Trichoderma harzianum Strain T22 Modulates Direct Defense of Tomato Plants in Response to Nezara viridula Feeding Activity.

Plant growth-promoting fungi belonging to genus Trichoderma are known to help plants when dealing with biotic stressors by enhancing plant defenses. While beneficial effects of Trichoderma spp. against plant pathogens have long been documented, fewer studies have investigated their effect on insect pests. Here, we studied the impact of Trichoderma root colonization on the plant defense responses against stink bug feeding attack. For this purpose, a model system consisting of tomato plant, Solanum lycopersicum cv Dwarf San Marzano, Trichoderma harzianum strain T22 and the southern green stink bug, Nezara viridula, was used. We firstly determined stink bug performance in terms of relative growth rate and survival on tomato plants inoculated by T. harzianum T22. Then, we evaluated relative expression of plant defense-related genes on inoculated plants induced by stink bug feeding. We found evidence that T. harzianum T22 affects tomato defense responses against N. viridula nymphs leading to reduction of growth rate. Our results also showed that T. harzianum T22 enhances plant direct defenses by an early increase of transcript levels of jasmonic acid marker genes. Yet this effect was time-dependent and only detected 8 h after herbivore induction. Taken together, our findings provide better understanding on the mechanisms underlying tomato induced resistance against herbivorous stink bugs.

Camphor and Borneol as the Male-Produced Sex Pheromone of the Shield Bug, Orsilochides leucoptera (Hemiptera: Scutelleridae).

The ability of stink bugs to release high amounts of strong-smelling and irritating defensive compounds is related to their metathoracic gland (MTG), which is an exocrine gland with defensive, sexual, alarm and aggregation signal functions. Orsilochides leucoptera (Scutelleridae) is a widespread species in the Neotropical region that feeds on plants of the families Malvaceae, Poaceae and Euphorbiaceae. A series of compounds (ketones, alcohols and esters) have been identified in the MTGs among the three species of Scutelleridae whose MTG secretions have been investigated thus far; however, no sex pheromone compounds have been described for any scutellerid species. The aim of this work was to study sex pheromone communication within this family of stink bugs, and identify the compounds present in the MTG of O. leucoptera. Analysis by gas chromatography/mass spectrometry (GC/MS) revealed two male specific compounds identified as (R)-camphor (1) and (R)-borneol (2), which were attractive to females in Y-tube olfactometer bioassays. Also, GC/MS analysis of secretions from MTG of males and females of O. leucoptera, identified eight additional compounds: 2-(E)-hexenal (3), (E)-4-oxo-2-hexenal (4), (E)-hex-2-enyl acetate (5), (R)-linalool (6), (R)-α-terpineol (7), dodecane (8), 1-tridecene (9) and n-tridecane (10). From these, (R)-α-terpineol was detected only in the gland of males, and is probably a biosynthetic intermediate of the pheromone components. Most of the MTG compounds identified in O. leucoptera have been identified in other heteropteran species. Camphor is often a toxic and repellent compound for insects. However, we report it, for the first time, as a sex pheromone component of an insect.

The role of Oryza sativa L. 'Milyang 44' husks on the resistance to two rice stink bugs.

To elucidate the resistance mechanisms of the rice (Oryza sativa L.) cultivar 'Milyang 44' against rice stink bugs, we compared the number of stylet sheaths, husk perforations, and feeding marks on the surface of the grains caused by Leptocorisa chinensis and Cletus punctiger on Milyang 44 and the control cultivar, i.e., 'Aichinokaori SBL'. We also examined the cross-sectional structure of the rice husks. We found that the number of stylet sheaths per panicle was higher in Milyang 44 than in Aichinokaori SBL for both rice stink bug species, except in one test involving C. punctiger. However, Milyang 44 had significantly less damage per number of stylet sheaths than Aichinokaori SBL, resulting in a lower percentage rates of pecky rice grains in Milyang 44. Interestingly, there was no difference in the percentage rates of pecky rice between the two cultivars after removing one third of the husks. Histological analysis showed that the sclerenchymatous cell wall containing lignin of husk was thicker in Milyang 44 than in Aichinokaori SBL, suggesting that the husk of Milyang 44 plays an important role in its resistance to these two rice stink bug species.

Insight into Salivary Gland Proteomes of Two Polyphagous Stink Bugs: Nezara viridula L. and Halyomorpha halys Stål.

The invasive brown marmorated stink bug (BMSB), Halyomorpha halys Stål, and the southern green stink bugs (SGSBs), Nezara viridula L., are widely distributed in Europe, even if the date of introduction and the diet differ. Saliva of Hemipteran pests plays essential roles in the interaction between insects and their host plants. The salivary proteomes of several aphid species have been studied and found to differ according to the species, while no comparative investigation between phytophagous stink bugs has been performed yet. Here, the salivary proteins from two bugs, BMSB and SGSB, are analyzed using LC-MS/MS. Data are available via ProteomeXchange with identifiers PXD011920 and PXD011976. A total of 238 and 305 proteins are identified in salivary glands of BMSB and SGSB, respectively. In comparison with salivary proteome from other Hemiptera, the most striking feature of the salivary gland proteomes of SGSB and BMSB is the similar pattern of protein functions between both species. Some of the proteins are speculated to play a significant role in plant-insect interactions. The results herein provide a framework for future research to elucidate the molecular basis of differential impact of piercing-sucking insects on host plants.

Exploring Phylogenetic Relationships of the Mecocephala Group (Hemiptera: Pentatomidae) Using Continuous and Discrete Characters, with Notes on Taxonomy.

The Mecocephala group comprises about 50 species, restricted to the Neotropics and with the highest species richness in Neotropical South America. Several species use rice as host plants and their identification is facilitated by the presence of exaggerated head proportions and a unique male genitalic morphology. The taxonomy of the group has been extensively explored, but inferring its monophyly and especially its internal phylogenetic relationships has been challenging. Here, we inferred the phylogenetic relationships for the group assembling the most complete taxonomic sampling to date, analyzing discrete and continuous morphological characters through equal and implied weighted parsimony analyses. The monophyly of the group was recovered, but internal relationships varied slightly according to the dataset tested. Thus, we propose internal arrangements for the group and provide a formal description of the Mecocephala group, diagnoses for each genus, a dichotomous key to identify its genera, and illustrations of the morphological characters and type species.

A Faboideae-Specific Floral Scent Betrays Seeds to an Important Granivore Pest.

Seed predation by insect herbivores reduces crop production worldwide. Foraging on seeds at pre-dispersal generally means that females need to find the suitable host plant within a relatively short timeframe in order to synchronize larval development with seed production. The mechanistic understanding of host finding by seed pests can be harnessed for more sustainable pest management strategies. We here studied the chemical communication between the bean bug Riptortus pedestris, a major pest of legumes, and several crop species and cultivars in the Fabaceae. Via a comparative chemical analysis, we found that 1-octen-3-ol is the principal constituent of the floral scents of most species tested in the subfamily Faboideae, including soybean and faba bean. With field trapping and laboratory bioassays, including electroantennography, we further revealed that this compound can be perceived, and stimulate attraction responses, by R. pedestris nymphs and adults. The addition of 1-octen-3-ol to pheromone traps might therefore improve trapping efficacy for controlling populations of this important granivore pest on legumes.

The Introduction and Establishment of Four Invasive Insect Species in Serbia.

Urban areas are often populated by specific species of insects, some colorful and appealing, such as ladybugs and butterflies, and others irritating as nuisance bitters or as vectors of pathogens of public health importance. Mosquitoes in urban areas often utilize habitats adjacent to human residences, while phytophagous insect species such as stink bugs often colonize ornamental plants and utilize human-made structures including houses as overwintering shelters. This article discusses the early detection and the current distribution of two invasive mosquito species, Aedes albopictus Skuse 1894 and Ae. japonicus (Theobald 1901), in Serbia, introduced in 2009 and 2018, respectively. From the first findings until today, regular monitoring has been carried out and the establishment of both species in the newly invaded areas has been confirmed. Both species can become nuisance species, especially at high population densities, but more importantly, they are capable of transmitting a wide variety of arboviruses of public health importance. This article also discusses two invasive stink bug species Halyomorpha halys Stål 1855 and Nezara viridula Linnaeus 1758, introduced in Serbia in 2015 and 2008, respectively. These two stink bug species have also been monitored, and the establishment of their populations in the country has been confirmed. Both species have caused damage to a wide range of crops and ornamental plants and sometimes become nuisance pests in urban areas.

The Predatory Stink Bug Arma custos (Hemiptera: Pentatomidae) Produces a Complex Proteinaceous Venom to Overcome Caterpillar Prey.

Predatory stink bugs capture prey by injecting salivary venom from their venom glands using specialized stylets. Understanding venom function has been impeded by a scarcity of knowledge of their venom composition. We therefore examined the proteinaceous components of the salivary venom of the predatory stink bug Arma custos (Fabricius, 1794) (Hemiptera: Pentatomidae). We used gland extracts and venoms from fifth-instar nymphs or adult females to perform shotgun proteomics combined with venom gland transcriptomics. We found that the venom of A. custos comprised a complex suite of over a hundred individual proteins, including oxidoreductases, transferases, hydrolases, ligases, protease inhibitors, and recognition, transport and binding proteins. Besides the uncharacterized proteins, hydrolases such as venom serine proteases, cathepsins, phospholipase A2, phosphatases, nucleases, alpha-amylases, and chitinases constitute the most abundant protein families. However, salivary proteins shared by and unique to other predatory heteropterans were not detected in the A. custos venom. Injection of the proteinaceous (>3 kDa) venom fraction of A. custos gland extracts or venom into its prey, the larvae of the oriental armyworm Mythimna separata (Walker, 1865), revealed insecticidal activity against lepidopterans. Our data expand the knowledge of heteropteran salivary proteins and suggest predatory asopine bugs as a novel source for bioinsecticides.

Predator-Pest Dynamics of Arthropods Residing in Louisiana Soybean Agroecosystems.

Over the past two decades, management practices within Louisiana soybean production have shifted. Successful application of an integrated pest management (IPM) strategy requires an understanding of how these changes have affected predator-pest dynamics. Surveys monitoring foliage-foraging arthropod populations in soybean took place across six locations within Louisiana over six years (2012-2014 and 2015-2018). Temporal associations of pest groups, defoliating and piercing-sucking, and predator groups relating to soybean phenology were observed. Additionally, soybean maturity groups (III, IV, and V) were also evaluated to delineate potential differences. Results indicated higher abundances of piercing-sucking pests compared to defoliating pests across both datasets (2012-2014 and 2015-2018). Pest groups were more abundant in later soybean reproductive periods, mainly attributed to Chrysodeixis includens and Piezodorus guildinii. Predator populations were mainly comprised of Araneae and Geocoridae throughout the survey periods. From 2015 to 2018, soybean growth had a significant effect on total predator abundance with more predators present at the pod-fill and soybean maturity stage. Correlations between total pest abundance and total predators exhibited a moderate positive linear relationship. Soybean maturity groups only influenced piercing-sucking pest abundance, with later maturing groups (IV and V) having higher numbers. Thus, control tools and tactics aimed at controlling late season pests should be modified to avoid reducing predator populations.

New Records and Updated Checklist of the Pentatomoidea (Hemiptera: Heteroptera) of Greece.

Eight species of the superfamily Pentatomoidea are recorded from Greece for the first time: Aelia germari Küster 1852, Eurygaster hottentotta (Fabricius 1775), Eysarcoris aeneus (Scopoli 1763), Neottiglossa lineolata (Herrich-Schaeffer 1830), Neottiglossa pusilla (Gallen 1789), Picromerus bidens (Linnaeus 1758), Podops (Podops) inunctus (Fabricius 1775) and Tarisa pallescens (Jakovlev 1871). A complete updated species checklist with distributional data notes for all the new species for Greece are provided.

Contrasting reproductive traits of competing parasitoids facilitate coexistence on a shared host pest in a biological control perspective.

BACKGROUND: Interspecific competition in insect parasitoids is an important ecological phenomenon that has relevant implications for biological pest control. To date, interspecific intrinsic (=larval) competition has been intensively studied, while investigations on extrinsic (=adult) competition have often lagged behind. In this study we examined the role played by parasitoid reproductive traits and host clutch size on the outcome of extrinsic competition between Trissolcus basalis (Wollaston) and Ooencyrtus telenomicida (Vassiliev), two egg parasitoids of the pest Nezara viridula (L). Laboratory experiments were conducted by allowing both parasitoid species to exploit an egg mass made of 10, 20, 30, or 40 hosts through single or simultaneous releases. Furthermore, under field conditions, egg masses consisting of 10 or 40 hosts were exposed in a tomato crop in order to validate laboratory investigation. RESULTS: The results show that the egg mass size is an important predictor of extrinsic competition in our study system as a higher proportion of T. basalis emerged from large egg masses, while O. telenomicida dominated in small egg masses. Analysis of reproductive traits of parasitoid species indicates that T. basalis has superior abilities in host exploitation compared with O. telenomicida. CONCLUSIONS: We found that contrasting reproductive traits of two competing egg parasitoid species facilitate coexistence on a shared stink bug host. This work also highlights the importance to consider extrinsic competitive interactions between parasitoid species in a biological control perspective. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Discrimination between Eggs from Stink Bugs Species in Europe Using MALDI-TOF MS.

In the current paper, we used a method based on stink bug egg-protein immobilization on filter paper by drying, followed by post-(storage and shipping) extraction in acidified acetonitrile containing matrix, to discriminate between nine different species using MALDI-TOF MS. We obtained 87 correct species-identifications in 87 blind tests using this method. With further processing of the unblinded data, the highest average Bruker score for each tested species was that of the cognate reference species, and the observed differences in average Bruker scores were generally large and the errors small except for Capocoris fuscispinus, Dolycoris baccarum, and Graphosoma italicum, where the average scores were lower and the errors higher relative to the remaining comparisons. While we observed clear discrimination between the nine species using this method, Halyomorpha halys and Piezodorus lituratus were more spectrally related than the other pairwise comparisons.

Lost and found-Discovery of the presumed lost type of emArma/em empallipes/em Dallas, new synonymy and description of two new species of emPodisus/em Herrich-Schäffer (Hemiptera: Pentatomidae: Asopinae).

The presumed lost type of Arma pallipes Dallas is found and Thomas's (1992) neotype is hereby set aside (ICZN 1999, Art. 75.8). A. pallipes is proposed as junior subjective synonym of Podisus ventralis (Dallas). A lectotype designation by inference of holotype (Gaedike 1971) is accepted for the female typical specimen of P. neniator deposited at Senckenberg Deutsches Entomologisches Institut (SDEI, Müncheberg, Germany) (ICZN 1999, Art. 74.6). Two new species of Podisus Herrich-Schäffer are described: Podisus graziae Brugnera, Roell Lemaître sp. nov. and Podisus thomasi Lemaître, Roell Brugnera sp. nov. Photographs of all types as well as of live specimens are provided.

Characterization and Comparison of Intestinal Bacterial Microbiomes of Euschistus heros and Piezodorus guildinii Collected in Brazil and the United States.

Background: Herbivorous insects are one of the main biological threats to crops. One such group of insects, stink bugs, do not eat large amounts of tissue when feeding on soybean, but are damaging to the quality of the seed yield as they feed on green developing seeds leading to poorly marketable harvests. In addition to causing physical damage during sucking-feeding activities, the insects can also transmit microbial pathogens, leading to even greater yield loss. Conducting surveys of the insect intestinal microbiome can help identify possible pathogens, as well as detail what healthy stink bug digestive systems have in common. Methods: We used the conserved V4 region of the 16S rRNA gene to characterize the bacterial microbiome of the red-banded stink bug Piezodorus guildinii collected in Brazil and the United States, as well as the neotropical brown stink bug Euschistus heros collected in Brazil. Results: After quality filtering of the data, 192 samples were kept for analyses: 117 samples from P. guildinii covering three sites in Brazil and four sites in the United States, and 75 samples for E. heros covering 10 sites in Brazil. The most interesting observations were that the diversity and abundance of some bacterial families were different in the different ecoregions of Brazil and the United States. Conclusion: Some families, such as Acetobacteraceae, Bacillaceae, Moraxellaceae, Enterobacteriaceae, and Rhodocyclaceae, may be related to the better adaptation in some localities in providing nutrients, break down cellulose, detoxify phytochemicals, and degrade organic compounds, which makes it difficult to control these species.

Potential of Ficus carica extracts against Euschistus heros: Toxicity of major active compounds and selectivity against beneficial insects.

BACKGROUND: Using plant-based extracts and their constituents has been suggested as an alternative tool to replace or integrate with the synthetic compounds used to manage insect pests. Here, we evaluated the potential of extracts obtained from Ficus carica Linn (Moraceae) branches and leaves against the Neotropical brown stink bug, Euschistus heros, one of the most prevalent insect pests in soybean fields. We further isolated and evaluated the toxicity of the extracts' major components against E. heros. Additionally, by using computational docking analysis and toxicological approaches, we assessed the physiological basis for the selectivity of these extracts against beneficial insects such as pollinator bees (i.e. Apis mellifera and the Neotropical stingless bee Partamona helleri), ladybeetles (Eriopis connexa and Coleomegilla maculata), and lacewings (Chrysoperla externa). RESULTS: Our results demonstrate that branch (LC50  = 5.9 [4.7-7.1] mg mL-1 ) and leaf (LC50  = 14.1 [12.5-15.4] mg mL-1 ) extracts exhibited similar toxicity against E. heros. Our phytochemical analysis revealed psoralen and bergapten furanocoumarins as the major components of the extract. Based on our computational predictions, these molecules' differential abilities to physically interact with the acetylcholinesterases of E. heros and beneficial insects play relevant roles in their selectivity actions. The estimated LC90 values of branch (30.0 mg mL-1 ) and leaf (30.0 mg mL-1 ) extracts killed less than 12% of the beneficial insects. CONCLUSION: Overall, our findings revealed that furanocoumarin-rich extracts obtained from F. carica extracts have the potential to be used as alternative tools in the integrated management of stink bug pests. © 2021 Society of Chemical Industry.

Illustrated guide to Pentatominae (Hemiptera: Pentatomidae) species associated with the four main grain crops in Rio Grande do Sul state, Brazil.

Several stink bugs in the subfamily Pentatominae are crop pests or have the potential to damage plants of economic importance. In the Brazilian state of Rio Grande do Sul, where agriculture plays a major role in the economy, the knowledge about pest stink bugs is fragmented and, in some instances, outdated. This work provides a summary of Pentatominae species recorded in Rio Grande do Sul feeding on the four most important grain crops for the state, i.e. soybean, rice, maize, and wheat, plus canola, an emerging crop. This survey is enhanced with new records from scientific collections, a short diagnosis for each species, distribution maps, an identification key, and carefully illustrated to allow for species recognition in the field. With this work, we aim to reunite the scattered knowledge of the group in one single revision, and provide a useful tool for identifying the pest stink bugs of Rio Grande do Sul.

Plant volatiles guide the new pest Dichelops furcatus to feed on corn seedlings.

BACKGROUND: Recently, in temperate and neotropical regions of South America the generalist stink bug Dichelops furcatus (Hemiptera: Pentatomidae) became a new pest of corn (Zea mays) seedlings. Implementation of no-tillage cultivation system left organic matter covering the soil, which shelters adults of stink bugs during winter. In spring, corn is sowed under soybean stubble and D. furcatus adults start to feed on seedlings. To determine corn-derived volatile organic compounds (VOCs) that attract this stink bug species, we evaluated stink bug preferences from two corn hybrids with contrast germplasm backgrounds, a temperate and a tropical hybrid. RESULTS: Stink bugs preferred to feed on temperate seedlings rather than on the tropical ones. GC-MS and PCA analysis of VOCs suggested that hybrids emitted contrasting blends. Linalool represented 68% of total VOCs emitted from temperate corn, while in the tropical hybrid this compound represented 48%. Olfactometer experiments demonstrated that linalool was attractive to stink bugs. However, 2 h of D. furcatus attack induced emission of 14 additional VOCs in temperate seedlings, and olfactometer bioassay and blend of VOCs emission suggested that perceived volatiles by stink bugs induced feeding avoidance. The increment of VOCs emission was associated with the induction of JA, JA-Ile, ABA, and IAA, and decreasing of SA concentrations. CONCLUSION: This is the first time showing a complete profile of defensive phytohormones induced by stink bugs feeding on corn, and further demonstrating that a blend of corn seedling-associated VOCs, mainly composed by linalool, modulates D. furcatus adults' behavior and feeding preferences. © 2021 Society of Chemical Industry.

Stink Bug Communication and Signal Detection in a Plant Environment.

Plants influenced the evolution of plant-dwelling stink bugs' systems underlying communication with chemical and substrate-borne vibratory signals. Plant volatiles provides cues that increase attractiveness or interfere with the probability of finding a mate in the field. Mechanical properties of herbaceous hosts and associated plants alter the frequency, amplitude, and temporal characteristics of stink bug species and sex-specific vibratory signals. The specificity of pheromone odor tuning has evolved through highly specific odorant receptors located within the receptor membrane. The narrow-band low-frequency characteristics of the signals produced by abdomen vibration and the frequency tuning of the highly sensitive subgenual organ vibration receptors match with filtering properties of the plants enabling optimized communication. A range of less sensitive mechanoreceptors, tuned to lower vibration frequencies, detect signals produced by other mechanisms used at less species-specific levels of communication in a plant environment. Whereas the encoding of frequency-intensity and temporal parameters of stink bug vibratory signals is relatively well investigated at low levels of processing in the ventral nerve cord, processing of this information and its integration with other modalities at higher neuronal levels still needs research attention.

The complete mitochondrial genome of Eysarcoris guttigerus (Hemiptera: Pentatomidae).

The mitochondrial genome of Eysarcoris guttigerus has been decoded using Illumina sequencing. The mitogenomic size is 15,368 bp with 42.97% A, 33.90% T, 13.22% C, and 9.90% G. It encoded 13 protein-coding genes, two ribosomal RNA, and 22 transfer RNAs. Phylogenetic tree showed that genus Eysarcoris had a closer relationship with the genus Carbula than the rest of 20 genera.

Comparative Morphology of the Mouthparts in Three Predatory Stink Bugs (Heteroptera: Asopinae) Reveals Feeding Specialization of Stylets and Sensilla.

Mouthpart structures were observed in three species of Asopinae using scanning electron microscopy to investigate their morphological disparity. The examined species attack mainly slow-moving, soft-bodied insects, primarily larval forms of the Lepidoptera, and are the natural enemies of many pests. This is the first detailed description of their external mouthparts. The triangular and elongated labrum and four-segmented tube-like labium are longer in Picromerus species (Picromerus bidens (Linnaeus, 1758) and Picromerus lewisi Scott, 1874 than in Cazira bhoutanica Schouteden, 1907. The labrum of P. lewisi and C. bhoutanica appear to be equipped with olfactory sensilla basiconica Sb3, a special type of sensilla with nanopores. The labium surface in all studied species bears 14 types of sensilla (St1-St4, Sb1-7, Sst, Sca1-2). A new characteristic of sensilla trichodea is represented in sensillum St1; in both Picromerus species, it is classified as an olfactory sensillum with nanopores. The tripartite apex of the labium consists of two lateral lobes and a central membranous lobe having microtrichial extensions. Each lobe has one sensory field, including sensilla basiconica (Sb7), sensilla styloconica (Sst), and sensilla trichodea (St4). In the three studied predatory stink bugs, each mandibular stylet tip has five irregular teeth and three long, pointed hooks. The two opposing maxillae, which are held together by a tongue-and-groove system, form a food canal and a salivary canal. The apices of the right maxilla have small teeth and few short barbs along the edge of the food canal. In P. bidens and P. lewisi, there are 5 teeth, while in C. bhoutanica there are 2. Based on structural differences, we inferred that the hook-shaped mandibular teeth, right maxilla with small teeth, and few short barbs along edge of the food canal are more adapted for a predatory lifestyle. Predatory stink bugs use sharp recurved hooks and irregular teeth penetrating, tearing, or filing devices that aid in the mechanical disruption of host tissue. Stiff bristles in the food canal may indicate their possible adaptation to feeding on insect larvae. The evolution of mouthpart morphology and the putative functional significance of sensilla are discussed, providing insight into the sensory mechanism.