Functional types of long trichoid sensilla responding to sex pheromone components in Plutella xylostella.

Sex pheromones, which consist of multiple components in specific ratios promote intraspecific sexual communications of insects. Plutella xylostella (L.) is a worldwide pest of cruciferous vegetables, the mating behavior of which is highly dependent on its olfactory system. Long trichoid sensilla on male antennae are the main olfactory sensilla that can sense sex pheromones. However, the underlying mechanisms remain unclear. In this study, 3 sex pheromone components from sex pheromone gland secretions of P. xylostella female adults were identified as Z11-16:Ald, Z11-16:Ac, and Z11-16:OH in a ratio of 9.4 : 100 : 17 using gas chromatography - mass spectrometry and gas chromatography with electroantennographic detection. Electrophysiological responses of 581 and 385 long trichoid sensilla of male adults and female adults, respectively, to the 3 components were measured by single sensillum recording. Hierarchical clustering analysis showed that the long trichoid sensilla were of 6 different types. In the male antennae, 52.32%, 5.51%, and 1.89% of the sensilla responded to Z11-16:Ald, Z11-16:Ac, and Z11-16:OH, which are named as A type, B type, and C type sensilla, respectively; 2.93% named as D type sensilla responded to both Z11-16:Ald and Z11-16:Ac, and 0.34% named as E type sensilla were sensitive to both Z11-16:Ald and Z11-16:OH. In the female antennae, only 7.53% of long trichoid sensilla responded to the sex pheromone components, A type sensilla were 3.64%, B type and C type sensilla were both 0.52%, D type sensilla were 1.30%, and 1.56% of the sensilla responded to all 3 components, which were named as F type sensilla. The responding long trichoid sensilla were located from the base to the terminal of the male antennae and from the base to the middle of the female antennae. The pheromone mixture (Z11-16:Ald : Z11-16:Ac : Z11-16:OH = 9.4 : 100 : 17) had a weakly repellent effect on female adults of P. xylostella. Our results lay the foundation for further studies on sex pheromone communications in P. xylostella.

Identification, expression profiles, and binding properties of chemosensory protein 18 in Plutella xylostella (Lepidoptera: Plutellidae).

Chemosensory proteins (CSPs) are highly efficient carry tools to bind and deliver hydrophobic compounds, which play an important role in the chemosensory process in insects. The diamondback moth, Plutella xylostella L. (Lepidoptera: Plutellidae), is a cosmopolitan pest that attacks cruciferous crops. However, the detailed physiological functions of CSPs in P. xylostella remain limited to date. Here, we identified a typical CSP, named PxylCSP18, in P. xylostella and investigated its expression patterns and binding properties of volatiles. PxylCSP18 was highly expressed in antennae and head (without antennae), and the expression level in the male antennae of P. xylostella was obviously higher than that in the female antennae. Moreover, PxylCSP18 has a relatively broad binding spectrum. Fluorescence competitive binding assays showed that PxylCSP18 had strong binding abilities with 14 plant volatiles (Ki < 10 µM) that were repellent or attractive to P. xylostella. Notably, PxylCSP18 had no significant binding affinity to (Z)-11-hexadecenal, (Z)-11-hexadecenyl acetate, and (Z)-11-hexadecenyl alcolol, which are the pheromone components of P. xylostella. The attractive effects of trans-2-hexen-1-ol and isopropyl isothiocyanate to male adults and the attractive effects of isopropyl isothiocyanate and the repellent effects of linalool to female adults were significantly decreased after knocked down the expression of PxylCSP18. Our results revealed that PxylCSP18 might play an important role in host plant detection, avoidance of unsuitable hosts, and selection of oviposition sites; however, it does not participate in mating behavior. Overall, these results extended our knowledge on the CSP-related functions, which provided insightful information about CSP-targeted insecticides.

Electrophysiological and Behavioral Responses of Plodia interpunctella (Hübner) Females to Aldehyde Volatiles from Dried Fruits.

Plodia interpunctella (Lepidoptera: Pyraloidea) is a notorious pest of stored grain globally. The dried fruits (Ziziphus jujuba, Malus pumila, and Fragaria ananassa) can strongly attract P. interpunctella. However, specific volatile compounds responsible for such effects have not been identified. Volatiles were analyzed by using headspace solid-phase microextraction (HS-SPME) and chromatography-mass spectrometry (GCMS) techniques. Five aldehyde compounds were selected for electroantennogram (EAG), single sensillum recording (SSR), and behavioral response assays. The three chemicals that elicited the strongest EAG responses to mated females at 100 µg/µL include hexanal (1.13 mV), heptanal (0.92 mV), and octanal (0.73 mV). In SSR experiments, the basiconic sensilla of the antennae responded to these aldehyde compounds. The results of behavioral responses showed that all aldehydes exhibited dose-dependent responses, with hexanal having the highest attractant rate of 74.56%. These compounds have the potential to be used for monitoring P. interpunctella and its integrated management program.

Identification and evaluation of cruciferous plant volatiles attractive to Plutella xylostella L. (Lepidoptera: Plutellidae).

BACKGROUND: The diamondback moth, Plutella xylostella, has developed resistance to almost all insecticides used for its control. The 'push-pull' method has been shown as an effective control strategy to address this resistance challenge of P. xylostella. The key focus of the strategy is the identification of attractive or repellent volatile components. The aim of this study was to identify attractive volatile compounds released from host plants. Identified compounds were applied in the biological control of this pest. RESULTS: Nine active compounds released into the headspace of seven cruciferous plant species were identified using gas chromatography-electroantennographic detection and gas chromatography-mass spectrometry. Electroantennographic detection-active compounds included five green leaf volatiles (hexanal, trans-2-hexen-1-ol, cis-3-hexen-1-ol, cis-3-hexenyl acetate, and 1-penten-3-ol), three isothiocyanates (isopropyl isothiocyanate, allyl isothiocyanate, and butyl isothiocyanate), and nonanal. Except for nonanal, all the identified green leaf volatiles and isothiocyanates elicited strong electrophysiological and behavioral responses in P. xylostella. The strongest attractive compounds, trans-2-hexen-1-ol and isopropyl isothiocyanate, were further evaluated in oviposition and field-trapping assays. Results showed that they both lured female moths to lay eggs, and were highly attractive to P. xylostella adults in field, especially when used in combination with yellow and green sticky boards. However, a blend of the two compounds showed no synergistic effect, but rather an antagonistic effect. CONCLUSIONS: Green leaf volatiles and isothiocyanates were identified as key olfactory cues for host selection of P. xylostella. Trans-2- hexen-1-ol and isopropyl isothiocyanate were identified as candidate attractive compounds to serve in a 'push-pull' strategy for P. xylostella control. © 2023 Society of Chemical Industry.

Behavioral, Electrophysiological, and Toxicological Responses of Plutella xylostella to Extracts from Angelica pubescens.

Plutella xylostella L. is a destructive pest affecting cruciferous vegetables, causing massive economic losses worldwide. Plant-based insecticides are considered promising insect control agents. The Angelica pubescens extract inhibited female oviposition, with an oviposition deterrence index (ODI) of 61.65% at 12.5 mg/mL. We aimed to identify the bioactive compounds in A. pubescens extract. The compounds from A. pubescens extract were analyzed using LC-MS techniques. The toxicity and behavioral responses of larvae and adults of P. xylostella to ten compounds were investigated. We found that the caryophyllene oxide and 3,4-dimethoxycinnamic acid inhibited female oviposition; the ODIs were 98.31% and 97.59% at 1.25 mg/mL, respectively. The A. pubescens extract, caryophyllene oxide, and 3,4-dimethoxycinnamic acid caused larval mortality, with LC50 values of 21.31, 4.56, and 5.52 mg/mL, respectively. The EAG response of females was higher than that of males under A. pubescens extract conditions, while the EAG response of males was higher than that of females in caryophyllene oxide and 3,4-dimethoxycinnamic acid conditions. The A. pubescens extract and caryophyllene oxide showed repellent activity against both female and male adults, while the 3,4-dimethoxycinnamic acid did not elicit any notable behavioral responses from P. xylostella adults. A. pubescens extract and caryophyllene oxide are potential insecticides, oviposition deterrents, and behavioral regulators against P. xylostella, and they could be potential candidates for the development of biological insecticides to control P. xylostella.

Nitric oxide synthase is required for the pea aphid's defence against bacterial infection.

Compared to other insects, the pea aphid Acyrthosiphon pisum has a reduced immune system with an absence of genes coding for a lot of immunity-related molecules. Notably, nitric oxide synthase (NOS), which catalyses the synthesis of nitric oxide (NO), is present in the pea aphid. However, the role of NO in the immune system of pea aphid remains unclear. In this study, we explored the role of NO in the defence of the pea aphid against bacterial infections and found that the NOS gene of the pea aphid responded to an immune challenge, with the expression of ApNOS observably upregulated after bacterial infections. Knockdown of ApNOS using RNA interference or inhibition of NOS activity increased the number of live bacterial cells in aphids and the mortality of aphids after bacterial infection. Conversely, the increase in NO level in aphids using NO donor inhibited the bacterial growth, increased the survival of bacteria-infected aphids, and upregulated immune genes, such as Toll pathway and phagocytosis related genes. Thus, NO promotes immune responses and plays an important role in the immune system of pea aphid.

Serpin-4 Negatively Regulates Prophenoloxidase Activation and Antimicrobial Peptide Synthesis in the Silkworm, Bombyx mori.

The prophenoloxidase (PPO) activation and Toll antimicrobial peptide synthesis pathways are two critical immune responses in the insect immune system. The activation of these pathways is mediated by the cascade of serine proteases, which is negatively regulated by serpins. In this study, we identified a typical serpin, BmSerpin-4, in silkworms, whose expression was dramatically up-regulated in the fat body and hemocytes after bacterial infections. The pre-injection of recombinant BmSerpin-4 remarkably decreased the antibacterial activity of the hemolymph and the expression of the antimicrobial peptides (AMPs) gloverin-3, cecropin-D, cecropin-E, and moricin in the fat body under Micrococcus luteus and Yersinia pseudotuberculosis serotype O: 3 (YP III) infection. Meanwhile, the inhibition of systemic melanization, PO activity, and PPO activation by BmSerpin-4 was also observed. Hemolymph proteinase 1 (HP1), serine protease 2 (SP2), HP6, and SP21 were predicted as the candidate target serine proteases for BmSerpin-4 through the analysis of residues adjacent to the scissile bond and comparisons of orthologous genes in Manduca sexta. This suggests that HP1, SP2, HP6, and SP21 might be essential in the activation of the serine protease cascade in both the Toll and PPO pathways in silkworms. Our study provided a comprehensive characterization of BmSerpin-4 and clues for the further dissection of silkworm PPO and Toll activation signaling.

The IMD pathway in Hemipteran: A comparative analysis and discussion.

The evolutionary patterns of the genes in the IMD pathway in hemipterans were characterized and compared. The hemipteran insects were clustered into two groups. One group that encompasses whitefly, plant lice, and scale insect partially lacks the IMD pathway and all antimicrobial peptide (AMP) genes, with the vast majority of IMD pathway and all AMP genes being absent in aphids. The reasons for the absence of the IMD pathway and AMP genes in hemipterans were analyzed based on aphids, in terms of fitness costs. In case of limited resources, aphids have to make a trade-off between the necessary costs such as clean food sources, the essential amino acids supplied by primary bacterial symbionts for survival, nutrients and/or protections against stress provided by secondary symbionts, and the high reproductive capacity, and the costs that do not increase the fitness. Obviously, aphids have to abandon the strong immune system, especially the AMPs and IMD pathway which is mainly against Gram-negative bacteria. The common ground shared with aphids may be the reason for the absence of the IMD pathway and AMP genes in other hemipteran insects.

The Impacts of Climate Change on the Potential Distribution of Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae) in China.

The Indian meal moth Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae) is a notorious stored-grain pest that can be found in most parts of China. The corpses, excretions, and other secretions of P. interpunctella larvae cause serious grain pollution, seriously affecting the nutritional and economic value of stored grain in China. To elucidate the potential distribution of P. interpunctella in China, we used the CLIMEX 4.0 model to project the potential distribution of the pest using historical climate data (1960-1990) and estimated future climate data (2030, 2050, and 2070). Under the historical climate situation, P. interpunctella was distributed in most areas of China, and its highly favorable habitats account for 48.14% of its total potential distribution. Because of temperature change in the future climate, suitable habitats will increase in the eastern part of Qinghai and will decrease in the mid-eastern, northeastern, and southeastern parts of China. Under these scenarios, the area of this pest's highly favorable habitat will be reduced by 1.24 million km2, and its proportion will decrease to about 28.48%. These predicted outcomes will help to distinguish the impact of climate change on the potential distribution of P. interpunctella, thereby providing important information to design early forecasting and strategies to prevent pest harm to stored grain.

Electrophysiological and Behavioral Responses of Plutella xylostella (Lepidoptera: Plutellidae) to Volatiles from a Non-host Plant, Geranium, Pelargonium × hortorum (Geraniaceae).

The diamondback moth (DBM), Plutella xylostella (Lepidoptera: Plutellidae), is a notorious cruciferous vegetable pest globally. Mechanically damaged geranium (Pelargonium × hortorum) can strongly repel DBM, but specific plant bioactive compounds responsible for such effects have not been identified. The headspace volatiles from wounded geranium were analyzed using gas chromatography-electroantennographic detection and gas chromatography-mass spectrometry. The electrophysiological and behavioral responses of DBM females to these chemicals and two blends were investigated. The results showed that five components myrcene, γ-terpinene, linalool, camphor, and terpinen-4-ol in geranium plants mediated the repellence of DBM mated females. These substances significantly repelled the oviposition of females, and blend-2 (a mixture of the five components with a ratio 1:5:3:4:3) was most effective. The slow-release blend-2 had a repellent range of 0.9 m and a repellent longevity of 26 days. These five substances are promising behavioral regulators of the destructive moths and could be potential candidates for "push" components in plant-based "push-pull" strategies.

Expression Profiles and Binding Properties of the Chemosensory Protein PxylCSP11 from the Diamondback Moth, Plutella xylostella (Lepidoptera: Plutellidae).

The diamondback moth, Plutella xylostella L. (Lepidoptera: Plutellidae) is one of the most destructive pests to cruciferous plants worldwide. The oligophagous moth primarily utilizes its host volatiles for foraging and oviposition. Chemosensory proteins (CSPs) are soluble carrier proteins with low molecular weight, which recognize and transport various semiochemicals in insect chemoreception. At present, there is limited information on the recognition of host volatiles by CSPs of P. xylostella. Here, we investigated expression patterns and binding characteristics of PxylCSP11 in P. xylostella. The open reading frame of PxylCSP11 was 369-bp encoding 122 amino acids. PxylCSP11 possessed four conserved cysteines, which was consistent with the typical characteristic of CSPs. PxylCSP11 was highly expressed in antennae, and the expression level of PxylCSP11 in male antennae was higher than that in female antennae. Fluorescence competitive binding assays showed that PxylCSP11 had strong binding abilities to several ligands, including volatiles of cruciferous plants, and (Z)-11-hexadecenyl acetate (Z11-16:Ac), a major sex pheromone of P. xylostella. Our results suggest that PxylCSP11 may play an important role in host recognition and spouse location in P. xylostella.

Glomerular Organization of the Antennal Lobes of the Diamondback Moth, Plutella xylostella L.

The antennal lobe of the moth brain is the primary olfactory center processing information concerning pheromones and plant odors. Plutella xylostella is a major worldwide pest of cruciferous vegetables and its behavior is highly dependent on their olfactory system. However, detailed knowledge of the anatomy and function of the P. xylostella olfactory system remains limited. In the present study, we present the 3-Dimentional (3-D) map of the antennal lobe of P. xylostella, based on confocal microscopic analysis of glomerular segmentation and Neurobiotin backfills of Olfactory Receptor Neurons (ORNs). We identified 74-76 ordinary glomeruli and a macroglomerular complex (MGC) situated at the entrance of the antennal nerve in males. The MGC contained three glomeruli. The volumes of glomeruli in males ranged from 305.83 ± 129.53 to 25440.00 ± 1377.67 µm3. In females, 74-77 glomeruli were found, with the largest glomerulus ELG being situated at the entrance of the antennal nerve. The volumes of glomeruli in females ranged from 802.17 ± 95.68 to 8142.17 ± 509.46 µm3. Sexual dimorphism was observed in anomalous supernumerary, anomalous missing, shape, size, and array of several of the identified glomeruli in both sexes. All glomeruli, except one in the antennal lobe (AL), received projections of antennal ORNs. The glomeruli PV1 in both sexes received input from the labial palp nerve and was assumed as the labial pit organ glomerulus (LPOG). These results provide a foundation for better understanding of coding mechanisms of odors in this important pest insect.

Distribution patterns and morphology of sensilla on the antennae of Plutella xylostella (L.)-A scanning and transmission electron microscopic study.

The antennal morphology, types of antennal sensilla, fine structures and distributions of the sensilla in Plutella xylostella L. (Lepidoptera: Plutellidae) were studied by scanning (SEM) and transmission (TEM) electron microscopy. The antenna, scape, pedicel and flagellum were all longer in males than in females. A total of seventeen types of sensilla were identified on the antennae: trichodea (two subtypes), basiconica, coeloconica (three subtypes), Böhm's bristles (two subtypes), styloconica (two subtypes), squamiformia, auricillica, furcatea (three subtypes), cupuliform organs and terminal sensory pegs. Their numbers and distributions were studied in both male and female, and we found some of the sensilla exhibited various degrees of sexual dimorphisms. Sensilla trichodea were the most abundant of all sensillum types whereas terminal peg was present only once per antenna. Sensilla trichodea in males were bigger (subtype I) and more abundant than in females, however, sensilla basiconica and squamiformia were significantly smaller and less abundant in males than in females. Sensilla styloconica II was only found in females. Seven common sensillum types were studied with TEM to reveal its fine internal structure providing morphological evidences of their sensory functions. Sensilla trichodea I, basiconica and coeloconica III have porous walls suggesting olfactory functions. Combined with the sexual dimorphism, sensilla trichodea male P. xylostella might be involved in detecting sexual pheromones and sensilla basiconica of female might respond to host plant volatiles. Whereas sensilla coeloconica (subtype I and II) and Böhm's bristles have nonporous walls suggesting non-olfactory functions. The study presented a thorough inventory of sensilla on the antennae and laid a solid foundation for future functional studies of these sensilla in this important economical pest.

MicroRNA-17-5p promotes gastric cancer proliferation, migration and invasion by directly targeting early growth response 2.

MicroRNA-17-5p (miR-17-5p) has previously been reported to play an important role in tumor development and progression. However, it functions differently regarding different kinds of malignant tumor, and its role and mechanism in gastric cancer (GC) still lacks investigation. In this study, we detected the relationship between miR-17-5p and the development of GC by qRT-PCR, and it turned out that the level of miR-17-5p was significantly higher in GC patients than that in normal controls, and the aberrant expression of miR-17-5p was correlated with lymph node metastasis. After that, we examined the effect of miR-17-5p taking on the proliferation, apoptosis, migration and invasion of GC cells and the underlying mechanism. Experiments indicated that knockdown of miR-17-5p inhibited the proliferation, invasion and migration, while promoting apoptosis of SGC7901 cells. Early Growth Response 2 (EGR2) protein or mRNA levels were downregulated or upregulated after overexpression or knockdown of miR-17-5p, respectively. By using dual luciferase assay and Western blot, we identified EGR2 as a functional target of miR-17-5p. As far as we know, this could be the first study to demonstrate that miR-17-5p is associated with tumor stage of GC and that it could possibly become a new therapeutic method for the treatment of GC.