Shifted phenology in the pine processionary moth affects the outcome of tree-insect interaction.

In the Mediterranean and temperate regions, an increase in the frequency and intensity of drought events has been recorded, probably due to climate change. In consequence, trees will more frequently experience hydric stress, a condition that can be expected to affect insect-tree interactions, while adaptation mechanisms may be further in course. The effect of tree water stress on the performance of two allochronic populations of Thaumetopoea pityocampa was here studied. Namely, we compared a unique population of this insect, in which the larvae develop in the summer (SP), with the typical population having winter larval development (WP), to test the adaptation hypothesis to host plant status. Larvae of each population were fed on needles of young potted Pinus pinaster plants under two water supply regimes: (i) well-watered (control) and (ii) subjected to 3 months of drought stress. Compared to control, stressed plants had higher amounts of soluble sugars, phenols, and higher C/N ratio, whereas water content and chlorophylls concentrations were lower. In general, T. pityocampa larvae had lower performances on water-stressed plants, as shown by lower survival rates, lower needle consumption, and longer development times. Yet, the detrimental effects of tree stress were only significant for the WP larvae, while SP larvae were able to overcome such conditions. Results demonstrate that tree water stress can negatively affect T. pityocampa populations. Furthermore, the evidence is also provided that responses to the physiological condition of the host trees may occur at the population level, as a result of adaptation mechanisms driven by climate change.

Draft genome and reference transcriptomic resources for the urticating pine defoliator Thaumetopoea pityocampa (Lepidoptera: Notodontidae).

The pine processionary moth Thaumetopoea pityocampa (Lepidoptera: Notodontidae) is the main pine defoliator in the Mediterranean region. Its urticating larvae cause severe human and animal health concerns in the invaded areas. This species shows a high phenotypic variability for various traits, such as phenology, fecundity and tolerance to extreme temperatures. This study presents the construction and analysis of extensive genomic and transcriptomic resources, which are an obligate prerequisite to understand their underlying genetic architecture. Using a well-studied population from Portugal with peculiar phenological characteristics, the karyotype was first determined and a first draft genome of 537 Mb total length was assembled into 68,292 scaffolds (N50 = 164 kb). From this genome assembly, 29,415 coding genes were predicted. To circumvent some limitations for fine-scale physical mapping of genomic regions of interest, a 3X coverage BAC library was also developed. In particular, 11 BACs from this library were individually sequenced to assess the assembly quality. Additionally, de novo transcriptomic resources were generated from various developmental stages sequenced with HiSeq and MiSeq Illumina technologies. The reads were de novo assembled into 62,376 and 63,175 transcripts, respectively. Then, a robust subset of the genome-predicted coding genes, the de novo transcriptome assemblies and previously published 454/Sanger data were clustered to obtain a high-quality and comprehensive reference transcriptome consisting of 29,701 bona fide unigenes. These sequences covered 99% of the cegma and 88% of the busco highly conserved eukaryotic genes and 84% of the busco arthropod gene set. Moreover, 90% of these transcripts could be localized on the draft genome. The described information is available via a genome annotation portal (http://bipaa.genouest.org/sp/thaumetopoea_pityocampa/).

Deciphering the demographic history of allochronic differentiation in the pine processionary moth Thaumetopoea pityocampa.

Understanding the processes of adaptive divergence, which may ultimately lead to speciation, is a major question in evolutionary biology. Allochronic differentiation refers to a particular situation where gene flow is primarily impeded by temporal isolation between early and late reproducers. This process has been suggested to occur in a large array of organisms, even though it is still overlooked in the literature. We here focused on a well-documented case of incipient allochronic speciation in the winter pine processionary moth Thaumetopoea pityocampa. This species typically reproduces in summer and larval development occurs throughout autumn and winter. A unique, phenologically shifted population (SP) was discovered in 1997 in Portugal. It was proved to be strongly differentiated from the sympatric "winter population" (WP), but its evolutionary history could only now be explored. We took advantage of the recent assembly of a draft genome and of the development of pan-genomic RAD-seq markers to decipher the demographic history of the differentiating populations and develop genome scans of adaptive differentiation. We showed that the SP diverged relatively recently, that is, few hundred years ago, and went through two successive bottlenecks followed by population size expansions, while the sympatric WP is currently experiencing a population decline. We identified outlier SNPs that were mapped onto the genome, but none were associated with the phenological shift or with subsequent adaptations. The strong genetic drift that occurred along the SP lineage certainly challenged our capacity to reveal functionally important loci.

Effect of heat waves on embryo mortality in the pine processionary moth.

Extreme climate events such as heat waves are predicted to become more frequent with climate change, representing a challenge for many organisms. The pine processionary moth Thaumetopoea pityocampa is a Mediterranean pine defoliator, which typically lays eggs during the summer. We evaluated the effects of heat waves on egg mortality of three populations with different phenologies: a Portuguese population with a classical life cycle (eggs laid in summer), an allochronic Portuguese population reproducing in spring, and a Tunisian population from the extreme southern limit of T. pityocampa distribution range, in which eggs are laid in fall. We tested the influence of three consecutive hot days on egg survival and development time, using either constant (CT) or daily cycling temperatures (DT) with equivalent mean temperatures. Maximum temperatures (T max) used in the experiment ranged from 36 to 48°C for DT and from 30 to 42°C for CT. Heat waves had a severe negative effect on egg survival when T max reached 42°C for all populations. No embryo survived above this threshold. At high mean temperatures (40°C), significant differences were observed between populations and between DT and CT regimes. Heat waves further increased embryo development time. The knowledge we gained about the upper lethal temperature to embryos of this species will permit better prediction of the potential expansion of this insect under different climate warming scenarios.

Contrasted invasion processes imprint the genetic structure of an invasive scale insect across southern Europe.

Deciphering the colonization processes by which introduced pests invade new areas is essential to limit the risk of further expansion and/or multiple introductions. We here studied the invasion history of the maritime pine bast scale Matsucoccus feytaudi. This host-specific insect does not cause any damage in its native area, but it devastated maritime pine forests of South-Eastern France where it was detected in the 1960s, and since then reached Italy and Corsica. We used population genetic approaches to infer the populations' recent evolutionary history from microsatellite markers and Approximate Bayesian Computation. Consistent with previous mitochondrial data, we showed that the native range is geographically strongly structured, which is probably due to the patchy distribution of the obligate host and the limited dispersal capacity of the scale. Our results show that the invasion history can be described in three successive steps involving different colonization and dispersal processes. During the mid-XXth century, massive introductions occurred from the Landes planted forest to South-Eastern France, probably due to transportation of infested wood material after World War II. Stepping-stone expansion, consistent with natural dispersal, then allowed M. feytaudi to reach the maritime pine forests of Liguria and Tuscany in Italy. The island of Corsica was accidentally colonized in the 1990s, and the most plausible scenario involves the introduction of a limited number of migrants from the forests of South-Eastern France and Liguria, which is consistent with an aerial dispersal due to the dominant winds that blow in spring in this region.

Permanent genetic resources added to Molecular Ecology Resources Database 1 August 2011-30 September 2011.

This article documents the addition of 299 microsatellite marker loci and nine pairs of single-nucleotide polymorphism (SNP) EPIC primers to the Molecular Ecology Resources (MER) Database. Loci were developed for the following species: Alosa pseudoharengus, Alosa aestivalis, Aphis spiraecola, Argopecten purpuratus, Coreoleuciscus splendidus, Garra gotyla, Hippodamia convergens, Linnaea borealis, Menippe mercenaria, Menippe adina, Parus major, Pinus densiflora, Portunus trituberculatus, Procontarinia mangiferae, Rhynchophorus ferrugineus, Schizothorax richardsonii, Scophthalmus rhombus, Tetraponera aethiops, Thaumetopoea pityocampa, Tuta absoluta and Ugni molinae. These loci were cross-tested on the following species: Barilius bendelisis, Chiromantes haematocheir, Eriocheir sinensis, Eucalyptus camaldulensis, Eucalyptus cladocalix, Eucalyptus globulus, Garra litaninsis vishwanath, Garra para lissorhynchus, Guindilla trinervis, Hemigrapsus sanguineus, Luma chequen. Guayaba, Myrceugenia colchagüensis, Myrceugenia correifolia, Myrceugenia exsucca, Parasesarma plicatum, Parus major, Portunus pelagicus, Psidium guayaba, Schizothorax richardsonii, Scophthalmus maximus, Tetraponera latifrons, Thaumetopoea bonjeani, Thaumetopoea ispartensis, Thaumetopoea libanotica, Thaumetopoea pinivora, Thaumetopoea pityocampa ena clade, Thaumetopoea solitaria, Thaumetopoea wilkinsoni and Tor putitora. This article also documents the addition of nine EPIC primer pairs for Euphaea decorata, Euphaea formosa, Euphaea ornata and Euphaea yayeyamana.

Incipient allochronic speciation in the pine processionary moth (Thaumetopoea pityocampa, Lepidoptera, Notodontidae).

A plausible case of allochronic differentiation, where barrier to gene flow is primarily attributable to a phenological shift, was recently discovered in Portugal for the pine processionary moth Thaumetopoea pityocampa. Previous results suggested that the observed 'summer population' (SP) originated from the sympatric winter population (WP). Our objectives were to finely analyse these patterns and test their stability in time, through field monitoring and genetic analyses of larvae and adults across different years. Reproductive activity never overlapped between SP and WP. Microsatellites showed a clear differentiation of the SP, consistent with a strong reduction in gene flow owing to the phenological shift. Assignment tests suggested that some individuals shift from the SP to the WP phenology, causing some hybridization. We discuss these patterns and their maintenance over time. This could be a first stage of allochronic speciation, and SP should be considered as a distinct phenological race.

Phylogeography of maritime pine inferred with organelle markers having contrasted inheritance.

Range-wide variation of maritime pine was studied at maternally inherited and paternally inherited markers (mitochondrial DNA and chloroplast DNA). While chloroplast DNA exhibits the highest diversity, phylogeographic inferences from this marker are blurred by homoplasy and extensive pollen flow. In contrast, the only three mitochondrial haplotypes found provide a clear picture of nonoverlapping areas colonized from different refugia, with no single population having a mixed composition (GST = 1). Comparison of the genetic structure inferred from both organelle genomes allows the investigation of differential seed and pollen dispersal, pointing to pollen, but not seed, dispersal across the Strait of Gibraltar (from Morocco into Iberia). A comparison with already available genetic information, especially that of one of the maritime pine's most threatening insect pests, the bast scale Matsucoccus feytaudi, further completes the picture.

Rangewide variation of the maritime pine bast scale matsucoccus feytaudi duc. (Homoptera: matsucoccidae) in relation to the genetic structure of its host

The bast scale Matsucoccus feytaudi is a specific pest of maritime pine, but the damage inflicted by the insect on the host trees is variable, ranging from no apparent effect to severe decline of the maritime pine stands. Rangewide variation of mitochondrial DNA among M. feytaudi populations was analysed by polymerase chain reaction-restriction fragment length-single-strand conformation polymorphism (PCR-RFLP-SSCP) analysis and the results compared with the genetic information already available for its host. Three main nonoverlapping lineages can be distinguished in M. feytaudi. The phylogeography of the pest population is clearly related to the history of its host. Most local associations could result from common evolution while others must be interpreted as intraspecific host shifts. Because the distribution of cultivated tree species is greatly influenced by humans, much may be learned concerning their genetic structure from the indirect study of their specific pests.

Terpene variations in maritime pine constitutive oleoresin related to host tree selection byDioryctria sylvestrella RATZ. (Lepidoptera: Pyralidae).

Gas chromatographic analysis of the volatile terpenes of the wood oleoresin of maritime pines showed that the relative proportions of terpenes within the same tree did not vary according to sampling height and date. Pruning and tree vigor did not affect terpene profiles, but oleoresin composition varied widely from tree to tree. The trees attacked byDioryctria sylvestrella contained a significantly higher percentage of limonene, longipinene, and copaene, and a discriminant analysis, using the relative percentage of six terpenes, significantly separated infested and uninfested trees. The olfactory sensitivity of adult moths to a series of 20 volatile chemicals, including maritime pine oleoresin components, was evaluated with the electroantennogram method. Both sexes were sensitive to monoterpenes, oxygenated terpenoid compounds, and oleoresin samples. The use of terpenes as host attractants and biochemical markers for resistance toD. sylvestrella is discussed.

[Spontaneous pregnancies in couples after failed or successful in vitro fertilization]. / Grossesses spontanées chez les couples après échec ou réussite de fécondation in vitro.

OBJECTIVE: To determinate the true incidence of treatment-independent pregnancy in an in vitro fertilization programme. To establish and to compare the characteristics of couples with and without spontaneous pregnancy. To analyze the outcome of pregnancies. TYPE OF STUDY: Retrospective. SETTING: In Vitro Fertilization Unit, Conception's Hospital, Marseille, France. SUBJECTS: 594 couples having attempted one or more IVF procedures. The study concerned low-fertility couples (484) ruling our true sterile couples (110). RESULTS: Spontaneous pregnancies occurred in 54 couples (11.2%). The characteristics of the two populations were not statistically different, except a shorter duration of infertility (p < 0.05) in spontaneous pregnancies. The rate of ectopic pregnancies in the spontaneous pregnancies was statistically higher than that observed in IVP pregnancies (12% versus 6.5%). The fertility rare of these couples was very low (0.38%). CONCLUSION: Spontaneous pregnancies in vitro fertilization programmes are not rare. The evaluation of the results of the in vitro fertilization requires taking the possible spontaneous pregnancies into account.