Tuta absoluta (Meyrick), a key pest of tomato, is quickly spreading over the world and biological control is considered as one of the control options. Worldwide more than 160 species of natural enemies are associated with this pest, and an important challenge is to quickly find an effective biocontrol agent from this pool of candidate species. Evaluation criteria for control agents are presented, with the advantages they offer for separating potentially useful natural enemies from less promising ones. Next, an aggregate parameter for ranking agents is proposed: the pest kill rate km. We explain why the predator's intrinsic rate of increase cannot be used for comparing the control potential of predators or parasitoids, while km can be used to compare both types of natural enemies. As an example, kill rates for males, females and both sexes combined of three Neotropical mirid species (Campyloneuropsis infumatus (Carvalho), Engytatus varians (Distant) and Macrolophus basicornis (Stål)) were determined, taking all life-history data (developmental times, survival rates, total nymphal and adult predation, sex ratios and adult lifespan) into account. Based on the value for the intrinsic rate of increase (rm) for T. absoluta and for the kill rate km of the predators, we predict that all three predators are potentially able to control the pest, because their km values are all higher than the rm of the pest. Using only km values, we conclude that E. varians is the best candidate for control of T. absoluta on tomato, with C. infumatus ranking second and M. basicornis last.

Immature fruit fly stages of the family Tephritidae are commonly intercepted on breadfruit from Pacific countries at the New Zealand border but are unable to be identified to the species level using morphological characters. Subsequent molecular identification showed that they belong to Bactrocera xanthodes, which is part of a species complex that includes Bactrocera paraxanthodes, Bactrocera neoxanthodes and an undescribed species. To establish a more reliable molecular identification system for B. xanthodes, a reference database of DNA barcode sequences for the 5’-fragment of COI gene region was constructed for B. xanthodes from Fiji, Samoa and Tonga. To better understand the species complex, B. neoxanthodes from Vanuatu and B. paraxanthodes from New Caledonia were also barcoded. Using the results of this analysis, real-time TaqMan polymerase chain reaction (PCR) assays for the detection of B. xanthodes complex and for the three individual species of the complex were developed and validated. The assay showed high specificity for the target species, with no cross-reaction observed for closely related organisms. Each of the real-time PCR assays is sensitive, detecting the target sequences at concentrations as low as ten copies µl−1 and can be used as either singleplex or multiplex formats. This real-time PCR assay for B. xanthodes has been successfully applied at the borders in New Zealand, leading to the rapid identification of intercepted Tephritidae eggs and larvae. The developed assays will be useful biosecurity tools for rapid detection of species in the B. xanthodes complex worldwide.

Parasitoids, released in augmentative biological control programmes, which display a rapid host-location capacity, have a higher likelihood of successfully controlling target pest species. By learning to associate sensory cues to a suitable oviposition site, might parasitoids used as biological control agents, locate hosts more rapidly, and perhaps increase the efficacity of e.g. Tephritidae fruit fly management. We studied associative learning of Fopius arisanus (Hymenoptera: Braconidae) and tested its range of learning in natural and conditional hosts and host fruits, i.e. Bactrocera dorsalis, Zeugodacus cucurbitae, Ceratitis capitata and Ceratitis cosyra (Diptera: Tephritidae) and on fruits (papaya, tomato, banana). Naïve female F. arisanus were compared with experienced wasps, which had been offered infested and non-infested fruit, and been allowed to oviposit. Preferences for olfactory cues from infested fruits were thereafter assessed in a two-choice olfactometer. Naïve and trained parasitoids preference differed in general and non-responders to infested fruits were higher among naïve parasitoids. The trained wasps preferred the fruit infested in the training more than the control fruit, for all combination, except when C. cosyra infested the fruits, hence avoidance behavioural response was observed towards the odour of the infested fruit. Fopius arisanus was capable of behaviourally respond to the learned information, e.g. associative odour learning was achieved, yet limited depending on interaction level, fruit fly and fruit combination. To create F. arisanus preference of an associated odour, it might hence be needed to ensure oviposition in perceived suitable host and host fruit, for the parasitoid learning to become favourable in a biological control setup.

Seedbank control has been the cornerstone of agricultural management practices. Regulating weeds by using their predators, as a weed control strategy, may be a prerequisite of decreasing herbicide use, and has thus attracted much research investigating the possible contribution of both vertebrates and invertebrates as weed seed predators. Carabid beetles are considered as one of the most important invertebrate seed predators in agroecosystems. We aimed at investigating carabid beetle preferences to a single prey type, seeds of Viola arvensis. We measured the consumption of seeds in 28 species of carabid beetles, under controlled experimental conditions. Two main tribes are identified in tested species, Harpalini with 12 species and Pterostichini with ten species. We found no relationships between species body mass and Viola’s seed consumption, nor with the ratio between mandible length and labrum width (ML/LW). However, trends became significant with the ratio ML/LW when restricting these analyses to species that ate at least five seeds. In addition, we detected a positive and significant relationship between consumption rate and occurrence in trapping sessions over the last 3 years. These results are in favor of weed seeds control by carabids. Clear limits of this study are the use of a single seed species and under control conditions. This experimentation calls for additional studies to check for consistencies in consumption against seed species.

The Ceratitis FAR complex (Diptera, Tephritidae) includes four economically important frugivorous flies (Ceratitis anonae, Ceratitis fasciventris, Ceratitis quilicii, Ceratitis rosa) whose immature stages and adult females cannot be properly resolved through morphological identification. In order to develop a simplified molecular tool for the identification of two of these species (C. rosa, C. quilicii), we selected a subset of six microsatellite markers out of a panel of 16 loci that were previously developed for the molecular differentiation of the taxa within the complex. These six markers were first tested in silico and then used for the actual genotyping of C. quilicii and C. rosa, resulting in the correct identification of all male reference specimens. Here, we propose an integrated morphological and molecular setup for the identification of the four species of the FAR complex. The decision map relies on preliminary DNA barcoding or morphological identification (when possible) to exclude species not belonging to the complex followed by (a) morphological identification of all adult male specimens and female C. anonae, (b) molecular identification via a panel of 16 microsatellite markers for immature stages, damaged vouchers and samples potentially including adult female C. fasciventris/C. quilicii/C. rosa and (c) molecular identification via a reduced panel of six microsatellite markers for samples including only C. quilicii and C. rosa. This simplified diagnostic setup was profitably implemented in the framework of the ERAfrica fruit fly project and will help correctly identify species within the FAR complex for their early detection and monitoring.

Populations of the recently described black oak gall wasp, Zapatella davisae Buffington (Hymenoptera: Cynipidae), have been identified as the cause of extensive tree damage and mortality to black oaks, Quercus velutina Lamarck (Fagales: Fagaceae), in the northeastern United States. Relatively little is known, however, about the distribution, phylogenetic placement, and lifecycle of this important tree pest. Therefore, we conducted next-generation sequencing using the Ion Torrent™ PGM (ThermoFisher Scientific, Inc.) platform to develop genomic resources for the study of Z. davisae and for other closely related species of oak gall wasps. Individual sequence reads were aligned, assembled into unique contigs, and the contigs were then utilized for the in silico isolation and development of microsatellite markers. In total, we screened 36 candidate microsatellite loci, of which 23 amplified consistently (five polymorphic and 18 monomorphic). We then examined whether the polymorphic loci could be used to infer whether populations of Z. davisae from Cape Cod and Nantucket are sexual or asexual by calculating several metrics of genetic diversity that might indicate the mode of reproduction. These included testing for statistical deviations from Hardy–Weinberg equilibrium (HWE) and for linkage disequilibrium (LD), observations for the presence of the Meselson effect, and by calculating the probability that clonal individuals are more prevalent than would be expected in a randomly mating population. While we found significant deviations from HWE and more clonal individuals than expected, our estimates of the Meselson effect were inconclusive due to limited sampling, and we found no evidence of LD. Therefore, the sexual/asexual status of Z. davisae populations remains uncertain.

Dietary delivery of bacterially expressed double-stranded RNA (dsRNA) has a great potential for management of Leptinotarsa decemlineata. An important first step is to discover possible RNA-interference (RNAi)-target genes effective against larvae, especially the old larvae. In the present paper, five putative Broad-Complex (BrC) cDNAs (Z1-Z4, and Z6) were identified in L. decemlineata. The expression of the five LdBrC isoforms was suppressed by juvenile hormone signaling, whereas the transcription was upregulated by 20-hydroxyecdysone signaling at the fourth (final) instar larval stage. Feeding of bacterially expressed dsBrC (derived from a common fragment of the five LdBrC variants) in the third- and fourth-instar larvae successfully knocked down the target mRNAs. For the fourth-instar LdBrC RNAi hypomorphs, they had a higher larval mortality compared with the controls. Moreover, most dsBrC-fed beetles did not pupate normally. After removal of the apolysed larval cuticle, a miniature adult was found. The adult head, compound eyes, prothorax, mesothorax, metathorax were found on the dorsal view. Distinct adult cuticle pigmentation was seen on the prothorax. The mouthparts, forelegs, midlegs, and hindlegs could be observed on the ventral view of the miniature adults. For the third-instar LdBrC RNAi specimens, around 20% moribund beetles remained as prepupae and finally died. Therefore, LdBrC is among the most attractive candidate genes for RNAi to control the fourth-instar larvae in L. decemlineata.

The sugar beet weevil (SBW), Bothynoderes punctiventris Germar, 1824, is a significant pest in most of Eastern Europe. Here, the SBW is described and its seasonal activity characterized, in terms of its different developmental stages in relation to Julian days (JDs), degree-day accumulations (DDAs), and precipitation, as a key to improving monitoring and forecasting of the pest. The phenology and population characteristics of SBW were investigated in sugar beet fields in eastern Croatia over a 4-year period (2012–2015). By using the degree-day model (lower development threshold of 5°C, no upper development threshold, biofix 1 January), the first emergence of overwintering adults was determined as becoming established when the DDA reached 20. The adult emergence was completed when the DDA reached 428. SBW males emerged first, following which the females dominated the adult population. Overwintering adults were present in the field until early July. In August, adults of the offspring generation began to appear. The eggs laid by the overwintering generation required, on average, 10–15 days to develop into larvae; however, eggs were found in soil samples over a period of 102 days (between JDs 112 and 214). Larvae were present in the soil samples over a period of a maximum of 143 days (the first larvae were established on JD 122 and the last one on JD 265), and pupae were established in the soil over a period of 102 days (between JDs 143 and 245). This study provides important data for understanding SBW population dynamics and developing potential population dynamic models for pest forecasting on a regional scale.

Phenoloxidase (PO) plays a key role in melanin biosynthesis during insect development. Here, we isolated the 2310-bp full-length cDNA of PPO1 from Zeugodacus tau, a destructive horticultural pest. qRT-polymerase chain reaction showed that the ZtPPO1 transcripts were highly expressed during larval–prepupal transition and in the haemolymph. When the larvae were fed a 1.66% kojic acid (KA)-containing diet, the levels of the ZtPPO1 transcripts significantly increased by 2.79- and 3.39-fold in the whole larvae and cuticles, respectively, while the corresponding PO activity was significantly reduced; in addition, the larval and pupal durations were significantly prolonged; pupal weights were lowered; and abnormal phenotypes were observed. An in vitro inhibition experiment indicated that KA was an effective competitive inhibitor of PO in Z. tau. Additionally, the functional analysis showed that 20E could significantly up-regulate the expression of ZtPPO1, induce lower pupal weight, and advance pupation. Knockdown of the ZtPPO1 gene by RNAi significantly decreased mRNA levels after 24 h and led to low pupation rates and incomplete pupae with abnormal phenotypes during the larval-pupal interim period. These results proved that PO is important for the normal growth of Z. tau and that KA can disrupt the development of this pest insect.

In social insects such as the honey bee, the quality of drones at the time of their emergence can affect their maintenance in the colony until maturity. Body mass, wing size and wing asymmetry of emerging honey bee drones were measured and correlated with their life span in the colony and compared between individuals reaching maturity or not. The life span of drones differed among colonies in which they were maintained after emergence but not between colonies in which they were reared. More drones heavier at emergence reached sexual maturity at 15 days and had a longer life span compared with light-weight drones of lower mass. The size and symmetry of drone forewings was not correlated with their life span. Our results suggest that body mass at emergence is a good predictor of drone survival in the colony.

Lysinibacillus sphaericus (Bacillales: Planococcaceae) is a spore-forming bacillus used for the biological control of mosquitoes (Diptera: Culicidae) due to its larvicidal activity determined by various toxins and S-layer protein produced either during sporulation or by the vegetative cell. Aedes aegypti and Culex quinquefasciatus are the vectors of arboviruses that cause tropical diseases representing a current public health problem. Both species may coexist in the same larval development sites and are susceptible to the larvicidal activity of L. sphaericus. In this study, we compared the larvicidal effects of L. sphaericus 2362 (WHO Reference strain) and native strains III(3)7 and OT4b.25 against Cx. quinquefasciatus and Ae. aegypti in single-species and mixed-culture bioassays. Findings showed that L. sphaericus spores, vegetative cells and a combination thereof possessed high larvicidal activity against Cx. quinquefasciatus larvae, whereas only the formulation of L. sphaericus vegetative cells was effective against Ae. aegypti larvae. Similar results were obtained for field-collected larvae. We propose that a formulation of vegetative cells of L. sphaericus 2362 or III(3)7 could be a good alternative to chemical insecticides for the in situ control of mixed populations of Ae. aegypti and Cx. quinquefasciatus.

The major component of aphid alarm pheromone is (E)-β-farnesene (EβF), but the molecular mechanisms of EβF synthesis are poorly understood. Here we established a biological model to study the modulation of EβF synthesis in the bird cherry-oat aphid Rhopalosiphum padi by using quantitative polymerase chain reaction, gas chromatography/mass spectrometry and RNA interference. Our results showed that the rearing conditions significantly affected the weight of adult and modulated EβF synthesis in a transgenerational manner. Specifically, the quantity of EβF per milligram of aphid was significantly reduced in the individually reared adult or 1st-instar nymphs derived from 1-day-old adult reared individually, but EβF in the nymph derived from 2-day-old adult that experienced collective conditions returned to normal. Further study revealed that the production of EβF started in embryo and was extended to early nymphal stage, which was modulated by farnesyl diphosphate synthase genes (RpFPPS1 and RpFPPS2) and rearing conditions. Knockdown of RpFPPS1 and RpFPPS2 confirmed the role played by FPPS in the biosynthesis of aphid alarm pheromone. Our results suggested that the production of EβF starts at the embryo stage and is modulated by FPPS and rearing conditions in R. padi, which sheds lights on the modulatory mechanisms of EβF in the aphid.

The annual bluegrass weevil Listronotus maculicollis requires chilling exposure to terminate reproductive diapause during overwintering, but the effects of temperature on its post-diapause development in spring remain unclear. To explore this effect, overwintering adults were transferred from cold conditions (6°C/4°C, L:D 10:14) to different warm-up temperatures at L:D 12:12. When weevils were transferred to 7, 14 and 21°C in December and late January, the sizes of male and female reproductive organs were significantly smaller at 7°C than at 14 and 21°C. When weevils were transferred to 7, 9, 11, 13 and 15°C in late January, higher temperatures facilitated the post-diapause development. In both sexes, the sizes of reproductive organs and developmental rate increased with temperature. Reproductive organs did not grow significantly at 7°C in males and at 7–9°C in females, at which the percentage of developing weevils remained low. The time required for 50% of individuals to resume development was 44, 18, 13 and 8 days at 9, 11, 13 and 15°C, respectively, in males and 19, 14 and 8 days at 11, 13 and 15°C, respectively, in females. The threshold temperature for post-diapause development was 7.8°C in males, based on which 61.7 degree-days coincided with 50% of individuals developing. Under field conditions, the percentage of male and female maturity and insemination rate were low until early March, but all reached 100% by late March.

Pheromones play an important role in mediating interspecific interactions in insects. In an insect community, pheromones can reveal information about the senders, which could be used by other members of the food web (competitor, natural enemies, etc.) to their own advantage. The aggregation pheromones of two closely related thrips species, Frankliniella occidentalis and Frankliniella intonsa, have been identified with the same major compounds, (R)-lavandulyl acetate and neryl (S)-2-methylbutanoate, but in different ratios. However, the roles of the aggregation pheromones in the interspecific interactions between these two closely related species are unknown. Here, we investigated the roles of major aggregation pheromone compounds in interspecific interactions between F. occidentalis and F. intonsa for both long and short ranges. The results showed that, at tested doses, neither aggregation pheromone-induced long range cross-attraction nor short range cross-mating was detected between F. occidentalis and F. intonsa. Field-trapping trials showed that the species-specificity in aggregation pheromones was regulated by the ratio of two major compounds. However, species-specific blends of the two major compounds had no effect on short-range interactions between these two species. Our data from the thrips species provide support for the ‘aggregation model of coexistence’, explaining the species-specific pheromone-mediated coexistence of closely related species. Thus, species-specific pheromones could be one of the factors affecting population dynamics and community structure in closely related insects with similar niches.

Although parasites and microbial pathogens are both detrimental to insects, little information is currently available on the mechanism involved in how parasitized hosts balance their immune responses to defend against microbial infections. We addressed this in the present study by comparing the immune response between unparasitized and parasitized pupae of the chrysomelid beetle, Octodonta nipae (Maulik), to Escherichia coli invasion. In an in vivo survival assay, a markedly reduced number of E. coli colony-forming units per microliter was detected in parasitized pupae at 12 and 24 h post-parasitism, together with decreased phagocytosis and enhanced bactericidal activity at 12 h post-parasitism. The effects that parasitism had on the mRNA expression level of selected antimicrobial peptides (AMPs) of O. nipae pupae showed that nearly all transcripts of AMPs examined were highly upregulated during the early and late parasitism stages except defensin 2B, whose mRNA expression level was downregulated at 24 h post-parasitism. Further elucidation on the main maternal fluids responsible for alteration of the primary immune response against E. coli showed that ovarian fluid increased phagocytosis at 48 h post-injection. These results indicated that the enhanced degradation of E. coli in parasitized pupae resulted mainly from the elevated bactericidal activity without observing the increased transcripts of target AMPs. This study contributes to a better understanding of the mechanisms involved in the immune responses of a parasitized host to bacterial infections.

In this study the host plant genotype effect on cabbage aphid, Brevicoryne brassicae (L.)(Hemiptera: Aphididae) preference and performance, the effect of aphid genotype on parasitoids performance, as well as the indirect effects of plant genotypes on aphid parasitoid performance, were tested using different population samples of the aphid and its primary endoparasitoid wasp, Diaeretiella rapae (M'Intosh) (Hymenoptera: Braconidae). Experiments were run as fully-factorial randomized block design in a greenhouse. Accordingly, host plant cultivar had significant effects on the total number of aphids and aphid-load whilst the fitness of the aphid genotypes were also influenced by plant cultivar. The effect of parasitism on cabbage aphids was significantly different between plant cultivars. Overall, the results revealed that cabbage aphid is under different selective pressures arising from both higher (parasitoid) and lower (host plant cultivar) trophic levels. The host plant cultivar had a significant effect on both aphid fitness and parasitism rate on particular aphid genotypes. This indicates that host-plant-adapted aphid species can create much context-dependency in the nature and strength of ‘fitness benefits parasitism’, which may in turn alter the costs and benefits of host specialization.

Variation in parasitism risk among hosts can arise from between-patch and within-patch factors, but considerably less information is known about the latter. This study investigated how distributions of the oriental fruit fly Bactrocera dorsalis influenced its parasitism by the pupal parasitoid Dirhinus giffardii in the laboratory. Because B. dorsalis larvae pupate underground, pupation depth was considered as an important factor that affects the risk of parasitism. When the density of B. dorsalis larvae was varied (1, 10, and 100 larvae per arena), average pupation depth increased with the density. When the depth of pupae was manipulated, the rate of parasitism differed by depths. Parasitism at 0 cm differed from the random parasitoid model expectation, but parasitism at 1 cm was not different from the model expectation. Few pupae at 2 cm were parasitized. In another experiment, when pupae were simultaneously presented at 0 cm and 1 cm depths, parasitism at 1 cm was weakened by the presence of puape at 0 cm. These results imply that the density of the host influences pupation depth as well as the distribution of parasitism and plays an important role in host-parasitoid dynamics.

Understanding the mechanisms underpinning spatiotemporal diversity patterns of biological communities is a major goal of ecology. We aimed to test two ecological hypotheses: (i) temporal patterns of β-diversity will mostly be driven by nestedness, with a loss of species from summer to winter, and (ii) nestedness values will correlate with climatic variables instead of turnover values, indicating either a loss of species during winter or a gain of species during summer. We sampled dung beetles using standardized sampling protocols along a year in four Atlantic forest sites: two at the northwest and two at the central region of Rio Grande do Sul state, southern Brazil. We partitioned temporal patterns of β-diversity into turnover and nestedness in order to investigate if community changes are driven by species substitution or gain/loss across time. Our results highlighted five main findings: (i) dung beetle composition varied more with sites than site geographic position; (ii) there was almost one and a half ‘true’ dung beetle assemblages regarding the spatial distribution of species weighed by abundance; (iii) we found a positive influence of mean temperature and a negative influence of relative humidity on both species richness and abundance; (iv) both spatial and temporal dissimilarity among sites were dominated by species replacement, while the relative importance of nestedness was higher in temporal than spatial patterns; (v) there was an effect of precipitation and relative humidity on temporal patterns of β-diversity components, but these effects were site-dependent. Contrary to our expectations, the β-diversity component of turnover dominated both spatial and temporal patterns in dung beetle dissimilarity among sites and months. Distinct climatic variables affected differently the α-diversity and β-diversity components of dung beetle assemblages. Partitioning β-diversity into temporal components is a promising approach to unveil patterns of the community dynamics and to produce insights on mechanisms underlying such patterns.

Instrumentation and software for the automated analysis of insect flight trajectories is described, intended for quantifying the behavioural dynamics of moths in the vicinity of artificial light. For its time, this moth imaging system was relatively advanced and revealed hitherto undocumented insights into moth flight behaviour. The illumination source comprised a 125 W mercury vapour light, operating in the visible and near ultraviolet wavelengths, mounted on top of a mobile telescopic mast at heights of 5 and 7.1 m, depending upon the experiment. Moths were imaged in early September, at night and in field conditions, using a ground level video camera with associated optics including a heated steering mirror, wide angle lens and an electronic image intensifier. Moth flight coordinates were recorded at a rate of 50 images per second (fields) and transferred to a computer using a light pen (the only non-automated operation in the processing sequence). Software extracted ground speed vectors and, by instantaneous subtraction of wind speed data supplied by fast-response anemometers, the airspeed vectors. Accumulated density profiles of the track data revealed that moths spend most of their time at a radius of between 40 and 50 cm from the source, and rarely fly directly above it, from close range. Furthermore, the proportion of insects caught by the trap as a proportion of the number influenced by the light (and within the field of view of the camera) was very low; of 1600 individual tracks recorded over five nights, a total of only 12 were caught. Although trap efficiency is strongly dependent on trap height, time of night, season, moonlight and weather, the data analysis confirmed that moths do not exhibit straightforward positive phototaxis. In general, trajectory patterns become more complex with reduced distance from the illumination, with higher recorded values of speeds and angular velocities. However, these characteristics are further qualified by the direction of travel of the insect; the highest accelerations tended to occur when the insect was at close range, but moving away from the source. Rather than manifesting a simple positive phototaxis, the trajectories were suggestive of disorientation. Based on the data and the complex behavioural response, mathematical models were developed that described ideal density distribution in calm air and light wind speed conditions. The models did not offer a physiological hypothesis regarding the behavioural changes, but rather were tools for quantification and prediction. Since the time that the system was developed, instrumentation, computers and software have advanced considerably, allowing much more to be achieved at a small fraction of the original cost. Nevertheless, the analytical tools remain useful for automated trajectory analysis of airborne insects.

The alarm behavior plays a key role in the ecology of aphids, but the site and molecular mechanism for the biosynthesis of aphid alarm pheromone are largely unknown. Farnesyl diphosphate synthase (FPPS) catalyzes the synthesis of FPP, providing the precursor for the alarm pheromone (E)-β-farnesene (EβF), and we speculate that FPPS is closely associated with the biosynthetic pathway of EβF. We firstly analyzed the spatiotemporal expression of FPPS genes by using quantitative reverse transcription-polymerase chain reaction, showing that they were expressed uninterruptedly from the embryonic stage to adult stage, with an obvious increasing trend from embryo to 4th-instar in the green peach aphid Myzus persicae, but FPPS1 had an overall significantly higher expression level than FPPS2; both FPPS1 and FPPS2 exhibited the highest expression in the cornicle area. This expression pattern was verified in Acyrthosiphon pisum, suggesting that FPPS1 may play a more important role in aphids and the cornicle area is most likely the site for EβF biosynthesis. We thus conducted a quantitative measurement of EβF in M. persicae by gas chromatography-mass spectrometry. The data obtained were used to perform an association analysis with the expression data, revealing that the content of EβF per aphid was significantly correlated with the mean weight per aphid (r = 0.8534, P = 0.0307) and the expression level of FPPS1 (r = 0.9134, P = 0.0109), but not with that of FPPS2 (r = 0.4113, P = 0.4179); the concentration of EβF per milligram of aphid was not correlated with the mean weight per aphid or the expression level of FPPS genes. These data suggest that FPPS1 may play a key role in the biosynthesis of aphid alarm pheromone.