Climate, host and geography shape insect and fungal communities of trees.

Non-native pests, climate change, and their interactions are likely to alter relationships between trees and tree-associated organisms with consequences for forest health. To understand and predict such changes, factors structuring tree-associated communities need to be determined. Here, we analysed the data consisting of records of insects and fungi collected from dormant twigs from 155 tree species at 51 botanical gardens or arboreta in 32 countries. Generalized dissimilarity models revealed similar relative importance of studied climatic, host-related and geographic factors on differences in tree-associated communities. Mean annual temperature, phylogenetic distance between hosts and geographic distance between locations were the major drivers of dissimilarities. The increasing importance of high temperatures on differences in studied communities indicate that climate change could affect tree-associated organisms directly and indirectly through host range shifts. Insect and fungal communities were more similar between closely related vs. distant hosts suggesting that host range shifts may facilitate the emergence of new pests. Moreover, dissimilarities among tree-associated communities increased with geographic distance indicating that human-mediated transport may serve as a pathway of the introductions of new pests. The results of this study highlight the need to limit the establishment of tree pests and increase the resilience of forest ecosystems to changes in climate.

First Report of the Pepper Fruit Fly Atherigona orientalis (Schiner 1968) (Diptera: Muscidae) Infesting Commercial Pepper Crops in Greece.

The pepper fruit fly Atherigona orientalis (Schiner 1968) (Diptera: Muscidae) is a cosmopolitan tropical pest which has been recently recorded in several European countries. The biology of the pest has been primarily associated not only with decomposing fruits and vegetables, but even vertebrate and invertebrate carrion, dung and faeces. Relatively recently, A. orientalis has been reported as a primary pest of pepper fruits as well. In this short communication, we report, for the first time in Greece and, to the best of our knowledge, in Europe, cases of pepper fruit fly damage to pepper fruits in commercial greenhouse crops (in Crete in 2022). In this direction, possible implications and concerns regarding the occurrence of this pest in Crete are discussed.

Uncovering the Male Presence in Parthenogenetic Marchalina hellenica (Hemiptera: Marchalinidae): Insights into Its mtDNA Divergence and Reproduction Strategy.

Marchalina hellenica (Hemiptera: Marchalinidae), an endemic species in Greece and Turkey, is a major contributor to the annual honey production in its native range. However, in the areas that it invades, lacking natural enemies, it has detrimental effects on pine trees and potentially contributes to tree mortality. Although it was originally reported as thelytokous, males were later reported in Turkey and on several of the islands of Greece. To further disambiguate the exact parthenogenetic reproduction strategy of M. hellenica, we studied the emergence pattern of male individuals in Greece for two consecutive years (2021 and 2022). Furthermore, we examined the genetic variation among 15 geographically distant populations of M. hellenica in Greece using a mitochondrial DNA marker and compared the results with data from Turkey. The findings of this study document the existence of an additional M. hellenica population in its native range that repeatedly produces males, apart from the areas of Greece and Turkey in which they were initially reported, suggesting that males play a major, so far unknown role in the reproduction of this species. The populations in Greece and Turkey exhibited a strong genetic affinity, while human-aided dispersal seems to have obscured the genetic pattern acquired.

Three new species of Afropselaphus Jeannel, 1950 (Staphylinidae: Pselaphinae) from Greece and a redescription of Pselaphogenius treskanus (Karaman, 1940).

Three new species of the genus Afropselaphus Jeannel, 1950, Afropselaphus taygetensis sp. n., Afropselaphus tymficus sp. n. and Afropselaphus euboicus sp. n. are described and illustrated. Pselaphogenius treskanus (Karaman, 1940) is redescribed and new data of its distribution are provided.

Photosynthetic and defensive responses of two Mediterranean oaks to insect leaf herbivory.

Insect herbivory is a dominant interaction across virtually all ecosystems globally and has dramatic effects on plant function such as reduced photosynthesis activity and increased levels of defenses. However, most previous work assessing the link between insect herbivory, photosynthesis and plant defenses has been performed on cultivated model plant species, neglecting a full understanding of patterns in natural systems. In this study, we performed a field experiment to investigate the effects of herbivory by a generalist foliar feeding insect (Lymantria dispar) and leaf mechanical damage on multiple leaf traits associated with defense against herbivory and photosynthesis activity on two sympatric oak species with contrasting leaf habit (the evergreen Quercus coccifera L. and the deciduous Quercus pubescens Willd). Our results showed that, although herbivory treatments and oak species did not strongly affect photosynthesis and dark respiration, these two factors exerted interactive effects. Insect herbivory and mechanical damage (vs control) decreased photosynthesis activity for Q. coccifera but not for Q. pubescens. Insect herbivory and mechanical damage tended to increase chemical (increased flavonoid and lignin concentration) defenses, but these effects were stronger for Q. pubescens. Overall, this study shows that two congeneric oak species with contrasting leaf habit differ in their photosynthetic and defensive responses to insect herbivory. While the evergreen oak species followed a more conservative strategy (reduced photosynthesis and higher physical defenses), the deciduous oak species followed a more acquisitive strategy (maintained photosynthesis and higher chemical defenses).

Worldwide diversity of endophytic fungi and insects associated with dormant tree twigs.

International trade in plants and climate change are two of the main factors causing damaging tree pests (i.e. fungi and insects) to spread into new areas. To mitigate these risks, a large-scale assessment of tree-associated fungi and insects is needed. We present records of endophytic fungi and insects in twigs of 17 angiosperm and gymnosperm genera, from 51 locations in 32 countries worldwide. Endophytic fungi were characterized by high-throughput sequencing of 352 samples from 145 tree species in 28 countries. Insects were reared from 227 samples of 109 tree species in 18 countries and sorted into taxonomic orders and feeding guilds. Herbivorous insects were grouped into morphospecies and were identified using molecular and morphological approaches. This dataset reveals the diversity of tree-associated taxa, as it contains 12,721 fungal Amplicon Sequence Variants and 208 herbivorous insect morphospecies, sampled across broad geographic and climatic gradients and for many tree species. This dataset will facilitate applied and fundamental studies on the distribution of fungal endophytes and insects in trees.

Phenology and Potential Fecundity of Neoleucopis kartliana in Greece.

Neoleucopis kartliana Tanasijtshuk (Diptera, Chamaemyiidae) is the most abundant predator of the giant pine scale (GPS), Marchalina hellenica (Hemiptera, Margarodidae) in Greece. GPS is native to Greece and Turkey, where it is not considered a pest of Pinus spp., but a valuable resource for pine honey production. However, its introduction to new areas leads to high population densities of the scale, linked to declines in tree health and insect biodiversity. To assess the potential use of N. kartliana for a classical biological control program in Australia, we studied selected life-history traits of the silver fly, namely its phenology in northern Greece, feeding preferences of adult flies on artificial food sources, and potential fecundity of female flies. The silver fly was present in every site in northern Greece studied and was found to have at least three generations per year in this area. The fly's overall sex ratio was 1:1, and adult females emerged with no or few mature eggs in their ovaries, but egg production was exponential until at least the eighth day after emergence. These findings increase our knowledge about the biology of N. kartliana and aided in the evaluation of the silver fly as a classical biological control agent against invasive GPS in Australia.

Nematodes in the Pine Forests of Northern and Central Greece.

In the context of plants or plant products protection by harmful organisms, measures have been taken by EU countries in order to prevent their introduction and establishment into the EU, and also limit their expansion in case they do enter. Such a case is Bursaphelenchus xylophilus (Parasitaphelenchidae, Nematoda), already recorded in Portugal and Spain. So, Member States should take all the appropriate steps in order to monitor and confine if necessary susceptible plants and/or plant products. Such measures include annual surveys even in countries where pine wilt disease does not occur yet. Therefore, national survey programs are widely established, sampling and examining samples from pine trees showing suspicious symptoms that could potentially be attributed to B. xylophilus. In this direction, such a network has also been established in Greece collecting and examining wood samples nationwide. In total, 123 wood samples were collected from conifer trees of Northern and Central Greece. Though B. xylophilus was absent from all samples examined, four other Bursaphelenchus species were identified. In addition, other nematode taxa were also recorded, including several phytophagous, microbivorous as well as predatory nematode species. This highlights the fact that besides preventing the introduction of B. xylophilus in Greece, national survey programs can significantly contribute to and enhance our knowledge of the indigenous nematode species.

Immediate and Delayed Mortality of Four Stored-Product Pests on Concrete Surfaces Treated with Chlorantraniliprole.

Chlorantraniliprole is an effective pesticide against a plethora of pests, but its efficacy against stored-product pests is very poorly explored. In this study we treated concrete surfaces with four different doses of chlorantraniliprole (0.01, 0.05, 0.1, and 0.5 mg a.i./cm2) against the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) adults and larvae, the lesser grain borer, Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae) adults, the rice weevil, Sitophilus oryzae (L.) (Coleoptera: Curculionidae) adults, and the flour mite, Acarus siro L. (Sarcoptiformes: Acaridae) adults and nymphs, to examine the immediate mortalities after 1, 2, 3, 4, and 5 days of exposure. Additionally, the delayed mortality of the individuals that survived the 5-day exposure was also evaluated after a further 7 days on untreated concrete surfaces. We documented high mortality rates for all tested species and their developmental stages. After 5 days of exposure to 0.5 mg a.i./cm2, T. castaneum larvae and A. siro adults exhibited the highest immediate mortality levels, reaching 96.7% and 92.2%, respectively. Delayed mortality was also very high for all tested species and their developmental stages. Nymphs of A. siro displayed a 96.3% delayed mortality followed by the adults of R. dominica (98.6%) after exposure to 0.5 mg a.i./cm2. All other tested species and their developmental stages reached complete (100.0%) delayed mortality, where even 0.01 mg a.i./cm2 caused ≥86.6% delayed mortality in all species and their developmental stages. Taking into consideration the effectiveness of chlorantraniliprole on this wide range of noxious arthropods, coupled with its low toxicity towards beneficial arthropods and mammals, this pesticide could provide an effective management tool for stored-product pests in storage facilities.

Alientoma, a Dynamic Database for Alien Insects in Greece and Its Use by Citizen Scientists in Mapping Alien Species.

Invasive alien species have been increasingly acknowledged as a major threat to native biodiversity and ecosystem services, while their adverse impacts expand to human health, society and the economy on a global scale. Insects represent one of the most numerous alien organismic groups, accounting for about one fifth of their total number. In Greece, a large number of alien insects have been identified, currently reaching 469 species. In recent decades, the contribution of citizen science towards detecting and mapping the distribution of alien insects has been steeply increasing. Addressing the need for up-to-date information on alien species as well as encouraging public participation in scientific research, the Alientoma website-derived from "alien" and the Greek word "entoma", meaning insects, is presented. The website aims towards providing updated information on alien species of insects to the public as well as the scientific community, raising awareness about biological invasions and addressing their distribution and impacts inter alia. By maintaining a dynamic online database alongside a strong social media presence since its launch, Alientoma has attracted individuals mainly from Greece and Cyprus, interacting with the website through a total of 1512 sessions. Alientoma intends to establish a constantly increasing network of citizen scientists and to supplement early detection, monitoring and management efforts to mitigate the adverse impacts of alien insects in Greece.

Biotic threats for 23 major non-native tree species in Europe.

For non-native tree species with an origin outside of Europe a detailed compilation of enemy species including the severity of their attack is lacking up to now. We collected information on native and non-native species attacking non-native trees, i.e. type, extent and time of first observation of damage for 23 important non-native trees in 27 European countries. Our database includes about 2300 synthesised attack records (synthesised per biotic threat, tree and country) from over 800 species. Insects (49%) and fungi (45%) are the main observed biotic threats, but also arachnids, bacteria including phytoplasmas, mammals, nematodes, plants and viruses have been recorded. This information will be valuable to identify patterns and drivers of attacks, and trees with a lower current health risk to be considered for planting. In addition, our database will provide a baseline to which future impacts on non-native tree species could be compared with and thus will allow to analyse temporal trends of impacts.

Hylotelephium spectabile, a New Host for Carnation Tortrix Moth (Cacoecimorpha pronubana) and Molecular Characterization in Greece.

Cacoecimorpha pronubana (Hübner) (Lepidoptera, Tortricidae) is a highly polyphagous pest of a wide range of crop and ornamental plants. It is of Mediterranean origin and widespread in European and Mediterranean Plant Protection Organization (EPPO) region. For the first time, infestations of Hylotelephium spectabile (Boreau) Ohba (syn.: Sedum spectabile Boreau) (Saxifragales, Crassulaceae) ornamental plants by C. pronubana larvae, in private gardens in urban area of Drama, Greece, were found. Species identification was conducted based on morphology of female genitalia. In addition, due to reports on occurrence of cryptic C. pronubana species within Europe, DNA barcoding was carried out to determine the molecular status of the pest. This communication reports a new host of C. pronubana and places the Greek pest population along with European species clade.

Bark and Wood Boring Insects-Past, Present, and the Future Knowledge We Need.

Bark and wood-boring insects represent a very diverse group of insects that includes bark and ambrosia beetles, cerambycids, weevils, jewel beetles, or even anobiids from the order of beetles (Coleoptera), but in the broader sense other insect orders like Lepidoptera (e [...].

Effective Defense of Aleppo Pine Against the Giant Scale Marchalina hellenica Through Ecophysiological and Metabolic Changes.

Aleppo pine (Pinus halepensis) is widely distributed in the Mediterranean region and in other areas of the world, where it has been introduced due to its adaptive capacity to xerothermic conditions. The giant pine scale Marchalina hellenica often infests Aleppo pine, as well as other pines, in several southeastern European countries, causing pine declines. When combined with the expected intensified heat and drought events in eastern Mediterranean, the impact of this biotic parameter on the host pines may be exacerbated. The importance of understanding the defense mechanisms of Aleppo pine is emphasized by the recent invasion of the pine scale in new regions, like Australia, lacking the insect's natural enemies, where more intense negative effects on pine species may occur. To date, Aleppo pine's physiological responses to the infestation by M. hellenica are largely unknown. This study aimed at assessing the responses of Aleppo pine to the giant pine scale attack, both on an ecophysiological and a metabolic level. For this purpose, gas exchange, needle water status, and carbon and nitrogen content were measured during 1 year on healthy and infested adult trees. M etabolic profiling of Aleppo pine needles was also performed before, during, and after the high feeding activity of the insect. The maintenance of stable relative water content, δ13C signatures, and chlorophyll fluorescence in the needles of infested pines indicated that infestation did not induce drought stress to the host pines. At the peak of infestation, stomatal closure and a pronounced reduction in assimilation were observed and were associated with the accumulation of sugars in the needles, probably due to impaired phloem loading. At the end of the infestation period, tricarboxylic acids were induced and phenolic compounds were enhanced in the needles of infested pines. These metabolic responses, together with the recovery of photosynthesis after the end of M. hellenica intense feeding, indicate that in the studied region and under the current climate, Aleppo pine is resilient to the infestation by the giant pine scale. Future research should assess whether these promising defense mechanisms are also employed by other host pines, particularly in regions of the world recently invaded by the giant pine scale, as well as under more xerothermic regimes.

New Data on the Range Expansion of the Thaumetopoea pityocampa (Lepidoptera: Notodontidae) 'ENA clade' in Greece: The Role of Bacterial Endosymbionts.

The pine processionary moth, Thaumetopoea pityocampa (Denis and Schiffermüller), is an important insect in the Mediterranean region, as it defoliates pines and its urticating hairs can cause allergic reactions in humans and animals. Moreover, this species exhibits an interesting genetic structure as recently a distinct East-North African mtDNA lineage ('ENA clade') has been described. This clade has been recently detected in Greek populations where it has currently expanded its range by replacing the 'endemic' T. pityocampa lineages. Here, we report new data on the rapid spread of 'ENA clade' in the Greek island Evoia in only a few years. As the underlying mechanisms of the 'ENA clade' range expansion has not been studied so far, we screened T. pityocampa for an infection with the heritable bacterial endosymbionts Wolbachia (Bacteria: Anaplasmataceae), Cardinium (Bacteria: Bacteroidaceae), Rickettsia (Bacteria: Rickettsiaceae) and Spiroplasma (Bacteria: Spiroplasmataceae). These bacteria can manipulate the reproduction of infected hosts, something that could potentially explain the rapid spread of 'ENA clade' lineage. Therefore, we screened 28 individuals that exhibited T. pityocampa 'ENA clade' and 'endemic' T. pityocampa haplotypes from nine populations scattered all over Greece. None of them was infected with any of the four endosymbionts, suggesting that these bacteria do not cause reproductive manipulations in T. pityocampa lineages and, thus, other factors should be explored in future research efforts.

The Apennines as a cryptic Pleistocene refugium of the bark beetle Pityogenes chalcographus (Coleoptera: Curculionidae).

The Apennine Mountains in Italy are an important biogeographical region and of particular interest in phylogeographical research, because they have been a refugium during Pleistocene glaciation events for numerous European species. We performed a genetic study on the Eurasian bark beetle Pityogenes chalcographus (Linnaeus, 1760), focusing on two Apennine (Italian) and two Central European (Austrian) locations to assess the influence of the Apennines in the evolutionary history of the beetle, particularly during the Pleistocene. We analysed a part of the mitochondrial COI gene and a set of 5470 informative genome-wide markers to understand its biogeography. We found 75 distinct mitochondrial haplotypes, which are structured in three main clades. In general, the Apennine locations harbour a higher number of mitochondrial clades than Central European sites, with one specific clade exclusively detected in the Apennines. Analysis of our genome-wide, multi-locus dataset reveals a clustering of P. chalcographus by geography, with Italian individuals clearly separated from Austrian samples. Our data highlight the significance of the Apennines for the genetic diversity of P. chalcographus and support the hypothesis that this area was an important refugium during unfavourable conditions in the Pleistocene. We discuss additional life-history traits and processes that shaped the evolution of this widespread beetle.

New species of cynipid inquiline, Saphonecrus kuriphilusi (Hymenoptera: Cynipidae: Synergini), from Dryocosmus kuriphilus galls in Greece.

Since Dryocosmus kuriphilus (Hymenoptera: Cynipidae) emerged worldwide as a dangerous pest of chestnuts, there is only one dubious record from Japan of a cynipid inquiline reared from its galls. This is the first comprehensively documented record of a cynipid inquiline, Saphonecrus kuriphilusi, new species reared from galls of D. kuriphilus in Greece. Morphological description, morphological and molecular diagnostic characters for the new species are given.

Limited Genetic Structure of Gypsy Moth Populations Reflecting a Recent History in Europe.

The gypsy moth, Lymantria dispar, a prominent polyphagous species native to Eurasia, causes severe impacts in deciduous forests during irregular periodical outbreaks. This study aimed to describe the genetic structure and diversity among European gypsy moth populations. Analysis of about 500 individuals using a partial region of the mitochondrial COI gene, L. dispar was characterized by low genetic diversity, limited population structure, and strong evidence that all extant haplogroups arose via a single Holocene population expansion event. Overall 60 haplotypes connected to a single parsimony network were detected and genetic diversity was highest for the coastal populations Croatia, Italy, and France, while lowest in continental populations. Phylogenetic reconstruction resulted in three groups that were geographically located in Central Europe, Dinaric Alps, and the Balkan Peninsula. In addition to recent events, the genetic structure reflects strong gene flow and the ability of gypsy moth to feed on about 400 deciduous and conifer species. Distinct genetic groups were detected in populations from Georgia. This remote population exhibited haplotypes intermediate to the European L. dispar dispar, Asian L. dispar asiatica, and L. dispar japonica clusters, highlighting this area as a possible hybridization zone of this species for future studies applying genomic approaches.

Pleistocene climate cycling and host plant association shaped the demographic history of the bark beetle Pityogenes chalcographus.

Historical climatic oscillations and co-evolutionary dependencies were key evolutionary drivers shaping the current population structure of numerous organisms. Here, we present a genome-wide study on the biogeography of the bark beetle Pityogenes chalcographus, a common and widespread insect in Eurasia. Using Restriction Associated DNA Sequencing, we studied the population structure of this beetle across a wide part of its western Palaearctic range with the goal of elucidating the role of Pleistocene glacial-interglacial cycling and its close relationship to its main host plant Norway spruce. Genetic distance among geographic sites was generally low, but clustering analysis revealed three genetically distinct groups, that is, southern, central/south-eastern, and north-eastern locations. Thus, three key P. chalcographus glacial refugia were identified: in the Italian-Dinaric region, the Carpathians, and the Russian plain, shared with its main host. The current phylogeographic signal was affected by genetic divergence among geographically isolated refugia during glacial periods and postglacial re-establishment of genetic exchange through secondary contact, reflected by admixture among genetic groups. Additionally, certain life history traits, like the beetle's dispersal and reproductive behaviour, considerably influenced its demographic history. Our results will help to understand the biogeography of other scolytine beetles, especially species with similar life history traits.

Revisiting the Distribution of Thaumetopoea pityocampa (Lepidoptera: Notodontidae) and T. pityocampa ENA Clade in Greece.

In the present work, we sampled individuals of the processionary pine moth, Thaumetopoea pityocampa (Denis and Schiffermüller; Lepidoptera: Notodontidae) from different areas of Greece between 2014 and 2016. These samples were sequenced for a 760-bp long mtDNA COI locus and the haplotypes retrieved clearly showed that the occurrence of T. pityocampa in Greece is being considerably restricted, with only 8 individuals out of the 221 exhibiting T. pityocampa haplotypes and the rest being identified as T. pityocampa ENA clade haplotypes. To that, one haplotype in particular exhibited the highest abundance and broadest geographic distribution, occurring both in mainland and on islands. Our data suggest a rather recent and rapid population expansion of the ENA clade in Greece and a concomitant recent displacement of T. pityocampa. It thus seems that the relation between T. pityocampa and T. pityocampa ENA clade needs to be further and thorough analyzed before the taxonomic status of T. pityocampa ENA clade can be concluded with confidence.