Chestnut Gall Wasp Dryocosmus Kuriphilus Research Articles

Torymus sinensis and its close relatives in Europe: a multilocus phylogeny, detailed morphological analysis, and identification key

The introduction of the biological control agent Torymus sinensis Kamijo (Hymenoptera, Chalcidoidea, Torymidae) to control the populations of the chestnut gall wasp Dryocosmus kuriphilus Yasumatsu (Hymenoptera, Cynipidae) is considered one of the successful programs in biological control. The species was involved in interspecific hybridisation in Japan and the specimens imported into Europe were derived from this hybrid lineage, showing signs of introgression. The discovery of mitochondrial haplotypes or possible Enolase haplotypes from T. beneficus or of specimens with shorter ovipositor does not necessarily imply that T. beneficus is present in Europe, only that the European specimens are of hybrid origin. Of the native European Torymus species associated with D. kuriphilus, the molecular and morphometric results indicate Torymus notatus (Walker) as the closest species to T. sinensis. The two are part of the same species-group (cyaneus group), are nested together in the multivariate ratio analysis and are the closest genetically based on all three nuclear markers: Enolase (1.5% divergence), Wingless (2%) and ITS2 (13%). However, on the mitochondrial marker COI the closest species is Torymus rubi (Schrank) at 9.9% divergence. As such, T. notatus is the most likely candidate for accidental interspecific hybridisation if this is to happen in Europe. We provide an illustrated identification key for the European species of Torymus associated with D. kuriphilus, an important but lacking tool for biological control programs.

Assessing the susceptibility levels of chestnut cultivars/genotypes to Asian chestnut gall wasp (Dryocosmus kuriphilus Yasumatsu)

The Asian chestnut gall wasp Dryocosmus kuriphilus is a major insect pest affecting chestnut trees worldwide. Due to the galls that it creates on shoots and leaves, this pest prevents shoot development and blooming and decreases the fruit yield. In Türkiye, this pest was first seen in Gacık village of Yalova province in the year 2014. Today, it is seen in orchards and forests in different provinces of Marmara, Aegean, and Black Sea regions. The medium- and long-term control of this pest can be improved by making use of resistant or less-susceptible cultivars and hybrid clones. The susceptibility levels of our cultivars to this pest are not known. For this purpose, by using three infestation indicators, the susceptibility to Asian chestnut gall wasp (ACGW) of 3 hybrids (Castanea sativa x Castanea crenate) and 20 cultivars/genotypes (C. sativa), which came to the forefront via selection studies, were assessed for five years (2017-2021). The susceptibility to ACGW varied between the tested genotypes/cultivars and, since no gall development was observed in ‘YD-1’, ‘Tülü’, and ‘Ertan’ cultivars of C. sativa species, they were determined to be completely resistant to ACGW. Furthermore, the numbers of ACGW eggs on the buds of cultivars were recorded and it was found that ACGW laid eggs on the buds of all the cultivars. In microscopic analyses, it was revealed that the resistance originated from the prevention of larva development within the bud. The finding that resistant cultivars belong to C. sativa species is thought to be important for future genetic breeding programs.

Torymus sinensis against the invasive chestnut gall wasp: Evaluating the physiological host range and hybridization risks of a classical biological control agent

BackgroundA common strategy to limit the negative impact of biological invasions is biological control through the release of specialized alien natural enemies. However, biological control plans are not without risks, which include parasitism of native hosts and hybridization with related native species, particularly those that are potential natural enemies of the invasive species. Here, we evaluate these potential risks resulting from the introduction of the parasitoid wasp Torymus sinensis (Hymenoptera: Torymidae) in Europe to control the invasive Asian chestnut gall wasp Dryocosmus kuriphilus (Hymenoptera: Cynipidae). ResultsWe found that, under laboratory conditions, the physiological host range of T. sinensis includes several native non-target species of Cynipidae, with oviposition observed on the galls of 8 of the 11 species tested. However, physiological host range of T. sinensis appears to be limited under field conditions, as we observed only one parasitized gall of Andricus curvator in the field. Regarding hybridization, inter-species mating between T. sinensis and its phylogenetically closest native Torymus species was not observed in the laboratory. Moreover, discordance between nuclear (ITS2) and mitochondrial (coxI) data does not support the presence of genetic introgression, suggesting that hybridization between T. sinensis and native Torymus species does not occur. In addition, we cite and discuss the unexpected presence of one individual of the related alien species Torymus beneficus in Spain. ConclusionOur results suggest that T. sinensis may negatively impact several non-target species, highlighting the need for careful monitoring of the extent of such undesired behaviour and its effects on the native fauna.

No loss of genetic divergence in Torymus sinensis following its release for Dryocosmus kuriphilus control in Europe

Abstract Arrival, establishment, and further dispersal of non‐native natural enemies are considered essential for a successful biological control programme, while among the factors that may determine the success of such a programme, genetic diversity of the introduced population plays an important role in the establishment of a non‐native species. The Chinese parasitoid wasp Torymus sinensis Kamijo (Hymenoptera: Torymidae) was initially released in Europe in Italy to control biologically the Asian chestnut gall wasp Dryocosmus kuriphilus Yasumatsu (Hymenoptera: Cynipidae), and reduce the damage induced on sweet chestnut (Castanea sativa Miller). In the following years, T. sinensis was then released in numerous other European countries as a biological control agent of D. kuriphilus. Its presence has also been reported beyond the countries of release due to rapid natural dispersal. To assess the post‐release genetic diversity of D. kuriphilus, we screened T. sinensis populations from six European countries and tested the possibility of these populations suffering from frequently observed genetic effects that could threaten its successful establishment in Europe. Our results exhibit that T. sinensis populations have suffered neither from the Allee effect nor from genetic bottleneck after their release and establishment in Europe, something that increases the possibility to effectively control D. kuriphilus in Europe.

Damage ratio of the Asian chestnut gall wasp, Dryocosmus kuriphilus Yasumatsu, 1951 (Hemiptera: Cynipidae) in Samsun Province of Türkiye: First Report

The Asian chestnut gall wasp Dryocosmus kuriphilus Yasumatsu (Hymenoptera, Cynipidae) is a critical pest that threatens chestnut cultivation. The chestnut gall wasp causes yield losses by deforming the buds of the infected trees. This pest, which was first detected in Italy in 2002 in Europe, was seen in Gacık village of Yalova province in 2014 in Türkiye. In the following years, it spread to Bursa, Istanbul, Sakarya, Kocaeli, Balıkesir, Bilecik, Düzce, Giresun, Bartın, Zonguldak, Sinop and İzmir provinces. In this study, the orchard of chestnut gene resources belonging to the Ali Nihat Gökyiğit research station in Samsun province was examined for the presence of the Asian chestnut gall wasp. As a result of the study, the presence of the Asian chestnut gall wasp was determined in the cultivars Marigoule' and 'Macit 55'. As a result of the counts made in 50 cm parts of the branches, it was determined that the presence of chestnut gall wasp is in a higher population in the 'Marigoule' cultivar than in the other cultivars. However, the invasion in Samsun is at the early stage.

Assessment of Chestnut Gall Toughness: Implications for a Biocontrol Agent.

(1) Torymus sinensis, the biocontrol agent of the Asian chestnut gall wasp Dryocosmus kuriphilus, is univoltine, but in NW Italy a small percentage of individuals exhibits a prolonged diapause, mainly as late instar larva. (2) In 2020, the diapause was investigated to evaluate its trend over the years. Due to the low survival rate of diapausing T. sinensis adults, the seasonal variation in the galls' toughness was evaluated, thus assuming that dry galls over time can negatively affect emergence. The window of vulnerability of the gall wasp galls was also evaluated in controlled conditions. (3) The results showed that the average number of second year T. sinensis emerging per 100 cells was 0.41 ± 0.05, and dead adults accounted for 4.1 ± 0.23 per 100 cells. Gall toughness resulted in lower values for galls collected in May and June, and then gradually increased over time. In general, no difference was detected in the wall toughness of galls formed during the previous year when compared to current-year dry galls. Oviposition was recorded on all the tested galls collected in May and June, and no difference in the number of oviposition events was detected. Conversely, no oviposition was observed in July. Comparing the number of oviposition events by T. sinensis and the gall toughness, a negative correlation was found (R2 = -0.99). (4) The present findings contribute descriptive information on this gall's structural traits, and the influence on gall wasp management is also discussed.

Dryocosmus kuriphilus Yasumatsu (Hymenoptera: Cynipidae) in Minho (Northern Portugal): Bioecology, Native Parasitoid Communities and Biological Control with Torymus sinensis Kamijo (Hymenoptera: Torymidae)

The Asian chestnut gall wasp Dryocosmus kuriphilus Yasumatsu, native to China, was first detected in Europe in Italy in 2002. In Portugal it was declared in 2014, and it has since affected the Portuguese chestnut production. The introduction of its natural parasitoid Torymus sinensis Kamijo started through inoculative releases according to the National Action Plan for the Control of Dryocosmus kuriphilus, established by the Direção Geral de Alimentação e Veterinária (DGAV), in 2015. This research was carried out during 2018 and 2019, in five chestnut orchards in the Minho region (Northern Portugal). Between January and March 2018, at each location, three monthly harvests of 100 buds were carried out to determine the infestation level. Between April and June 2018, 240 galls were harvested every two weeks, with the purpose of studying the bioecology of D. kuriphilus. In each sampling site, 160 galls were placed in emergence boxes to follow adults of the D. kuriphilus population, and 80 galls were dissected with a binocular magnifying glass, to count and record the evolution of the larval stages of D. kuriphilus and of the parasitoids. The identification of T. sinensis and native parasitoids was carried out. The parasitism rates and Simpson diversity indices were determined. The percentage of buds infested by D. kuriphilus was high in all studied places, varying between 67% and 80%, regardless of the number of T. sinensis releases made and of the chestnut cultivars present in each orchard. The highest rate of parasitism by T. sinensis was 0.5044% and it was recorded in an orchard where there were two releases of T. sinensis in 2016 and 2017 (Arcos de Valdevez—Rio Cabrão—P1). Considering that the exotic parasitoid has already been released in the region since 2016, its establishment in all the chestnut orchards studied was confirmed, although with very low parasitism rates. The natural parasitism associated with the native parasitoid species of D. kuriphilus was confirmed in this work, recording values between 34% and 51% of the parasitized chestnut galls, with 11 species of native parasitoids having been identified: Eupelmus azureos, Eupelmus uruzonus, Eurytoma brunnivientris, Eurytoma pistaciae, Megastigmus dorsalis, Mesopolobus sp., Ormyrus pomaceus, Sycophila biguttata, Sycophila iracemae, Sycophila variegatta and Torymus flavipes.

Transcriptomics and Antioxidant Analysis of Two Chinese Chestnut (Castanea mollissima BL.) Varieties Provides New Insights Into the Mechanisms of Resistance to Gall Wasp Dryocosmus kuriphilus Infestation.

Chinese chestnut is a popular fruit tree with a high nutritional value of its nuts, which can suffer from infestation by the chestnut gall wasp Dryocosmus kuriphilus (GWDK) that results in gall formation and resultant loss of production and profitability. The physiological and molecular mechanisms of GWDK resistance found in certain genotypes currently remains elusive. To gain new insights into this phenomenon, a series of RNA-Seq integrated with metabolomic profiling experiments were executed to investigate the chemical and transcriptional differences in response to GWDK infestation in two contrasting chestnut varieties grown in China (the susceptible “HongLi,” HL and the partially resistant “Shuhe_Wuyingli,” SW). Three time points were selected for comparison: The initiation stage (A), growth stage (B), and maturation stage (C). Results showed that concentrations of hydrogen peroxide (H2O2) and the activities of peroxidase (POD) and superoxide dismutase (SOD) enzyme were elevated in the resistant SW leaves compared with those in HL leaves at all three developmental stages, while catalase (CAT) and polyphenol oxidase (PPO) activities were mostly higher in HL leaves. RNA-Seq transcriptomic analyses of HL and SW leaves revealed that various metabolic pathways involved in GWDK stress responses, such as plant hormone signal transduction, MAPK signaling, and the peroxisome pathway, were enriched in the contrasting samples. Moreover, the weighted gene co-expression network analysis (WGCNA) of differentially expressed genes in the POD pathway combined with transcription factors (TFs) indicated that the expression of TF members of bHLH, WRKY, NAC, and MYB family positively correlated with POD pathway gene expression. The TFs CmbHLH130 (EVM0032437), CmWRKY31 (EVM0017000), CmNAC50 (EVM0000033), and CmPHL12 (EVM0007330) were identified as putative TFs that participate in the regulation of insect-induced plant enzyme activities in chestnut, which may contribute to GWDK resistance in SW. Expression levels of 8 random differentially expressed genes (DEGs) were furthermore selected to perform quantitative reverse transcription PCR (qRT-PCR) to validate the accuracy of the RNA-Seq-derived expression patterns. This study guides the functional analyses of further candidate genes and mechanisms important for GWDK resistance in chestnuts in the future as well as can help in identifying the master transcriptional regulators and important enzyme steps that support major insect defense pathways in chestnut.

Host Gall Size and Temperature Influence Voltinism in an Exotic Parasitoid

Insect phenology is highly temperature-dependent. Higher temperatures can lead to earlier emergence and lengthening of the active period, which enable many insect groups to complete more generations. Studies on the effects of climate change on insect populations are providing concerning evidence supporting this relationship. These kind of shifts in phenology and voltinism also occur in agricultural and forest insect pests and their natural enemies, with potential implications for biological control. The consequences derived from changing temperature regimes on tritrophic interactions remain poorly studied, particularly in gall-inducing insects and their parasitoids. Here we detail the occurrence of bivoltinism in the exotic parasitoid Torymus sinensis, previously categorized as univoltine, a widely introduced species to fight against the invasive Asian chestnut gall wasp Dryocosmus kuriphilus wherever this pest spread. This plasticity in voltinism has been observed in the southernmost European distribution of D. kuriphilus, and appears to be mediated by both temperature and gall traits, namely size or the number of gall chambers. Bivoltinism was most common at annual mean temperatures around 13.5°C and in galls with more chambers. Through this work, we intend to unravel the factors behind this phenomenon and discern the possible consequences on host-parasitoid interactions.

Local-scale dispersal patterns and susceptibility to Dryocosmus kuriphilus in different Castanea species and hybrid clones: insights from a field trial

The chestnut gall wasp Dryocosmus kuriphilus is a major insect pest affecting chestnut trees worldwide. Medium and long-term control of this pest can be improved by using resistant or less susceptible cultivars and hybrid clones. In addition, little is known about the local patterns of dispersal of the pest. We obtained data from trees of 3 chestnut species and 27 hybrid clones in a field trial in NW Spain with the aim of evaluating the susceptibility of the material to the gall wasp and identifying possible drivers of local spatial dispersal. In the first 3 years of the invasion by D. kuriphilus, the number of trees attacked and the number of galls on each tree were spatially clustered. Tree height significantly predicted both variables, suggesting that gall wasps may use visual cues to locate suitable host trees, at least in the early stage of invasion. Assessment of the susceptibility of hybrid clones/pure species must take concurrent indicators of infestation levels into account. We suggest the use of indices involving galls on shoots because these enable good assessment of the damage to chestnut trees. The study findings add to existing knowledge on the susceptibility of hybrid chestnut clones. We report, for the first time, two hybrid clones resistant to the pest and one hybrid clone which exhibited consistently low values for all of the indicators of infestation level. The results have important implications regarding selection of plant material for use in afforestation in Spain, where the current high rate of chestnut planting is expected to continue.

Could Gnomoniopsis castaneae Be Used as a Biological Control Agent against Insect Pests?

Gnomoniopsis castaneae is the cause of the chestnut brown rot but has been also regarded as an important mortality factor for the chestnut gall wasp Dryocosmus kuriphilus. The question to whether G. castaneae could serve as a natural biocontrol agent against insect pests is investigated in the present study. We used three serious insect pests as experimental model insects: Plodia interpuctella and Trogoderma granarium, which are important pests of stored products, and Myzus persicae, a cosmopolitan, serious pest of annual and perennial crop plants. Although chemical pesticides represent effective control means, they are also related to several environmental and health risks. In search for alternative pest management methods, scientific interest has been focused, inter alia, on the use of entomopathogenic fungi. While Isaria fumosorosea has long been recognized as an effective control agent against several pests, G.castaneae has been very little studied. The present study examined whether and to what extent G. castaneae and I. fumosorosea exhibit insecticidal activity against fourth-instar larvae of P. interpunctella and T. granarium and adults of M. persicae. Mortality was examined in interrelation with dosage and time exposure intervals. Both fungi exhibited pesticidal action. However, G. castaneae induced noteworthy mortality only at very high doses. In general, we concluded that G. castaneae failed to cause high insect pathogenicity at normal doses and may not be an efficient biocontrol agent compared with other entomopathogens. On the other hand, our study reiterates the pathogenic potential of I. fumosorosea. More studies are needed to further our insight into the potential of EF species as a component of IPM.

Impact of the Asian gall wasp Dryocosmus kuriphilus on the radial growth of the European chestnut Castanea sativa

Abstract The invasion of the Asian chestnut gall wasp Dryocosmus kuriphilus (ACGW) in Europe has caused serious biological and economic impacts on chestnut stands that have been partially solved with the introduction of the biological control agent Torymus sinensis. However, information concerning tree‐ring growth during the ACGW epidemic has been lacking so far. Using dendrochronology techniques, we analysed the impact of the ACGW on tree‐ring growth and the capacity of the chestnut tree to recover when biocontrol was achieved in seven sites covering the area of first detection and subsequent spread of the ACGW in Europe. In each site, a non‐target control species (i.e. trees not attacked by the ACGW) has been included as a reference in the analysis. Results show a reduction in the tree‐ring increment by 60% on average during the ACGW epidemic. Such effects were higher in the magnitude and longer in the duration when compared to other stresses such as insect defoliation or extreme summer droughts. Synthesis and applications. Marked reductions in radial growth were evident in the years of ACGW attack regardless of the age of the chestnut coppices. Since most of the trees only recovered to an almost normal growth rate after 3–4 years of biocontrol by T. sinensis, an immediate release of the antagonist at the first ACGW appearance is highly recommended. The consequences and perspectives of ACGW's attacks on the quality of the timber produced are also discussed.

Short-Term Cold Stress Affects Parasitism on the Asian Chestnut Gall Wasp Dryocosmus kuriphilus.

Simple SummaryThe Asian cynipid gall wasp (ACGW) “Dryocosmus kuriphilus” has become widespread in Europe. In all invaded areas, it is parasitized by native parasitoids associated with oak galls, for which the ACGW represents a non-saturated adaptation space. Considering the increase in the frequency of extreme climatic events over the last twenty years (e.g., low temperatures during the vegetative period of the chestnut tree), this study aimed to elucidate the effects of cold stress on both ACGW biology and parasitism by native and introduced parasitoids. The ACGW–parasitoid system represents an ideal subject in which to evaluate the effect of sudden cold stress events due to the wasps’ biological characteristics, which include the ability to complete development even in galls detached from plants. We show that parasitism on and the mortality of ACGWs in three chestnut fields were affected by a cold treatment. Our results reveal species-specific differences in the abundance and performance of parasitoids associated with the ACGW in response to cold stress. For example, the frequency of Eupelmus spp. and Mesopolobus tibialis doubled as a result of the cold treatment in all three chestnut fields in both study years. Therefore, the plasticity in response to short-term temperature variation is associated with individual fitness in some parasitoid species.Temperature variation affects interactions involving plants, herbivores, and parasitoids, causing a mismatch between their phenological cycles. In the context of climate change, climatic factors can undergo profound and sudden changes, such as sudden hot or cold snaps. Herein, we show that the number of episodes of short but sustained low temperatures has increased, mainly during May, over the last two decades. We subjected galls induced by the Asian chestnut gall wasp (ACGW) Dryocosmus kuriphilus to cold stress to assess whether and, if so, how it affected the pest and its parasitoids. Over the course of two years, we measured seasonal parasitism, parasitism rates, the relative abundance of each parasitoid species, and ACGW mortality. We found that the cold treatment affected both the pest and the parasitoids, resulting in a reduction in the emergence of ACGWs and differing ratios of species within the parasitoid community. The most striking example was the change in the relative frequency of three species of Eupelmus spp. and Mesopolobus tibialis, which doubled in cold-stressed galls in all chestnut fields. The effects of temperature on the development of the host and the direct effects of cold temperatures on the surface of galls (in terms of the humidity or hardness of the galls) warrant further research in this direction.

Recruitment of native parasitoids by an introduced gall wasp Dryocosmus kuriphilus Yasumatsu, 1951 (Hymenoptera: Cynipidae) in Britain and France

The invasive chestnut gall wasp Dryocosmus kuriphilus Yasumatsu (Hymenoptera: Cynipidae) is a major pest species of Castanea spp. worldwide. It only arrived in Britain in 2015 and the parasitoid community associated with it was studied in 2019. Eight native species of chalcidoid, previously known to be associated with oak gall wasps, were identified. These results are compared to a similar study (2016–2019) in France where four parasitoids were reared. However, this community included the deliberately introduced Torymus sinensis Kamijo, a specific biological control agent of D. kuriphilus. The presence of T. sinensis and the differences between the two communities is discussed.

Hot water treatment: an effective method for disinfecting Castanea sativa mill. dormant scions against Dryocosmus kuriphilus Yasumatsu.

The Asian chestnut gall wasp (ACGW) Dryocosmus kuriphilus Yasumatsu is the most severe pest of the genus Castanea worldwide. After its arrival in Europe, the insect dispersed rapidly from Italy to other European countries, as well as spreading over long distances within each country because of the unintentional transport of infested chestnut material owing to the fact that infestation inside the dormant buds is visually undetectable. Hot water treatment (HWT) has been tested in terms of pest and diseases control in propagation processes with a wide range of plant material and species, including the Asian chestnut Castanea mollissima cv 'Qing'. Taking into account the different thermal tolerances of plant species, an experiment to disinfect Castanea sativa scions from ACGW by HWT was performed. The effects of the immersion of dormant scions for 10min at two water temperatures (49 and 52 °C), with and without a previous period of water hydration, were tested. A 10min soak at 49 °C was effective in killing ACGW larvae while retaining an excellent percentage of successful grafts. The pre-HWT hydration stage did not influence the success of the treatment. Positive shoot growth was also recorded after HWT. We report for the first time the effectiveness of a simple and environmentally friendly protocol based on hot water for the disinfection of European chestnut (C. sativa) scions against D. kuriphilus. This will enable nurseries to supply plants free from this pest, preventing the insect dispersion, which will have a positive socio-economic impact to the chestnut tree sector. Also positive environmental impact will be favoured as gall wasp proliferation will decrease. © 2019 Society of Chemical Industry.

The Asian Chestnut Gall Wasp in Italy: Surveys on its Native and Exotic Parasitoids as Well as on Chestnut Cultivar Susceptibility

In the context of biological control against the Asian chestnut gall wasp Dryocosmus kuriphilus Yasumatsu (Hymenoptera: Cynipidae) in Italy, the aim of this study was to survey its native parasitoids as well as the exotic Torymus sinensis Kamijo which had been introduced and released as a biological control agent. Furthermore, both parasitism and pest susceptibility of local chestnut trees were investigated. Surveys were carried out in 2014 in the Municipality of Sambuca Pistoiese (Pistoia), in a chestnut forest which included three cultivars: Nerattino, Carpinese, and Pastinese. On a total of 35 trees, selected among the three cultivars, 24 current-year shoots were randomly collected and examined in the laboratory. After the damage by the cynipid had been assessed, leaf samples were used for molecular analyses. Galls were also collected on the 35 selected trees both for dissection and to rear parasitoid specimens in the laboratory. The three cultivars were confirmed by genetic tests. The Carpinese cultivar resulted to be the most damaged, with highest percentage of attacked shoots (84.38%). Moreover it was also the cultivar with the lower parasitism rate (about 16%). From reared galls seven parasitoid species emerged, all belonging to the superfamily of Chalcidoidea. Exception made for the non-native T. sinensis, the other six were native species associated to oak gall wasps. Native parasitoids were more scarcely present, while the exotic T. sinensis amounted for the 84.27% of all emerged parasitoids. Our results confirm how chestnut susceptibility to D. kuriphilus varies depending on the cultivar. Overall, the Nerattino cultivar resulted as the best among the three examined: it was subject to a less intense attack than the other cultivars, and the resulting smaller galls were more effectively exploited by parasitoids.

Fungal community associated with adults of the chestnut gall wasp Dryocosmus kuriphilus after emergence from galls: Taxonomy and functional ecology

The diversity of the fungal community associated with adults of Dryocosmus kuriphilus following emergence was examined using HTS analysis. Ascomycota dominated the fungal core-biome community. The functional guilds of the 90 taxa forming the core-biome were assessed, demonstrating three main groups: saprotrophs, plant pathogens and entomopathogens. Twenty-nine OTUs out of 90 were resolved to species level identifying 26 different fungal species. Among these species, many were cosmopolitan or previously recorded in Europe. Ten taxa were previously recorded on chestnut, including some recognized plant pathogens associated with foliage and green tissues such as Epicoccum nigrum, Gnomoniopsis castanea, Colletotrichum acutatum, Stromatoseptoria castaneicola, Ramularia endophylla. Beauveria bassiana; within the core microbiome, Fusarium larvarum represented the most abundant entomopathogenic species. Some of these species are known to impact directly or indirectly the vitality of the insects in the galls. The chestnut blight pathogen, Cryphonectria parasitica, was never found associated with D. kuriphilus. Based on the present study, an active role for D. kuriphilus as a vector of chestnut fungal endophyte/pathogens cannot be demonstrated but neither ruled out.

Torymus sinensis Kamijo, a biocontrol agent against the invasive chestnut gall wasp Dryocosmus kuriphilus Yasumatsu in Spain: its natural dispersal from France and the first data on establishment after experimental releases

Aim of study: The globally invasive gall wasp, Dryocosmus kuriphilus Yasumatsu, 1951 (Cynipidae: Cynipini), reached Spain seven years ago and is already regarded as an important pest of chestnuts (Castanea spp.) in this country as well as worldwide. In this paper, we present comprehensive data on the establishment in Spain of Torymus sinensis Kamijo, 1982 (Chalcidoidea: Torymidae), an effective non-native natural enemy of this pest, as a result of both natural dispersal and settlement after controlled releases since 2015.Area of study: Sites of the Spanish autonomous communities of Galicia, Asturias, Basque Country, Catalonia, Andalusia and Madrid where D. kuriphilus is present.Material and methods: To study the natural dispersal of T. sinensis from France, we selected two sampling sites in Catalonia, six in the Basque Country and two in Navarra known for their heavy Asian chestnut gall wasp (ACGW) infestation; to study T. sinensis establishment after authorized controlled releases by the concerned authorities, the field samplings were done mainly in Galicia (35 sites) and Andalusia (8 sites). Additionally an experimental release study was made in Madrid.Main Results: Our results showed that T. sinensis has spread throughout Spain by natural dispersal across the French border and now occurs in Catalonia (two sites), the Basque Country (three sites) and Navarra (one site) but not in the neighbouring region of Cantabria. The percentage of parasitism by T. sinensis on D. kuriphilus is higher in the Basque Country sites, which are close to the French border, thus indicating that its establishment in these localities is not recent. After controlled releases, T. sinensis has been successfully established in five release sites in Andalusia (Valle del Genal and Sierra Blanca, Málaga Province) and one release site in Madrid. However, in the region of Galicia (NW Spain), where the number of authorized releases has been higher, the establishment of T. sinensis still appears to be very low.Research highlights: Established populations of T. sinensis may exert a positive buffer against D. kuriphilus-driven chestnut infestation in Spain, similar to what is observed in other invaded European countries.Additional Keywords: Controlled releases, Torymidae, invasive species, Cynipidae, natural spread, biological control.Abbreviations used: Asian chestnut gall wasp (ACGW); Torymus sinensis parasitism rate (TsPR); mean of ACGW larval chambers per gall (MACGW).

Environmental thermal levels affect the phenological relationships between the chestnut gall wasp and its parasitoids

AbstractStudies of thermal level‐related asynchrony in a host–parasitoid relationship are necessary to understand the effects of climate change on new host–parasitoid interactions. In the Asian chestnut gall wasp Dryocosmus kuriphilus (Hymenoptera: Cynipidae) and its Chalcidoidea parasitoids, phenological synchrony is assumed to be weather‐dependent in a new area of expansion. To evaluate the effects of environmental thermal regimes on the host, a phenology model for different cynipid stages (larvae, pupae, adults, and adult emergence) and a host–parasitoid phenological estimator are developed in three chestnut fields during two successive growth seasons and subsequently validated in areas with chestnut fields at two different altitudes. Comparisons of the timings of the juvenile and adult stages with those of the parasitoid complex demonstrate that the shortest period of occurrence for cynipids within galls has negative effects on the host–parasitoid relationships at higher temperature levels, thereby increasing phenological asynchrony for some parasitoids species. Reducing the development time of pupae and adults decreases the likelihood of success for some parasitoid species at higher temperature levels. We also record the extension of the gall wasp development time (approximately 15 days) at higher altitudes (linked to a lower mean temperature of approximately 1.5 °C). These results highlight how parasitization on the new hosts is dependent on the host phenology and, in the present study, is limited by the short duration of the presence of the host in galls, which could explain the considerable differences in cynipid gall wasp parasitization recorded at different altimeters.

Seasonal flight activity and genetic relatedness of Torymus species in Italy

Abstract Torymus sinensis Kamijo was released into Europe to control the Asian chestnut gall wasp Dryocosmus kuriphilus Yasumatsu, proving to be an effective biocontrol agent. In Japan, hybrids between this exotic parasitoid and the native congeneric Torymus beneficus Yasumatsu and Kamijo have been recorded, whereas no case of hybridization with natives has been reported for Europe. The seasonal phenology of the Torymus species was investigated in north‐west Italy and 172 997 chestnut galls (Castanea spp.), 7866 oak galls (Quercus spp.), 29 wild rose galls (Rosa spp.) and 44 bramble galls (Rubus spp.) were collected over a 5‐year period. The seasonal flight activity was recorded for eight native Torymus species [Torymus affinis (Fonscolombe), Torymus auratus (Müller), Torymus bedeguaris (L.), Torymus calcaratus Nees, Torymus cyaneus Walker, Torymus flavipes (Walker), Torymus geranii (Walker) and Torymus notatus (Walker)] and five of them were found to overlap with T. sinensis flight, namely T. affinis, T. auratus, T. flavipes, T. geranii and T. notatus. Among the Torymus native species, T. notatus revealed to be morphologically and phylogenetically closely related to T. sinensis, thus suggesting hybridization potential.