Modelling diapause termination and phenology of the Japanese beetle, Popillia japonica

We developed a mechanistic, stage-structured model simulating the phenology of Popillia japonica. The model simulates the influence of soil temperature on the larval diapause termination and on the development rate function of post-overwintering larvae and pupae. Model parameters are estimated based on literature evidence for pupae development and on a parameterisation process that allows estimating parameters for larval diapause termination and for the development rate function (and the related uncertainty) of post-overwintering larvae. Data used for model parameterisation and validation refer to time-series adult trap catches collected during the P. japonica monitoring programme performed by the Phytosanitary Service of Lombardy Region within the infested area in Lombardy (Italy) from 2015 to 2019. A total of 12 randomly selected locations are used to estimate biologically realistic model parameters (parameterisation dataset). We applied a Jackknife nonparametric resampling procedure on the parameterisation dataset to quantify uncertainty associated with parameters’ estimates. Parameterised model is then validated on time-series adult trap catches data referring to a different set of 12 randomly selected locations (validation dataset) surveyed in Lombardy. The model successfully predicted the beginning of adult emergence and the overall curve of adult emergence in the validation dataset. The model presented can support the definition of the best timing for the implementation of monitoring and control activities for the local and the area-wide management of P. japonica.

Insulin-Like Peptide and FoxO Mediate the Trehalose Catabolism Enhancement during the Diapause Termination Period in the Chinese Oak Silkworm (Antheraea pernyi)

In insects, trehalose accumulation is associated with the insulin/insulin-like growth factor signalling (IIS) pathway. However, whether insulin-like peptide is involved in the regulation of the trehalose metabolism during diapause termination remains largely unknown. This study assessed whether insulin-like peptide (ApILP) enhances the trehalose catabolism in the pupae of Antheraea pernyi during their diapause termination process. Injection of 10 μg of bovine insulin triggered diapause termination and synchronous adult eclosion in diapausing pupae. Moreover, treatment with bovine insulin increased the expression of trehalase 1A (ApTre-1A) and trehalase 2 (ApTre-2), as well as the activity of soluble and membrane-bound trehalase, resulting in a decline in trehalose levels in the haemolymph. Silencing ApILP via RNA interference significantly suppressed the expression of ApTre-1A and ApTre-2, thus leading to an increase in the trehalose concentration during diapause termination. However, neither injection with bovine insulin nor ApILP knockdown directly affected trehalase 1B (ApTre-1B) expression. Moreover, overexpression of the transcription factor forkhead box O (ApFoxO) induced an increase in trehalose levels during diapause termination; however, depletion of ApFoxO accelerated the breakdown of trehalose in diapausing pupae by increasing the expression of ApTre-1A and ApTre-2. The results of this study help to understand the contributions of ApILP and ApFoxO to the trehalose metabolism during diapause termination.

Transcriptomic analysis elucidates the molecular processes associated with hydrogen peroxide-induced diapause termination in Artemia-encysted embryos

Treatment with hydrogen peroxide (H2O2) raises the hatching rate through the development and diapause termination of Artemia cysts. To comprehend the upstream genetic regulation of diapause termination activated by exterior H2O2 elements, an Illumina RNA-seq analysis was performed to recognize and assess comparative transcript amounts to explore the genetic regulation of H2O2 in starting the diapause termination of cysts in Artemia salina. We examined three groupings treated with no H2O2 (control), 180 μM H2O2 (low) and 1800 μM H2O2 (high). The results showed a total of 114,057 unigenes were identified, 41.22% of which were functionally annotated in at least one particular database. When compared to control group, 34 and 98 differentially expressed genes (DEGs) were upregulated in 180 μM and 1800 μM H2O2 treatments, respectively. On the other hand, 162 and 30 DEGs were downregulated in the 180 μM and 1800 μM H2O2 treatments, respectively. Cluster analysis of DEGs demonstrated significant patterns among these types of 3 groups. GO and KEGG enrichment analysis showed the DEGs involved in the regulation of blood coagulation (GO: 0030193; GO: 0050818), regulation of wound healing (GO:0061041), regulation of hemostasis (GO: 1900046), antigen processing and presentation (KO04612), the Hippo signaling pathway (KO04391), as well as the MAPK signaling pathway (KO04010). This research helped to define the diapause-related transcriptomes of Artemia cysts using RNA-seq technology, which might fill up a gap in the prevailing body of knowledge.

Diapause Regulation in Newly Invaded Environments: Termination Timing Allows Matching Novel Climatic Constraints in the Box Tree Moth, Cydalima perspectalis (Lepidoptera: Crambidae)

The association between indirect environmental cues that modulate insect diapause and the actual stressors is by no means granted when a species encounters new environments. The box tree moth, Cydalima perspectalis, is an Asian pest whose rapid invasion in Europe causes considerable economic and ecological impacts. Larvae enter a winter diapause induced by the photoperiod in both native and invaded ranges, but factors that trigger the return to an active phase are still unknown. Yet, identifying them is crucial to understand how diapause end synchronizes with the end of the winter stress encountered in Europe. To test whether activity resumption is regulated by thermal and/or photoperiodic thresholds, or additive effects between these factors often involved in diapause termination, diapausing caterpillars from an invaded area were exposed to crossed treatments at the laboratory. The evolution of diapause rate was monitored over time and compared to that of nearby field sites invaded. A strong positive effect of increasing temperature was found on the rate and dynamics of diapause termination, whereas no compelling effect of photoperiod appeared. Resuming development directly when main stressors fade, not in response to indirect photoperiodic cues that could be mismatched outside native areas, likely contributes to the good match observed between diapause and the new climates that this pest encountered in the invaded range.

The Modulation of Trehalose Metabolism by 20-Hydroxyecdysone in Antheraea pernyi (Lepidoptera: Saturniidae) During its Diapause Termination and Post-Termination Period

Trehalose plays a crucial role in the diapause process of many insects, serving as an energy source and a stress protectant. Trehalose accumulation has been reported in diapause pupae of Antheraea pernyi; however, trehalose metabolic regulatory mechanisms associated with diapause termination remain unclear. Here, we showed that the enhanced trehalose catabolism was associated with an increase in endogenous 20-hydroxyecdysone (20E) in hemolymph of A. pernyi pupae during their diapause termination and posttermination period. Injection of 20E increased the mRNA level of trehalase 1A (ApTre-1A) and trehalase 2 (ApTre-2) of A. pernyi diapause pupae in a dose-dependent manner but did not affect the mRNA level of trehalase 1B (ApTre-1B). Meanwhile, exogenous 20E increased the enzyme activities of soluble and membrane-bound trehalase, leading to a decline in hemolymph trehalose. Conversely, the expression of ApTre-1A and ApTre-2 were down-regulated after the ecdysone receptor gene (ApEcRB1) was silenced by RNA interference or by injection of an ecdysone receptor antagonist cucurbitacin B (CucB), which inhibits the 20E pathway. Moreover, CucB treatment delayed adult emergence, which suggests that ApEcRB1 might be involved in regulating pupal-adult development of A. pernyi by mediating ApTre-1A and ApTre-2 expressions. This study provides an overview of the changes in the expression and activity of different trehalase enzymes in A. pernyi in response to 20E, confirming the important role of 20E in controlling trehalose catabolism during A. pernyi diapause termination and posttermination period.

Diapause regulation in newly invaded environments: termination timing allows matching novel climatic constraints in the box tree moth, Cydalima perspectalis (Lepidoptera: Crambidae)

The association between indirect environmental cues that modulate insect diapause and the actual stressors is by no means granted when a species encounters new environments. The box tree moth, Cydalima perspectalis, is an Asian pest whose rapid invasion in Europe cause considerable economic and ecological impacts. Larvae enter a winter diapause induced by photoperiod in both native and invaded ranges, but factors that trigger the return to an active phase are still unknown. Yet, identifying them is crucial to understand how diapause end synchronizes with the end of the winter stress encountered in Europe. To test whether activity resumption is regulated by thermal and/or photoperiodic thresholds, or additive effects between these factors often involved in diapause termination, diapausing caterpillars from an invaded area were exposed to crossed treatments at the laboratory. The evolution of diapause rate was monitored over time and compared to that of nearby field sites invaded. A strong positive effect of increasing temperature was found on the rate and dynamics of diapause termination, whereas no compelling effect of photoperiod appeared. Resuming development directly when main stressors fade, not in response to indirect photoperiodic cues that could be mismatched outside native areas, likely contributes to the good match observed between diapause and the new climates this pest encountered in the invaded range.

The genomics of trait combinations and their influence on adaptive divergence

ABSTRACTUnderstanding rapid adaptation requires quantifying natural selection on traits and elucidating the genotype-phenotype relationship for those traits. However, recent studies have often failed to predict the direction of adaptive allelic variation in natural populations from laboratory studies. Here, we test for genomic signatures of genetic correlations to illustrate how multifarious, correlational selection may drive counterintuitive patterns of population divergence in the apple maggot fly, Rhagoletis pomonella (Diptera: Tephritidae). Apple-infesting populations with relatively early emerging adults have recently evolved from hawthorn-infesting populations consisting of relatively later emerging adults. Multiple studies have reported a paradoxical relationship between population differentiation and seasonal timing, as determined by the timing of diapause termination; alleles associated with late termination occur at higher frequencies in the earlier emerging apple-infesting populations compared to hawthorn-infesting populations. We present evidence that historical selection on diapause termination and another trait, initial diapause intensity, over geographic gradients generated genetic correlations between the traits in a direction antagonistic to contemporary selection on apple-infesting populations. Single nucleotide polymorphism in genomic regions of high linkage disequilibrium associated strongly with diapause termination and intensity, population divergence, geography, and evolutionary responses in laboratory selection experiments. These associations were consistent with geographically variable selection and with correlated evolutionary responses driving higher frequencies of late-associated alleles in the early emerging apple race. In contrast, loci associated only with diapause termination showed the expected pattern (more early-associated alleles in the apple race) in half of the population pairs. Our results suggest that selection on loci demonstrating antagonistic pleiotropy may often shape genomic footprints of rapid adaptation.

Effects of photoperiod and relative humidity on diapause termination and post-winter development of Rhagoletis cerasi pupae

The European cherry fruit fly, Rhagoletis cerasi (Diptera: Tephritidae), is a univoltine species that undergoes obligatory summer–winter diapause at pupal stage in the soil (2–5 cm) beneath host trees. To study the effects of photoperiod and relative humidity on diapause termination and post-winter developmental duration of R. cerasi, pupae collected from Dossenheim (Germany) were exposed to different photoperiod or relative humidity regimes during a chilling period ranging from 2 to 8.5 months. Specifically, pupae were exposed to four photoperiod regimes: (a) light conditions (24L:00D), (b) dark conditions (00L:24D), (c) short photoperiod (08L:16D) and (d) long photoperiod (16L:08D), as well as to three relative humidity regimes: (a) low (40% RH), (b) medium (60% RH) and (c) high (70–80% RH). Data revealed that relative humidity is not a significant predictor of diapause termination, but it affects the post-winter developmental period. Higher relative humidity promotes post-winter pupae development. On the other hand, photoperiod significantly affected both diapause termination and post-winter development of R. cerasi pupae. Light conditions (24L:00D) accelerate adult emergence, particularly for females. Regardless of the photoperiod (24L:00D, 00L:24D, 08L:16D), rates of adult emergence were high (>75%) for chilling intervals longer than 6.5 months. Nonetheless, exposure to a long day photoperiod (16L:08D), during chilling, dramatically reduced the proportion of adult emergence following 6 months exposure to chilling. Our findings broaden the understanding of factors regulating diapause responses in European cherry fruit fly, local adaptation and synchronization of adult emergence with the ripening period of major hosts.