Typical Insect Research Articles

A tiny sample rapid visual detection technology for imidacloprid resistance in Aphis gossypii by CRISPR/Cas12a

Insecticide resistance monitoring is essential for guiding chemical pest control and resistance management policies. Currently, rapid and effective technology for monitoring the resistance of tiny insects in the field is absent. Aphis gossypii Glover is a typical tiny insect, and one of the most frequently reported insecticide-resistant pests. In this study, we established a novel CRISPR/Cas12a-based rapid visual detection approach for detecting the V62I and R81T mutations in the β1 subunit of the nAChR in A. gossypii, to reflect target-site resistance to imidacloprid. Based on the nAChR β1 subunit gene in A. gossypii, the V62I/R81T-specific RPA primers and crRNAs were designed, and the ratio of 10 μM/2 μM/10 μM for ssDNA/Cas12a/crRNA was selected as the optimal dosage for the CRISPR reaction, ensuring that Cas12a only accurately recognizes imidacloprid-resistance templates. Our data show that the field populations of resistant insects possessing V62I and R81T mutations to imidacloprid can be accurately identified within one hour using the RPA-CRISPR/Cas12a detection approach under visible blue light at 440–460 nm. The protocol for RPA-CRISPR detection necessitates a single less than 2 mm specimen of A. gossypii tissues to perform RPA-CRISPR detection, and the process only requires a container at 37 °C and a portable blue light at 440–460 nm. Our research represents the first application of RPA-CRISPR technology in insecticide resistance detection, offers a new method for the resistance monitoring of A. gossypii or other tiny insects, helps delay the development of resistance to imidacloprid, improves the sustainability of chemical control, and provides theoretical guidance for managing pest resistance.

Identification and expression analysis of four sHsp genes in Sogatella furcifera under abiotic stress

Sogatella furcifera is a typical r-strategist insect with strong adaptability and one of the major pests in rice cultivation. Small heat shock proteins (sHsp) play a vital role in insects responding to external abiotic stress. However, little is known about the role of sHsp in the defense against abiotic stress in S. furcifera. In this study, four sHsp genes (SofHsp20.7, SofHsp21.6, SofHsp21.7, and SofHsp21.8) were identified from S. furcifera, and their gene expression patterns were analyzed at different developmental stages, temperature stress, insecticide exposure, and UV-A irradiation. The results indicated that four sHsp were expressed at various developmental stages of the S. furcifera, with the highest transcription levels in the first-instar nymphs, fourth day fifth instar nymphs, third-day adult females, and eggs, respectively. High-temperature stress (30 °C and 40 °C) significantly induced the expression of all four sHsp genes, while low-temperature stress (10 °C) reduced their expression. Sublethal concentrations (LC10 and LC25) of buprofezin increased the expression levels of SofHsp20.7 and SofHsp21.8. Under UV-A irradiation, the S. furcifera also significantly induced the expression of all four sHsp genes, with the highest expression observed at 90 min after exposure to the stress. These results suggest that sHsp genes are essential for the resistance or tolerance of S. furcifera to high temperature, buprofezin, and UV-A irradiation. This research provides a foundation for understanding the molecular mechanisms underlying the S. furcifera adaptation to abiotic stress.

A chromosome-level haplotype-resolved genome assembly of oriental tobacco budworm (Helicoverpa assulta)

Oriental tobacco budworm (Helicoverpa assulta) and cotton bollworm (Helicoverpa armigera) are two closely related species within the genus Helicoverpa. They have similar appearances and consistent damage patterns, often leading to confusion. However, the cotton bollworm is a typical polyphagous insect, while the oriental tobacco budworm belongs to the oligophagous insects. In this study, we used Nanopore, PacBio, and Illumina platforms to sequence the genome of H. assulta and used Hifiasm to create a haplotype-resolved draft genome. The Hi-C technique helped anchor 33 primary contigs to 32 chromosomes, including two sex chromosomes, Z and W. The final primary haploid genome assembly was approximately 415.19 Mb in length. BUSCO analysis revealed a high degree of completeness, with 99.0% gene coverage in this genome assembly. The repeat sequences constituted 38.39% of the genome assembly, and we annotated 17093 protein-coding genes. The high-quality genome assembly of the oriental tobacco budworm serves as a valuable genetic resource that enhances our comprehension of how they select hosts in a complex odour environment. It will also aid in developing an effective control policy.

Response of the serine/threonine kinase AKT and phosphoinositide-dependent kinase PDK in Frankliniella occidentalis (Thysanoptera: Thripidae) to three kinds of foods and their regulation of reproductive function.

Frankliniella occidentalis (Pergande) is a typical omnivorous insect that feeds on host plants, pollens and mite eggs, and poses a threat to crops worldwide. The insulin signalling pathway (ISP) is a typical nutrient-sensitive pathway that participates in the regulation of various functions in insects. Serine/threonine kinases (AKTs) and phosphoinositide-dependent kinases (PDKs) are key components of the ISP. In this study, the FoAKT and FoPDK genes in F. occidentalis were cloned, and the effects of three foods on their expression were determined. The expression of FoAKT and FoPDK in the thrips fed on kidney bean leaves supplemented with pine pollen or mite eggs was higher than in those primarily fed on leaves alone. Meanwhile, the fecundity of thrips fed on leaves supplemented with pine pollen was highest. In addition, RNA interference-mediated knockdown of FoAKT and FoPDK decreased vitellogenin (Vg) content and Vg expression in females, shortened ovariole length, delayed egg development and reduced fecundity and offspring hatching rates. Furthermore, the synthesis of juvenile hormone (JH) was reduced, and the contents of glucose, trehalose, glycogen and trehalase were affected. These results suggest that FoAKT and FoPDK regulate the reproduction of F. occidentalis by regulating Vg and JH production as well as carbohydrate metabolism.

A comprehensive sampling of mitogenomes shows the utility to infer phylogeny of termites (Blattodea: Termitoidae).

The mitogenome sequence data have been widely used in inferring the phylogeny of insects. In this study, we determined the complete mitogenome for Macrotermes sp. (Termitidae, Macrotermitinae) using next-generation sequencing. Macrotermes sp. possesses a typical insect mitogenome, displaying an identical gene order and gene content to other existing termite mitogenomes. We present the first prediction of the secondary structure of ribosomal RNA genes in termites. The rRNA secondary structures of Macrotermes sp. exhibit similarities to closely related insects and also feature distinctive characteristics in their helical structures. Together with 321 published mitogenomes of termites as ingroups and 8 cockroach mitogenomes as outgroups, we compiled the most comprehensive mitogenome sequence matrix for Termitoidae to date. Phylogenetic analyses were conducted using datasets employing different data coding strategies and various inference methods. Robust relationships were recovered at the family or subfamily level, demonstrating the utility of comprehensive mitogenome sampling in resolving termite phylogenies. The results supported the monophyly of Termitoidae, and consistent relationships within this group were observed across different analyses. Mastotermitidae was consistently recovered as the sister group to all other termite families. The families Hodotermitidae, Stolotermitidae, and Archotermopsidae formed the second diverging clade, followed by the Kalotermitidae. The Neoisoptera was consistently supported with strong node support, with Stylotermitidae being sister to the remaining families. Rhinotermitidae was found to be non-monophyletic, and Serritermitidae nested within the basal clades of Rhinotermitidae and was sister to Psammotermitinae. Overall, our phylogenetic results are largely consistent with earlier mitogenome studies.

Per- and polyfluoroalkyl substance (PFAS) mixtures induce gut microbiota dysbiosis and metabolic disruption in silkworm (Bombyx mori L.)

Mixed legacy and emerging per- and polyfluoroalkyl substances (PFASs) are commonly found in soil and dust; however, the potential toxicity of PFAS mixtures (mPFASs) in insects is unknown. Using 16S rRNA gene sequencing and transcriptome sequencing (RNA-Seq), we evaluated the adverse effects of mPFASs on silkworms, a typical lepidopteran insect. After exposure to mPFASs, the silkworm midgut was enriched with high levels of PFASs, which induced histopathological changes. The composition of the midgut microbiota was significantly affected by mPFAS exposure, and functional predictions revealed significant disruption of some metabolic pathways. RNA-seq analysis revealed that mPFASs significantly changed the transcription profiles. Functional enrichment analysis of the differentially expressed genes also revealed that biological processes related to metabolic pathways and the digestive system were significantly affected, similar to the results of the gut microbiota analysis, suggesting that mPFAS exposure had an adverse effect on the metabolic function of silkworms and may further affect their normal growth. Finally, the significant correlation between abundance changes in the gut microbiota and metabolism/digestion-related genes further highlighted the role of the gut microbiota in mPFAS-related processes affecting the metabolic functions of silkworms. To our knowledge, this study is the first to evaluate the toxic effects of mPFASs in insects and provide basic data for further PFAS toxicity investigations in insects and comprehensive ecological risk assessments of mPFASs.

Molecular identification and functional analysis of chitinase genes reveal their importance in the metamorphosis of Sarcophaga peregrina (Diptera: Sarcophagidae).

Chitinases play a crucial role in insect metamorphosis by facilitating chitin degradation. Sarcophaga peregrina (Robineau-Desvoidy, 1830) (Diptera: Sarcophagidae) is a typical holometabolous insect and an important hygiene pest that causes myiasis in humans and other mammals and acts as a vector for various parasitic agents, including bacteria, viruses, and parasites. Enhancing the understanding of the metamorphosis in this species has significance for vector control. In this study, we identified a total of 12 chitinase genes in S. peregrina using bioinformatic analysis methods. Based on transcriptome data, SpIDGF2 and SpCht10 were selected for further functional investigation. The down-regulation of these genes by RNA interference led to developmental delays, disruptions in molting, and differences in cuticle composition during the pupal stage. These findings underscore the pivotal role of chitinase genes in the metamorphic process and offer valuable insights for effective control strategies.

Microbiome of Zoophytophagous Biological Control Agent Nesidiocoris tenuis.

Many insects are associated with endosymbionts that influence the feeding, reproduction, and distribution of their hosts. Although the small green mirid, Nesidiocoris tenuis (Reuter) (Hemiptera: Miridae), a zoophytophagous predator that feeds on plants as well as arthropods, is a globally important biological control agent, its microbiome has not been sufficiently studied. In the present study, we assessed the microbiome variation in 96 N. tenuis individuals from 14 locations throughout Japan, based on amplicon sequencing of the 16S ribosomal RNA gene. Nine major bacteria associated with N. tenuis were identified: Rickettsia, two strains of Wolbachia, Spiroplasma, Providencia, Serratia, Pseudochrobactrum, Lactococcus, and Stenotrophomonas. Additionally, a diagnostic PCR analysis for three typical insect reproductive manipulators, Rickettsia, Wolbachia, and Spiroplasma, was performed on a larger sample size (n = 360) of N. tenuis individuals; the most prevalent symbiont was Rickettsia (69.7%), followed by Wolbachia (39.2%) and Spiroplasma (6.1%). Although some symbionts were co-infected, their prevalence did not exhibit any specific tendency, such as a high frequency in specific infection combinations. The infection frequency of Rickettsia was significantly correlated with latitude and temperature, while that of Wolbachia and Spiroplasma was significantly correlated with host plants. The predominance of these bacteria and the absence of obligate symbionts suggested that the N. tenuis microbiome is typical for predatory arthropods rather than sap-feeding insects. Rickettsia and Wolbachia were vertically transmitted rather than horizontally transmitted from the prey. The functional validation of each symbiont would be warranted to develop N. tenuis as a biological control agent.

Different Tea Germplasms Distinctly Influence the Adaptability of Toxoptera aurantii (Hemiptera: Aphididae).

Aphids are typical phloem-sucking insect pests. A good understanding regarding their feeding behavior and population dynamics are critical for evaluating host adaptation and screening of aphid-resistant resources. Herein, the adaptability of Toxoptera aurantii (Boyer) (Hemiptera: Aphididae) to different hosts was evaluated via electropenetrography and an age-stage, two-sex life table on six tea germplasms: Zikui (ZK), Zhongcha108 (ZC108), Zhongcha111 (ZC111), Qianmei419 (QM419), Meitan5 (MT5), and Fudingdabaicha (FD). Our findings revealed that the feeding activities of T. aurantii differed considerably among the host plants. T. aurantii exhibited significantly more pathway activities on ZK and FD than on the other hosts. However, the duration of feeding of T. aurantii on ZK phloem considerably decreased compared with those of the other germplasms. Life parameters indicated that T. aurantii exhibited the highest intrinsic rate of increase (r), net reproductive rate (R0), and finite rate of increase (λ) on MT5, and the maximum values of total longevity and oviposition period were recorded on FD; these variables were reduced significantly on ZK. The results of our study demonstrate that T. aurantii can successfully survive on the six tea germplasms; however, ZK was less suitable for T. aurantii and should be considered as a potential source of resistance in breeding and Integrated Pest Management.

Microrheology of haemolymph plasma of the bumblebee Bombus terrestris.

Viscosity, which impacts the rate of haemolymph circulation and heat transfer, is one of the transport properties that affects the performance of an insect. Measuring the viscosity of insect fluids is challenging because of the small amount available per specimen. Using particle tracking microrheology, which is well suited to characterise the rheology of the fluid part of the haemolymph, we studied the plasma viscosity in the bumblebee Bombus terrestris. In a sealed geometry, the viscosity exhibits an Arrhenius dependence with temperature, with an activation energy comparable to that previously estimated in hornworm larvae. In an open to air geometry, it increases by 4-5 orders of magnitude during evaporation. Evaporation times are temperature dependent and longer than typical insect haemolymph coagulation times. Unlike standard bulk rheology, microrheology can be applied to even smaller insects, paving the way to characterise biological fluids such as pheromones, pad secretions or cuticular layers.

Ingestion and egestion of microplastic by aquatic insects in Egypt wastewater

AbstractMicroplastics (MPs) are frequently regarded as environmental and biota contaminants. Yet, research on the accumulation of MPs in living entities, particularly aquatic insects that serve as food resources in the aquatic food chain, is limited. This study to investigate the accumulation of MPs in aquatic insects from water and sediment in an Egyptian wastewater basin. Four typical freshwater insect groups were used. The highest MP load per gram wet weight was reported by collector‐gatherers (Chironomus sp. and Hydrophilus sp.), followed by collector‐filterers (Culex sp.) with the second highest MP load. However, Predators (Aeshna sp.) had the lowest values. Also, the present results showed a reduction in the number of MPs in all insect taxa tested after a 24 h depuration time, with differences in the observed egestion ability. The mean number of MPs per individual significantly reduced after 24 h in both Chironomus sp. and Culex sp. larvae, where 53% and 40% of MPs particles were ejected from them, respectively. However, the ability of MP egestion decreases in Aeshna sp. nymph (25%), and the lowest proportion of ejection was observed in Hydrophilus sp. adults (9%). Polyethylene terephthalate fibers were the most abundant type of MP in both sediment and water, followed by fragments (polyethylene and polypropylene). Yet, only polyester fibers were detected in the various insect species. The average length of fibers in the various insects was somewhat shorter than in the surrounding environment. The current study reveals that MP ingestion by aquatic insects is not always related to levels of pollution in the environment, since other factors such as feeding strategies may play a role in MP ingestion. Based on these observations, further studies should be carried out on studies on toxicological impacts of MPs on freshwater/aquatic biota.

Phylogeography of alpine and subalpine adapted Pseudostenophylax caddisflies (Limnephilidae: Trichoptera): a strong relationship with mountain formation

Abstract Organisms that inhabit the alpine zone tend to have distribution areas that are isolated and scattered in patches, in effect ‘sky islands’. We focused on Pseudostenophylax caddisflies, a typical alpine aquatic insect group. Phylogenetic analyses were conducted on all eight Japanese species, with the monophyletic lineage of each strongly supported. The traditional classification system dividing Pseudostenophylax into ‘ondakensis’ and ‘adlimitans’ species groups was also strongly supported. For the first time, the existence of a cryptic species in Hokkaido was revealed in this study. Two species of the adlimitans species group live in western Honshu, Shikoku and Kyushu, but those distribution areas are difficult to explain in terms of the current geography. The phylogenetic analyses results also closely reflected the related geological history and palaeogeography. Interestingly, Pseudostenophylax amurensis, recently recorded on the Korean Peninsula, was positioned within the Japanese lineage of the ondakensis species group. Thus, some Japanese lineages may have reversely dispersed to the Korean Peninsula. Many interesting insights can be gained from the close relationship between the genetic structure of aquatic insects adapted to alpine and subalpine regions and their palaeogeographical history.

Biomonitoring of DEET and DCBA in Canadian children following typical protective insect repellent use

N,N-diethyl-m-toluamide (DEET) is an ingredient found in many consumer insect repellents and its use is recommended to Canadians by government agencies, including Health Canada, for protection against insect bites including mosquitos and ticks. The majority of research on DEET exposure and toxicokinetics in humans has focused on adult populations with little information from vulnerable populations, including children. We aimed to fill this knowledge gap by examining real-world exposure data for DEET and its metabolite 3-diethylcarbamoyl benzoic acid (DCBA) in a sample population of Canadian children. We conducted a 24-h observational exposure human biomonitoring study at three overnight summer camps in Ontario, Canada through July and August 2019. Participating children aged 7-13 years provided multiple spot urine samples over a 24-h period and completed a journal to document insect repellent use and factors that could influence absorption of DEET. Children were instructed to use insect repellent as they usually would while attending a summer camp. Exposure was quantified using the information from the participant's journal and the change in the mass of their insect repellent containers over the course of the study. A total of 389 urine samples were collected from 124 children. Among participants using insect repellent, urinary levels of DEET were elevated between 2 and 8h post-application and decreased thereafter but remained qualitatively higher than concentrations in participants who did not use insect repellent on the study day, even at 18-22h post-application. DCBA was the predominant metabolite of DEET exposure in urine. DCBA was elevated between 8 and 14h post-application, and declined thereafter, but not to the level observed among those who did not use insect repellent on the study day. Children who used more insect repellent, or used higher concentration insect repellent (10%-30% DEET) excreted higher levels of DEET and DCBA. Excreted DEET and DCBA accounted for 0.001% (median) and 1.3% (median) of the estimated applied DEET, respectively. Children did not reach an undetectable level of DEET or DCBA in urine, even among those not using insect repellent during the study day, indicating a potentially complex multi-route exposure to insect repellents in a real world scenario. This work provides targeted biomonitoring data for children intentionally using DEET-based insect repellents for normal protective use, and will support the risk re-evaluation of DEET by Health Canada.

Ab-GALFA, A bioassay for insect gall formation using the model plant Arabidopsis thaliana

Insect galls are abnormal plant organs formed by gall-inducing insects to provide shelter and nutrients for themselves. Although insect galls are spatialized complex structures with unique shapes and functions, the molecular mechanism of the gall formation and the screening system for the gall inducing effectors remains unknown. Here, we demonstrate that an extract of a gall-inducing aphid, Schlechtendalia chinensis, induces an abnormal structure in the root-tip region of Arabidopsis seedlings. The abnormal structure is composed of stem-like cells, vascular, and protective tissues, as observed in typical insect galls. Furthermore, we confirm similarities in the gene expression profiles between the aphid-treated seedlings and the early developmental stages of Rhus javanica galls formed by S. chinensis. Based on the results, we propose a model system for analyzing the molecular mechanisms of gall formation: the Arabidopsis-based Gall-Forming Assay (Ab-GALFA). Ab-GALFA could be used not only as a model to elucidate the mechanisms underlying gall formation, but also as a bioassay system to isolate insect effector molecules of gall-induction.

The first complete mitochondrial genome sequences for Ulidiidae and phylogenetic analysis of Diptera.

Tetanops sintenisi is a pest that mainly damages the root of quinoa (Chenopodium quinoa) and it is first discovered in China in 2018. Here, the complete mitochondrial genome (mitogenome) of T. sintenisi was sequenced and compared with the mitogenomes of other Diptera species. The results revealed that the mitogenome of T. sintenisi is 15,763bp in length (GenBank accession number: MT795181) and is comprised of 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA) genes, 2 ribosomal RNA genes, and a non-coding A + T-rich region (959bp). The highly conserved gene arrangement of the mitogenome of T. sintenisi was identical to that of other Diptera insects. Twelve PCGs contained the typical insect start codon ATN, while cox1 had CGA as the start codon. The genes cox2, nad4, and nad1 contained an incomplete termination codon T; nad3, nad5, and cob contained the complete termination codon TAG; and the remaining seven PCGs contained the termination codon TAA. All tRNA genes were predicted to fold into the typical cloverleaf secondary structure. Phylogenetic analysis of 48 species based on the mitogenome sequence revealed that T. sintenisi clustered with the Tephritidae family, indicating that T. sintenisi and Tephritidae have a close phylogenetic relationship. The phylogenetic relationship of T. sintenisi based on the mitogenome is consistent with the traditional morphological taxonomy, according to which T. sintenisi belongs to the family Otitidae, which is closely related to the family Muscidae.

Host plant-mediated effects on Buchnera symbiont: implications for biological characteristics and nutritional metabolism of pea aphids (Acyrthosiphon pisum).

The pea aphid, Acyrthosiphon pisum, is a typical sap-feeding insect and an important worldwide pest. There is a primary symbiont-Buchnera aphidicola, which can synthesize and provide some essential nutrients for its host. At the same time, the hosts also can actively adjust the density of bacterial symbiosis to cope with the changes in environmental and physiological factors. However, it is still unclear how symbionts mediate the interaction between herbivorous insects' nutrient metabolism and host plants. The current study has studied the effects of different host plants on the biological characteristics, Buchnera titer, and nutritional metabolism of pea aphids. This study investigated the influence of different host plants on biological characteristics, Buchnera titer, and nutritional metabolism of pea aphids. The titer of Buchnera was significantly higher on T. Pretense and M. officinalis, and the relative expression levels were 1.966±0.104 and 1.621±0.167, respectively. The content of soluble sugar (53.46±1.97µg/mg), glycogen (1.12±0.07µg/mg) and total energy (1341.51±39.37µg/mg) of the pea aphid on V. faba were significantly higher and showed high fecundity (143.86±11.31) and weight (10.46±0.77µg/mg). The content of total lipids was higher on P. sativum and T. pretense, which were 2.82±0.03µg/mg and 2.92±0.07µg/mg, respectively. Correlation analysis found that the difference in Buchnera titer was positively correlated with the protein content in M. officinalis and the content of total energy in T. pratense (P < 0.05). This study confirmed that host plants not only affected the biological characteristics and nutritional metabolism of pea aphids but also regulated the symbiotic density, thus interfering with the nutritional function of Buchnera. The results can provide a theoretical basis for further studies on the influence of different host plants on the development of pea aphids and other insects.

Invertebrate herbivory accelerates shift towards forbs caused by warming in a sedge‐dominated alpine meadow

AbstractInvertebrate herbivory is suggested to modulate community shifts caused by climate warming in grasslands. However, experimental evidence is lacking, especially in high elevation areas. We examined the interactive effect of experimental warming and the grassland caterpillar Gynaephora alpherakjj, a typical invertebrate herbivorous insect with sedge preference, on the structure of plant community in a sedge‐dominated alpine meadow in the Tibetan Plateau. We manipulated both herbivore and temperature in a full factorial field experiment using passive warming chambers over one plant‐growing season. Results show that simulated warming significantly increased the aboveground plant biomass (APB) of forbs and total APB, while the presence of caterpillars significantly decreased sedge APB and total APB. The increase in consumption of sedges by caterpillars under warmed conditions can largely be ascribed to the increase in feeding time of caterpillars. Importantly, the presence of caterpillars significantly decreased the APB ratio of sedges to forbs in the warmed but not in the unwarmed chambers. These results suggest that with a warming climate, invertebrate herbivory could accelerate community shifts from sedge dominance to forb dominance in only one plant‐growing season in the Tibetan alpine meadow. This shift would reduce the biomass of high‐quality forage and further threaten animal husbandry in this area. Considering the harm of G. alpherakjj to grasslands, its population should be controlled more strictly under the global warming scenario.

Increasing the activities of protective enzymes is an important strategy to improve resistance in cucumber to powdery mildew disease and melon aphid under different infection/infestation patterns.

Powdery mildew, caused by Sphaerotheca fuliginea (Schlecht.) Poll., and melon aphids (Aphis gossypii Glover) are a typical disease and insect pest, respectively, that affect cucumber production. Powdery mildew and melon aphid often occur together in greenhouse production, resulting in a reduction in cucumber yield. At present there are no reports on the physiological and biochemical effects of the combined disease and pest infection/infestation on cucumber. This study explored how cucumbers can regulate photosynthesis, protective enzyme activity, and basic metabolism to resist the fungal disease and aphids. After powdery mildew infection, the chlorophyll and free proline contents in cucumber leaves decreased, while the activities of POD (peroxidase) and SOD (superoxide dismutase) and the soluble protein and MDA (malondialdehyde) contents increased. Cucumber plants resist aphid attack by increasing the rates of photosynthesis and basal metabolism, and also by increasing the activities of protective enzymes. The combination of powdery mildew infection and aphid infestation reduced photosynthesis and basal metabolism in cucumber plants, although the activities of several protective enzymes increased. Aphid attack after powdery mildew infection or powdery mildew infection after aphid attack had the opposite effect on photosynthesis, protective enzyme activity, and basal metabolism regulation. Azoxystrobin and imidacloprid increased the contents of chlorophyll, free proline, and soluble protein, increased SOD activity, and decreased the MDA content in cucumber leaves. However, these compounds had the opposite effect on the soluble sugar content and POD and CAT (catalase) activities. The mixed ratio of the two single agents could improve the resistance of cucumber to the combined infection of powdery mildew and aphids. These results show that cucumber can enhance its pest/pathogen resistance by changing physiological metabolism when exposed to a complex infection system of pathogenic microorganisms and insect pests.

Investigation on phytoplasma diseases, their potential insect vectors and other hosts in pepper (Capsicum annuum L.) growing areas of Hatay-Turkey

Aims: This study was conducted to determine the phytoplasma diseases of pepper plants in Hatay province between 2016 and 2019.Methods and Results: Samples of pepper (Capsicum annuum), sesame (Sesamum indicum), basil (Ocimum basilicum), tomato (Solanum lycopersicum) and bindweed (Convolvulus arvensis) which had typical phytoplasma symptoms and insect (Cicadellidae spp.) samples were collected from the pepper fields. Empoasca sp., Cicadulina bipunctata, Psammotettix sp., Balchutha hebe, Euscelidius sp., Anaceratagallia laevis and Exitianus capicola were found as potential insect vectors of the phytoplasmas as descending order of population densities. T-budding grafting technique was successful for phytoplasma transmission from infected sesame and basil plants to healthy periwinkle (Catharanthus roseus) plants in controlled conditions. Phytoplasmas were detected in pepper, tomato, basil, sesame and bindweed and from some insect samples. Total nucleic acid isolation was accomplished by the CTAB method. Direct and Nested PCR were used employing R16F1/R16R0 and R16F2n/R16R2 primer pairs respectively.Conclusions: DNA of positive samples were sequenced, and uploaded to Genbank, and were identified as Ca. Phytoplasma trifolii on pepper (MT993358), sesame (MT994434), tomato (MT992754), basil (MT994432), Empoasca sp. (MT994430), Exitianus capicola (MT994433), Euscelidius sp. (MT994431); and as Ca. Phytoplasma solani on binweed (MT993422) and tomato (MT992796).Significance and Impact of the Study: According to our knowledge, this study is the first to identify and upload to Genbank of Ca. Phytoplasma trifolii on basil and Empoasca sp., Exitianus capicola and Euscelidius sp. as insect vectors of the diseases in Turkey.

The complete mitochondrial genome of Crematogaster matsumurai (Forel 1901) (Hymenoptera: Formicidae) and its phylogenetic relationship

Crematogaster matsumurai (Forel 1901) is an important arboreal ant species commonly found on Phyllostachys heterocycla (Carr.) in Lishui, Zhejiang, China. This study analyzed the mitochondrial genome sequence of C. matsumurai and discussed its phylogenetic relationship in Hymenoptera. The circular mitochondrial genome was 16,028 bp long, including a standard set of 22 transfer RNAs (tRNAs), two ribosomal RNAs (rRNAs), and 13 protein-coding genes (PCGs), which showed the typical insect mitochondrial genome arrangement. The AT and GC contents of the mitochondrial genome sequence were 76.92% and 23.08%, respectively. The maximum-likelihood (ML) phylogenetic analysis based on whole mitochondrial genome sequences showed that C. matsumurai is closest to Crematogaster teranishii.