A PCR-based method for estimating parasitism rates in the olive fly parasitoids Psyttalia concolor and P. lounsburyi (Hymenoptera: Braconidae)

International audienceSeveral parasitoids of the genus Psyttalia have been repeatedly introduced as biological control agents against the principal pest of olive, the fly Bactrocera oleae. However, few of the parasitoids released have become established and proved effective against B. oleae. It may however still be possible to find effective biological control agents adapted to local environmental conditions among the highly diverse Psyttalia species and populations infesting B. oleae worldwide. For this purpose, we have developed a rapid, sensitive molecular method based on the polymerase chain reaction (PCR) for estimating and comparing the parasitism success of Psyttalia parasitoids through the detection of eggs and larvae within the host. This method was tested and shown to be appropriate for two Psyttalia species (Psyttalia concolor and Psyttalia lounsburyi). The possible detection of DNA was also demonstrated for several populations of these species and for other Psyttalia species, namely Psyttalia humilis and Psyttalia ponerophaga. For P. concolor and P. lounsburyi, a strong correlation was observed between the parasitism rates estimated by PCR, host larva dissection and counts of emerging parasitoids. No significant difference was found between the rates of parasitism estimated by host larva dissection and PCR, whereas the rates of parasitism estimated by PCR were significantly higher than those estimated from emergence, suggesting occurrence of mortality during the parasitoid development. This PCR method is thus highly reliable and provides an objective criterion for estimating the efficacy of biological control agent candidates from diverse taxa and populations of Psyttalia. ⇑ Corresponding author

Bridgehead Effect in the Worldwide Invasion of the Biocontrol Harlequin Ladybird

Recent studies of the routes of worldwide introductions of alien organisms suggest that many widespread invasions could have stemmed not from the native range, but from a particularly successful invasive population, which serves as the source of colonists for remote new territories. We call here this phenomenon the invasive bridgehead effect. Evaluating the likelihood of such a scenario is heuristically challenging. We solved this problem by using approximate Bayesian computation methods to quantitatively compare complex invasion scenarios based on the analysis of population genetics (microsatellite variation) and historical (first observation dates) data. We applied this approach to the Harlequin ladybird Harmonia axyridis (HA), a coccinellid native to Asia that was repeatedly introduced as a biocontrol agent without becoming established for decades. We show that the recent burst of worldwide invasions of HA followed a bridgehead scenario, in which an invasive population in eastern North America acted as the source of the colonists that invaded the European, South American and African continents, with some admixture with a biocontrol strain in Europe. This demonstration of a mechanism of invasion via a bridgehead has important implications both for invasion theory (i.e., a single evolutionary shift in the bridgehead population versus multiple changes in case of introduced populations becoming invasive independently) and for ongoing efforts to manage invasions by alien organisms (i.e., heightened vigilance against invasive bridgeheads)

Utilization of microcosms to test invasion biology hypotheses

Comprendre les facteurs déterminant le succès ou l’échec des processus invasifs est un objectif majeur en biologie de l’invasion. De nombreux travaux théoriques se sont intéressés aux composantes écologiques et évolutives de ces facteurs. Cependant, les tests d’hypothèses associés à une démarche expérimentale restent rares. La plupart des résultats empiriques sont issus de l’analyse a posteriori d’invasions fortuites et ne permettent donc, au mieux, que des approches corrélatives. Dans cet article, nous discutons de la pertinence des microcosmes, i.e. des environnements simplifiés, contrôlés et reproductibles, comme alternatives aux introductions expérimentales en milieu naturel. En nous basant sur une synthèse de la littérature, nous présentons les avantages et limites des approches en microcosmes pour l'étude des invasions biologiques. Notre analyse se concentre sur les publications impliquant des populations en dynamique transitoire après un goulot d’étranglement et/ou soumises à un challenge adaptatif, deux caractéristiques clés des processus invasifs. Malgré le nombre encore réduit de telles études, leur intérêt a été montré pour explorer les influences des caractéristiques de l’aire envahie (les conditions environnementales ainsi que leur hétérogénéité spatiale ou temporelle). Dans une moindre mesure, les microcosmes ont également permis de tester l’influence des caractéristiques des populations introduites et de la communauté envahie. Cependant, ils doivent être utilisés avec précaution car ils ne permettent pas de reproduire la complexité des milieux naturels. Les expériences en microcosmes sont donc complémentaires aux études théoriques et à celles menées en populations naturelles et contribuent à renforcer la valeur prédictive de la biologie de l’invasion en liant théorie et expérimentation.Understanding the factors underlying establishment and spread of exotic species in order to predict invasion risks is a major goal in invasion biology. Many theoretical studies investigated the ecological and evolutionary components of these factors and their impact on the invasive process. Yet, hypothesis tests through experimental approaches are still scarce because of the practical and ethical difficulties associated with the introduction of exotic species in nature. Thus, most empirical results come from a posteriori analyses of fortuitous invasions, which allow correlative approaches at best and give no information about invasion failures. In this paper, we propose microcosms, i.e. reproducible controlled simplified environments, as an alternative to experimental introductions in natura. From a review of the literature, we discuss the distinctive features of microcosms to test theoretical predictions about invasion. Our analysis focuses on studies involving populations in transitory dynamics after a demographic bottleneck and/or subject to an adaptive challenge, two key characteristics of invasive processes. Despite their small number, these studies have been used successfully to explore the influences of various factors, mainly related to the introduction site characteristics (its abiotic conditions and their spatial and temporal heterogeneity), and to a lesser extent to the introduced individuals themselves (propagule pressure, genetic diversity and adaptations in the introduced population) or the invaded community. We argue that microcosms, as model systems, can be powerful tools to test theoretical hypotheses. They must however be used with care, as they do not account for the same complexity as natural systems. They are thus complementary to theoretical studies and field surveys, and contribute to reinforce the predictive value of invasion biology by linking theory and experimentation

Rapid and dfferential evolution of the venom composition of a parasitoid wasp depending on the host strain

Abstract: Parasitoid wasps rely primarily on venom to suppress the immune response and regulatethe physiology of their host. Intraspecific variability of venom protein composition has beendocumented in some species, but its evolutionary potential is poorly understood. We performed anexperimental evolution initiated with the crosses of two lines of Leptopilina boulardi of differentvenom composition to generate variability and create new combinations of venom factors. Theoffspring were maintained for 10 generations on two strains of Drosophila melanogaster differing inresistance/susceptibility to the parental parasitoid lines. The venom composition of individuals wascharacterized by a semi-automatic analysis of 1D SDS-PAGE electrophoresis protein profiles whoseaccuracy was checked by Western blot analysis of well-characterized venom proteins. Results madeevident a rapid and differential evolution of the venom composition on both hosts and showed thatthe proteins beneficial on one host can be costly on the other. Overall, we demonstrated the capacityof rapid evolution of the venom composition in parasitoid wasps, important regulators of arthropodpopulations, suggesting a potential for adaptation to new hosts. Our approach also proved relevantin identifying, among the diversity of venom proteins, those possibly involved in parasitism successand whose role deserves to be deepened

Development of 55 novel polymorphic microsatellite loci for the critically endangered L. (Actinopterygii: Perciformes: Percidae) and cross-species amplification in five other percids

International audienceBy combining biotin-enrichment protocol and next generation pyrosequencing, through 454 GS-FLX Titanium technology, 55 polymorphic microsatellites loci with perfect motif were isolated from the Rhone streber (), a critically endangered European fish species. Eight multiplex PCR kits were optimised in order to genotype a total of 58 polymorphic loci, including three previously published loci. The level of genetic diversity was assessed for 68 , 30 , 33 and four individuals. Amplification success was also assessed on and using single individuals. These markers will be useful to investigate the population structure of the highly fragmented Rhone streber. They represent a powerful tool for conservation issues and evolutionary approaches of this endemic species. Moreover, part of our markers demonstrated applicability to other percid species, allowing for potential applications to fisheries and aquaculture management

Parasitoids (Hymenoptera) of Mealybug Pests (Hemiptera: Pseudococcidae) from Southern Brazil: Molecular and Morphological Characterization

Parasitoids of three mealybug pests (Hemiptera: Pseudococcidae), Planococcus ficus (Signoret), Pseudococcus sociabilis Hambleton, and Pseudococcus viburni (Signoret) have been identified for the first time in Brazil. Mealybugs were collected in fruit-growing areas along southern Brazil during 2013-2016. An integrative approach, combining morphological and molecular methods, was used to identify the Brazilian parasitoids to the species level. Fifteen species were recorded, including 14 primary parasitoids belonging to Encyrtidae and Platygastridae and a single secondary parasitoid species belonging to Signiphoridae. The encyrtid parasitoids Acerophagus flavidulus (Brethes), Anagyrus calyxtoi Noyes and Zaplatycerus sp., and the signiphorid secondary parasitoid Chartocerus axillaris De Santis are reported for the first time in Brazil.Centro de Estudios Parasitológicos y de VectoresCentro de Investigación en Sanidad Vegeta

Guiding Classical Biological Control of an Invasive Mealybug Using Integrative Taxonomy

The analysed sequences were deposited in Genbank under accession numbers KP771926-KP771972. Mealybug slides are available at the Polytechnic University of Valencia (Valencia, Spain). Parasitoid slides are deposited at Università degli Studi di Napoli Federico II (Portici, Italy).[EN] Delottococcus aberiae De Lotto (Hemiptera: Pseudococcidae) is a mealybug of Southern African origin that has recently been introduced into Eastern Spain. It causes severe distortions on young citrus fruits and represents a growing threat to Mediterranean citrus production. So far, biological control has proven unsatisfactory due to the absence of efficient natural enemies in Spain. Hence, the management of this pest currently relies only on chemical control. The introduction of natural enemies of D. aberiae from the native area of the pest represents a sustainable and economically viable alternative to reduce the risks linked to pesticide applications. Since biological control of mealybugs has been traditionally challenged by taxonomic misidentification, an intensive survey of Delottococcus spp. and their associated parasitoids in South Africa was required as a first step towards a classical biological control programme. Combining morphological and molecular characterization (integrative taxonomy) a total of nine mealybug species were identified in this study, including three species of Delottococcus. Different populations of D. aberiae were found on wild olive trees, in citrus orchards and on plants of Chrysanthemoides monilifera, showing intra-specific divergences according to their host plants. Interestingly, the invasive mealybug populations from Spanish orchards clustered together with the population on citrus from Limpopo Province (South Africa), sharing COI haplotypes. This result pointed to an optimum location to collect natural enemies against the invasive mealybug. A total of 14 parasitoid species were recovered from Delottococcus spp. and identified to genus and species level, by integrating morphological and molecular data. A parasitoid belonging to the genus Anagyrus, collected from D. aberiae in citrus orchards in Limpopo, is proposed here as a good biological control agent to be introduced into Spain.This work was supported by the European Union Seventh Framework Programme FP7-IRSES #269196 "IPRABIO" and FP7-IAPP #324475 "COLBICS" and FP7-IRSES #612566 "BIOMODICS". The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.Beltrà Ivars, A.; Addison, P.; Ávalos Masó, JA.; Crochard, D.; García Mari, F.; Guerrieri, E.; Giliomee, JH.... (2015). Guiding Classical Biological Control of an Invasive Mealybug Using Integrative Taxonomy. PLoS ONE. 10(6):1-14. https://doi.org/10.1371/journal.pone.0128685S11410

Accuracy and quality assessment of 454 GS-FLX Titanium pyrosequencing

<p>Abstract</p> <p>Background</p> <p>The rapid evolution of 454 GS-FLX sequencing technology has not been accompanied by a reassessment of the quality and accuracy of the sequences obtained. Current strategies for decision-making and error-correction are based on an initial analysis by Huse <it>et al. </it>in 2007, for the older GS20 system based on experimental sequences. We analyze here the quality of 454 sequencing data and identify factors playing a role in sequencing error, through the use of an extensive dataset for Roche control DNA fragments.</p> <p>Results</p> <p>We obtained a mean error rate for 454 sequences of 1.07%. More importantly, the error rate is not randomly distributed; it occasionally rose to more than 50% in certain positions, and its distribution was linked to several experimental variables. The main factors related to error are the presence of homopolymers, position in the sequence, size of the sequence and spatial localization in PT plates for insertion and deletion errors. These factors can be described by considering seven variables. No single variable can account for the error rate distribution, but most of the variation is explained by the combination of all seven variables.</p> <p>Conclusions</p> <p>The pattern identified here calls for the use of internal controls and error-correcting base callers, to correct for errors, when available (e.g. when sequencing amplicons). For shotgun libraries, the use of both sequencing primers and deep coverage, combined with the use of random sequencing primer sites should partly compensate for even high error rates, although it may prove more difficult than previous thought to distinguish between low-frequency alleles and errors.</p

Representativeness of microsatellite distributions in genomes, as revealed by 454 GS-FLX Titanium pyrosequencing

<p>Abstract</p> <p>Background</p> <p>Microsatellites are markers of choice in population genetics and genomics, as they provide useful insight into patterns and processes as diverse as genome evolutionary dynamics and demographic processes. The acquisition of microsatellites through multiplex-enriched libraries and 454 GS-FLX Titanium pyrosequencing is a promising new tool for the isolation of new markers in unknown genomes. This approach can also be used to evaluate the extent to which microsatellite-enriched libraries are representative of the genome from which they were isolated. In this study, we deciphered potential discrepancies in microsatellite content recovery for two reference genomes (<it>Apis mellifera </it>and <it>Danio rerio</it>), selected on the basis of their extreme heterogeneity in terms of the proportions and distributions of microsatellites on chromosomes.</p> <p>Results</p> <p>The <it>A. mellifera </it>genome, in particular, was found to be highly heterogeneous, due to extremely high rates of recombination, with hotspots, but the only bias consistently introduced into pyrosequenced multiplex-enriched libraries concerned sequence length, with the overrepresentation of sequences 160 to 320 bp in length. Other deviations from expected proportions or distributions of motifs on chromosomes were observed, but the significance and intensity of these deviations was mostly limited. Furthermore, no consistent adverse competition between multiplexed probes was observed during the motif enrichment phase.</p> <p>Conclusions</p> <p>This approach therefore appears to be a promising strategy for improving the development of microsatellites, as it introduces no major bias in terms of the proportions and distribution of microsatellites.</p