main conclusionsThe lack of phylogeographical structure in C. soldanella, in comparison with the clear phylogeographical patterns observed in other coastal plant species analysed previously, is postulated to be the result of the specific biology of C. soldanella. The combination of high seed longevity, high dispersability of seeds by sea water and clonal growth and probable high clone age are likely to be responsible for the observed absence of phylogeographical structure. This implies that extreme biological properties such as those shown by C. soldanella can either erase or prevent the formation of historical patterns of genetic variation.

Broad-scale phylogenetic studies give first insights in numbers, relationships, and ages of C4 lineages. They are, however, generally limited to a model that treats the evolution of the complex C4 syndrome in different lineages as a directly comparable process. Here, we use a resolved and well-sampled phylogenetic tree of Camphorosmeae, based on three chloroplast and one nuclear marker and on leaf anatomical traits to infer a more detailed picture of C4 leaf-type evolution in this lineage. Our ancestral character state reconstructions allowed two scenarios: (i) Sedobassia is a derived C3/C4 intermediate, implying two independent gains of C4 in Bassia and Camphorosma; or (ii) Sedobassia is a plesiomorphic C3/C4 intermediate, representing a syndrome ancestral to the Bassia/Camphorosma/Sedobassia lineage. In Bassia, a kochioid leaf type (Bassia muricata and/or Bassia prostrata type) is ancestral. At least three independent losses of water-storage tissue occurred, resulting in parallel shifts towards an atriplicoid leaf type. These changes in leaf anatomy are adaptations to different survival strategies in steppic or semi-desert habitats with seasonal rainfall. In contrast, Camphorosma shows a fixed C4 anatomy differing from Bassia types in its continuous Kranz layer, which indeed points to an independent origin of the full C4 syndrome in Camphorosma, either from an independent C3 or from a common C3/C4 intermediate ancestor, perhaps similar to its C3/C4 intermediate sister genus Sedobassia. The enlarged bundle sheath cells of Sedobassia might represent an important early step in C4 evolution in Camphorosmeae.

Aim  Natural and human‐induced differences in frugivore assemblages can influence the seed dispersal distances of trees. An important issue in seed dispersal systems is to understand whether differences in seed dispersal distances also affect the genetic structure of mature trees. One possible approach to test for a relationship between seed dispersal and the genetic structure of mature trees is to compare the genetic structure of two closely related tree species between two biogeographical regions that differ in frugivore assemblages and seed dispersal distances. Previous studies on two Commiphora species revealed that Commiphora guillauminii in Madagascar has a much lower seed dispersal distance than Commiphora harveyi in South Africa. We tested whether the lower seed dispersal distance might have caused decreased gene flow, resulting in a stronger genetic structure in Madagascar than in South Africa. Location  Madagascar and South Africa. Methods  Using amplified fragment length polymorphism markers we investigated the genetic structure of 134 trees in Madagascar and 158 trees in South Africa at a local and a regional spatial scale. Results  In concordance with our hypothesis, kinship analysis suggests that gene flow was restricted mostly to 3 km in Madagascar and to 30 km in South Africa. At the local spatial scale, the genetic differentiation among groups of trees within sample sites was marginally significantly higher in Madagascar (FST = 0.069) than in South Africa (FST = 0.021). However, at a regional spatial scale genetic differentiation was lower in Madagascar (FST = 0.053) than in South Africa (FST = 0.163). Main conclusions  Our results show that lower seed dispersal distances of trees were linked to higher genetic differentiation of trees only at a local spatial scale. This suggests that seed dispersal affects the genetic population structure of trees at a local, but not at a regional, spatial scale.

Palestine has a rich and prestigious heritage of herbal medicines. To investigate the impact of variable extraction techniques on the cytotoxic effects of medicinal plant extracts, 5 well-known medicinal plants from Palestine were extracted with 90% ethanol, 80% methanol, acetone, coconut water, apple vinegar, grape vinegar or 5% acetic acid. The resulting 35 extracts were screened for cytotoxic activities against three different cancer cell lines (B16F10, MCF-7 and HeLa) using a standard resazurin-based cytotoxicity assay and Nile Blue A as the positive control. Highly variable toxicities and tissue sensitivity were observed, depending upon the solvent used for extraction. The acetone extract of Salvia officinalis L. exhibited the most potent cytotoxicity (IC50 = 14 -36 ug/ml), but very little sensitivity between the three cell lines. More moderate cytotoxicity with improved tissue sensitivity was observed with coconut water extract of Salvia officinalis L (IC50 = 114 µg/ml) and methanolic extract of Teucrium polium L (IC50 = 104 µg/ml). In this study, acetone consistently gave lower extraction yields but higher cytotoxicity, whereas other solvent systems gave much higher extraction yields with lower cytotoxicity. These results demonstrate how the cytotoxicity of plant extracts can be inversely proportional to the yield, and that solvent selection plays an important role in both factors. (IC 50 = 104 µg/ml). In this study, acetone consistently gave lower extraction yields but higher cytotoxicity, whereas other solvent systems gave much higher extraction yields with lower cytotoxicity. These results demonstrate how the cytotoxicity of plant extracts can be inversely proportional to the yield, and that solvent selection plays an important role in both factors.

The Atlas Pistachio tree, Pistacia atlantica Desf., has great importance in the ecological landscape of North Africa, due to its adaptive plasticity, as well as its use as a rootstock in the cultivation of the economically important species, Pistacia vera L. The conservation and valuation of this species require sampling and an assessment of its genetic variability. For the first time in North Africa, the inter-simple sequence repeats (ISSR) molecular marker has been used in genetic-diversity assessment and in the population relationships of P. atlantica subsp. atlantica. The ISSR markers tested showed 74.1% polymorphism, while molecular variance (AMOVA) analysis revealed a high percentage of the total genetic diversity of 55.7% among the four populations studied. Cluster analysis with neighbor-joining (NJ) and principal coordinate analysis (PCO) divided the study sites into four distinct groups according to their geographical locations (Tiaret, Batna, Djelfa, and Bechar). Isolation by distance or Mantel test gave a positive correlation of r = 0.86 between geographical and genetic distances. The results in this study indicate an absence of gene flow, implying that conservation efforts should be taken separately for each population.