Interleg coordination in free-walking bug Erthesina fullo (Hemiptera: Pentatomidae).

Insects can adapt their walking patterns to complex and varied environments and retain the ability to walk even after significant changes in their physical attributes, such as amputation. Although the interleg coordination of intact insects has been widely described in previous studies, the adaptive walking patterns in free-walking insects with amputation of 1 or more legs are still unclear. The pentatomid bug Erthesina fullo exhibits a tripod gait, when walking freely on horizontal substrates, like many other insects. In this study, amputations were performed on this species to investigate changes in interleg coordination. The walking parameters were analyzed, such as the locations of touchdown and liftoff, cycle period, walking speed, and head displacement of intact and amputated insects. The results show that E. fullo displays adaptive interleg coordination in response to amputations. With 1 amputated leg, bugs changed to a 3-unit gait, whereas with 2 amputated legs they employed a wave gait. These data are helpful in exploring the motion mode control in walking insects and provide the theoretical basis for the gait control strategy of robots, when leg failure occurs.

Insecticide Susceptibilities and Enzyme Activities of Four Stink Bug Populations in Mississippi, USA.

In Mississippi, the Pentatomidae complex infesting soybean is primarily composed of Euschistus servus, Nezara viridula, Chinavia hilaris, and Piezodorus guildinii. This study employed spray bioassays to evaluate the susceptibilities of these stink bugs to seven commonly used formulated insecticides: oxamyl, acephate, bifenthrin, λ-cyhalothrin, imidacloprid, thiamethoxam, and sulfoxaflor. Stinks bugs were collected from soybeans in Leland, MS, USA during 2022 and 2023, as well as from wild host plants in Clarksdale, MS. There was no significant difference in the susceptibility of C. hilaris to seven insecticides between two years, whereas P. guildinii showed slightly increased susceptibility to neonicotinoids in 2023. Among all four stink bug species, susceptibility in 2022 was ranked as P. guildinii ≤ C. hilaris ≈ N. viridula, while in 2023, it was ranked as P. guildinii ≤ C. hilaris ≤ E. Servus. Additionally, populations of E. servus and P. guildinii collected from Clarksdale exhibited high tolerance to pyrethroids and neonicotinoids. Moreover, populations of E. servus and P. guildinii from SIMRU-2022 and Clarksdale-2023 showed elevated esterase and cytochrome P450 activity, respectively. These findings from spray bioassays and enzyme activity analyses provide a baseline for monitoring insecticide resistance in Pentatomidae and can guide insecticide resistance management strategies for Mississippi soybean.

Optimizing the Release Pattern of Telenomus podisi for Effective Biological Control of Euschistus heros in Soybean.

An augmentative biological control program using the egg parasitoid Telenomus podisi Ashmead (Hymenoptera: Platygastridae) is a promising tool for the management of the brown stink bug, Euschistus heros (Fabricius) (Hemiptera: Pentatomidae) in soybeans. The T. podisi are released as adults or pupae within lab-reared E. heros eggs. Because of the small size of the parasitoid and potentially limited dispersal ability, determining the optimal release pattern is critical for biological control of the target pest. This study used sentinel E. heros eggs to investigate T. podisi dispersal within soybean crops during two distinct phenological stages: the beginning of flowering (Vn-R1) and the grain-filling phase (R5-R6). Data were analyzed using semi-variograms and kriging maps. The results indicate significant differences in parasitism rates between the two plant growth stages and among different matrices. The maximum dispersal range for T. podisi was calculated at 39.0 m in the Vn-R1 stage with a maximum parasitism rate of 42%, while in the R5-R6 stage, the maximum dispersal range was calculated to be 30.9 m with a maximum parasitism of 73%. Therefore, it is recommended that release points for T. podisi be spaced no further than 30 m apart. These results provide valuable insights for future research and applications in biological control strategies, including adjustment of the logistics and release technique depending upon the crop phenological stage.

Effects of photoperiod and temperature on the developmental duration and diapause in Dolycoris baccarum (Heteroptera: Pentatomidae) from Hohhot, Inner Mongolia, China.

The shield bug, Dolycoris baccarum (L.) (Heteroptera: Pentatomidae), is widely distributed across Asia and Europe. At high latitudes, it overwinters, as adult in diapause, which then becomes the insect source for the following year. To fully understand the developmental duration and diapause characteristics of D. baccarum, the effects of photoperiod and temperature were studied in a population from Hohhot, Inner Mongolia, China. The results indicated that the developmental duration was significantly prolonged at temperatures of 20 or 25 °C, with a prolonged light period; however, when the light period was prolonged to 16L:8D and 18L:6D, the developmental duration was shortened significantly. Furthermore, the developmental duration was also shortened significantly with increasing temperature, when the photoperiod was 12L:12D for short days and 16L:8D for long days. All individuals entered diapause under short-day conditions of 10L:14D and 12L:12D at a temperature of 20 °C; however, the diapause rate decreased significantly under 14L:10D and 16L:8D photoperiods, and the diapause rate decreased significantly at a temperature of 25 °C with prolonged photoperiod. Interestingly, when the photoperiod was fixed at 12L:12D, the diapause rates at different temperatures (20, 25, 28, and 30 °C) exceeded 95%; while the effect of temperature on diapauses was nonsignificant under this photoperiod, it was still sensitive to the photoperiod; at a photoperiod of 16L:8D, the effect of temperature on the diapause rate was noticeable, and the diapause rate decreased significantly with increasing temperature.

Metabolic Response of Peach Fruit to Invasive Brown Marmorated Stink Bug (Halyomorpha halys Stål.)'s Infestation.

The brown marmorated stink bug (BMSB; Halyomorpha halys Stål.) is a highly destructive and polyphagous invasive pest that poses a serious threat to more than a hundred reported host plants. In the current study, the metabolic response of peach fruit of two cultivars-'Maria Marta' and 'Redhaven'-to BMSB infestation was studied using high-performance liquid chromatography (HPLC) and mass spectrometry (MS). In general, a strong phenolic response to BMSB infestation in peach flesh in the injury zone was observed, with flavanol content increasing by 2.4-fold, hydroxycinnamic acid content by 5.0-fold, flavonol content by 3.2-fold, flavanone content by 11.3-fold, and dihydrochalcones content by 3.2-fold compared with the undamaged tissue in the cultivar 'Maria Marta'. The phenolic response in the 'Redhaven' cultivar was even stronger. Consequently, the total phenolic content in the injured flesh also increased, 3.3-fold in 'Maria Marta' and 6.9-fold in 'Redhaven', compared with the uninjured flesh. Infestation with BMSB induced the synthesis of cyanidin-3-glucoside, which is not normally present in peach flesh. In comparison, the phenolic response was lower in peach peel, especially in the cultivar 'Maria Marta'. The study showed that both peach cultivars reacted to BMSB infestation with an increase in phenolic content in the peach flesh, but in a limited area of injury.

Protective Geometry and Reproductive Anatomy as Candidate Determinants of Clutch Size Variation in Pentatomid Bugs.

AbstractMany animals lay their eggs in clusters. Eggs on the periphery of clusters can be at higher risk of mortality. We asked whether the most commonly occurring clutch sizes in pentatomid bugs could result from geometrical arrangements that maximize the proportion of eggs in the cluster's interior. Although the most common clutch sizes do not correspond with geometric optimality, stink bugs do tend to lay clusters of eggs in shapes that protect increasing proportions of their offspring as clutch sizes increase. We also considered whether ovariole number, an aspect of reproductive anatomy that may be a fixed trait across many pentatomids, could explain observed distributions of clutch sizes. The most common clutch sizes across many species correspond with multiples of ovariole number. However, there are species with the same number of ovarioles that lay clutches of widely varying size, among which multiples of ovariole number are not overrepresented. In pentatomid bugs, reproductive anatomy appears to be more important than egg mass geometry in determining clutch size uniformity. In addition, our analysis demonstrates that groups of animals with little variation in ovariole number may nonetheless lay a broad range of clutch shapes and sizes.

Distribution and prevalence of viral genomes in Italian populations of the invasive brown marmorated stink bug Halyomorpha halys.

Halyomorpha halys (Stål), the brown marmorated stink bug, is a highly invasive insect species that causes significant agricultural losses, especially to orchard fruits, vegetables, herbaceous and ornamental plants. It is also a nuisance pest that seeks shelter in indoor spaces during the winter months. Harnessing the H. halys virome can result in new environmentally sustainable approaches to contain its populations and its relatated agricultural damages. In this study, RNA-Seq data were used to explore the virome associated to ten field populations collected in the Lombardy region in Northern Italy. We identified six complete viral genomes, three of which were previously unknown, belonging to the orders Reovirales, Articulavirales, Ghabrivirales, Durnavirales, and Picornavirales. The prevalence of the six viruses was evaluated by Real-time reverse transcription-quantitative PCR on eighty individuals. Halyomorpha halys ifla-like virus 2 turned out to be the most geographically widespread virus, as it was found in more than 50% of the analyzed insects and in nine out of the ten sampling locations. Moreover, in some individuals, this iflavirus was found in association with each of the other viruses in various combinations that involved up to four viruses. Further studies on such virus-virus interactions and their relationships with the insect host may open the possibility to exploit these naturally occurring viruses as specific and targeted biocontrol agents of H. halys.

First report of Colletotrichum fioriniae infections in brown marmorated stink bugs, Halyomorpha halys.

An epizootic caused by fungal pathogens occurred among Halyomorpha halys, brown marmorated stink bugs, while they were overwintering, with infections also occurring after overwintering. We report that one of the two pathogens responsible was Colletotrichum fioriniae (Marcelino & Gouli) Pennycook; a species well known as a plant pathogen and endophyte and which has only previously been reported naturally infecting elongate hemlock scales, Fiorinia externa. To prove pathogenicity, H. halys adults challenged with conidia died from infections and the fungus subsequently produced conidia externally on cadavers.

First record of the predatory stink bug species Picromerusgriseus (Dallas) (Hemiptera, Heteroptera, Pentatomidae, Asopinae) in Japan, with an illustrated key to the Japanese species of the genus Picromerus Amyot & Serville.

Background: The predatory stink bug genus Picromerus Amyot & Serville, 1843 (Hemiptera, Heteroptera, Pentatomidae, Asopinae) comprises 11 species found in the Northern Hemisphere. In Japan, two species have been recorded to date. However, an easy-to-understand identification method, such as an illustrated key, is lacking. Currently, Picromerusgriseus (Dallas, 1851) has been recorded in Bangladesh, Bhutan, China, Indonesia, Myanmar, Pakistan and Taiwan, but not in Japan. New information: Picromerusgriseus was recorded in Japan for the first time, based on a single individual collected from grasslands around the fields of Ishigaki Island of the Ryukyu Islands, which belong to the Oriental Region. This discovery represents the easternmost record of the species. An illustrated key to the species of Picromerus occurring in Japan is also provided.

Diversity of stink bugs (Hemiptera: Heteroptera: Pentatomoidea) in the Brazilian Pampa and the Parque Estadual de Itapuã, Rio Grande do Sul.

Biological surveys represent important contributions to the knowledge of species diversity, conservation, taxonomy, and biogeography. Few surveys of stink bugs and allied groups (Pentatomoidea) were produced in Brazil, especially regarding the Brazilian Pampa, an overlooked biome. Here, a list of Pentatomoidea species of the Brazilian Pampa is presented for the first time, composed of seven families and 152 species. The results of five years of sampling in the Parque Estadual de Itapuã (PEI) are also presented. A total of 693 individuals were sampled, representing 41 species, 29 genera, and five families of Pentatomoidea. Pentatomidae presented a higher richness (28 species), followed by Cydnidae, Scutelleridae, Dinidoridae, and Megarididae. The most abundant species collected in the PEI was Brachystetus geniculatus (Fabricius) sampled in Bromelia balansae Mez (32.76%), followed by Oebalus poecilus (Dallas) found in Persicaria hydropiper (Linnaeus) (11.99%), Pallantia macula (Dallas) collected in B. balansae (10.92%), Dinidor saucius Stål (8.57%) in Smilax cf. campestris Griseb., and Caonabo pseudoscylax (Bergroth) (5.35%) found in Homolepis glutinosa (Sw.) Zuloaga & Soderstrom. This is the first time that a list of species is presented for the Brazilian Pampa and Parque Estadual de Itapuã, providing a baseline to develop further studies with Pentatomoidea in the biome.

Temperature-Dependent Life Table Parameters of Brown Marmorated Stink Bug, Halyomorpha halys (Stål) (Hemiptera: Pentatomidae) in the United States.

Brown marmorated stink bug, Halyomorpha halys (Stål) (Hemiptera: Pentatomidae), is a generalist pest that causes serious injury to a variety of crops around the world. After the first detection in the USA, H. halys became a serious threat to growers resulting in significant crop damage. Understanding the effect of temperature on H. halys development will help to achieve successful control by predicting the phenological timing of the pest. Here, life table parameters (survival, development, reproduction, and daily mortality) of H. halys were evaluated for New Jersey and Oregon populations in the US. Parameters were determined from field-collected and laboratory-reared individuals. The results indicated that New Jersey populations had higher levels of egg-laying than Oregon populations and exhibited higher and earlier fecundity peaks. Survival levels were similar between populations. Linear and nonlinear fit were used to estimate the minimum (14.3 °C), optimal (27.8 °C), and maximum (35.9 °C) temperatures where development of H. halys can take place. An age-specific fecundity peak (Mx = 36.63) was recorded at 936 degree days for New Jersey populations, while maximum fecundity (Mx = 11.85) occurred at 1145 degree days in Oregon. No oviposition was recorded at the lowest (15 °C) or highest (35 °C) trialed temperatures. Developmental periods increased at temperatures above 30 °C, indicating that such higher temperatures are suboptimal for H. halys development. Altogether the most optimal temperatures for population increase (rm) ranged from 25 to 30 °C. Survival rates of H. halys at suboptimal low temperatures of 8 °C (i.e., 61%) is comparable to previous reports. The present paper provides additional data and context from a range of experimental conditions and populations. Such temperature-related H. halys life table parameters can be used to provide determine the risk to susceptible crops.

Efficacy of Kairomone Lures to Attract Parasitoids of Halyomorpha halys.

In its native range, Halyomorpha halys (Stål) is suppressed by parasitoids in the genus Trissolcus (Hymenoptera: Scelionidae). Trissolcus native to Utah have demonstrated low parasitism of H. halys, while adventive Trissolcus japonicus (Ashmead) have shown parasitism of up to 20%. Custom rubber septa lures containing stink bug kairomones, n-tridecane (attractant), and (E)-2-decenal (repellent), at 100%, 90%, and 80% levels of attractant (10 mg load rate), were placed adjacent to sentinel H. halys egg masses in northern Utah field trials. Egg masses were evaluated for the presence and intensity (proportion of parasitized eggs) of parasitism. Parasitism by T. japonicus and T. euschisti (Ashmead) was low; however, the 100% lure showed double the parasitism of the control and more than three times that of the 90% and 80%. Two-way choice mesocosm trials in the laboratory evaluated previous lures and a lower load rate of 5 mg-100% attractant treatment. Lures of 10 mg at 100% and 80% were more attractive to T. japonicus than the control, while 5 mg at 100% and 10 mg at 90% showed no significant attraction. Our results support a proof-of-concept of rubber septa as release devices for kairomones to attract T. japonicus and provide a baseline for future field-based studies.

Citizen Science Meet South American Tachinids: New Records of Feather-Legged Fly Trichopoda (Galactomyia) pictipennis Bigot (Diptera: Tachinidae) from Chile.

The species of Gymnosomatini (Diptera: Tachinidae) are specialized to attack stink bugs (Hemiptera: Pentatomidae). Some of these flies, such as those in the genus Trichopoda Berthold, are conspicuous and represent target species to study with citizen science. Here we report for the first time the distribution range of T. (Galactomyia) pictipennis Bigot, a newly introduced biological control species of tachinid for use against stink bugs in Chile using data obtained through the citizen science method. Additionally, we update the distribution of T. arcuata (Bigot), a native biological control species of tachinid, and assess the grade of overlap between the flies and their main stink bug hosts. We obtained data regarding the occurrence of flies and stink bugs from the citizen science program "Moscas Florícolas de Chile," from Facebook groups, and from iNaturalist. We standardized our data to avoid temporal, spatial, and geographic biases. We report the extent of occurrence (EOO) and area of occupancy (AOO) for each fly and stink bug species. The EOOs for T. pictipennis and T. arcuata are 85,474 km2 and 20,647 km2 through central Chile ecosystems, respectively. The EOO of T. arcuata is overlapped by 53% with the introduced species. Trichopoda pictipennis has a high percentage of overlap with the EOO of Nezara viridula Linnaeus (61%), whereas T. arcuata has low percentages of overlap with different native Acledra species and with N. viridula. We discuss the potential of citizen science to detect (1) areas with higher diversity or gaps of occurrences, (2) new biological control agents, (3) prey records, (4) negative impacts on non-target species, (5) changes in composition at long-term, and (6) areas to promote conservation biological control in agricultural landscapes.

Bioactivity of essential oils from Croton grewioides and its major compounds: toxicity to soybean looper Chrysodeixis includens and selectivity to the predatory stink bug Podisus nigrispinus.

Natural biological control is a key factor that ensures the regulation of insect pest populations in agroecosystems. However, the indiscriminate use of pesticides has compromised this environmental service. Thus, the search for environmentally safe pesticides is an increasing requirement for sustainable food production. In this study, we analyzed the toxicity of essential oils from two accessions (CGR112 and CGR126) of Croton grewioides and its major compounds, methyl eugenol and eugenol, on the soybean pest Chrysodeixis includens. In addition, we investigated the sublethal effects of these compounds on the predatory bug Podisus nigrispinus, analyzing its developmental, reproduction and life table parameters. Essential oils and their major compounds were toxic to C. includens and P. nigrispinus. In general, the presence of eugenol made the essential oil more toxic to the pest and selective to the natural enemy. Eugenol was the most toxic compound for 2nd instar larvae of C. includens at LD50, followed by CGR126 essential oil from C. grewioides which was equally toxic at higher doses. The estimated lethal times for essential oils to cause mortality in 50% of the population of C. includens were less than 15 h. There was selectivity of the essential oil of CGR126 accession of C. grewioides at lethal doses above 90%. Although the treatments showed little effect on the development of P. nigrispinus, body mass and reproductive parameters were negatively affected, with the exception of the essential oil of CGR126 accession of C. grewioides. The essential oil of C. grewioides may be a promising active ingredient for the synthesis of new insecticides, which are efficient against C. includens and at the same time are safer for the natural enemy P. nigrispinus.

Diversity and composition of the microbiome associated with eggs of the Southern green stinkbug, Nezara viridula (Hemiptera: Pentatomidae).

Although microbial communities of insects from larval to adult stage have been increasingly investigated in recent years, little is still known about the diversity and composition of egg-associated microbiomes. In this study, we used high-throughput amplicon sequencing and quantitative PCR to get a better understanding of the microbiome of insect eggs and how they are established using the Southern green stinkbug Nezara viridula (L.) (Hemiptera: Pentatomidae) as a study object. First, to determine the bacterial community composition, egg masses from two natural populations in Belgium and Italy were examined. Subsequently, microbial community establishment was assessed by studying stinkbug eggs of different ages obtained from laboratory strains (unlaid eggs collected from the ovaries, eggs less than 24 h old, and eggs collected 4 days after oviposition). Both the external and internal egg-associated microbiomes were analyzed by investigating egg washes and surface-sterilized washed eggs, respectively. Eggs from the ovaries were completely devoid of bacteria, indicating that egg-associated bacteria were deposited on the eggs during or after oviposition. The bacterial diversity of deposited eggs was very low, with on average 6.1 zero-radius operational taxonomic units (zOTUs) in the external microbiome and 1.2 zOTUs in internal samples of egg masses collected from the field. Bacterial community composition and density did not change significantly over time, suggesting limited bacterial growth. A Pantoea-like symbiont previously found in the midgut of N. viridula was found in every sample and generally occurred at high relative and absolute densities, especially in the internal egg samples. Additionally, some eggs harbored a Sodalis symbiont, which has previously been found in the abdomen of several insects, but so far not in N. viridula populations. We conclude that the egg-associated bacterial microbiome of N. viridula is species-poor and dominated by a few symbionts, particularly the species-specific obligate Pantoea-like symbiont.

Spiroplasma as facultative bacterial symbionts of stinkbugs.

Many insects are associated with facultative symbiotic bacteria, and their infection prevalence provides an important clue to understand the biological impact of such microbial associates. Here we surveyed diverse stinkbugs representing 13 families, 69 genera, 97 species and 468 individuals for Spiroplasma infection. Diagnostic PCR detection revealed that 4 families (30.8%), 7 genera (10.1%), 11 species (11.3%) and 21 individuals (4.5%) were Spiroplasma positive. All the 21 stinkbug samples with Spiroplasma infection were subjected to PCR amplification and sequencing of Spiroplasma's 16S rRNA gene. Molecular phylogenetic analysis uncovered that the stinkbug-associated Spiroplasma symbionts were placed in three distinct clades in the Spiroplasmataceae, highlighting multiple evolutionary origins of the stinkbug-Spiroplasma associations. The Spiroplasma phylogeny did not reflect the host stinkbug phylogeny, indicating the absence of host-symbiont co-speciation. On the other hand, the Spiroplasma symbionts associated with the same stinkbug family tended to be related to each other, suggesting the possibility of certain levels of host-symbiont specificity and/or ecological symbiont sharing. Amplicon sequencing analysis targeting bacterial 16S rRNA gene, FISH visualization of the symbiotic bacteria, and rearing experiments of the host stinkbugs uncovered that the Spiroplasma symbionts are generally much less abundant in comparison with the primary gut symbiotic bacteria, localized to various tissues and organs at relatively low densities, and vertically transmitted to the offspring. On the basis of these results, we conclude that the Spiroplasma symbionts are, in general, facultative bacterial associates of low infection prevalence that are not essential but rather commensalistic for the host stinkbugs, like the Spiroplasma symbionts of fruit flies and aphids, although their impact on the host phenotypes should be evaluated in future studies.

Comparative analysis and phylogeny of mitochondrial genomes of Pentatomidae (Hemiptera: Pentatomoidea).

The Phyllocephalini is a group of herbivorous insects in Pentatomidae, which lack distinctive morphological characteristics and systematic studies. Up to now, there are only two complete mitochondrial genomes of Phyllocephalini have been reported. In this study, we sequenced and analyzed the complete mitochondrial genomes of three Phyllocephalini species, Gonopsis coccinea, Gonopsimorpha nigrosignata, and Chalcopis glandulosus, which were 16,534, 16,531, and 16,534 bp in length, respectively. The mitochondrial genomes contained 37 genes, including 13 protein-coding genes, two rRNA genes, 22 tRNA genes, and a control region. The gene arrangement was consistent with that of the putative ancestral insect, with no rearrangement. The cox1 gene of Pentatomidae showed the lowest evolutionary rate among the protein-coding genes, the mean genetic distance of species, genera, and subfamilies of Pentatomidae increased hierarchically based on cox1 gene. The 16S rRNA of Pentatomidae was more conserved than 12S rRNA in sequence and secondary structure. All tRNAs could be folded into a typical cloverleaf structure except trnS1. The stem region was more conserved than the loop region in the secondary structure of tRNAs within Pentatomidae. Gonopsis coccinea and Gonopsimorpha nigrosignata had one type of tandem repetition unit in the control region, while C. glandulosus had two types. The heterogeneity analysis of Pentatomidae showed that Phyllocephalinae was the most heterogeneous. Phylogenetic trees based on the newly obtain mitochondrial genomes along with other 50 mitochondrial genomes of Pentatomidae using Bayesian Inference and Maximum Likelihood strongly supported the following three relationships: (((Anaxilaus + (Plautia + Glaucias)) + (Nezara + Palomena)) + (Eysarcorini + Carpocorini)), (Hoplistoderini + (Menidini + Asopinae)), and ((Sephelini + Halyini) + (Caystrini + (Cappaeini + (Placosternum + Phyllocephalini)))). The relationships within Phyllocephalini were (Chalcopis + (Dalsira + (Gonopsimorpha + Gonopsis))). Our results provide valuable molecular data for further phylogenetic analyses of Pentatomidae.

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.

Diversity and Molecular Barcoding of Stink Bugs (Hemiptera: Pentatomidae) Associated with Macadamia in South Africa.

Stink bugs are major pests of macadamia in South Africa. Accurate identification and knowledge of species composition are important to inform management practices. The overall aims of this study were to identify stink bug species from macadamia orchards in South Africa using morphology, and to establish a DNA database based on the cytochrome c oxidase subunit 1 gene region. A total of 21 stink bug species were found in macadamia orchards in KwaZulu-Natal, Limpopo and Mpumalanga provinces. Bathycoelia distincta Distant, 1878, was the dominant species throughout all three growing regions. Two unidentified species of Boerias Kirkaldy, 1909, here designated as Boerias sp. 1 and Boerias sp. 2, were the second and third most abundant species found in KwaZulu-Natal. No species of Boerias has previously been reported in association with macadamia. Evidence of a cryptic third species of Boerias was also found. Species composition fluctuated over three growing seasons in Limpopo and differed between the three growing regions during the 2019-2020 season, highlighting the need for ongoing monitoring of these important pest species. The DNA barcode database developed in this study will be valuable for future monitoring and identifications, including cryptic or polymorphic stink bug species and different life stages.

Determination of the Absolute Configuration of the Male-Produced Sex Pheromone of the Stink Bug Pellaea stictica, (2R,4R,8R)-2,4,8,13-Tetramethyltetradecan-1-ol by Stereoselective Synthesis Coupled with Enantiomeric Resolution.

In a previous study, we reported the identification and synthesis of a male-specific sex pheromone component of the stink bug, Pellaea stictica, as the alcohol 2,4,8,13-tetramethyltetradecan-1-ol (1). To establish the correlation between the stereochemistry of the pheromone and its bioactivity, it first was necessary to determine its absolute configuration. For this purpose, a series of syntheses were designed to: (a) furnish a mixture of all possible stereoisomers; (b) a narrowed down group of diastereomers, and (c) one specific enantiomer. A crucial step in the syntheses involved a coupling reaction between two key intermediates: a phosphonium salt and an aldehyde, through a Wittig olefination. Nuclear magnetic resonance data of a mixture of the synthetic pheromone diastereomers and further comparison of GC retention times with that of the natural product by gas chromatography suggested that the methyl branches at C2 and C4 were in a syn relationship, reducing the possibilities to only four of the eight possible stereoisomers. Employing GC analysis, chiral derivatization reagents and synthetic (8R)-2,4-syn-1 it was possible to confirm the configuration of the methyl branch at C8 as R, reducing the number of possible stereoisomers to two. After enantioselective synthesis of (2R,4R,8R)-1, the absolute configurations of all methyl branches of the natural compound were confirmed as R, fully identifying the male-produced sex pheromone of P. stictica as (2R,4R,8R)-2,4,8,13-tetramethyltetradecan-1-ol.