Exploring the history of Serapias politisii (Orchidaceae), a narrow endemic on the opposite coasts of the Otranto channel: insights from molecular investigations

Knowledge of genetic spatial structure may provide insights into the causes of population disjunctions in plants. Serapias politisii is a narrow endemic with only a few populations scattered along the opposite coasts of the Otranto strait (southern Adriatic Sea). It was originally considered to be of hybrid origin between S. vomeracea ssp. laxiflora and S. parviflora, a possibility suggested also by a DNA phylogenetic study that grouped Italian and Greek populations in two distinct clades. In this study we have carried out additional plastid DNA sequencing and an AFLP analysis of the three taxa. Whereas the geographical distribution of four plastid DNA haplotypes supports the likelihood of a double hybrid origin or of a plastid capture, AFLP data do not support such a hypothesis, because S. politisii shows several private alleles, some of which are shared by Italian and Greek populations. In light of the floristic specificity of the coasts bordering the Otranto strait, we consider that the present‐day disjunction of S. politisii could have originated either by a long‐distance seed dispersal or by a fragmentation of an old polymorphic population. The pairwise mismatch distribution excludes a recent expansion of the populations examined. In light of recent evidence concerning the Mediterranean Lago‐Mare period and the westward haplotype evolution detected in some Euro‐Mediterranean trees, we argue that fragmentation of a previously continuous population could be an intriguing possibility. © 2010 The Linnean Society of London, Botanical Journal of the Linnean Society, 2010, 162 , 572–580.     

Crossing barriers between the unrewarding Mediterranean orchids Serapias vomeracea and Serapias cordigera

Understanding the genetic architecture of admixed hybridizing populations helps in evaluating the nature of species boundaries and the levels of gene exchange between co-occurring species. In the present study, we examined a contact zone between Serapias vomeracea and Serapias cordigera , two unrewarding Mediterranean orchid species with a non-specific pollination strategy. Fruit production and seed viability from interspecific hand-pollination treatments pointed out the weaknesses of post-pollination barriers. The occurrence of hybridization was molecularly confirmed in the genus Serapias for the first time, as parts of plants with a transitional morphology were observed in both alleles of the parental LEAFY intron. Amplified fragment length polymorphism analysis revealed that plants with uncertain morphology and classified as one of the other parental species are actually backcrosses, attesting to an extensive interspecific gene exchange. Overall, the contact zone is more similar to a hybrid zone of Ophrys species, well known for their highly specialized pollination, than to a hybrid zone of unspecialized food-deceptive orchids. Therefore, species boundaries in Serapias are maintained by pre-pollination mechanisms that need to be better investigated. In light of the intriguing similarities between Serapias and Ophrys underlined by the present study, we hypothesize that the emission of floral scents could be involved in the maintenance of species boundaries in Serapias .     

Asymmetric dispersal of southern bull‐kelp (Durvillaea antarctica) adults in coastal New Zealand: testing an oceanographic hypothesis

Coastal populations are often connected by unidirectional current systems, but the biological effects of such asymmetric oceanographic connectivity remain relatively unstudied. We used mtDNA analysis to determine the phylogeographic origins of beach‐cast bull‐kelp (Durvillaea antarctica) adults in the Canterbury Bight, a 180 km coastal region devoid of rocky‐reef habitat in southern New Zealand. A multi‐year, quantitative analysis supports the oceanographically derived hypothesis of asymmetric dispersal mediated by the north‐flowing Southland Current. Specifically, 92% of beach‐cast specimens examined had originated south of the Bight, many drifting north for hundreds of kilometres, and some traversing at least 500 km of ocean from subantarctic sources. In contrast, only 8% of specimens had dispersed south against the prevailing current, and these counter‐current dispersers likely travelled relatively small distances (tens of kilometres). These data show that oceanographic connectivity models can provide robust estimates of passive biological dispersal, even for highly buoyant taxa. The results also indicate that there are no oceanographic barriers to kelp dispersal across the Canterbury Bight, indicating that other ecological factors explain the phylogeographic disjunction across this kelp‐free zone. The large number of long‐distance dispersal events detected suggests drifting macroalgae have potential to facilitate ongoing connectivity between otherwise isolated benthic populations.     

No evidence for Z‐chromosome rearrangements between the pied flycatcher and the collared flycatcher as judged by gene‐based comparative genetic maps

Revealing the genetic basis of reproductive isolation is fundamental for understanding the speciation process. Chromosome speciation models propose a role for chromosomal rearrangements in promoting the build up of reproductive isolation between diverging populations and empirical data from several animal and plant taxa support these models. The pied flycatcher and the collared flycatcher are two closely related species that probably evolved reproductive isolation during geographical separation in Pleistocene glaciation refugia. Despite the short divergence time and current hybridization, these two species demonstrate a high degree of intrinsic post‐zygotic isolation and previous studies have shown that traits involved in mate choice and hybrid viability map to the Z‐chromosome. Could rearrangements of the Z‐chromosome between the species explain their reproductive isolation? We developed high coverage Z‐chromosome linkage maps for both species, using gene‐based markers and large‐scale SNP genotyping. Best order maps contained 57–62 gene markers with an estimated average density of one every 1–1.5 Mb. We estimated the recombination rates in flycatcher Z‐chromosomes to 1.1–1.3 cM/Mb. A comparison of the maps of the two species revealed extensive co‐linearity with no strong evidence for chromosomal rearrangements. This study does therefore not provide support the idea that sex chromosome rearrangements have caused the relatively strong post‐zygotic reproductive isolation between these two Ficedula species.     

Functional bases of host-acceptance behaviour in the aphid parasitoid Aphidius ervi

Abstract.  The host acceptance behaviour in Aphidius ervi is investigated, assessing the role of both external and internal host-associated cues, offered to the experimental parasitoids with parafilm-made aphid dummies. The reaction to internal cues present in the host haemolymph is clearly evident, and its intensity is enhanced by external cues. Parasitoid females lay few eggs in aphid dummies filled with host haemolymph. A significant increase in the number of both oviposition reactions (host stinging) and egg laying is observed only when these dummies are coated with cornicle secretion. However, this enhancement is not observed when the aphid dummies contain distilled water. Thus, the host acceptance behaviour of A. ervi females appears to be controlled by the integration of both external and internal chemical cues. The physiological basis of this behavioural response is investigated with a detailed study on the anatomy and ultrastructure of A. ervi ovipositor. The detection of chemical cues present in the host haemolymph that act as kairomones is made possible by the presence of gustatory sensilla on the tip of the ovipositor. These sensilla consist of porous areas, reached by unbranched dendrites running inside both the lower valves (i.e. first valvulae) and the unpaired upper valve (i.e. second valvulae). The mechanosensory function during oviposition appears to be provided mostly by the basiconic sensilla found on the tip of external valves (i.e. third valvulae). A tentative functional model accounting for the observed oviposition behaviour of A. ervi is proposed.     

Development of Zn‐related necrosis in tobacco is enhanced by expressing AtHMA4 and depends on the apoplastic Zn levels

AtHMA4 was previously shown to contribute to the control of Zn root‐to‐shoot translocation and tolerance to high Zn. However, heterologous expression of 35S::AtHMA4 in tobacco (Nicotiana tabacum cv. Xanthi) results in enhanced Zn sensitivity. This study provides a better understanding of the development of this Zn‐sensitive phenotype and demonstrates that substantial modifications of Zn homeostasis occur due to AtHMA4 expression. We show that ectopically expressing AtHMA4 in tobacco results in overloading the root and leaf apoplast with Zn. The tissue and cellular distribution of Zn, monitored using Zinpyr‐1, was altered in the AtHMA4‐expressing plants compared with wild type. Increased loading of the leaf apoplast with Zn in AtHMA4 transformants induced necrosis; this appeared at lower levels of Zn supply in the transgenics compared with wild type. This study suggests that Zn concentration may be sensed in the apoplast of leaves, and if concentrations are above a certain threshold then particular groups of cells accumulate Zn and necrosis is initiated. Therefore, this could be considered as a mechanism for protecting the other parts of the photosynthetically active leaf from Zn toxicity.     

Nine new polymorphic microsatellite loci for the amplification of archived otolith DNA of Atlantic cod,Gadus morhua L.

Nine out of 22 microsatellite primers tested were successfully amplified on three samples of cod Gadus morhua L. (two contemporary and one archived otolith samples). All loci were polymorphic (5–23 alleles/locus). The average observed heterozygosity across loci and samples was 0.625, ranging from 0.294 to 0.895 at each locus. All loci were under Hardy–Weinberg equilibrium, except PGmo56 that showed significant excess of heterozygotes in all studied samples. The isolated loci were suitable for degraded DNA and therefore useful for conducting a long‐term temporal study with DNA obtained from archived otoliths of cod.     

Molecular phylogeny of the Mediterranean species of Philaenus (Hemiptera: Auchenorrhyncha: Aphrophoridae) using mitochondrial and nuclear DNA sequences

The phylogenies of all eight European species of Philaenus were estimated from cytochrome oxidase subunit I, cytochrome B and internal transcribed spacer 2 (ITS2) fragments of DNA using phylogenetic reconstruction methods: maximum parsimony (MP), maximum likelihood (ML) and Bayesian inference (BI) analyses. Based on the topologies of all obtained phylogenetic trees, the monophyly of Philaenus is well supported, being congruent with morphological, ecological and chromosomal data. Three phylogenetic lineages were distinguished in the mitochondrial and combined (mtDNA with ITS2) trees. The first lineage is represented by only one species, Philaenus maghresignus, which inhabits Maghreb and southern Spain. Clade A includes three species: P. tarifa (Southern Iberia), P. italosignus (Sicily and Southern Italy) and P. signatus (the Balkans and Middle East). In clade B two subclades were recognized: B1 represented by P. loukasi (Southern Balkans) and P. arslani (Middle East), and B2 comprising P. spumarus (the most widespread Palaearctic species) and P. tesselatus (from Southern Iberia and Maghreb). These clades were also retrieved in trees reconstructed from nuclear sequences. However, four species (P. maghresignus, P. tarifa, P. italosignus and P. signatus) showed unresolved polytomy at the base of the nuclear tree. Clade A together with P. maghresignus clustered with the ‘signatus’ group defined from morphology, and clade B with the ‘spumarius’ group; these might be considered separate subgenera. Genetic distances in mitochondrial DNA between ingroup species ranged from 14.0% between P. signatus and P. spumarius to 2.4% between P. tesselatus and P. spumarius. By contrast, genetic divergence of ITS2 between ingroup species was very low, at most 2.1%. The divergence of Philaenus species is estimated to have occcurred between 7.9 and 0.6 Ma. Possibly three main speciation events occurred: the first at the Miocene/Pliocene boundary (c. 5.5 Ma) for deeper splits; the second between 4.2 and 2.5 Ma in the Pliocene, when pairs of more closely related species diverged; and the most recent during the Pleistocene glaciations, when the separation of P. tesselatus and P. spumarius took place. The species status of all Philaenus species is confirmed except for P. tesselatus.