Project description:Linking omics and ecology to dissect interactions between the apple proliferation phytoplasma and its psyllid vector Cacopsylla melanoneura
Project description:Crop protection requires the accurate identification of disease vectors, a task that can be made difficult when these vectors encompass cryptic species. Here we developed a rapid molecular diagnostic test to identify individuals of Cacopsyllapruni (Scopoli, 1763) (Hemiptera: Psyllidae), the main vector of the European stone fruit yellows phytoplasma. This psyllid encompasses two highly divergent genetic groups that are morphologically similar and that are characterized by genotyping several microsatellite markers, a costly and time-consuming protocol. With the aim of developing species-specific PCR primers, we sequenced the Internal Transcribed Spacer 2 (ITS2) on a collection of C. pruni samples from France and other European countries. ITS2 sequences showed that the two genetic groups represent two highly divergent clades. This enabled us to develop specific primers for the assignment of individuals to either genetic group in a single PCR, based on ITS2 amplicon size. All previously assigned individuals yielded bands of expected sizes, and the PCR proved efficient on a larger sample of 799 individuals. Because none appeared heterozygous at the ITS2 locus (i.e., none produced two bands), we inferred that the genetic groups of C. pruni, whose distribution is partly sympatric, constitute biological species that have not exchanged genes for an extended period of time. Other psyllid species (Cacopsylla, Psylla, Triozidae and Aphalaridae) failed to yield any amplicon. These primers are therefore unlikely to produce false positives and allow rapid assignment of C. pruni individuals to either cryptic species.
Project description:Chemosensory systems play an important role in insect behavior, and some key associated genes have potential as novel targets for pest control. Cacopsylla chinensis is an oligophagous pest and has become one of the main pests of pear trees, but little is known about the molecular-level means by which it locates its hosts. In this study, we assembled the head transcriptome of C. chinensis using Illumina sequencing, and 63,052 Unigenes were identified. A total of 36 candidate chemosensory genes were identified, including five different families: 12 odorant binding proteins (OBPs), 11 chemosensory proteins (CSPs), 7 odorant receptors (ORs), 4 ionotropic receptors (IRs), and 2 gustatory receptors (GRs). The number of chemosensory gene families is consistent with that found in other Hemipteran species, indicating that our approach successfully obtained the chemosensory genes of C. chinensis. The tissue expression of all genes using quantitative real-time PCR (qRT-PCR) found that some genes displayed male head, female head, or nymph-biased specific/expression. Our results enrich the gene inventory of C. chinensis and provide valuable resources for the analysis of the functions of some key genes. This will help in developing molecular targets for disrupting feeding behavior in C. chinensis.
Project description:BackgroundCacopsylla pruni is a psyllid that has been known since 1998 as the vector of the bacterium 'Candidatus Phytoplasma prunorum', responsible for the European stone fruit yellows (ESFY), a disease that affects species of Prunus. This disease is one of the major limiting factors for the production of stone fruits, most notably apricot (Prunus armeniaca) and Japanese plum (P. salicina), in all EU stone fruit-growing areas. The psyllid vector is widespread in the Western Palearctic and evidence for the presence of the phytoplasma that it transmits to species of Prunus has been found in 15 of the 27 EU countries.Recent studies showed that C. pruni is actually composed of two cryptic species that can be differentiated by molecular markers. A literature review on the distribution of C. pruni was published in 2012, but it only provided presence or absence information at the country level and without distinction between the two cryptic species.Since 2012, numerous new records of the vector in several European countries have been published. We ourselves have acquired a large amount of data from sampling in France and other European countries. We have also carried out a thorough systematic literature review to find additional records, including all the original sources mentioning C. pruni (or its synonyms) since the first description by Scopoli in 1763. Our aim was to create an exhaustive georeferenced occurrence catalogue, in particular in countries that are occasionally mentioned in literature with little detail. Finally, for countries that seem suitable for the proliferation of C. pruni (USA, Canada, Japan, China etc.), we dug deeper into literature and reliable sources (e.g. published checklists) to better substantiate its current absence from those regions.Information on the distribution ranges of these vector psyllids is of crucial interest in order to best predict the vulnerability of stone fruit producing countries to the ESFY threat in the foreseeable future.New informationWe give free access to a unique file of 1975 records of all occurrence data in our possession concerning C. pruni, that we have gathered through more than twenty years of sampling efforts in Europe or through intensive text mining.We have made every effort to retrieve the source information for the records extracted from literature (1201 records). Thus, we always give the title of the original reference, together with the page(s) citing C. pruni and, if possible, the year of sampling. To make the results of this survey publicly available, we give a URL to access the literature sources. In most cases, this link allows free downloads of a PDF file.We also give access to information extracted from GBIF (162 exploitable data points on 245 occurrences found in the database), which we thoroughly checked and often supplemented to make the information more easily exploitable.We give access to our own unpublished georeferenced and genotyped records from 612 samples taken over the last 20 years in several European countries (Switzerland, Belgium, Netherlands, Spain etc.). These include two countries (Portugal and North Macedonia), for which the presence of C. pruni had not been reported before. As our specimens have been genotyped (74 sites with species A solely, 202 with species B solely and 310 with species A+B), our new data enable a better overview of the geographical distribution of the two cryptic species at the Palaearctic scale.
Project description:The psyllid Cacopsylla melanoneura is considered one of the vectors of 'Candidatus Phytoplasma mali', the causal agent of apple proliferation disease. In Northern Italy, overwintered C. melanoneura adults reach apple and hawthorn around the end of January. Nymph development takes place between March and the end of April. The new generation adults migrate onto conifers around mid-June and come back to the host plant species after overwintering. In this study we investigated behavioural differences, genetic differentiation and gene flow between samples of C. melanoneura collected from the two different host plants. Further analyses were performed on some samples collected from conifers. To assess the ecological differences, host-switching experiments were conducted on C. melanoneura samples collected from apple and hawthorn. Furthermore, the genetic structure of the samples was studied by genotyping microsatellite markers. The examined C. melanoneura samples performed better on their native host plant species. This was verified in terms of oviposition and development of the offspring. Data resulting from microsatellite analysis indicated a low, but statistically significant difference between collected-from-apple and hawthorn samples. In conclusion, both ecological and genetic results indicate a differentiation between C. melanoneura samples associated with the two host plants.