Epidemiology of tomato spotted wilt virus in Chrysanthemum morifolium in South Korea and its management using a soil-dwelling predatory mite (Stratiolaelaps scimitus) and essential oils.

Tomato spotted wilt virus (TSWV) is one of most destructive viruses in vegetable and ornamental crop production worldwide. A greenhouse survey to determine the incidence of TSWV in Chrysanthemummorifolium Ramat. was conducted during the 2018 and 2019 growing seasons in South Korea. TSWV was detected using a double antibody sandwich-enzyme-linked immunosorbent assay, and positive results were confirmed using reverse transcription-polymerase chain reaction (RT-PCR). A total of 1569 chrysanthemum plants (70.77 %) tested positive for TSWV among 2217 symptomatic chrysanthemum plants collected from 16 greenhouses. In addition, 116 thrips (72.96 %; Frankliniella occidentalis Pergande) that contained TSWV were identified using RT-PCR from a total of 159 thrips collected from the greenhouses during the survey. A high incidence of viruliferous thrips may have played a role in TSWV occurrence in the chrysanthemum greenhouse. To develop a novel approach for thrips management, the effectiveness of a soil-dwelling predatory mite (Stratiolaelaps scimitus Berlese) and 45 essential oils (as bio-insecticides applied via foliar treatment) was assayed. Four essential oils (cinnamon oil, cinnamon bark oil, oregano oil, and thyme oil) were shown to be significantly toxic to eggs, larvae, and adults of F. occidentalis. For the combined treatment, individuals of S. scimitus (60/m2) were placed on the soil in the chrysanthemum greenhouses. Then, a mixture of the four essential oils was applied as foliar treatment at 4-day intervals. A very low incidence of thrips emerged as adults from the soil (1.2-8.5 %) in the combined treatment in the chrysanthemum greenhouses when surveyed twice per month, compared with the non-treated control or when conventional insecticide sprays were applied. The incidence of TSWV (0.93 %) in chrysanthemum treated with S. scimitus in conjunction with the mixture of four essential oils decreased significantly compared with that treated with chemical insecticides (32.05 %) and in the non-treated controls (84.85 %). Our findings contribute to the development of novel strategies to control TSWV disease in chrysanthemum plants; notably, the control of F. occidentalis using eco-friendly insecticides appears promising.

Identification and field bioassays of the sex pheromone of Eurytoma maslovskii (Hymenoptera: Eurytomidae).

Long-range sex pheromones are generally considered to be a main cue for mate recognition in the order Hymenoptera. Although considerable attention has been given to the identification of semiochemicals in the superfamily Chalcidoidea, which comprises 19 families, no study has identified active components in a field bioassay. We herein report the sensitive and selective pheromone communication system of the Korean apricot wasp, Eurytoma maslovskii (Eurytomidae), whose larvae feed on Prunus mume seeds. Using gas chromatography coupled with electroantennographic detection (GC-EAD) and mass spectrometry (GC-MS), we detected 2,10-dimethyldodecyl propionate and 2,8-dimethyldecyl propionate in thoracic extracts of female E. maslovskii at a ratio of 8:2 as the active pheromone components. Field experiments showed that the attractive effect of the two compounds is highly enantioselective. Racemic 2,10-dimethyldodecyl propionate and 2,8-dimethyldecyl propionate were not attractive to E. maslovskii males. In bioassays with single enantiomers, the (2 S,10 R)-enantiomer was highly attractive to male wasps, and the (2 S,8 S)-enantiomer was also attractive, although to a lesser degree. No synergistic effect between (2 S,10 R)- and (2 S,8 S)-enantiomers was identified, and the (2 S,10 R)-enantiomer alone caught significantly more males than the natural pheromone extracts. The addition of other enantiomers to the (2 S,10 R)-isomer significantly decreased the attraction of conspecific males. In addition, a very low dose of synthetic pheromone attracted conspecific males, showing that both female signaling and male response traits may have evolved to contribute to species-specific sexual communication in this species.

Male-Produced Aggregation Pheromone of the Sloe Bug, Dolycoris baccarum L. (Hemiptera: Heteroptera: Pentatomidae).

The sloe bug, Dolycoris baccarum L. (Hemiptera: Heteroptera: Pentatomidae), is a Palearctic species and an important polyphagous pest. Previous studies have reported that male D. baccarum produce an aggregation pheromone, to which conspecific adults of both sexes respond. We used solid-phase microextraction (SPME) to collect volatiles released by sexually mature virgin males and females, and identified potential components of the male-produced aggregation pheromone. Analysis of the SPME samples by gas chromatography-mass spectrometry (GC-MS) revealed three male-specific components, α-bisabolol, trans-α-bergamotene, and ß-bisabolene, at a ratio of 100:10:3. A field trapping experiment showed that (±)-α-bisabolol alone attracted D. baccarum adults, and it was synergized by trans-α-bergamotene, but not (S)-ß-bisabolene. However, neither trans-α-bergamotene nor (S)-ß-bisabolene was attractive individually. A binary blend of (±)-α-bisabolol and trans-α-bergamotene in a 10:1 ratio was less attractive than caged live males, suggesting that further study is necessary to determine the absolute configuration or steroisomeric composition of the natural pheromone components. This is the first report of α-bisabolol and trans-α-bergamotene as semiochemicals among Pentatomidae, and the first to identify α-bisabolol from insects.

Safflower (Carthamus tinctorius L.) seed attenuates memory impairment induced by scopolamine in mice via regulation of cholinergic dysfunction and oxidative stress.

Cholinergic dysfunction and oxidative stress are the most common causes of Alzheimer's disease (AD). Safflower seed contains various anti-oxidant and cholinergic improvement compounds, such as serotonin and its derivatives. In the present study, we investigated the protective effects and mechanisms of a safflower seed extract on scopolamine-induced memory impairment in a mouse model. The safflower seed extract was orally administered at a dose of 100 mg kg-1 day-1, and then behavior tests (such as T-maze and novel object recognition tests) were conducted. Acetyl cholinesterase (AChE) activity, reactive oxygen species (ROS) production, and antioxidant enzymes in the brain were measured. In behavior tests, the novel route exploration and object recognition were improved by the administration of the safflower seed extract, which suggests that the safflower seed extract improves memory function in the scopolamine-treated mouse model. In addition, the safflower seed extract-administered group showed inhibition of the AChE activity and improved cholinergic dysfunction. Furthermore, the administration of the safflower seed extract resulted in lower ROS production and higher antioxidant enzyme levels as compared to the scopolamine-treated group, suggesting the protective role of the safflower seed extract against oxidative stress. The results of the present study suggest that the safflower seed extract improves scopolamine-induced memory deficits via the inhibition of cholinergic dysfunction and oxidative stress. Therefore, safflower seeds might become a promising agent for memory improvement in AD patients.

Safflower seed extract synergizes the therapeutic effect of cisplatin and reduces cisplatin-induced nephrotoxicity in human colorectal carcinoma RKO cells and RKO-transplanted mice.

Safflower seed is effective against oxidative stress, mediating the activation of the apoptotic signaling pathway in the renal tissues of cisplatin-treated mice. The anticancer activity of safflower in various cancer cell lines has also been reported. The present study was conducted to evaluate the potential synergistic anticancer effects of the co-treatment of safflower seed extracts and cisplatin in RKO cells and in BALB/c mice bearing RKO cell-derived human colorectal tumors. In the cellular system, RKO cells were treated with safflower seed extract in the presence or absence of cisplatin for 48 h and the cytotoxicity was evaluated by using microscopy. In the in vivo system, mice were injected with RKO cells and subsequently orally administered 100 or 200 mg/kg body weight safflower seed extract plus cisplatin-treated or untreated mice for 3 days to examine the inhibitory effect on the tumor. Treatment with safflower seed extract or cisplain to RKO cells resulted in a greater cell death than in with untreated cells. In the RKO cells co-treated with both safflower seed extract and cisplatin, greater cell damage was observed. In addition, mice co-administered safflower seed extract and cisplatin had lower concentrations of serum creatinine, which were indicative of less damage to the kidney, and had a lower solid tumor mass and higher expression of the caspase-3 protein. The results showed that safflower seed extract was highly toxic to RKO cells and inhibited tumor growth in cisplatin-treated mice through renoprotective effects.

Benzaldehyde Synergizes the Response of Female Xyleborinus saxesenii (Coleoptera: Curculionidae, Scolytinae) to Ethanol.

The ambrosia beetle, Xyleborinus saxesenii Ratzeburg (Coleoptera: Curculionidae, Scolytinae), infests physiologically stressed apple and peach trees in Korea. Dispersing females utilize the degradation product ethanol and host-related volatiles to locate and colonize new host trees. We examined the extent to which 12 chemicals emitted from fruit trees act synergistically with ethanol to attract X. saxesenii. The addition of benzaldehyde to ethanol significantly increased beetle attraction, although benzaldehyde was not attractive by itself. The addition of (-)-α-pinene, ethyl butyrate, ethyl isovalerate, (R)-(+)-limonene, 3-methyl-1-butanol, ethyl tiglate, (+)-aromadendrene, vanillin, 2-butanol, styrene, or ethyl 3,3-dimethylacrylate to ethanol had no effect on beetle attraction. In a dose-response test, the addition of 5-50% benzaldehyde doses synergistically increased the number of beetle captures; however, trap catches did not increase as the benzaldehyde dosage increased. The synergistic influence of benzaldehyde on beetle response to ethanol was lower in early spring than in late summer to early fall, probably because synthetic benzaldehyde emissions from field lures were overwhelmed by background levels of natural benzaldehyde emitted from peach twigs in the flowering stage.

Sex pheromones and reproductive isolation in five mirid species.

Mate location in many mirid bugs (Heteroptera: Miridae) is mediated by female-released sex pheromones. To elucidate the potential role of the pheromones in prezygotic reproductive isolation between sympatric species, we investigated differences in the pheromone systems of five mirid species, Apolygus lucorum, Apolygus spinolae, Orthops campestris, Stenotus rubrovittatus and Taylorilygus apicalis. GC/MS analyses of metathoracic scent gland extracts of virgin females showed that all five species produced mixtures of hexyl butyrate, (E)-2-hexenyl butyrate and (E)-4-oxo-2-hexenal, but in quite different ratios. (E)-2-hexenyl butyrate was the major component of A. spinolae, while hexyl butyrate was the most abundant component in the pheromone blends of the other four species. In addition to the three compounds, a fourth component, (E)-2-octenyl butyrate, was present in the gland extracts of A. lucorum and T. apicalis females. Field tests suggest that the ternary blends of hexyl butyrate, (E)-2-hexenyl butyrate and (E)-4-oxo-2-hexenal as found in the extracts of the females of each species do not inhibit attraction of conspecific males but ensure species-specificity of attraction between A. lucorum, O. campestris and T. apicalis. Furthermore, (E)-2-octenyl butyrate was essential for attraction of A. lucorum and T. apicalis males, but strongly inhibited attraction of male A. spinolae, O. campestris and S. rubrovittatus. The combined results from this study and previous studies suggest that the minor component and pheromone dose in addition to the relative ratio of the major components play an important role in reproductive isolation between mirid species.

Identification of the female-produced sex pheromone of the plant bug Apolygus spinolae.

Apolygus spinolae (Meyer-Dür) (Heteroptera: Miridae) is an important pest of fruit and tea trees in Korea and Japan. Analyses of extracts of metathoracic scent glands revealed that those of female bugs contained hexyl butyrate, (E)-2-hexenyl butyrate, and (E)-4-oxo-2-hexenal in a ratio of 20:100:7. The glands of males contained the same three compounds, but the ratio of the components was quite different, with hexyl butyrate being the most abundant. Field trapping tests with various blends of the synthetic compounds dispensed from high-density polyethylene tubes showed that (E)-2-hexenyl butyrate and (E)-4-oxo-2-hexenal were essential for attraction of male A. spinolae, and catches with a wide range of ratios of these two compounds did not differ significantly. However, adding hexyl butyrate at 50 % or more of the (E)-2-hexenyl butyrate to the binary blend strongly inhibited attraction of males. Trap catches increased with increasing amounts of a 10:1 blend of (E)-2-hexenyl butyrate and (E)-4-oxo-2-hexenal from 0.011 to 11 mg loaded into the tube. Catches of males in traps baited with lures containing 1.1 mg of the binary blend were not significantly different from catches in traps baited with live virgin females.

(E)-5-hexadecenyl acetate: a novel moth sex pheromone component from Stathmopoda auriferella.

The sex pheromone of Stathmopoda auriferella (Walker), an important pest of kiwifruit in Korea, was studied. Two candidate pheromone components detected in the gland extracts of females were identified as (E)-5-hexadecenyl acetate (E5-16:OAc) and (E)-5-hexadecenol (E5-16:OH) in a ratio of 75:25 by mass spectral analysis of natural pheromone components and dimethyldisulfide adducts, and retention index comparisons with synthetic standards. In the kiwifruit orchards, E5-16:OAc alone was attractive to S. auriferella males and caught significantly more males than live virgin females. However, addition of E5-16:OH strongly inhibited attraction to E5-16:OAc. These results suggest that the major component of the female-produced sex pheromone of S. auriferella is E5-16:OAc. This hexadecenyl acetate is a novel moth sex pheromone component.

Sex pheromone of the smaller clearwing moth Synanthedon tenuis (Butler).

The smaller clearwing moth, Synanthedon tenuis (Butler) (Lepidoptera: Sesiidae), is a major pest of persimmon in northeast Asia. A previous study reported attraction of S. tenuis males to Z3,Z13-18:OAc, but this compound had no effect on male catch in the persimmon orchards in Korea. In this study, we analyzed pheromone gland extracts of S. tenuis females and identified Z3,Z13-18:OH as the main component. In field trapping trial, Z3,Z13-18:OH alone was attractive to S. tenuis males and competitive with live virgin females. These results indicate that the pheromone of this species consists of a single component, Z3,Z13-18:OH. However, Z3,Z13-18:OAc, a previously reported attractant, was not detected in the gland extracts of females. Furthermore, the addition of Z3,Z13-18:OAc to the main pheromone component strongly inhibited attraction for males, suggesting that the diene acetate is not a pheromone component. This is the first report of an octadecadienol as female-produced sex pheromone from the genus Synanthedon.

Female sex pheromone of the Gelechiid moth Scrobipalpa salinella (Zeller).

The sex pheromone system of Scrobipalpa salinella (Zeller), an important pest of the halophyte Salicornia europaea in the tidal salt marshes, was studied. Z3-12:OAc and Z5-12:OAc were identified from both female pheromone glands and female emissions, but in quite different ratios. Field trapping tests demonstrated that Z3-12:OAc and Z5-12:OAc are essential for optimal attraction of male moths, and a 100:5 blend found in gland extracts is significantly more attractive to males than a 100:50 ratio similar to that found in SPME samples. Small amounts of E3-12:OAc and Z5-14:OAc also were present in pheromone gland extracts. A blend of E3-12:OAc with Z3-12:OAc attracted a few males, but was not as attractive as the binary blend of Z3-12:OAc and Z5-12:OAc. Moreover, addition of E3-12:OAc did not affect captures of males to the primary binary blend. Another glandular component, Z5-14:OAc, had no behavioral activity in field bioassays. Therefore, a synthetic mixture of Z3-12:OAc and Z5-12:OAc in a 100:5 ratio can be used as an effective tool for monitoring and control of this species.

Identification and field evaluation of the sex pheromone of Synanthedon bicingulata (Staudinger).

The sex pheromone of Synanthedon bicingulata (Staudinger), a major pest of Prunus species in many regions of northeast Asia, was identified. Two major components from the pheromone gland extracts of female moths are (E,Z)-3,13-octadecadienyl acetate (E3,Z13-18:OAc) and (Z,Z)-3,13-octadecadienyl acetate (Z3,Z13-18:OAc), and the average ratio of these components is about 4:6, respectively. In addition to the major components, four minor components, (Z)-13-octadecenyl acetate (Z13-18:OAc), (E,Z)-2,13-octadecadienyl acetate (E2,Z13-18:OAc), (E,Z)-3,13-octadecadien-1-ol (E3,Z13-18:OH), and (Z,Z)-3,13-octadecadien-1-ol (Z3,Z13-18:OH) also were identified from pheromone gland extracts. Field tests showed that E3,Z13-18:OAc and Z3,Z13-18:OAc are essential for attraction of male S. bicingulata moths, and males are optimally attracted to the blend ratio found in pheromone gland extracts of conspecific females. Addition of the minor glandular components (Z13-18:OAc, E2,Z13-18:OAc, E3,Z13-18:OH, and Z3,Z13-18:OH) did not affect captures of males to the primary binary blend. Thus, the blend of E3,Z13-18:OAc and Z3,Z13-18:OAc at the natural ratio can be used for monitoring populations of this species.

RNA interference of pheromone biosynthesis-activating neuropeptide receptor suppresses mating behavior by inhibiting sex pheromone production in Plutella xylostella (L.).

Sex pheromone production is regulated by pheromone biosynthesis-activating neuropeptide (PBAN) in many lepidopteran species. We cloned a PBAN receptor (Plx-PBANr) gene from the female pheromone gland of the diamondback moth, Plutella xylostella (L.). Plx-PBANr encodes 338 amino acids and has conserved structural motifs implicating in promoting G protein coupling and tyrosine-based sorting signaling along with seven transmembrane domains, indicating a typical G protein-coupled receptor. The expression of Plx-PBANr was found only in the pheromone gland of female adults among examined tissues and developmental stages. Heterologous expression in human uterus cervical cancer cells revealed that Plx-PBANr induced significant calcium elevation when challenged with Plx-PBAN. Female P. xylostella injected with double-stranded RNA specific to Plx-PBANr showed suppression of the receptor gene expression and exhibited significant reduction in pheromone biosynthesis, which resulted in loss of male attractiveness. Taken together, the identified PBAN receptor is functional in PBAN signaling via calcium secondary messenger, which leads to activation of pheromone biosynthesis and male attraction.

Identification and field bioassays of the sex pheromone of Synanthedon haitangvora.

Two major components from pheromone gland extracts of Synanthedon haitangvora females were identified as (Z,Z)-3,13-octadecadienyl acetate (Z3,Z13-18:OAc) and (E,Z)-2,13-octadecadienyl acetate (E2,Z13-18:OAc), and the average ratio of these components was about 1:1. Seven minor components, (Z)-9-hexadecenyl acetate (Z9-16:OAc), (Z)-11-hexadecenyl acetate (Z11-16:OAc), (Z)-9-octadecenyl acetate (Z9-18:OAc), (Z)-13-octadecenyl acetate (Z13-18:OAc), (E,Z)-3,13-octadecadienyl acetate (E3,Z13-18:OAc), (Z,Z)-3,13-octadecadien-1-ol (Z3,Z13-18:OH), and (E,Z)-2,13-octadecadien-1-ol (E2,Z13-18:OH), also were identified from gland extracts. Field tests showed that male S. haitangvora were attracted to Z3,Z13-18:OAc alone, but the maximum number of males was attracted to the binary blend of Z3,Z13-18:OAc and E2,Z13-18:OAc mimicking the blend found in female extracts. The addition of minor components to a 1:1 blend of Z3,Z13-18:OAc and E2,Z13-18:OAc did not increase the numbers of moths captured. The only significant effect of minor components was the strong inhibitory effect of adding Z3,Z13-18:OH to the primary binary blend. Increasing doses of the optimum pheromone blend in the lures from 0.1 to 2.0 mg increased trap catches of male S. haitangvora.

Sex pheromones and reproductive isolation of three species in genus Adoxophyes.

We tested differences in female pheromone production and male response in three species of the genus Adoxophyes in Korea. Females of all three species produced mixtures of (Z)-9-tetradecenyl acetate (Z9-14:OAc) and (Z)-11-tetradecenyl acetate (Z11-14:OAc) as major components but in quite different ratios. The ratio of Z9-14:OAc and Z11-14:OAc in pheromone gland extracts was estimated to be ca. 100:200 for Adoxophyes honmai, 100:25 for Adoxophyes orana, and 100:4,000 for Adoxophyes sp. Field tests showed that males of each species were preferentially attracted to the two-component blends of Z9-14:OAc and Z11-14:OAc mimicking the blends found in pheromone gland extracts of conspecific females. The effects of minor components identified in gland extracts on trap catches varied with species. Addition of 10-methyldodecyl acetate (10me-12:OAc) or (E)-11-tetradecenyl acetate (E11-14:OAc) to the binary blend of Z9-14:OAc and Z11-14:OAc significantly increased captures of A. honmai males, whereas E11-14:OAc exhibited a strongly antagonistic effect on catches of Adoxophyes sp. males. Moreover, (Z)-9-tetradecen-1-ol (Z9-14:OH) or (Z)-11-tetradecen-1-ol (Z11-14:OH) added to the binary blends increased attraction of male A. orana but not A. honmai and Adoxophyes sp. males, suggesting that these minor components, in addition to the relative ratios of the two major components, play an important role in reproductive isolation between Adoxophyes species in the southern and midwestern Korea where these species occur sympatrically.