Application of Direct Immersion Solid-Phase Microextraction (DI-SPME) for Understanding Biological Changes of Mediterranean Fruit Fly (Ceratitis capitata) During Mating Procedures.

Samples from three different mating stages (before, during and after mating) of the Mediterranean fruit fly Ceratitis capitata were used in this experiment. Samples obtained from whole insects were subjected to extraction with the two mixtures of solvents (acetonitrile/water (A) and methanol/acetonitrile/water (B)) and a comparative study of the extractions using the different solvents was performed. Direct immersion-solid phase microextraction (DI-SPME) was employed, followed by gas chromatographic-mass spectrometry analyses (GC/MS) for the collection, separation and identification of compounds. The method was validated by testing its sensitivity, linearity and reproducibility. The main compounds identified in the three different mating stages were ethyl glycolate, α-farnesene, decanoic acid octyl ester, 2,6,10,15-tetramethylheptadecane, 11-tricosene, 9,12-(Z,Z)-octadecadienoic acid, methyl stearate, 9-(Z)-tricosene, 9,11-didehydro-lumisterol acetate; 1,54-dibromotetrapentacontane, 9-(Z)-hexadecenoic acid hexadecyl ester, 9-(E)-octadecenoic acid and 9-(Z)-hexadecenoic acid octadecyl ester. The novel findings indicated that compound compositions were not significantly different before and during mating. However, new chemical compounds were generated after mating, such as 1-iodododecane, 9-(Z)-tricosene and 11,13-dimethyl-12-tetradecen-1-acetate which were extracted with both (A) and (B) and dodecanoic acid, (Z)-oleic acid, octadecanoic acid and hentriacontane which were extracted with (A) and ethyl glycolate, 9-hexadecenoic acid hexadecyl ester, palmitoleic acid and 9-(E)-octadecenoic acid, which were extracted with solvent (B). This study has demonstrated that DI-SPME is useful in quantitative insect metabolomics by determining changes in the metabolic compounds in response to mating periods. DI-SPME chemical extraction technology might offer analysis of metabolites that could potentially enhance our understanding on the evolution of the medfly.

Evaluation of the Synergistic Effect Between Ethyl Formate and Phospine for Control of Aphis gossypii (Homoptera: Aphididae).

Cotton aphid, Aphis gossypii Glover, is known as a quarantine pest that is hard to control with short periods of fumigation with phosphine (PH(3)) or low concentrations of ethyl formate. Moreover, low-temperature fumigation with ethyl formate can lead to phototoxic damage of some perishable commodities. Therefore, a laboratory study was conducted to evaluate the synergistic effect of mixing ethyl formate and PH(3) for the treatment of adults and nymphs of A. gossypii. Combined toxicity was observed and compared with a single dose of eitherrethyl formate or PH(3). When insects were exposed to 0.5 g/m(3) of PH(3) combined with different levels of ethyl formate from 1.6 to 16.3 g/m(3) at 5 and 20C for 2 h, L(Ct)(50) and L(Ct)(99) values were greatly reduced in comparison with a single dose of either ethyl formate or PH(3). The synergistic ratio (SR) is described as L(Ct) of ethyl formate alone/L(Ct) of ethyl formate + PH(3). The SR values of L(Ct)(50) and L(Ct)(99) for adult A. gossypii at 5C were 4.55 and 2.33, respectively. However, at 20C the SR levels of L(Ct)(50) and L(Ct)(99) were 2.22 and 1.45, respectively, but still showed significant synergism (significant difference, P<0.5). This new technology could meet quarantine and preshipment requirements for shorter exposure times and less damage of perishable commodities, and could also be extended for controlling other quarantine pests and thereby be a useful alternative to methyl bromide for fruit and vegetable applications.

Synergistic Effects of Oxygen on Phosphine and Ethyl Formate for the Control of Phthorimaea operculella (Lepidoptera: Gelechiidae).

Phosphine (PH3) and ethyl formate (EF) are two potentially powerful postharvest fumigant insecticides. We investigated the effectiveness of both PH3 and EF as fumigants at all developmental stages of the potato tuber moth Phthorimaea operculella Zeller, and we also studied the synergistic effects of these fumigants under controlled atmospheres of 50 and 80% oxygen (O2). The larval stage of P. operculella was the most susceptible to fumigation with PH3 at both 5°C and 20°C. All of the developmental stages showed greater susceptibility to PH3 at 20°C than at 5°C, whereas the susceptibility of adult P. operculella to this fumigant was not affected by temperature. The toxicity of EF did not differ with temperature for any of the P. operculella developmental stages. The atmospheric oxidation of PH3 increased the toxicity of this fumigant toward all developmental stages at both temperatures. In contrast, no differences in toxicity were observed for oxidized EF compared with EF alone at any developmental stage. In conclusion, using fumigation tests, we showed that atmospherically oxidized PH3 was much more effective against P. operculella than PH3 alone, demonstrating a synergistic effect for this fumigant and O2. Therefore, treatment with PH3 and high concentrations of O2, as described in this study, could be useful for managing the postharvest pest P. operculella.

Structural determination of elicitors in Sogatella furcifera (Horváth) that induce Japonica rice plant varieties (Oryza sativa L.) to produce an ovicidal substance against S. furcifera eggs.

Certain Japonica rice plant varieties (Oryza sativa L.) show resistance to the eggs of the white backed planthopper (Sogatella furcifera Horváth) by the formation of watery lesions and production of benzyl benzoate, an active ovicidal compound. Benzyl benzoate results in high ovicidal activity of S. furcifera eggs and reduces egg hatchability. The production of benzyl benzoate by the rice plants is only induced by an unknown elicitor(s) from the female S. furcifera. Therefore, we tried to isolate and identify these elicitors. An active 80% MeOH extract of S. furcifera was separated on a reverse-phase ODS MPLC, and the elicitor(s) was eluted in 100% MeOH and 100% EtOH fractions. Further separation of the active 100% MeOH fraction using a HPLC led to the isolation of four active compounds. The structures of each compound were determined by using NMR, LC-MS, and GC-MS spectra. The compounds were 1,2-dilinoleoyl-sn-glycero-3-phosphocholine, 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine, 1-palmitoyl-2-oleoyl-X-glycero-3-phosphoethanolamine, and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine.

Ethyl Formate Fumigation for Controlling Two Major Aphid Pests, Aphis spiraecola and Aphis gossypii, on Passion Fruit, from Cultivation to Post-Harvest Storage.

Tropical and subtropical crops are being increasingly cultivated in South Korea, leading to an increase in damage by exotic insect pests. Consequently, ethyl formate (EF) is currently being considered for quarantine and pre-shipment fumigation. In this study, we evaluated the effectiveness of EF fumigation for controlling Aphis spiraecola Patch and Aphis gossypii Glover, two representative quarantine pests on passion fruit ("Pink Bourbon") during greenhouse cultivation and post-harvest storage. The efficacy of EF against both aphids in terms of the lethal concentration causing 50% mortality (LCt50%) and LCt99% was 1.36-2.61 g h/m3 and 3.73-7.55 g h/m3 under greenhouse conditions (23 °C), and 1.37-2.02 g h/m3 and 3.80-14.59 g h/m3 post-harvest (5 °C), respectively. EF at 4 g/m3 for 4 h resulted in 100% mortality of A. spiraecola, which was more resistant to EF, without causing phytotoxic damage to the trees in a 340 m3 greenhouse. Post-harvest fruit fumigation at 10 g/m3 for 4 h in a mid-size (0.8 m3) fumigation chamber resulted in complete disinfection. Moreover, the EF level decreased below the EF threshold within 10 min after natural ventilation in the greenhouse. Therefore, our results suggest EF fumigation as an effective method for controlling A. spiraecola and A. gossypii.

Elicitor(s) in Sogatella furcifera (Horváth) causing the Japanese rice plant (Oryza sativa L.) to induce the ovicidal substance, benzyl benzoate.

We elucidate the mechanism for inducing the production of ovicidal benzyl benzoate by Japonica rice varieties to kill eggs of the whitebacked planthopper, Sogatella furcifera (Horváth), lying in the rice plant. Even when subjected to physical damage by a needle or damage with water, the rice plant produced no benzyl benzoate. However, significant benzyl benzoate was produced when the plant was damaged with a methanol extract or homogenate of S. furcifera. The extract of the male did not induce the production of benzyl benzoate, but that of the female did. We concluded from these results that benzyl benzoate was induced by some elicitor(s) in the female of S. furcifera.

Development of a Concurrent Treatment Technique of Ethyl Formate and Mixtures (Nitrogen, Phosphine) to Control Citrus Mealybug (Planococcus citri).

Currently, the fumigant ethyl formate (EF) is stored as a liquified gas in metal cylinders mixed with carbon dioxide (CO2), but this product type is expensive to manufacture, transport, and maintain in cylinders. To address these problems, we developed a new EF fumigation technique with a nitrogen (N2) carrier. In this report, the susceptibility of citrus mealybugs, one of the most resistant mealybugs to fumigants, to EF was assessed; the phytotoxicity of an EF + N2 concurrent treatment applied to banana fruit was examined to evaluate the efficacy compared to the current EF + CO2 product; and the increased efficacy with a phosphine (PH3) addition to EF + N2 was also assessed. Concurrent treatment of EF and N2 was performed at an LC50 concentration of EF. N2 was applied in seven doses from concentrations of 79% to 95%. The phytotoxicity of EF to bananas was assessed by applying EF at 35 mg/L with N2 at 79%, and the color, sugar content, and weight loss of bananas were measured for 14 days after treatment. The EF with N2 treatment resulted in more than 50% mortality at all growth stages of the mealybug, and there was no significant difference between the untreated and treated banana fruits. EF mixed with PH3 showed a higher efficacy than treatment with EF alone, but only a slight increase in efficacy was observed when the PH3 concentration increased. These results indicate that concurrent treatment with EF and N2 can be used to control mealybugs on banana fruits, and combined treatment with EF and PH3 can also enhance the efficacy against mealybugs.

Synergistic Effect of Cold Treatment Combined with Ethyl Formate Fumigation against Drosophila suzukii (Diptera: Drosophilidae).

Drosophila suzukii is a quarantine pest that is rapidly spreading in berries. This study evaluated the synergistic effect of combination treatment with ethyl formate (EF) and cold temperature for D. suzukii control on imported grapes. A higher insecticidal effect was observed at 1 °C than at 5 °C at all developmental stages, and the pupal stage showed the strongest tolerance to cold temperature. After EF fumigation alone, eggs showed the highest tolerance at 216.67 mg·h/L (LCT99 value), and adults showed the highest susceptibility at <27.24 mg·h/L. Among the combination treatment methods, cold temperature after fumigation resulted in the best synergistic effect. The effect of this combination was significant, with 23.3% higher mortality for eggs, 22.4% for larvae, and 23.4% for pupae than observed with EF fumigation alone. Furthermore, the period of complete D. suzukii control in the 12 L desiccator was shorter in the combination treatment group at the LCT80 value than at the LCT50 value of the egg stage. EF showed a very high sorption rate (24%) after 4 h of exposure at a grape loading ratio of 15% in a 0.65 m3 fumigation chamber. As the grape loading ratio for combination treatment decreased, D. suzukii mortality increased, but when EF was administered at the LCT80 value, there was little difference in the mortalities of the eggs and larvae but not the pupae. All D. suzukii developmental stages were completely controlled within 7 days after combination treatment, and phytotoxicity was not observed in grapes. These results suggest that the combination of cold-temperature treatment and EF fumigation could be used for D. suzukii control.

Efficacy and Phytotoxicity Assessment of Successive Application of Methyl Bromide and Cold Treatment on Export Strawberry Fruits.

Recently, spotted wing Drosophila, Drosophila suzukii, is globally prevalent and causes agricultural losses to many fruits. To export Korean strawberry, methyl bromide fumigation is required to remove D. suzukii infestations, but Korean strawberry farmers are worried about fruit damage because methyl bromide can cause phytotoxicity on fresh commodities. In this report, we assessed the efficacy and phytotoxicity of single and successive application of methyl bromide and cold treatment on an export variety of strawberry to reduce fruit damage. The currently recommended dosage of methyl bromide, 40 g/m3 for 3 h at 18 °C, was enough to control all stages of D. suzukii without phytotoxicity. A dosage of 20 g/m3 of methyl bromide treatment for 3 h, followed by 1 d of cold (0 °C) treatment, showed 100% mortality in all growth stages of D. suzukii without fruit damage. Successive application of methyl bromide and cold treatment shows potential as a method of decreasing phytotoxicity and reducing the use of methyl bromide for environmental considerations.

Transcriptomic evaluation on methyl bromide-induced phytotoxicity in Arabidopsis thaliana and its mode of phytotoxic action via the occurrence of reactive oxygen species and uneven distribution of auxin hormones.

The increase in worldwide trade has caused the quality maintenance of commercialized agriproducts to be crucial in keeping its economic value. In recent years, methyl bromide (MB) has been used dominantly during quarantine and pre-shipment, despite it being an environmental hazard with global repercussions. Through this study, it was shown that Arabidopsis thaliana's 2 h exposure to the MB treatment displayed no signs of phytotoxicity, whereas its 4 h exposure significantly interfered with growth. The transcriptomic analysis found the molecular modifications in A. thaliana after the MB fumigation with the up-regulation of genes specifically relative to the abiotic and oxidative stress, and the down-regulation of auxin transporter genes. Some important gene expressions were verified by RT-qPCR and their expression patterns were similar. Oxidative stresses via the reactive oxygen species (ROS) in relation to MB phytotoxicity were confirmed with the increased malondialdehyde in MB-4h-treated A. thaliana. Uneven distribution of auxins via lower expression of auxin transporter genes was also determined using UPLC-ESI-QqQ MS. Application of two ROS scavengers such as N-acetyl-cysteine and L-glutathione minimized MB phytotoxic effect in A. thaliana. Therefore, MB caused severe oxidative stress, and alternatives regarding the use of MB should be considered.

Ethyl Formate as a Methyl Bromide Alternative for Fumigation of Citrus: Efficacy, Fruit Quality, and Workplace Safety.

Ethyl formate (EF) was evaluated as a potential alternative to methyl bromide (MB) for phytosanitary treatment of imported citrus fruit in the Republic of Korea. Planococcus citri (Risso) (Hemiptera: Pseudococcidae), a mealybug with known tolerance against EF and MB, was used as a representative pest to test efficacy of the two fumigants against eggs. In nine commercial-scale refrigerated container (67.5 m3) trials using imported orange, lemon and grapefruit, EF applied at the currently approved dose for citrus (70 g·m-3 at 5°C for 4 h, developed for Aspidiotus excisus Green (Hemiptera: Diaspididae), a species less EF tolerant than P. citri) resulted in 76.9-98.3% mortality of P. citri eggs. The EF treatment did not affect the sugar content or the color of peel and pulp of the treated fruit. When oranges were treated according to the current MB (64 g·m-3 at >5°C for 2 h) or EF treatment guidelines, the concentration of fumigant around the fruit fluctuated between 9.4 and 185.1 ppm for EF and 9.5-203.0 ppm for MB during the 72-h post-fumigation processes (venting [0-2 h], transportation to storage [2-24 h], and storage periods [24-72 h]) with both EF and MB maintained between 10 and 100 ppm during the storage period. Considering the efficacy of EF, its apparent lack of phytotoxicity, and its more manageable threshold limit value for humans (100 ppm EF compared to 1 ppm MB for an 8-h time weighted average exposure), our results suggest that EF may be a promising alternative to MB for the phytosanitary treatment of imported citrus in Korea.

Postharvest treatment of mandarin fruit using a combination of methyl bromide and phosphine against Bactrocera dorsalis (Diptera: Tephritidae).

BACKGROUND: Chinese Nanfeng mandarin (Citrus reticulate Blanco '2-6') fruit is host to the oriental fruit fly Bactrocera dorsalis Hendel, and therefore requires phytosanitary treatment before exporting abroad. The use of methyl bromide (MB) should be reduced because it has phytotoxic and ozone depleting properties. In the present study, fumigations with mixtures of gaseous phosphine (PH3 ) and MB were conducted to disinfest B. dorsalis. The effect of combined fumigation on postharvest quality of Nanfeng mandarin fruit was then evaluated. RESULTS: There was a clear synergistic effect between MB and PH3 against B. dorsalis at 20 °C. The third-instar larvae of B. dorsalis were the most tolerant to treatment with both MB alone and combined with PH3 . Toxicity assay of third instars indicated that higher PH3 concentrations increased MB toxicity, and 1.42-2.84 g m-3 of PH3 was optimal when combined with MB. Probit analysis showed that, combined with 2.13 g m-3 PH3 , similar mortality was achieved with 50% of the MB required when applied alone. Furthermore, the fruit quality test showed that MB fumigation resulted in a higher respiration rate and >40% fruit rot after storage. The combined treatment reduced these effects and did not have adverse effects on fruit firmness, soluble solid content, titratable acidity, or vitamin C content. CONCLUSION: Our findings demonstrate the synergistic effect between PH3 and MB and indicate that this treatment has potential as a novel strategy for postharvest control of B. dorsalis, especially in MB-sensitive fruit. © 2019 Society of Chemical Industry.

Susceptibility of the Cigarette Beetle Lasioderma serricorne (Fabricius) to Phosphine, Ethyl Formate and Their Combination, and the Sorption and Desorption of Fumigants on Cured Tobacco Leaves.

The susceptibility of Lasioderma serricorne to phosphine (PH3), ethyl formate (EF) and their combination (PH3 + EF) was evaluated in this study. Eggs, larvae, pupae and adults were subjected to treatment with fumigants to determine the 90% lethal concentration time (LCt90) values. Treatment with PH3 for 20 h resulted in LCt90 values of 1.15, 1.39, 14.97 and 1.78 mg h/L while treatment with EF resulted in values of 157.96, 187.75, 126.06 and 83.10 mg h/L, respectively. By contrast, the combination of PH3 + EF resulted in LCt90 values of 36.05, 44.41, 187.17 and 35.12 mg h/L after 4 h. These results show that, through treatment with PH3 + EF, control can be achieved at lower concentrations than for treatment with EF alone and at lower exposure times than for treatment with PH3 alone. The sorption rates of the fumigants on cured tobacco leaves were determined for filling ratios of 2.5%, 5.0% and 10.0% (w/v). Cured tobacco leaves were treated with either 2 mg/L PH3, 114 mg/L EF or 0.5 mg/L PH3 + 109 mg/L EF. Treatment with PH3 showed sorption rates of 0.0%, 7.1% and 14.3%. EF, however, showed higher sorption rates of 64.9%, 68.5% and 75.5%, respectively, for the indicated filling ratios. When PH3 and EF were combined, the sorption rate of PH3 was 0.0%, while the sorption rates of EF were lower (9.1%, 12.0% and 23.2%) than treatment with only EF. EF required a ventilation time of longer than 22 h to desorb from cured tobacco leaves. Therefore, PH3 + EF can effectively control L. serricorne in cured tobacco leaves, with sufficient ventilation time required after treatment for the safety of workers.

Fumigation Activity against Phosphine-Resistant Tribolium castaneum (Coleoptera: Tenebrionidae) Using Carbonyl Sulfide.

Phosphine resistance is occurring among stored-grain pests worldwide. This study investigated the fumigation activity of phosphine (PH3) and carbonyl sulfide (COS) against domestic strain (d-strain) Tribolium castaneum, resistance strain (r-strain) T. castaneum and Oryzaephilus surinamensis. All developmental stages of the pests were exposed to two fumigants (PH3 and COS), and the fumigation activity according to the dose and exposure time was evaluated in a 12-L desiccator and 0.5 m3 fumigation chamber. The rice sorption rate and quality following exposure to thetwofumigants were evaluated. The mortality was 2.9% in r-strain T. castaneum, 49.5% in d-strain T. castaneum and 99.2% in O. surinamensis when 2 mg/L PH3 was used in a 12-L desiccator for 4 h. However, all pest developmental stages showed 100% mortality after 24 h of exposure in the 0.5 m3 fumigation chamber, except for the r-strain T. castaneum. A mortalityof 100% was observed in all of the r-strain T. castaneum developmental stages at an exposure time of 192 h. For COS applied at 40.23 mg/L and 50 g/m3 in the 12-L desiccator and the 0.5 m3 fumigation chamber, respectively, 100% mortality was observed across all developmental stages regardless of species and strain. The sorption of COS was 10% higher than that of PH3, but there was no significant difference in rice quality compared to that in the control. Therefore, this study suggests that COS can be used for controlling T. castaneum resistant to PH3.

Ethyl Formate as a Methyl Bromide Alternative for Phytosanitary Disinfestation of Imported Banana in Korea With Logistical Considerations.

Methyl bromide (MB) use for quarantine and phytosanitary purposes is being phased out. Several effective MB alternatives have been identified depending on the target system. Industry commitment for the shift to new technologies will depend not only on the efficacy of alternatives but also on their compatibility and integrability into existing operations. We compared the efficacy of MB and ethyl formate (EF) for disinfestation of Planococcus citri (Risso) (Hemiptera: Pseudococcidae) as a representative surface pest on banana and evaluated whether EF fumigation is compatible with the current packaging materials and loading ratio used by the South Korean banana import industry. Fumigation trials with P. citri adults and eggs showed that EF was at least as effective as MB at the EF and MB concentrations recommended under the current phytosanitary disinfestation guidelines. From a logistical standpoint, EF gas could not effectively penetrate plastic bagging typically used in banana cartons during commercial shipping. Also, EF sorption to bananas and packing materials (used as cushioning inside bagging) further lowered realized EF concentrations around bananas. These reductions in EF concentration translated into reduced mortality of P. citri eggs, indicating that despite similar efficacy of MB and EF for controlling P. citri, further consideration and optimization of other industry logistics such as packaging and loading ratio is necessary to enhance the adoption of this MB alternative.

Plant Growth Regulators Improve the Production of Volatile Organic Compounds in Two Rose Varieties.

The study focused on the influence of the plant growth regulators (PGRs) benzyladenine (BA) and naphthalene acetic acid (NAA) on the production of volatile organic compounds (VOCs) from the flowers of two modern rose varieties, Hybrid Tea and Floribunda. Thirty-six plants of Hybrid Tea and Floribunda were tested. Benzyladenine and naphthalene acetic acid were applied at 0, 100 and 200 mg/L to both rose varieties. Gas chromatography, coupled with flame ionization detection and mass spectrometry, was used to analyze and identify the volatile organic compounds from the flowers. A three-phase fiber 50/30 µm divinylbenzene/carboxen/polydimethylsiloxane was used to capture VOCs, at 2, 4 and 8 weeks, and 4 weeks was selected as it had the highest peak area. In total, 81 and 76 VOCs were detected after treatment of both rose varieties with BA and NAA, respectively. In addition, 20 compounds, which had significant differences between different treatments, were identified from both rose varieties. The majority of VOCs were extracted after the application of 200 mg (BA and NAA) /L of formulation, and four important compounds, cis-muurola-4(141)5-diene, y-candinene, y-muurolene and prenyl acetate, increased significantly compared to the controls. These compounds are commercially important aroma chemicals. This study used the rapid and solvent-free SPME method to show that BA and NAA treatments can result in significant VOC production in the flowers of two rose varieties, enhancing the aromatic value of the flowers. This method has the potential to be applied to other valuable aromatic floricultural plant species.

Comparison of the Efficacy and Phytotoxicity of Phosphine and Ethyl Formate for Controlling Pseudococcus longispinus (Hemiptera: Pseudococcidae) and Pseudococcus orchidicola on Imported Foliage Nursery Plants.

The fumigation activity of phosphine (PH3) and ethyl formate (EF) and their phytotoxicity to 13 imported foliage nursery plant species were evaluated. The lethal concentration and time (LCT99) values of the PH3 indicated that the susceptibility of the nymphs (3.95 and <0.45 mg·h/liter, respectively) was higher than that of the adults (5.29 and 3.66 mg·h/liter, respectively) of two mealybugs [Pseudococcus longispinus (Targioni-Tozzetti) and P. orchidicola Takahashi]. The highest concentration reduction rate of PH3 and EF on the 13 foliage nursery plants in the 12-liter desiccator was 41.5% for Heteropanax fragrans and 71.7% for Schefflera arboricola, respectively, which indicates that PH3 has a lower sorption rate than EF. The phytotoxicities of PH3-treated foliage nursery plants did not significantly differ from those of the nontreated plants, but EF caused phytotoxicity in 11 foliage nursery plants a week after treatment. When the exposure time of PH3 increased to 24 h, the adults and nymphs of both mealybug species showed 100% mortality in the 0.5 m3 fumigation chamber. In the 10 m3 fumigation container used in the field, there was 100% mortality of both mealybugs after treatment with 2 g/m3 PH3 for 24 h at 16°C. These results indicate that EF is not a suitable mealybug fumigant due to its high sorption and phytotoxicity to foliage nursery plants, despite fumigation activity against the two species. However, PH3 seems to be suitable for mealybug fumigation in foliage nursery plants and can be used as a substitute for methyl bromide.

Minimization of energy transduction confers resistance to phosphine in the rice weevil, Sitophilus oryzae.

Infestation of phosphine (PH3) resistant insects threatens global grain reserves. PH3 fumigation controls rice weevil (Sitophilus oryzae) but not highly resistant insect pests. Here, we investigated naturally occurring strains of S. oryzae that were moderately resistant (MR), strongly resistant (SR), or susceptible (wild-type; WT) to PH3 using global proteome analysis and mitochondrial DNA sequencing. Both PH3 resistant (PH3-R) strains exhibited higher susceptibility to ethyl formate-mediated inhibition of cytochrome c oxidase than the WT strain, whereas the disinfectant PH3 concentration time of the SR strain was much longer than that of the MR strain. Unlike the MR strain, which showed altered expression levels of genes encoding metabolic enzymes involved in catabolic pathways that minimize metabolic burden, the SR strain showed changes in the mitochondrial respiratory chain. Our results suggest that the acquisition of strong PH3 resistance necessitates the avoidance of oxidative phosphorylation through the accumulation of a few non-synonymous mutations in mitochondrial genes encoding complex I subunits as well as nuclear genes encoding dihydrolipoamide dehydrogenase, concomitant with metabolic reprogramming, a recognized hallmark of cancer metabolism. Taken together, our data suggest that reprogrammed metabolism represents a survival strategy of SR insect pests for the compensation of minimized energy transduction under anoxic conditions. Therefore, understanding the resistance mechanism of PH3-R strains will support the development of new strategies to control insect pests.

Efficacy and Phytotoxicity of Phosphine as Fumigants for Frankliniella occidentalis (Thysanoptera: Thripidae) on Asparagus.

The insecticidal activity of phosphine (PH3) and ethyl formate (EF) toward Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) and their phytotoxicity to asparagus were evaluated. Both the PH3 and EF fumigants showed higher lethal concentration and time (LCT) values at lower temperatures. The LCT99 values of PH3 and EF at 5°C in a 12 liters desiccator for 4 h showed the following ranking: eggs (64.69 mg·h/liter for PH3 and EF indicating phytotoxicity to asparagus), nymphs (5.54 and 17.48 mg·h/liter, respectively), and adults (3.83 and 14.67 mg·h/liter, respectively). The adsorption of PH3 was approximately 11% at 2°C and 13% at 5°C, whereas the adsorption of EF increased sharply to 88% at 2°C and 97% at 5°C. The hatching rate of F. occidentalis eggs was approximately 95% at all locations (top, middle, and bottom) in the presence of 4 mg/liter PH3 at 5°C in a 0.65-m3 fumigation chamber for 24 h. However, extension of the treatment to 48 h resulted in 100% inhibition of egg hatching. The atmospheric level of PH3 decreased below the threshold limit value after 80 min, and phytotoxicity was not observed. The results revealed that EF is highly absorbed by asparagus and is not suitable as a fumigant, but PH3 is a suitable alternative to the fumigant methyl bromide for the control of western flower thrips in asparagus.

Fumigation Activity of Ethyl Formate and Phosphine Against Tetranychus urticae (Acari: Tetranychidae) on Imported Sweet Pumpkin.

This study investigated an alternative fumigant for imported sweet pumpkin. Laboratory fumigations with ethyl formate (EF) and phosphine (PH3) alone and in combination were conducted at different temperatures in 12-liter desiccators to determine their efficacy and synergism against adults and eggs of two-spotted spider mites (Tetranychus urticae Koch), a major pest of sweet pumpkins; larger scale fumigations with EF were also performed to evaluate the impact of EF fumigation on the postharvest quality of sweet pumpkin and EF absorption. EF fumigation of 4 h achieved 100% control of both adults and eggs at 10°C. Efficacy of EF also increased with temperature. Conversely, PH3 fumigation for 4 and 6 h was not effective against either adults or eggs, and no synergism between EF and PH3 was observed. In larger scale fumigation tests, EF at 60 g/m3 for 4 h resulted in 100% control of both adults and eggs at 10°C, without causing injury to the sweet pumpkins. These results suggest that EF has the potential to be a safe and effective alternative to methyl bromide fumigation for controlling two-spotted spider mites on sweet pumpkin.