Genetic structure and hybridization in the species group of Ficus auriculata: can closely related sympatric Ficus species retain their genetic identity while sharing pollinators?

Obligate mutualistic nursery pollination systems between insects and plants have led to substantial codiversification involving at least some parallel cladogenesis, as documented in Yucca, Ficus and Phyllanthaceae. In such systems, pollinators are generally species specific thus limiting hybridization and introgression among interfertile host species. Nevertheless, in the three systems, cases of one insect pollinating several plant species are reported. In most cases, host plants sharing pollinators are allopatric. However, in the case of the species group of Ficus auriculata, forms may co-occur over large parts of their range. We show here that the species group of F. auriculata is constituted by four well-defined genetic entities that share pollinators. We detected hybrids in nature mainly when both parental forms were growing nearby. Controlled crosses showed that F1 offspring could be successfully backcrossed. Hence, despite sharing pollinators and despite hybrid viability, the different forms have preserved their genetic and morphological identity. We propose that ecological differentiation among forms coupled with limited overlap of reproductive season has facilitated the maintenance of these interfertile forms. As such, establishment of pollinator host specificity may not be a prerequisite for sympatric diversification in Ficus.

Floral volatiles, pollinator sharing and diversification in the fig-wasp mutualism: insights from Ficus natalensis, and its two wasp pollinators (South Africa).

Combining biogeographic, ecological, morphological, molecular and chemical data, we document departure from strict specialization in the fig-pollinating wasp mutualism. We show that the pollinating wasps Elisabethiella stuckenbergi and Elisabethiella socotrensis form a species complex of five lineages in East and Southern Africa. Up to two morphologically distinct lineages were found to co-occur locally in the southern African region. Wasps belonging to a single lineage were frequently the main regional pollinators of several Ficus species. In South Africa, two sister lineages, E. stuckenbergi and E. socotrensis, pollinate Ficus natalensis but only E. stuckenbergi also regularly pollinates Ficus burkei. The two wasp species co-occur in individual trees of F. natalensis throughout KwaZulu-Natal. Floral volatile blends emitted by F. natalensis in KwaZulu-Natal were similar to those emitted by F. burkei and different from those produced by other African Ficus species. The fig odour similarity suggests evolutionary convergence to attract particular wasp species. The observed pattern may result from selection for pollinator sharing among Ficus species. Such a process, with one wasp species regionally pollinating several hosts, but several wasp species pollinating a given Ficus species across its geographical range could play an important role in the evolutionary dynamics of the Ficus-pollinating wasp association.

Ficus racemosa is pollinated by a single population of a single agaonid wasp species in continental South-East Asia.

High specificity in the Ficus-agaonid wasp mutualism has lead to the assumption of a mostly 'one-to-one' relationship, albeit with some exceptions. This view has been challenged by new molecular data in recent years, but surprisingly little is known about local and spatial genetic structuring of agaonid wasp populations. Using microsatellite markers, we analysed genetic structuring of Ceratosolen fusciceps, the fig wasp pollinating Ficus racemosa, a fig tree species widely distributed from India to Australia. In sampling stretching from the south of China to the south of Thailand we found evidence for only a single pollinating wasp species in continental South-East Asian mainland. We found no evidence for the co-occurrence of cryptic species within our subcontinent sampling zone. We observed no spatial genetic structure within sites and only limited structuring over the whole sampling zone, suggesting that F. racemosa is pollinated by a single population of a single agaonid wasp species all over continental South-East Asia. An additional sample of wasps collected on F. racemosa in Australia showed clear-cut genetic differentiation from the Asian continent, suggesting allopatric divergence into subspecies or species. We propose that the frequent local co-occurrence of sister species found in the literature mainly stems from contact zones between biogeographic regions, and that a single pollinator species over wide areas might be the more common situation everywhere else.

When assumptions on visual system evolution matter: nestling colouration and parental visual performance in birds.

Comparative studies in visual ecology of birds often rely on several assumptions on the evolution of avian vision. In this study, we show that when these assumptions are not upheld, conclusions may be strongly affected. To illustrate this purpose, we reanalysed the data of Avilés & Soler (J. Evol. Biol.22: 376-386, 2009) who demonstrated that nestling gape colouration in altricial birds is associated with visual system. We show that a slight change in analysis methodology leads to opposite conclusions. Such conflicting result raises the problem of applying powerful methods developed for continuous variables to a small sample and a small number of independent events of qualitative visual system shift in comparative analyses. Further, we show that the current trend to assume strong phylogenetic inertia of avian visual systems is contradicted by data and that the sequencing of the SWS1 opsin gene should be considered as an alternative approach.

Isolation and characterization of microsatellite primers for Ceratosolen fusciceps, the fig-pollinating wasp of Ficus racemosa, and amplification in two populations.

Ten microsatellite loci were isolated and characterized for Ceratosolen fusciceps, the pollinating wasp of Ficus racemosa, based on individuals from Xishuangbanna (China) and Chiangmai (Thailand). High levels of variation were found among loci for both populations; allelic number varied from two to 18 (respectively from two to 22), expected heterozygosities ranged from 0.362 to 0.890 (respectively from 0.430 to 0.930).

Mutualism, hybrid inviability and speciation in a tropical ant-plant.

Although biotic interactions are particularly intricate in the tropics, few studies have examined whether divergent adaptations to biotic interactions lead to speciation in tropical organisms. Ant-plant mutualisms are widespread in the tropics. Within Leonardoxa africana, two subspecies present contrasting defences against herbivores. Young leaves of subsp. africana are defended by mutualistic ants, whereas subsp. gracilicaulis satiates herbivores by synchronized leaf production. Subsp. africana possesses hollow internodes and many large foliar nectaries, housing and feeding ants. We detected no genetic introgression between the two subspecies in the contact zone between them. F1 hybrids were present. They were intermediate in phenotype, expressing reduced, nonfunctional but costly myrmecophilic traits. However, they suffered more herbivory than their parents. Hybrids remained small, failing to reach reproductive size, probably due to their maladapted defence phenotype. Hence, there could be a direct link between adaptation to mutualism and reproductive isolation: biotic interactions could be a driver of tropical diversity.

Reduced density due to logging and its consequences on mating system and pollen flow in the African mahogany Entandrophragma cylindricum.

In tropical forests, selective logging removes large trees that are often the main contributors to pollination. We studied pollination patterns of the African mahogany, Entandrophragma cylindricum (Sapelli). We investigated two plots in Cameroon corresponding to three tree densities: unlogged forest (Ndama 2002), a mildly logged forest 1 year after logging (Ndama 2003) and a severely logged forest 30 years after logging (Dimako). We used four microsatellite markers to perform paternity analysis. Selfing remained below 2% in all treatments. Pollen flow was mainly long distance but with some proximity effects. Average observed within-plot pollination distances were 338, 266 and 385 m, and pollination by trees outside the plots was 70% (Ndama 2002), 74% (Ndama 2003) and 66% (Dimako). Despite sampling a limited number of seeds from a limited number of mother trees, we obtained seeds sired by 35.6-38.3% of the potential within-plot pollen donors. While trees 20 cm in diameter contributed to pollination, results in Dimako suggest that individual larger trees contribute more to pollination than small ones. This effect was not detected in the other treatments. The results suggest extensive pollen flow in Sapelli. Hence, in Sapelli, the main limiting factor for regeneration after logging may be a reduction in the number of trees capable of producing seeds rather genetic effects due to limits to pollen dispersal.

Pharmacological control of opioid-induced pruritus: a quantitative systematic review of randomized trials.

BACKGROUND AND OBJECTIVE: Numerous drugs have been used to prevent or to treat opioid-induced pruritus in the surgical setting. Their relative efficacy is not well understood. METHODS: The methods employed involved the systematic search (MEDLINE, EMBASE, Cochrane library, bibliographies, without language restriction, up to June 2000) for full reports of randomized comparisons of any intervention which is thought to be anti-pruritic (active) compared with placebo or no treatment (control) in surgical (including labour) patients receiving opioids. The number of patients who had no pruritus were analysed using relative risk and number-needed-to-treat with 95% confidence interval. RESULTS: Twenty-two trials (1477 patients) were analysed. Two trials (66 patients), both with low-dose propofol, were on treatment of established pruritus; propofol had no anti-pruritic effect compared with Intralipid. In prophylaxis trials, the average incidence of pruritus with control was 59% (range, 10% to 100%). Most mu-receptor antagonists were efficacious: intravenous naloxone 0.25-2.4 microg kg-1 h-1, relative risk 2.31 (95% confidence interval, 1.5 to 3.54), number-needed-to-treat to prevent pruritus compared with control 3.5; oral naltrexone 9 mg, relative risk 2.80 (1.35-5.80), number-needed-to-treat 1.7; naltrexone 6 mg was less effective and 3 mg did not work; different intravenous and epidural nalbuphine regimens, relative risk 1.71 (1.12-2.62), number-needed-to-treat 4.2. Intravenous nalmefene 0.5 or 1 mg was not anti-pruritic. Intravenous (but not epidural) droperidol 2.5 mg was efficacious, relative risk, 1.71 (1.28-2.29), number-needed-to-treat 4.9. There was a lack of evidence for any anti-pruritic efficacy with prophylactic propofol, epidural or intrathecal epinephrine, epidural clonidine, epidural prednisone, intravenous ondansetron, or intramuscular hydroxyzine. CONCLUSION: Naloxone, naltrexone, nalbuphine and droperidol are efficacious in the prevention of opioid-induced pruritus; minimal effective doses remain unknown. There is a lack of valid data on the efficacy of interventions for the treatment of established pruritus.

Pollination mode in fig wasps: the predictive power of correlated traits.

The over 700 species of Ficus are thought to have co-speciated with their obligate pollinators (family Agaonidae). Some of these wasp species pollinate figs actively, while others are passive pollinators. Based on direct observations of mode of pollination in 88 species, we show that mode of pollination can confidently be predicted from fig traits only (anther-to-ovule ratio) or from wasp traits only (presence of coxal combs). The presence of pollen pockets is not a predictor of mode of pollination. Data, direct and indirect, on 142 species, demonstrate numerous cases of the loss of active pollination and suggest one or few origins of active pollination. Hence, active pollination, an impressive example of the sophisticated traits that may result from mutualistic coevolution, depends on selective forces that can be overcome in some species, allowing reversions. Despite frequent loss, active pollination remains the predominant mode of pollination in Ficus.

Phylogenetic relationships, historical biogeography and character evolution of fig-pollinating wasps.

Nucleotide sequences from the cytochrome oxidase I (COI) gene were used to reconstruct phylogenetic relationships among 15 genera of fig-pollinating wasps. We present evidence supporting broad-level co-cladogenesis with respect to most but not all of the corresponding groups of figs. Using fossil evidence for calibrating a molecular clock for these data, we estimated the origin of the fig-wasp mutualism to have occurred ca. 90 million years ago. The estimated divergence times among the pollinator genera and their current geographical distributions corresponded well with several features of the break-up of the southern continents during the Late Cretaceous period. We then explored the evolutionary trajectories of two characteristics that hold profound consequences for both partners in the mutualism: the breeding system of the host (monoecious or dioecious) and pollination behaviour of the wasp (passive or active). The fig wasp mutualism exhibits extraordinarily long-term evolutionary stability despite clearly identifiable conflicts of interest between the interactors, which are reflected by the very distinct variations found on the basic mutualistic theme.

Plasmodium falciparum: population genetic analysis by multilocus enzyme electrophoresis and other molecular markers.

Abderrazak, S. B., Oury, B, Lal, A. A., Bosseno, M.-F., Force-Barge, P., Dujardin, J.-P., Fandeur, T., Molez, J.-F., Kjellberg, F., Ayala, F. J., and Tibayrenc, M. 1999. Plasmodium falciparum: Population genetic analysis by multilocus enzyme electrophoresis and other molecular markers. Experimental Parasitology 92, 232-238. The population structure of Plasmodium falciparum, the agent of malignant malaria, is uncertain. We have analyzed multilocus enzyme electrophoresis (MLEE) polymorphisms at 7-12 gene loci in each of four populations (two populations in Burkina Faso, one in Sudan, one in Congo), plus one "cosmopolitan" sample consisting of parasite cultures from 15 distant localities in four different continents. We have also performed random amplified polymorphic DNA analysis (RAPD) and restriction fragment length polymorphism (RFLP) and characterized gene varia tion at four antigen genes in the Congo population. All genetic assays show abundant genetic variability in all populations analyzed. With the isoenzyme assays, strong linkage disequilibrium is apparent in at least two local populations, the Congo population and one population from Burkina Faso, as well as in the cosmopolitan sample, and less definitely in the other Burkina Faso population. However, no linkage disequilibrium is detected in the Congo population with the molecular assays. We failed to detect any nonrandom association between the different kinds of genetic markers; that is, MLEE with RAPD or RFLP, RAPD with RFLP, and so on. Although isoenzyme data show statistical departures from panmictic expectations, these results suggest that in the areas under survey, P. falciparum populations do not undergo predominant clonal evolution and show no clear-cut subdivisions, un like Trypanosoma cruzi, Leishmania sp., and other major parasitic species. We discuss the epidemiological and taxonomical significance of these results.

A gamma detector probe with ex vivo detection of carcinoid tumors superior to intraoperative palpation.

BACKGROUND: Despite the generally successful scintigraphic detection of endocrine tumors with [(111)In]-DTPA-D-[Phe1]-octreotide (OctreoScan, Mallinckrodt Medical, Petten, The Netherlands), its intraoperative application awaits the development of suitable gamma detectors. This study describes a novel probe (H-probe2) and its ex vivo efficacy for the detection of midgut carcinoid tumors. METHODS: The probe measures 180 mm x 24 mm, and contains a bismuth germinate crystal connected to a photomultiplicator tube, a lead shield, and a tantalum collimator with an angled, 3-mm wide opening. It was characterized in a test bench utilizing solublilized (111)In and 99mTc, and 8 fresh operative specimens containing 26 histologically verified midgut carcinoid tumors (2-40 mm) from patients exposed to OctreoScan. Measurements were made at 2.5-mm intervals over the entire specimens and presented three dimensionally. RESULTS: The test bench analysis supported efficient shielding, and a total collimation of 27 degrees for (111)In and 20 degrees for 99mTc. In addition to 18 palpable tumors, 4 of 6 surgically occult tumors (2-3 mm in dimension) could be discovered with H-probe2. Small tumors in close apposition to a large tumor could not be separated. Wellcounter examination showed that tumors detected with the H-probe2 exhibited 1.7-84.1 times higher radionuclide uptake than the surrounding normal tissue. CONCLUSION: Ex vivo analysis of H-probe2 supported detection of tumors inaccessible to surgical palpation, but its clinical efficiency awaits intraoperative evaluation.

Figs and fig pollinators: evolutionary conflicts in a coevoled mutualism.

Figs and fig wasps form one of the best known examples of species-specific mutualism and coevolution. Recent experiments and observations have led to a better understanding of the evolutionary processes involved in the origin and maintenance of species interactions. The observed fine-tuned traits involve not only coevolution but also selection acting on only one of the partners. Furthermore, some of the 'fine-tuned traits' appear to be be preadaptions - traits that existed before the mutalism was establised.

Are eukaryotic microorganisms clonal or sexual? A population genetics vantage.

We argue that the mode of reproduction of microorganisms in nature can only be decided by population genetic information. The evidence available indicates that many parasitic protozoa and unicellular fungi have clonal rather than sexual population structures, which has major consequences for medical research and practice. Plasmodium falciparum, the agent of malaria, is a special case: the scarce evidence available is contradictory, some suggesting that uniparental lineages may exist in nature. This is puzzling (because P. falciparum is known to have a sexual stage) and poses a challenge that can be readily settled by ascertaining the frequency distribution of genotypes in natural populations.

A clonal theory of parasitic protozoa: the population structures of Entamoeba, Giardia, Leishmania, Naegleria, Plasmodium, Trichomonas, and Trypanosoma and their medical and taxonomical consequences.

We propose a general theory of clonal reproduction for parasitic protozoa, which has important medical and biological consequences. Many parasitic protozoa have been assumed to reproduce sexually, because of diploidy and occasional sexuality in the laboratory. However, a population genetic analysis of extensive data on biochemical polymorphisms indicates that the two fundamental consequences of sexual reproduction (i.e., segregation and recombination) are apparently rare or absent in natural populations of the parasitic protozoa. Moreover, the clones recorded appear to be stable over large geographical areas and long periods of time. A clonal population structure demands that the medical attributes of clones be separately characterized; ubiquitous clones call for priority characterization. Uniparental reproduction renders unsatisfactory Linnean taxonomy; this needs to be supplemented by the "natural clone" as an additional taxonomic unit, which is best defined by means of genetic markers.

THE STABILITY OF THE SYMBIOSIS BETWEEN DIOECIOUS FIGS AND THEIR POLLINATORS: A STUDY OF FICUS CARICA L. AND BLASTOPHAGA PSENES L.

Each Ficus species depends on a specific mutualistic wasp for pollination. The wasp breeds on the fig, each larva destroying a female flower. It is, however, not known why the wasps have not evolved the ability to use all female flowers. In "dioecious" figs, the wasp can only breed in the female flowers of the "male" trees, so that pollination of a female tree is always lethal. The wasps should therefore be selected to avoid female trees. Field data is presented showing that the fruiting phenology of the dioecious fig Ficus carica is such that this selection does not occur: syconia are not receptive at the same time on "male" and female trees. Most wasps are forced to emerge from the syconia of "male" trees at a time when they will not be able to reproduce, whether they avoid female trees or not. This aspect of the life cycle of the wasp, although noticed, has been obscured in most previous studies. It is shown that the fruiting phenology of Ficus carica, which stabilizes the symbiosis, is the result of short-term selective pressures on the male function of the trees. Such selective pressures suggest a possible pathway from monoecy to dioecy in Ficus under seasonal climates.