Lapo Ragionieri

Phylogenetic and morphometric differentiation reveal geographic radiation and pseudo-cryptic speciation in a mangrove crab from the Indo-West Pacific

The presence of boundaries to dispersal has been recently documented for many Indo-West Pacific (IWP) species with planktonic propagules and a widespread distribution. We studied the phylogeography of the mangrove crab Neosarmatium meinerti (Brachyura: Sesarmidae) and the phylogenetic relationship to its presumed sister species N. fourmanoiri in the IWP in order to compare intraspecific with interspecific diversity. Portions of the mitochondrial genes 16S and CoxI were sequenced for 23 specimens of N. meinerti and 5 N. fourmanoiri, while a fragment of the 28S was obtained for a subset of specimens. Genetic data are supplemented by morphometric and based on 37 adult males of N. meinerti and 9 males of N. fourmanoiri. The conserved nuclear 28S reveals the existence of a genetic break between the Indian and Pacific oceans. Otherwise, mitochondrial genes as well as morphometry clearly support the presence of a species complex within N. meinerti composed by four well structured and geographically defined lineages: East African coast; western Indian Ocean islands; South East Asia; and Australia.

Demographic History and Reproductive Output Correlates with Intraspecific Genetic Variation in Seven Species of Indo-Pacific Mangrove Crabs.

The spatial distribution and the amount of intraspecific genetic variation of marine organisms are strongly influenced by many biotic and abiotic factors. Comparing biological and genetic data characterizing species living in the same habitat can help to elucidate the processes driving these variation patterns. Here, we present a comparative multispecies population genetic study on seven mangrove crabs co-occurring in the West Indian Ocean characterized by planktotrophic larvae with similar pelagic larval duration. Our main aim was to investigate whether a suite of biological, behavioural and ecological traits could affect genetic diversities of the study species in combination with historical demographic parameters. As possible current explanatory factors, we used the intertidal micro-habitat colonised by adult populations, various parameters of individual and population fecundity, and the timing of larval release. As the genetic marker, we used partial sequences of cytochrome oxidase subunit I gene. Genetic and ecological data were collected by the authors and/or gathered from primary literature. Permutational multiple regression models and ANOVA tests showed that species density and their reproductive output in combination with historical demographic parameters could explain the intraspecific genetic variation indexes across the seven species. In particular, species producing consistently less eggs per spawning event showed higher values of haplotype diversity. Moreover, Tajima’s D parameters well explained the recorded values for haplotype diversity and average γst. We concluded that current intraspecific gene diversities in crabs inhabiting mangrove forests were affected by population fecundity as well as past demographic history. The results were also discussed in terms of management and conservation of fauna in the Western Indian Ocean mangroves.

Establishing the eradication unit of Molara Island: a case of study from Sardinia, Italy.

Molara is a small island belonging to the Marine protected Area Tavolara—Punta Coda Cavallo, in Sardinia. During 2006–2007, a bio-monitoring program reported a strong presence of the black rat, Rattus rattus, on Molara island. Rat predation has detrimentally affected the unique biodiversity of this island, thus, in 2008 an eradication campaign was conducted. Our eradication protocol included a pre-eradication genetic investigation, using 8 microsatellite loci, on a rat population of Molara as well as on neighbour islands within the Marine Protected Area (MPA). The main goal of this genetic investigation was to establish the correct borders of the eradication unit of Molara island. As several recent eradication campaigns have been unsuccessful, due to incomplete and unstable eradication, we also aimed to assess possible hidden sources of reinvasion. Specimens were also collected during post- eradication monitoring on Molara for genetic screening to establish their origin, and thus validate the effectiveness of our eradication campaign. According to our genetic analysis, within the MPA there are four different eradication units, corresponding to the islands of Molara, Tavolara, Piana and to the Sardinia mainland. Gene flow among these four units is more or less absent. The assignment and clustering tests performed on pre and post-eradication samples seem to indicate that the population of Sardinia mainland is a possible source of re-invasion for the Piana and Molara populations.

Phylogeography of the marbled crab Pachygrapsus marmoratus (Decapoda, Grapsidae) along part of the African Mediterranean coast reveals genetic homogeneity across the Siculo-Tunisian Strait versus heterogeneity across the Gibraltar Strait

ABSTRACT We investigate the influence of previously postulated biogeographic barriers in the Mediterranean Sea on the population genetic structure of a highly dispersive and continuously distributed coastal species. In particular, we examine nuclear and mitochondrial genetic variation in the marbled crab, Pachygrapsus marmoratus, across part of the African Mediterranean coast in order to assess the influence of the Siculo-Tunisian Strait on its population genetic structure. Four polymorphic microsatellite loci were genotyped for 110 individuals, collected from eight locations covering parts of the Algerian, Tunisian and Libyan coasts. In addition, mtDNA corresponding to the Cox1 gene was sequenced for 80 samples. The corresponding results show contrasting patterns of genetic differentiation. While mtDNA results revealed a homogeneous haplotype composition in our study area, microsatellite data depicted genetic differentiation among populations, but not associated with any geographic barrier. This pattern, already recorded for this species from different geographic regions, may hint at the involvement of a complex series of abiotic and biotic factors in determining genetic structure. Demographic history reconstruction, inferred from mtDNA data, supports demographic and spatial expansion for the North African metapopulation dating back to the Mid-Pleistocene and following an historical bottleneck. Comparison of these African mitochondrial sequences with new sequences from a Turkish population and previously published sequences revealed a weak but significant separation of Atlantic and Mediterranean populations across the Gibraltar Strait, which was not recorded in previous studies of this grapsid species.

Identification of mature peptides from pban and capa genes of the moths Heliothis peltigera and Spodoptera littoralis

HIGHLIGHTSPBAN and CAPA peptides were identified from two moth species.pban and capa genes each encode for a sequence‐related tryptopyrokinin.The diapause hormone is two amino acids longer than expected.Release sites of CAPA peptides in the abdomen show unique structural features. ABSTRACT By transcriptome analysis, we identified PBAN and CAPA precursors in the moths Spodoptera littoralis and Heliothis peltigera which are among the most damaging pests of agriculture in tropical and subtropical Africa as well as in Mediterranean countries. A combination of mass spectrometry and immunocytochemistry was used to identify mature peptides processed from these precursors and to reveal their spatial distribution in the CNS. We found that the sites of expression of pban genes, the structure of PBAN precursors and the processed neuropeptides are very similar in noctuid moths. The sequence of the diapause hormone (DH; tryptopyrokinin following the signal peptide), however, contains two N‐terminal amino acids more than expected from comparison with already published sequences of related species. Capa genes of S. littoralis and H. peltigera encode, in addition to periviscerokinins, a tryptopyrokinin showing sequence similarity with DH, which is the tryptopyrokinin of the pban gene. CAPA peptides, which were not known from any noctuid moth so far, are produced in cells of abdominal ganglia. The shape of the release sites of these hormones in H. peltigera represents an exceptionally derived trait state and does not resemble the well‐structured abdominal perisympathetic organs which are known from many other insects. Instead, axons of CAPA cells extensively ramify within the ventral diaphragm. The novel information regarding the sequences of all mature peptides derived from pban and capa genes of H. peltigera and S. littoralis now enables a detailed analysis of the bioactivity and species‐specificity of the native peptides, especially those from the hitherto unknown capa genes, and to explore their interactions with PBAN/DH receptors.

Temporal patterns of megalopal settlement in different areas of an East African mangrove forest (Gazi Bay, Kenya)

Most intertidal brachyurans produce planktonic larvae which develop pelagically and, after a certain time in the ocean, migrate towards the habitats that they will eventually settle in. One of the main physical processes affecting larval release and settlement in species inhabiting estuaries and mangroves is the tidal regime. In this study, we investigated whether patterns of settlement of brachyuran larvae at four sites (differing in tidal inundation and crab zonation) of a Kenyan mangrove were affected by the diurnal and lunar cycle of the tide. We collected megalopae at the four sites twice a day throughout two lunar months. Settlement differed at the four sites: at the subtidal site (the main creek within the forest) megalopae arrived during diurnal and nocturnal neap and spring tides, while at the three sites within the forest settlement occurred only at spring tide periods. Specific differences among these latter sites existed in terms of full versus new moon spring tides and, to a smaller extent, with diurnal period. Our results show that larval settlement in mangrove forests takes place at both landward and seaward belts and is a temporally complex event, driven by tidal cycles, but also in synergy with other factors.

Coordinated RNA-Seq and peptidomics identify neuropeptides and G-protein coupled receptors (GPCRs) in the large pine weevil Hylobius abietis, a major forestry pest.

Hylobius abietis (Linnaeus), or large pine weevil (Coleoptera, Curculionidae), is a pest of European coniferous forests. In order to gain understanding of the functional physiology of this species, we have assembled a de novo transcriptome of H. abietis, from sequence data obtained by Next Generation Sequencing. In particular, we have identified genes encoding neuropeptides, peptide hormones and their putative G-protein coupled receptors (GPCRs) to gain insights into neuropeptide-modulated processes. The transcriptome was assembled de novo from pooled paired-end, sequence reads obtained from RNA from whole adults, gut and central nervous system tissue samples. Data analysis was performed on the transcripts obtained from the assembly including, annotation, gene ontology and functional assignment as well as transcriptome completeness assessment and KEGG pathway analysis. Pipelines were created using Bioinformatics tools and techniques for prediction and identification of neuropeptides and neuropeptide receptors. Peptidomic analysis was also carried out using a combination of MALDI-TOF as well as Q-Exactive Orbitrap mass spectrometry to confirm the identified neuropeptide. 41 putative neuropeptide families were identified in H. abietis, including Adipokinetic hormone (AKH), CAPA and DH31. Neuropeptide F, which has not been yet identified in the model beetle T. castaneum, was identified. Additionally, 24 putative neuropeptide and 9 leucine-rich repeat containing G protein coupled receptor-encoding transcripts were determined using both alignment as well as non-alignment methods. This information, submitted to the NCBI sequence read archive repository (SRA accession: SRP133355), can now be used to inform understanding of neuropeptide-modulated physiology and behaviour in H. abietis; and to develop specific neuropeptide-based tools for H. abietis control.