Diabrotica speciosa (Germar) (Coleoptera: Chrysomelidae) is a major pest of common bean (Phaseolus vulgaris L.; Fabales: Fabaceae), and adults can defoliate plants during the whole crop cycle. This study was conducted to evaluate the resistance to D. speciosa in 16 common bean genotypes (14 landraces and 2 cultivars), through three different experiments. In the laboratory, choice and no-choice feeding tests were performed to evaluate the percentage of leaf consumption. In the greenhouse, plant height, numbers of leaves, percentage of injured leaves, percentage of injury per leaf, weight of seeds, and D. speciosa survival were evaluated. Furthermore, trichome density, levels of peroxidase (POD), superoxide dismutase (SOD), and protein content in common bean leaves were assessed. In the laboratory, the genotypes Chumbinho Branco, Dobalde, Manteigado, IPR Tuiuiú, and 90D Mouro were the least consumed by D. speciosa. In the greenhouse, the genotypes Dobalde, Manteigado, and IPR Tuiuiú expressed tolerance to the pest, which was associated with a higher plant height and/or unchanged POD and SOD levels and protein content following insect feeding, and no reduction in seed production. The landrace 90D Mouro exhibited antixenosis and tolerance to D. speciosa, observed as a lower leaf injury, higher trichome density, lower protein contents, higher SOD level and no reduction in seed weight. Overall, we have shown that antixenosis and tolerance can help overcome damages resulting from D. speciosa feeding, with emphasis on four common bean genotypes that may be useful in plant breeding programs aimed at controlling D. speciosa in common bean crops.

Habitat selection of Culicoides spp. (Diptera: Ceratopogonidae) is influenced by the physicochemical factors such as temperature, pH, salinity, moisture, conductivity, organic and inorganic compounds of substrates. These factors determine the life history traits of the vectors. We studied the influence of substrate salinity (0–40 parts per thousand, ppt) and pH (pH 1–13) on oviposition, egg hatching, larval survivability, and adult emergence of Culicoides peregrinus Kieffer under laboratory conditions. Most eggs (80.74%) were laid in 0 ppt and 95% in pH 7 but lowered with increased salinity and pH levels. It was observed that the females did not lay eggs in 30 ppt to 40 ppt salinity; pH 1 and pH 13 but interestingly up to 95% of the eggs were retained within the abdomen. Little effect of salinity and pH on egg hatching was observed up to 5 ppt and 10 ppt except at the extreme values of 40 ppt and pH 1, pH 13. Pupation did not occur in rearing plates with high salinities, 30 ppt and 40 ppt, although the few eggs hatched when exposed to such salinity. In low salinity (0 to 2 ppt), occurrence of adult emergence was more and then decreased with increasing salinity. Maximum emergence was seen when the rearing media was alkaline. This study deals with the suitability of breeding substrate of C. peregrinus when exposed to salinity and pH ranges. Our study suggests the ambient salinity and pH ranges to be maintained during laboratory rearing of this vector species.

Mung bean is highly susceptible to insect attack during storage. Hermetic storage is an effective technique to control insect damage. This study investigated the potential of the hermetic SuperGrain bag (SGB) for controlling bruchids during storage. The dry samples were packed in SGB infested with adult bruchids (SGB-I), SGB natural field infested (SGB-N), woven polypropylene bags (WPP-I and WPP-N) and kept at room temperature for 180 days. Oxygen (O2) and carbon dioxide (CO2) concentrations were measured at 15 days intervals. Moisture content, infestation level, seed damage and weight loss were determined at 60 days intervals. Seed colour, hardness, crude protein and fat contents were analysed before and after storage. The O2 level decreased to 10.09%, whereas the CO2 level increased to 8.87% in both SGB-I and SGB-N treatments. The moisture content of mung bean was maintained as onset storage in both SGB-N and SGB-I treatments, whereas reduced in WPP-N (9.26% db) and WPP-I (9.21% db). In SGB treatments, no significant bruchids were detected, but they increased drastically in WPP-N (52 ± 9) and WPP-I (377 ± 14). Seed damage (2–3%) and weight loss (0.8–1.0%) were recorded in both SGB-N and SGB-I. Conversely, seed damage reached 26.67 and 54.17%, corresponding to weight losses of 12.33 and 20.82% in WPP-N and WPP-I, respectively. Seed colour, hardness, crude protein and fat contents in SGBs showed no significant changes than in the WPP bags. The study illustrated that the SGB is an efficient hermetic device in protecting mung beans against bruchids attacks compared to the WPP bags.

Lipaphis erysimi pseudobrassicae (Davis) and Myzus persicae (Sulzer) are important pests of brassica crops, causing significant yield losses on cabbage in Ghana. To inform the development of ecologically sound and sustainable pest management strategies for these pests, their biological and population growth parameters were studied on three cabbage varieties (Oxylus, Fortune, and Leadercross). The study was conducted in a screen house under ambient conditions at 30 ± 1°C and 75 ± 5% RH and 12:12 h photoperiod from September to November 2020. The parameters of the preadult developmental period, survival rates, longevity, reproduction, and life table were evaluated following the female age-specific life table. There were significant differences in the nymphal developmental time, longevity, and fecundity on the cabbage varieties for both aphid species. The highest population growth parameters, net reproductive rate (R0), intrinsic rate of increase r, and finite rate of increase (λ) were recorded on Oxylus variety for both L. e. pseudobrassicae and M. persicae. The lowest was recorded on Leadercross variety for L.e pseudobrassicae and Fortune for M. persicae. The results from this study suggest that Leadercross is a less suitable host for L. e. pseudobrassicae and Fortune for M. persicae, thus, should be considered as less susceptible varieties for use in primary pest management by small-scale farmers or as a component of an integrated pest management strategy for these pests on cabbage.

The role of bees in the environment, economic, biodiversity and pharmaceutical industries is due to its social behavior, which is oriented from the brain and hypopharyngeal gland that is the center of royal jelly (RJ) production. Limited studies have been performed on the head gene expression profile at the RJ production stage. The aim of this study was to compare the gene expressions in 9 and 1-day-old (DO) honeybee workers in order to achieve better understanding about head gene expression pattern. After sequencing of RNAs, transcriptome and their networks were compared. The head expression profile undergoes various changes. 1662 gene transcripts had differential expressions which 1125 and 537 were up and down regulated, respectively, in 9_DO compared with 1_DO honey bees. The day 1th had more significant role in the expression of genes related to RJ production as major RJ protein 1, 2, 3, 5, 6 and 9 encoding genes, but their maximum secretion occurred at day 9th. All process related to hypopharyngeal glands activities as CYP450 gene, fatty acid synthase gene, vitamin B6 metabolism and some of genes involved in fatty acid elongation and degradation process had an upward trend from 1_DO and were age-dependent. By increasing the age, the activity of pathways related to immune system increased for keeping the health of bees against the chemical compound. The expression of aromatic amino acid genes involved in Phenylalanine, tyrosine and tryptophan biosynthesis pathway are essential for early stage of life. In 9_DO honeybees, the energy supplying, reducing stress, protein production and export pathways have a crucial role for support the body development and the social duties. It can be stated that the activity of honeybee head is focused on energy supply instead of storage, while actively trying to improve the level of cell dynamics for increasing the immunity and reducing stress. Results of current study identified key genes of certain behaviors of honeybee workers. Deeper considering of some pathways will be evaluated in future studies.

Major efforts to control the population of Aedes aegypti mosquitoes involve the use of synthetic insecticides, which can be harmful to the environment. Most plant compounds are eco-friendly and some of them have biocontrol potential, whereas a fraction of these compounds is released into the environment through the leaf-leaching process. We evaluated the effects of secondary compounds from Ateleia glazioviana and Eucalyptus grandis senescent leaf leachates on Ae. aegypti larval mortality, adult emergence time, and wing size using a microcosm approach. The microcosms consisted of 10 larvae kept in water (control) and under four treatments with leachates from a combination of plant species and leaching time (7 or 14 days). Chemical analyses of the leachates showed the presence of carboxaldehyde and Heptatriocotanol, which have antimicrobial properties, potentially reducing the food available for larvae. β-Sitosterol, Stigmasterol, α-Amyrin, and Lupeol are compounds with inhibitory, neurotoxic, and larvicidal effects. Both plant species’ leachates increased larval mortality and decreased emergence time due to the presence of compounds toxic to the larvae. Larger organisms emerged in treatments with 7-days leachates, likely due to the high concentration of dissolved organic matter in the leachates. The higher mortality in 7-days leachates may also increase the organic matter from co-specific decomposition, improving adult size. Therefore, if the mosquito population is not locally extinct, compounds present in leaf leachates may act as a resource enhancing larvae growth, potentially increasing survivors’ fitness. In conclusion, biocontrol attempts using urban green spaces may have unexpected outcomes, such as resulting in larger pest organisms.

We quantified the life table parameters and predation capacity of a generalist predatory mite, Typhlodromus bagdasarjani Wainstein and Arutunjan on five monotypic diets, including Tetranychus urticae Koch (TSSM) eggs in the presence (SW) and absence (SN) of webs, Trialeurodes vaporariorum Westwood (GHWF) eggs (G), honeydew (H), and maize pollen (M) as well as three mixed diets, including SN + M, SN + G, and G + M. Our results showed that the individuals fed on the mixed diets had a considerably shorter developmental time and pre-oviposition period (APOP), higher oviposition days, higher fecundity and population growth rate than those raised on the monotypic diets. Furthermore, we found that the mixed diet of TSSM and GHWF eggs was the most favorable diet, resulted in the highest fecundity and population growth rate, shortest developmental time and APOP. While TSSM eggs alone in the presence of webs and honeydew were the worst diets resulted in the longest developmental time, lower oviposition day, higher fecundity and population growth rate. Our data determined that TSSM has more nutritional benefits than GHWF for T. bagdasarjani. We observed the positive effects of pollen addition to prey on the predatory mite's immature and adult life-history characters; however, it reduced the predation rate. Overall, maize pollen could enhance ecosystem services provided against spider mites and whiteflies by positively impacting the increase of T. bagdasarjani population. This predator may be more effective when two prey species are available than when only one species is present.

Two thrips, Megalurothrips usitatus (Bagnall) and Frankliniella intonsa (Trybom) are major pests of cowpea in South China. To realistically compare the growth, development and reproductive characteristics of these two thrips species, we compared their age-stage, two-sex life tables on cowpea pods under summer and winter natural environmental regimes. The results showed that the total preadult period of M. usitatus was 8.09 days, which was significantly longer than that of F. intonsa (7.06 days), while the adult female longevity of M. usitatus (21.14 days) was significantly shorter than that of F. intonsa (25.77 days). Significant differences were showed in male adult longevity (10.68 days for F. intonsa and 16.95 days for M. usitatus) and the female ratio of offspring (0.67 for F. intonsa and 0.51 for M. usitatus), and the total preadult period of M. usitatus (16.20 days) was significantly longer than that of F. intonsa (13.66 days) in the winter regime. The net reproductive rate (summer: R0 = 85.62, winter: R0 = 105.22), intrinsic rate of increase (summer: r = 0.3020 day−1, winter: r = 0.2115 day−1), finite rate of increase (summer: λ = 1.3526 day−1, winter: λ = 1.2356 day−1) and gross reproduction rate (summer: GRR = 139.34, winter: GRR = 159.88) of F. intonsa were higher than those of M. usitatus (summer: R0 = 82.91, r = 0.2741, λ = 1.3155, GRR = 135.71; winter: R0 = 80.62, r = 0.1672, λ = 1.1820, GRR = 131.26), and the mean generation times (summer: T = 14.73 days, winter: T = 22.01 days) of F. intonsa were significantly shorter than those of M. usitatus (summer: T = 16.11 days, winter: T = 26.25 days). These results may contribute to a better understanding of the bioecology of different thrips species, especially the interspecific competition between two economically important cowpea thrips with the same ecological niche in a changing environment.

Understanding the development and reproduction of insects surviving controlled atmosphere treatment may help in developing sound pest management strategies. The developmental duration, survival percentage, and oviposition of Plodia interpunctella and its F1 generation were determined after the fifth instar larvae (the last-stage larvae) were exposed to 98% N2 for different exposure times. The survival percentage of the last-stage larvae treated with 98% N2 for 6, 4, 1.5, and 0 day was 70, 80, 91, and 100%, respectively when measured 24 h after treatment. The survival percentage of the last-stage larvae that developed to pupae was 37, 55, 73, and 96%, corresponding to the different exposure times. The developmental time needed to pass from pupa to adult emergence of specimens treated as the last-stage larvae were 8, 7, 6, and 6 days corresponding respectively to high N2 treatment after 6, 4, 1.5, and 0 day of exposure. The mean number of eggs laid by the subsequent females developed from the treated last-stage larvae was 35, 66, 81, and 123, respectively. The oviposition inhibition ratio of the F1 generation decreased by more than 33% compared with that of the parental generation. When the last-stage larvae were exposed to 98% N2 for longer than 4 days, the immature developmental time of surviving individuals in the F1 generation was delayed more than 6 days due to slower egg hatching and longer development of the first and second instar larvae stages. The population trend index of the F1 generation was lower when raised from the treated last-stage larvae than those from untreated controls.

The honey bee is associated with a diverse community of microbes (viruses, bacteria, fungi, and protists), commonly known as the microbiome. Here, we present data on honey bee microbiota from two localities having different surrounding landscapes – mountain (the Rhodope Mountains) and lowland (the Danube plain). The bacterial communities of abdomen of adult bees were studied using amplicon sequencing of the 16S rRNA gene. The composition and dominance structure and their variability within and between localities, alpha and beta diversity, and core and differential taxa were compared at different hierarchical levels (operational taxonomic units to phylum). Seven genera (Lactobacillus, Gilliamella, Bifidobacterium, Commensalibacter, Bartonella, Snodgrassella, and Frischella), known to include core gut-associated phylotypes or species clusters, dominated (92–100%) the bacterial assemblages. Significant variations were found in taxa distribution across both geographical regions and within each apiary. Lactobacillus (Firmicutes) prevailed significantly in the mountain locality followed by Gilliamella and Bartonella (Proteobacteria). Bacteria of four genera, core (Bartonella and Lactobacillus) and non-core (Pseudomonas and Morganella), dominated the bee-associated assemblages of the Danube plain locality. Several ubiquitous bacterial genera (e.g., Klebsiella, Serratia, and Providencia), some species known also as potential and opportunistic bee pathogens, had been found in the lowland locality. Beta diversity analyses confirmed the observed differences in the bacterial communities from both localities. The occurrence of non-core taxa contributes substantially to higher microbial richness and diversity in bees from the Danube plain locality. We assume that the observed differences in the microbiota of honey bees from both apiaries are due to a combination of factors specific for each region. The surrounding landscape features of both localities and related vegetation, anthropogenic impact and land use intensity, the beekeeping management practices, and bee health status might all contribute to observed differences in bee microbiota traits.

Fruit fly infestation is one of the main obstacles to the exportation of fresh agricultural produce. Films of mineral particles and biomaterials have the potential to protect fruits against tephritid fruit fly infestation. The present study evaluated the effects of particle films on the tritrophic interactions of grape (Vitis vinifera L.), the fruit fly Ceratitis capitata (Wiedemann) and the parasitoid Diachasmimorpha longicaudata (Ashmead) under semi-field conditions. Grapes were biometrically characterised (i.e. colour, firmness, mass, length and diameter), treated with mineral particles, biomaterials or distilled water (control), and then used in oviposition and parasitism bioassays. In the oviposition bioassay, the treated grapes were exposed to 50 C. capitata pairs in field cages, and after 48 h, the punctures and eggs on each fruit were counted. In the parasitism bioassay, treated grapes were artificially infested with third-instar C. capitata larvae (two per fruit), exposed (2 h) to 50 D. longicaudata pairs in field cages to determine parasitism index, larval and pupal viabilities and number of flies and parasitoids emerged. Treatment with the mineral film affected fruit colour and reduced C. capitata oviposition but failed to significantly affect the parasitism capacity of D. longicaudata. The ability of the parasitoid to locate and parasitise C. capitata larvae in kaolin-coated fruits suggests that kaolin films could be used in conjunction with biological agents to control fruit flies.

Cytochrome P450 proteins (CYPs) in insects can encode various detoxification enzymes and catabolize heterologous substances, conferring tolerance to insecticides. This study describes the identification of a P450 gene (CYP6BQ8) from Tribolium castaneum (Herbst) and investigation of its spatiotemporal expression profile and potential role in the detoxification of terpinen-4-ol, a component of plant essential oils. The developmental expression profile showed that TcCYP6BQ8 expression was relatively higher in early- and late-larval stages of T. castaneum compared with other developmental stages. Tissue expression profiles showed that TcCYP6BQ8 was mainly expressed in the head and integument of both larvae and adults. The expression profiling of TcCYP6BQ8 in developmental stages and tissues is closely related to the detoxification of heterologous substances. TcCYP6BQ8 expression was significantly induced after exposure to terpinen-4-ol, and RNA interference against TcCYP6BQ8 increased terpinen-4-ol-induced larval mortality from 47.78 to 66.67%. This indicates that TcCYP6BQ8 may be involved in T. castaneum's metabolism of terpinen-4-ol. Correlation investigation between the CYP6BQ8 gene and terpinen-4-ol resistance in T. castaneum revealed that the TcCYP6BQ8 gene was one of the factors behind T. castaneum's resistance to terpinen-4-ol. This discovery may provide a new theoretical foundation for future regulation of T. castaneum.

The silkworm Bombyx mori (Lepidoptera: Bombycidae) is a lepidopteran model insect of great economic importance. The parasitoid Exorista sorbillans (Diptera, Tachinidae) is the major pest of B. mori and also a promising candidate for biological control. However, the molecular interactions between hosts and dipteran parasitoids have only partially been studied. Gene expression analysis by reverse-transcription quantitative real-time polymerase chain reaction (RT-qPCR) is indispensable to characterise their interactions. Accurate normalisation of RT-qPCR-based gene expression requires the use of reference genes that are constantly expressed irrespective of experimental conditions. In this study, the expression stability of 13 traditionally used reference genes was estimated by five statistical algorithms (ΔCt, geNorm, Normfinder, BestKeeper, and RefFinder) to determine the best reference genes for gene expression studies in different tissues of B. mori under E. sorbillans parasitism. Specifically, TATA-box-binding protein was the best reference gene in epidermis and testis, while elongation factor 1α was the most stable gene in prothoracic gland and midgut. Elongation factor 1γ, ribosomal protein L3, actin A1, ribosomal protein L40, glyceraldehyde-3-phosphate dehydrogenase and eukaryotic translation initiation factor 4A were the most suitable genes in head, silk gland, fat body, haemolymph, Malpighian tubule and ovary, respectively. Our study offers a set of suitable reference genes for gene expression normalisation in B. mori under the parasitic stress of E. sorbillans, which will benefit the in-depth exploration of host-dipteran parasitoid interactions, and also provide insights for further improvements of B. mori resistance against parasitoids and biocontrol efficacy of dipteran parasitoids.

DNA Barcoding is an important tool for disciplines such as taxonomy, phylogenetics and phylogeography, with Barcode of Life Data System (BOLD) being the largest database of partial cytochrome c oxidase subunit I (COI) sequences. We provide the first extensive revision of the information available in this database for the insect order Thysanoptera, to assess: how many COI sequences are available; how representative these sequences are for the order; and the current potential of BOLD as a reference library for specimen identification and species delimitation. The COI database at BOLD currently represents only about 5% of the over 6400 valid thrips species, with a heavy bias towards a few species of economic importance. Clear Barcode gaps were observed for 24 out of 33 genera evaluated, but many outliers were also observed. We suggest that the COI sequences available in BOLD as a reference would not allow for accurate identifications in about 30% of Thysanoptera species in this database, which rises to 40% of taxa within Thripidae, the most sampled family within the order. Thus, we call for caution and a critical evaluation in using BOLD as a reference library for thrips Barcodes, and future efforts should focus on improving the data quality of this database.

Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), is a major global pest of fruits. Currently, the sequential male annihilation technique, followed by the sterile insect technique has been used to significantly reduce the population of feral males in this species. However, issues with sterile males being killed by going to male annihilation traps have reduced the efficacy of this approach. The availability of males that are non-methyl eugenol-responding would minimize this issue and increase the efficacy of both approaches. For this, we recently established two separate lines of non-methyl eugenol-responding males. These lines were reared for 10 generations and in this paper, we report on the assessment of males from these lines in terms of methyl eugenol response and mating ability. We saw a gradual decrease in non-responders from ca. 35 to 10% after the 7th generation. Despite that, there were still significant differences until the 10th generation in numbers of non-responders over controls using laboratory strain males. We did not attain pure isolines of non-methyl eugenol-responding males, so we used non-responders from the 10th generation of those lines as sires to initiate two reduced-responder lines. Using these reduced responder flies, we found that there was no significant difference in mating competitiveness when compared with control males. Overall, we suggest that it may be possible to establish lines of low or reduced responder males to be used for sterile release programs, that could be applied until the 10th generation of rearing. Our information will contribute to the further development of an increasingly successful management technique incorporating the use of SIT alongside MAT to contain wild populations of B. dorsalis.

Liriomyza trifolii is a significant pest of vegetable and ornamental crops across the globe. Microwave radiation has been used for controlling pests in stored products; however, there are few reports on the use of microwaves for eradicating agricultural pests such as L. trifolii, and its effects on pests at the molecular level is unclear. In this study, we show that microwave radiation inhibited the emergence of L. trifolii pupae. Transcriptomic studies of L. trifolii indicated significant enrichment of differentially expressed genes (DEGs) in ‘post-translational modification, protein turnover, chaperones’, ‘sensory perception of pain/transcription repressor complex/zinc ion binding’ and ‘insulin signaling pathway’ when analyzed with the Clusters of Orthologous Groups, Gene Ontology and the Kyoto Encyclopedia of Genes and Genomes databases, respectively. The top DEGs were related to reproduction, immunity and development and were significantly expressed after microwave radiation. Interestingly, there was no significant difference in the expression of genes encoding heat shock proteins or antioxidant enzymes in L. trifolii treated with microwave radiation as compared to the untreated control. The expression of DEGs encoding cuticular protein and protein takeout were silenced by RNA interference, and the results showed that knockdown of these two DEGs reduced the survival of L. trifolii exposed to microwave radiation. The results of this study help elucidate the molecular response of L. trifolii exposed to microwave radiation and provide novel ideas for control.

Different species of Cyclocephala scarab beetles (Scarabaeidae, Dynastinae) perform key functional roles in both natural and agricultural systems, such as the cycling of organic matter and pollination, while also being known as destructive pests both as immatures and adults. Therefore, the identification of biological parameters is crucial for defining strategies for their conservation and efficient pest management. In a forest fragment within the Brazilian Atlantic Forest biodiversity hotspot, we field-captured adult individuals of Cyclocephala cearae, C. celata, and C. paraguayensis then reared and bred them under controlled temperature and humidity conditions. On a daily basis, we individually weighted eggs of all three species, from oviposition until hatching, and monitored egg development parameters (i.e., incubation duration, viability, and egg weight increase). Our findings provide novel empirical evidence showing (i) a positive correlation between egg weight and incubation duration, (ii) idiosyncratic characteristics on egg development, and (iii) a negative (involuntary) effect of manipulation on egg development and viability. Thus, the successful breeding and rearing of Cyclocephala spp. is correlated with egg integrity and the targeted species. Our analyses present a quantitative understanding of the egg phase and can assist in refining strategies for ovicidal activity and pest management of Cyclocephala spp. in agriculture systems. Moreover, they can provide a basis for new studies related to captivity breeding, pollinator management, and developmental biology for biodiversity conservation.

Arthropod species diversity enhances ecosystem productivity and sustainability by increasing pollination and biological control services. Although, it is declining rapidly due to conventional agricultural intensification, organic agriculture with reduced reliance on agronomic inputs can regenerate ecosystems' resilience and restore them. Here, we report whether hexapod communities differ on both types of farming systems in small-scale field plot experiments, wherein Maize variety AG-589 was grown organically and conventionally in the 2020 and 2021 seasons. Livestock manure was applied in organic fields, whereas nitrogen and phosphorous were used as synthetic fertilizers in conventional fields. Hexapods were sampled three weeks after sowing once a week from the middle rows of subplots from both organically and conventionally grown maize. Twelve species of herbivores and four species of predators were recorded. Hexapod abundance overall and that of herbivores only was higher in conventionally cultivated maize, while predator abundance was higher in organic maize. Herbivores species diversity and evenness were significantly higher in conventional maize. Predator species diversity and evenness were significantly higher in organic maize fields. We noted predator abundance, diversity, and evenness as strong predictors to lower herbivore populations. These findings suggest that organic farming conserves natural enemies’ biodiversity and regulates herbivores with increased provision of suitable habitats and prey resources for natural enemies, leading to enhanced relative abundance in their specialized niches. Thus, organic agriculture can potentially mediate better ecosystem services.

Insect response to cold stress is often associated with adaptive strategies and chemical variation. However, low-temperature domestication to promote the cold tolerance potential of Bactrocera dorsalis and transformation of main internal substances are not clear. Here, we use a series of low-temperature exposure experiments, supercooling point (SCP) measurement, physiological substances and cryoprotectants detection to reveal that pre-cooling with milder low temperatures (5 and 10°C) for several hours (rapid cold hardening) and days (cold acclimation) can dramatically improve the survival rate of adults and pupae under an extremely low temperature (−6.5°C). Besides, the effect of rapid cold hardening for adults could be maintained even 4 h later with 25°C exposures, and SCP was significantly declined after cold acclimation. Furthermore, content of water, fat, protein, glycogen, sorbitol, glycerol and trehalose in bodies were measured. Results showed that water content was reduced and increased content of proteins, glycogen, glycerol and trehalose after two cold domestications. Our findings suggest that rapid cold hardening and cold acclimation could enhance cold tolerance of B. dorsalis by increasing proteins, glycerol, trehalose and decreasing water content. Conclusively, identifying a physiological variation will be useful for predicting the occurrence and migration trend of B. dorsalis populations.