The spotted wing drosophila, Drosophila suzukii Matsumura (Diptera: Drosophilidae), distributes in most soft‐skinned fruit areas in China, is an economically important pest of fresh cherries during Sino‐European trade and is considered a quarantine pest in A2 list by European and Mediterranean Plant Protection Organization. Loop‐mediated isothermal amplification (LAMP) is an accurate, quick and convenient molecular identification method, applied to distinguish D. suzukii from other fruit flies. This method can be used for inspection in the field, and at the points of entry (POEs), because its results can be detected with the naked eye due to colour changes. In our study, first, we reported a simple and fast LAMP colorimetric detection method for molecular identification of D. suzukii. We designed primer sets for D. suzukii based on mitochondrial cytochrome oxidase I (COI) sequences. The specificity and sensitivity of the primers were tested by using target and non‐target fruit flies in the family Drosophilidae and Tephritidae, usually intercepted during the Sino‐European cherry trade. Second, for detection in the field and at POEs, the adoption of a quick DNA extraction method could save the total time of LAMP identification to about 90 min. Taken together, this accurate, quick and convenient LAMP‐based colorimetric identification assay combined with a quick DNA extraction method could visually detect clearly with just one portable heating device, which will be useful for rapid on‐site identification and inspection for D. suzukii by the quarantine department.
The spotted wing drosophila, Drosophila suzukii Matsumura, which is widely spread in the main soft-skinned fruits production areas in China, presents a threat to importing countries. In order to develop a phytosanitary cold treatment measure for preventing the movement of this drosophila fly, cold tolerance of six immature life stages of D. suzukii was compared followed by time-mortality and large-scale confirmatory tests on the most tolerant stage in grape fruit. Egg was defined as the most cold-tolerant stage by comparing the mortality of all the immature stages (egg, first, second, and third instars, early and late pupa) treated at 0 and 2°C. The minimal lethal time (LT) for 99.9968% mortality (95% confidence level [CL]) estimated by the probit model was 10.47 d at 0°C and 11.92 d at 2°C, respectively. Hence, 11 d (at 0°C) and 12 d (at 2°C) were chosen as the target time to conduct the confirmatory tests. No survivors were found among the estimated 50,385 and 57,366 treated eggs, which resulted in the efficacy of 99.9941 and 99.9948% mortality (95% CL) at 0 and 2°C, respectively. Our study suggests a technical basis for cold disinfestation on D. suzukii in cage-infested Chinese ‘Red Globe’ (Vitis vinifera L.) grape, which could provide flexible phytosanitary treatment for control of D. suzukii in the international trade of grape.
BACKGROUND Drosophila suzukii (Matsumura) is considered a quarantine pest in the A2 list because it causes serious infection and huge economic losses. Cold and controlled atmosphere treatments have been used to control immature stage pests in fresh fruits. Herein, the basal tolerance response of D. suzukii egg, larva and pupa to cold and hypoxia stress were studied, and underlying transcriptome mechanisms in the larva were pinpointed. RESULTS The third instar was more tolerant than 12‐h‐old egg and 8‐day‐old pupa when treated at 3 °C + 1% O2 for 7 days, with 34.00% ± 5.22% larval survival. Hypoxia influenced the effect of cold treatment on D. suzukii. Larval survival decreased at 3 °C + 1% O2, but increased at 0 °C + 1% O2. Survival increased with temperature between 0 and 5 °C + 1% O2, but decreased significantly at 25 °C + 1% O2. RNA‐sequencing results showed that the Tweedle (Twdl) family was upregulated and uniquely enriched in larvae treated at 3 °C + 1% O2. In addition, RNA interference‐mediated silencing of a key Twdl gene reduced the survival rate after cold and hypoxia treatment. CONCLUSION Hypoxia was able to influence the effect of cold treatment on the survival of D. suzukii positively or negatively. Structural constituents of the chitin‐based cuticle, in particular Twdl genes, body morphogenesis, and ATP synthesis‐coupled proton transport were involved in the tolerance to cold and hypoxia. In future, the Twdl gene could be used as a nanocarrier delivering RNA pesticides to control D. suzukii in the field and so prevent its worldwide spread. © 2023 Society of Chemical Industry.
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