Overwintering at multiple life stages in Schizotetranychus shii (Acari: Tetranychidae), a specialist of evergreen chinquapin

Overwintering at multiple life stages in Schizotetranychus shii (Acari: Tetranychidae), a specialist of evergreen chinquapin | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Overwintering at multiple life stages in Schizotetranychus shii (Acari: Tetranychidae), a specialist of evergreen chinquapin Kohei Nagata, Yamato Negoro, Katsura Ito This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4084840/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 12 Dec, 2024 Read the published version in Experimental and Applied Acarology → Version 1 posted You are reading this latest preprint version Abstract Host availability in winter affects the overwintering strategies of herbivorous arthropods. Spider mites (Acari: Tetranychidae) overwinter as eggs or adult females, but some do so as multiple life stages on evergreen hosts. For example, in Schizotetranychus brevisetosus , adult females and their eggs stay on host leaves in mid-winter. However, few studies have focused on proximate factors generating such overwintering stages. Here, we investigated photoperiodic responses and life-stage compositions in winter in a population of Schizotetranychus shii , a specialist of Japanese chinquapin (Fagaceae). The proportion of non-ovipositing females at 20°C followed a sigmoid curve with increasing photoperiod, and the critical day length (CDL) was estimated as 11.8L, which corresponds to the environments from late September to early October. Although females grown under 10–11L conditions never oviposited within 7 days, 90–96% of them started oviposition within only 30 days without chilling (n = 23–31). In the field, all life stages were observed to occur throughout winter, but their proportions varied drastically. The proportion of eggs declined from early October (62%) to early December (12%), as predicted by CDL, but steeply increased toward late February (96%), during which only adult females and eggs remained. In summary, a short photoperiod in October arrests oviposition in emerging females, but they soon commence oviposition in November while immature stages are still growing, and individuals at all life stages (including a new generation) coexist until all immature stages mature in February. This novel pattern suggests that evergreen hosts allow spider mites to evolve overwintering strategies with little phylogenetic constraint. Castanopsis sieboldii (Fagaceae) critical day length (CDL) diapause evergreen host plants photoperiodic response reproductive arrest Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 INTRODUCTION Host-plant phenology exerts strong selection pressure on overwintering strategies in herbivorous arthropods (Tauber et al. 1986 ; Danks 1987 , 2006 ). The timing of leaf extension and defoliation affects the timing of herbivore development and reproduction. Their overwintering patterns are shaped so that life stages vulnerable to adverse conditions such as food deprivation, low temperature, or frost do not encounter such harsh environments. The importance of host availability in life-cycle formation is strongly supported by the associations between host phenology and winter life-stage composition in diverse herbivorous taxa (Gotoh 1983 ; Morimoto and Takafuji 1983 ; Carey 1994 ; Hunter and McNeil 1997 ; Kurota and Shimada 2002 ). The success of development and reproduction in spider mites (Acari: Tetranychidae) depends on the spatio-temporal stability of hosts. Using mathematical models incorporating patchy habitats of different quality, Tsuda et al. ( 1997 ) found that the existence of winter hosts favours non-diapausing females, or females with phenotypic plasticity whose phenotypes (diapausing or non-diapausing) match the habitat quality (i.e., females avert diapause in good habitats). Because the spider mites parasitize various hosts of different phenology and have diverse life-history strategies, they are suitable model organisms to evaluate the effects of host phenology on life-cycle formation. The significance of host availability on life cycles is suggested in Tetranychus (Morimoto and Takafuji 1983 ), Schizotetranychus (Gotoh 1983 ; Ito and Yamanishi 2019 ), Sasanychus (Gotoh 1986a , b ; Gotoh 1986b ), Stigmaeopsis (Saito and Ueno 1979) and Panonychus (Morimoto and Takafuji 1983 ; Koveos and Broufas 1999 ). These studies have traditionally associated overwintering patterns with diapause induction. In most species, short day length and low temperatures during development (Veerman 1985 ) or adulthood (Lees 1953 ; Saito et al. 2005 ) induce diapause, and host conditions (Ito 2003 ; Ito and Saito 2006 ), maternal effects (Danilevskii 1965 ; Oku et al. 2003 ) and predation risks (Kroon et al. 2004 , 2005 , 2008 ) modify the primary response of day length. Despite the progress of these studies, however, the relationship between diapausing stages and overwintering stages realized in the field is still confusing (Saito 2010 ); only diapausing individuals can overwinter in cold regions or on deciduous hosts, but this restriction is relaxed in warmer regions, where winter hosts are plentiful. In temperate populations of Tetranychus urticae and T. kanzawai , all life stages occur throughout winter, even though populations retain sensitivity to adult diapause during short photoperiods (Takafuji and Kamibayashi 1984 ; So and Takafuji 1991 ; Takafuji et al. 1991 ; Takafuji and Morishita 2001 ). Adaptation to evergreen hosts, which provide temporally stable habitats, may relax the constraint of having a single overwintering stage (Gotoh 1986a , b ; Ito and Yamanishi 2019 ). Overwintering as adult females and their eggs are found in Yezonychus sapporensis (H. Yanagida, unpublished data, Saito 2010 ) and Sasanychus akitanus (Gotoh 1986a , b ) on Sasa bamboo, which grow in cool regions and are covered by snow in winter. Gotoh ( 1986a , b ) reports that S. akitanus females continue to lay diapausing eggs until early winter. Although the author did not explicitly relate the scarcity of predators to the oviposition patterns, the inactivity of midge larvae and phytoseiid mites might underlie the production of winter eggs until late in the season. On the other hand, in Schizotetranychus brevisetosus , which parasitizes Quercus glauca L. (Fagaceae) (Ehara 1989 ; Ehara and Gotoh 2009 ), adult females occupy the population in December, produce winter eggs until early March and die out (Ito and Hamada 2018 ; Ito and Yamanishi 2019 ; Oda and Ito 2021 ). This style is distinct from S. akitanus , whose females cease producing diapausing eggs and overwinter themselves and adults of different generations interbreed in spring (Gotoh 1986a ). Predators are almost absent during winter in S. brevisetosus (Ito 2020 ), and this low predation pressure enables the overwintering of winter eggs. This inter-specific difference in overwintering styles may be partly explained by climatic conditions; S. brevisetosus individuals can act even during winter. Despite these differences in the overwintering pattern, photoperiodism underlies reproductive arrest in both species (Gotoh 1986b ; Oda and Ito 2021 ). Schizotetranychus shii (Acari: Tetranychidae) is the specialist of evergreen chinquapin Castanopsis sieboldii (Ehara 1965). Individuals live gregariously in silk-web nests constructed in the depressions along leaf veins or edges. This species overwinters in various life stages and is therefore considered a non-diapausing species (Ehara and Gotoh 2009 ). It has been assigned to the same clade as S. brevisetosus in phylogenetic trees based on 18S and 28S rDNA gene sequences, whereas S. akitanus is in a separate clade (Matsuda et al. 2014 , Figs. 4 and 5 ). This makes S. shii a suitable species for studying evolutionary changes in overwintering patterns through comparison with those in S. brevisetosus . Here, we focused on the temporal changes in life-stage composition during winter in S. shii . We tested the hypothesis that photoperiodic response for reproductive arrest is responsible for the formation of overwintering patterns. Based on the results, we discuss the evolution of overwintering patterns in spider mites on evergreen hosts. MATERIALS AND METHODS Photoperiodic response A laboratory strain of S. shii was established from 50 individuals at various life stages collected from C. sieboldii trees in Monobe Campus, Kochi University, Nankoku, Kochi Prefecture, Japan (33° 33' 05"N, 133° 40' 30" E, WGS84, 8 m altitude) on 8 October 2021. The strain was maintained on detached C. sieboldii leaves using the leaf-culture method (Helle and Overmeer 1985 ). Briefly, a detached leaf was pressed on water-soaked cotton pads piled on a polyurethane mat in an insect breeding dish (100(d) × 40(h) mm, SPL Life Sciences, Gyeonggi-do, Korea). The leaf edge was surrounded by Kimwipe® strips to prevent mite escape and leaf curling. Each dish was covered with a mesh lid to mitigate a temperature decrease on the leaf surface due to evaporation (Saito and Suzuki 1987 ). The culture was maintained under 25 ± 0.5°C (mean ± SD) and a photoperiod of 15 h light and 9 h dark (15L) conditions in a MIR-154 chamber (SANYO, Osaka, Japan) for two months. Relative humidity in the chamber was not controlled, but data loggers (RTR-503, T&D Corporation, Nagano, Japan) showed that it typically ranged between 60 and 80%. We established photoperiodic treatments from 10L to 13L at one-hour intervals in temperature gradient incubator chambers (TG-300WLED-5LE, NK System, Osaka, Japan). We also included a 10.5L treatment, which approximated the day length excluding civil twilight in mid-November (NAOJ 2023 ), when the life-stage compositions of a nearby S. brevisetosus population underwent significant change (Ito and Yamanishi 2019 ). Temperatures in the chambers were maintained at 20 ± 1°C according to the TR-51i, 52i and RTR-503 data loggers (T&D Corporation). Two leaf cultures were prepared per day length treatment. Ten adult females collected from the rearing culture were introduced onto each culture using a fine brush and allowed to oviposit for 3 days at 25 ± 0.5°C under a photoperiod of 15L in the MIR-154 chamber. Subsequently, 37–76 eggs were obtained per leaf. The females were removed and the dishes were maintained at their respective treatment conditions. Emerged females were separated onto C. sieboldii leaf squares (1.5 cm × 1.5 cm) daily, maintained under the same photoperiodic treatment, and the onset of oviposition from emergence was monitored (n = 33–36 per treatment). The oviposition status of each female was assessed on the seventh day from isolation, considering that oviposition occurred within 7 days at 13L (see Results), which was a long day length at the end of August (NAOJ 2023 ) when mites are highly reproductive. Females that died within seven days were excluded from the analysis. The effects of day length on the proportions of non-ovipositing females were analysed using generalized linear models (GLM), assuming a binomial distribution with a logit link function. The glm command in the software R version 4.2.2 (R Core Team 2022 ) was used for this purpose. The critical day length (CDL), at which 50% of females oviposited, was estimated as \(-a/b\) , where \(a\) and \(b\) represent the intercept and the slope of the fitted model, respectively (Oda and Ito 2021 ). The model assumptions, especially the distribution and deviance of residuals, were assessed visually. To assess the depth of reproductive arrest, we also counted the number of females starting oviposition within 30 days of isolation in the 10L, 10.5L and 11L treatments. This threshold was selected based on previous studies on the duration of diapause termination in temperate regions (Mochizuki and Takafuji 1996 ; Ito 2004 ; Ito et al. 2013 ). The proportions of ovipositing females among the day length treatments were compared using Chi-square tests. Field survey of life stages Approximately 30 leaves (ranging from 12 to 32) with web nests were collected from the Monobe Campus using a 10× magnifying grass. These leaves were immediately observed using a stereomicroscope (SZ-61 and SZX-7, Olympus, Tokyo, Japan), and the life stages of S. shii were divided into four categories: egg , immature (larvae and nymphs), adult female and male . Sampling was conducted weekly from 8 October 2021 to 28 January 2022 and then biweekly until 22 April 2022, when colonies were disturbed by predators. The periodic changes in stage composition were tested via Chi-square tests with Yates’ correction using the chisq.test function in R. Adult sexes were pooled because males were rare. The association between the observation date and the density of each life stage was analysed by a generalised additive model (GAM) using the gam function in the mgcv package version 1.9.0 (Wood 2011 ) in R. Parameter estimation followed the procedure outlined by Crawley ( 2005 ). The statistical model was as follows: $$\text{log }\left(y\right)={\beta }_{0}+s\left(x\right)+\text{l}\text{o}\text{g}\left(n\right)$$ 1 where \(y\) is the number of individuals at each life stage, \({\beta }_{0}\) is the model intercept, \(s\left(x\right)\) is the smooth function against the day x and \(\text{l}\text{o}\text{g}\left(n\right)\) is the offset term expressed as the logarithm of leaf number. The best spline function was selected using the residual maximum likelihood (REML) method (Wood 2011 ). For each life stage, the GAM model and a simple linear model were compared by likelihood ratio (LR) tests using the anova function in R. Days required for hatching in the field (transplant experiment) A new laboratory strain was established from 15 adult females from the Monobe Campus on 13 September 2022 and maintained under 15L conditions at 25°C for a month. Three sets of experiments commenced on 15 October, 19 November and 24 December 2022. In each set, 10 (October) or 15 (November and December) adult females from the laboratory strain were placed on each of two leaves (November) or one leaf (December) prepared as above (the leaf culture), and allowed to oviposit for 3 days. Following the removal of females, the leaf cultures were enclosed in a mesh cage (internal dimension: 30 cm × 30 cm × 30 cm, BugDorm, Taichung, Taiwan) to prevent predator intrusion and exposure to rain. The cage was fixed to a plastic stand (10 cm in height) set on the forest floor using vinyl ropes. The leaf cultures were checked for egg hatching on-site using a 10× magnifying grass every 3 days, and subsequently daily after hatching began. Deformed or discoloured eggs were regarded as dead. For cases where precise counting was difficult, the leaves were assessed in the laboratory with a stereomicroscope. The average days required for hatching among groups were compared via Welch’s ANOVA using the oneway.test function in R. The survival rates of eggs were compared via Chi-square tests with Yates’ correction, and pairwise comparisons were conducted via Tukey’s multiple test for proportions (Zar 2010 ). Hatchability of autumnal eggs in the field Eggs were collected from the Monobe Campus on (I) 24 October and (II) 20–26 November 2023 to measure the days required for hatching described as below. Each month, several field-collected leaves with eggs were arbitrarily excised with a razor blade, and these pieces were placed on wet cotton pads in a breeding dish. The leaf pieces were maintained under 15L conditions at 20°C, and hatching was assessed daily, with larvae being removed each time they were observed. As a control, 20 females from the new laboratory strain (derived from 70 individuals at various life stages from the Monobe Campus on 23 July 2023 and maintained at 25°C/16L) were allowed to oviposit for 2 days on a detached leaf. Eggs that were laid on the first day were removed with a fine brush to adjust the age of eggs, and the occurrence of hatching was recorded as above. In other words, newly laid eggs were obtained for this strain. The data were structured as a two-way design (months × strain) and were analysed by GLM. However, the data were heavily underdispersed from Poisson distribution due to the mixing of data obtained in different environments (see Results), and established analytical methods are not applicable to such cases (Sáez-Castillo et al. 2022 ; Seck et al. 2022 ). Accordingly, we adopted a GLM with a two-way design (months × groups), assuming a COM-Poisson distribution with a log link function and a constant dispersion parameter, using the glm.CMP in the DGLMExtPoison package version 0.2.3 (Sáez-Castillo et al. 2022 ). The significance of each term was assessed by comparing it with the full model using likelihood ratio (LR) tests by lrtest in R. RESULTS Photoperiodic response The developmental period of adult females under 13L was 5.4 ± 1.2 days, with a range of 4 to 8 days, and a 97% oviposition rate within 7 days (n = 33). Accordingly, females ovipositing within 7 days were defined as reproducing. Figure 1 shows the response curve of oviposition rates against photoperiod. All females under 10L, 10.5L and 11L never oviposited within seven days (0%, n = 34–36), but most females laid eggs under 12L (80.6%, n = 36) and 13L. The logistic analysis estimated a positive intercept and a negative slope defined by y = 57.73–4.91 x , with residual deviance = 5.56 and degrees of freedom (df) = 3. The CDL was estimated as 11.8L. Though there were no apparent outliers, residual analysis suggested that the 12L datapoint is slightly deviated from the prediction line of the normal Q–Q plot and had a higher leverage. Even when kept under short-day conditions (10–11L), 64–86% of the non-ovipositing females survived 30 days after emergence, and 90–96% of them oviposited within that period (Table 1 ). Except for survival rates, no differences were found between 10L and the other two daylength treatments. Table 1 The proportions of surviving females 30 days after emergence and their ovipositing rate ( n = 36 for all treatments). The results of Chi-square test and, if significant, Tukey’s multiple comparisons for proportions (Zar 2010 ) are indicated by different lower-case letters. Daylength 10L 10.5L 11L χ 2 df P Survival rate 0.64 a 0.86 b 0.86 b 7.07 2 0.029 Oviposition rate 0.96 0.90 0.90 0.63 2 0.730 Field survey of life stages The proportion of life stages changed significantly during the field survey (Fig. 2A, χ 2 = 566.27, df = 42, P < 0.001). In October, the proportion of life stages did not significantly vary ( χ 2 = 5.06, df = 6, P = 0.537) and the proportions of eggs remained moderate (45.1–64.0%). The proportion of eggs gradually decreased to 11.8% by 3 December ( χ 2 = 22.86, df = 7, P = 0.002). Conversely, the proportion of immature stages increased from 23.1% on 8 October to 52.4% on 26 November ( χ 2 = 23.56, df = 7, P = 0.001). The proportion of adults increased from 15.4% on 8 October to 41.2% on 3 December ( χ 2 = 16.00, df = 8, P = 0.042), slowly decreased to 1.2% on 7 April (1.2%) ( χ 2 = 109.77, df = 11, P < 0.001) with a concurrent increase in the proportion of eggs and rose again to 20.4% on 22 April ( χ 2 = 62.70, df = 1, P < 0.001). The proportion of eggs rose from 3 December to 25 February ( χ 2 = 122.89, df = 9, P < 0.001), whereas the proportions of immature stages, including larvae (Table 2 ), declined during this period (0.7–47.1%, χ 2 = 93.78, df = 9, P < 0.001). Table 2 The number of larvae (L) and nymphs (N) of Schizotetranychus shii between 2021–2022. The number of teleiochrysalis females (Tf) is shown separately. Date Month 12 N 19 N 26 N 3 D 10 D 17 D 24 D 7 J 14 J 21 J 28 J 11 F 25 F 11 M 25 M L 15 12 13 1 4 1 1 0 0 1 0 0 0 0 13 N 10 9 19 7 6 7 16 4 7 3 1 3 3 5 0 (Tf) 2 1 3 1 The mean density of each life stage per sampled leaf is shown in Fig. 2 B. The total density remained below 3.0 until 7 January but then increased to approximately 15.5 on 22 April, with an increase in egg density after December (representing 70.8–96.4% of the population). The density of immature stages peaked on 26 November (1.1) and gradually decreased toward 25 February (0.0). The immature stages reappeared on 11 March (0.3) and increased towards 7 April (7.6). The adults of the next generation emerged on 22 April (3.2), and some were observed moving onto newly extended leaves. The density of adults did not show a clear seasonal trend. Sex ratios fluctuated between 8–50% until 28 January (except for 0% on 24 December), and males were absent between 11 February to 7 April (Fig. 2 C). However, the change in sex ratios over the survey period was not significant due to the small number of males ( χ 2 = 26.05, df = 21, P = 0.205). The results of GAM models are summarised in Fig. 3 and Table 3 . The response curve in adult females showed no conspicuous trend until March, although density increased in late April (Fig. 3 A). The curves for immature stages decreased toward late February and began to increase thereafter (Fig. 3 B). The curve for eggs fluctuated until November and gradually increased after December (Fig. 3 C). The smooth term for adult females was not significant ( P = 0.576, Table 3 ), which suggests that spline fitting is unnecessary. However, the smooth terms were significant for immature stages and eggs ( P < 0.001). Also, LR tests revealed that spline fitting was no better than a simple linear model for females (df = 2.32, deviance = 3.47, P = 0.220), but improved fit for immature stages (df = 3.72, deviance = 29.84, P < 0.001) and eggs (df = 8.03, deviance = 180.74, P < 0.001). Table 3 The summary of GAM analyses for each life stage in 2021–22 (n = 22). F: adult females, LN: larvae and nymphs, and E: eggs. The data of adult males were not analysed because of insufficient sample size. The results of LR tests based on REML methods are shown. Stage Estimated df χ 2 P Deviance (%) * –REML F 2.19 1.33 0.576 11.9 53.94 LN 3.43 54.41 < 0.001 74.7 56.74 E 7.41 231.51 < 0.001 81.9 97.18 * Deviance explained by the smooth term \(s\left(x\right)\) in Eq. ( 1 ) Days required for hatching in the field (transplant experiment) The hatchability of eggs placed in the field on October 15 (n = 21) and November 19 (n = 85) were 100.0% and 51.8%, respectively, but all eggs set on 24 December died during development. The survival rates of the eggs were significantly differed among months ( χ 2 = 70.353, df = 2, P < 0.001), and pairwise comparisons revealed differences between December and the other two months (Tukey’s multiple test for proportions, α = 0.05). Days required for hatching were 13.9 ± 1.8 (n = 21) and 20.1 ± 7.9 days (n = 44) for the eggs set in October and November, respectively (Welch’s ANOVA, F = 24.446, df 1 = 1, df 2 = 51.689, P < 0.001). The last eggs to hatch did so on 2 November and 5 January, respectively. Hatchability of autumnal eggs in the field Figure 4 shows the days for field and laboratory eggs to hatch under 15L conditions at 20°C. The hatch times in the field were highly variable, especially in November, and one egg needed more than 10 days to hatch. A COM-Poisson GLM showed underdispersion (i.e., higher average with lower variance, Fig. 4 ) as suggested by the dispersion parameter of 1.31 (> 1). The hatch times in November were significantly longer than those in October (Table 4A, Month), and the hatch times in culture were also longer than those in the field (Treatment). The interaction between treatments was highly significant (Month:Treatment). The LR tests revealed significance in all terms (Table 4B). Table 4. (A) Estimated coefficients of a COM-Poisson GLM on the days required for egg hatching (Fig. 4). (B) LR tests (compared with the full model; df = 57). (A) Source Estimate SE z P Fixed effects Intercept 1.04 0.10 10.42 < 0.001 Month (Nov) 0.78 0.13 6.06 < 0.001 Treatment (Culture) 1.44 0.11 13.15 < 0.001 Month:Treatment –0.79 0.14 –5.66 < 0.001 df 57 Dispersion coefficient ν (logit link) Intercept 1.31 0.18 7.22 < 0.001 (B) Null model df Statistic* P Treatment 2 29.81 < 0.001 Month 2 112.03 < 0.001 Treatment + Month 1 26.44 < 0.001 *–2(logL 0 /L 1 ), where L 0 and L 1 are the maximum likelihoods of the null (reduced) model and the full model, respectively. Discussion We demonstrated that various stages of S. shii coexist during winter and that stage compositions changed with time. Although this pattern appears to be consistent with the traditional view that S. shii is a non-diapausing species (Ehara and Gotoh 2009 ), with overwintering patterns being regulated by photoperiod like S. brevisetosus or common diapausing species (Ito and Yamanishi 2019 ), our findings indicate that short photoperiod strongly induced reproductive arrest in adult females, and that the photoperiodic response followed a sigmoid curve ranging from 0–100% of the population (Fig. 1 ). This study represents the first to reveal a tight relationship between population dynamics of species overwintering in multiple life stages and photoperiodism. The females of S. shii arrested reproduction in response to short photoperiods, as reported in S. brevisetosus on evergreen oak (Oda and Ito 2021 ). Females completely arrested oviposition under 10–11L conditions, with a CDL of 11.8L (Fig. 1 ), which corresponds with the day length in Kochi city at the beginning of October (NAOJ 2023 ). Consequently, immature stages that developed in late September (12L) or early October (11.5L) may cease reproduction, which explains the observed reduction of the egg proportion after late October (Fig. 2 A, B, Fig. 3 C). Although this CDL is slightly shorter than the consensus value of 10 geographic populations of S. brevisetosus (12.2L, Ito and Yamanishi 2019 ), it is close to CDL of a population at a nearby site (Noichi). Therefore, the season of oviposition arrest is likely to be similar between the two species, but the overwintering patterns of the two species differ significantly. In S. brevisetosus , adult females that emerge after early October do not oviposit, and the proportion of eggs steeply declined from October to November (Ito and Yamanishi 2019 ). Most of these females produce winter eggs from December to March, and only females and eggs are found throughout winter (Ito and Yamanishi 2019 ; Oda and Ito 2021 ). In contrast, in S. shii , the season for the two life stages occur much later in February and is short-lived (Fig. 2 A). This interspecific difference may be caused by differences in the time that it takes for females to resume activity after reproductive arrest, and earlier reactivation in S. shii may complicate its overwintering pattern. More than 90% of non-reproductive S. shii females oviposit within 30 days at 20°C without chilling treatment (Table 1 ), whereas S. brevisetosus females that had developed under 10L and 11L conditions take 49.5 ± 6.9 (n = 5) and 45.5 ± 6.5 days (n = 16), respectively, to oviposit under the same experimental conditions (N. Yamanishi and K. Ito, unpubl. data). Although statistical tests between the two species are not conducted due to the difference in data structure, these results predict that S. shii females would resume oviposition sooner than S. brevisetosus under short-day conditions. Indeed, eggs collected in the field in October all consisted of old eggs, which were soon to hatch, but eggs in November included a higher proportion of new eggs, which needed a longer time to hatch (Fig. 4 , Table 4). In addition, all eggs successfully hatched in both months, and the hatching time was short. Diapausing eggs of spider mites usually need a long period of chilling to hatch (Lees 1953 ; Shinkaji 1975 ; Koveos and Broufas 1999 ), such as in S. sapporensis eggs on evergreen bamboos, which require more than 40 days at 5°C (Gotoh 1986b ); given this, the S. shii eggs studied here may not be in diapause. Furthermore, the transplant experiments of laboratory eggs to the field conditions suggest that eggs in mid-October and November successfully hatched within the year (13.9 ± 1.8 and 20.1 ± 7.9 days, respectively; see Results). Taken together, females emerging in October stop oviposition but soon begin to lay non-diapausing eggs in November, when immature stages of the preceding generation are still growing. These eggs would hatch within the year and constitute the source of immature stages after December (Fig. 2 A, B, Fig. 3 B). The field survey demonstrated that immature stages (likely derived from new eggs above) develop throughout winter. The proportion of immature stages reached 46.7–52.4% from 19 November to 3 December, caused by of a gradual decrease in the egg density by hatching and increase in the nymph density (Figs. 2 B, 3 B). Although the period (degree-days) from hatching to adult emergence is unknown in S. shii , it takes approximately 20 days at 25°C/15L in the closely related S. brevisetosus (M. Muto-Yamawaki, unpubl. data) and appears to be similar from the present daylength experiments (data not shown). If so, immature stages emerged early in November may mature within or across the year, because the daily highest temperatures in November ranged from 16.4 to 22.3°C (Fig. 5 ) (Table 2 ). Furthermore, immature stages that emerge in the later season have a slow developmental rate in January and February. The mean temperature of these months drops to 6.0°C and 7.1°C on average, respectively (Fig. 4 , Japan Meteorological Agency 2024 ), but the highest temperature rises to approximately 15°C in both months, which permits development. Furthermore, the development of immature stages during winter is supported by the fact that immature stages of various ages occur in December, but only nymphs, including teleiochrysalis stages, are present from January to March (Table 2 ). Furthermore, adult males were found until the end of January (Fig. 1 C). These males may be composed of new adults or survivors from previous years, and their ability to survive through the coldest season suggests that temperatures in the study site are nonlethal. The arrest of oviposition despite the availability of winter habitats by evergreen hosts of S. shii may be attributed to a physiological shift that confers cold hardiness to their eggs, which is known as trans-generational effects in insects (Mousseau and Fox 1998 ; Magiafoglou and Hoffmann 2003 ; Zhou et al. 2013 ). Sub-zero temperatures over several months are stressful for spider mites in the summer form (Veerman 1985 ). Indeed, in the transplant experiment, all eggs that were collected from constant 25°C/15L conditions and placed in the field in mid-December did not survive (see Results). Still, this result does not agree with the pattern of wild eggs, which increased after December, the coldest season (Fig. 1 B). That is, cold hardiness in wild eggs in December is evidently higher than in laboratory-derived eggs (Fig. 2 A, B). Living on evergreen hosts provides the reproductive advantage of earlier initiation of reproduction than on deciduous hosts, especially where predation pressure in winter is minimal (Ito and Hamada 2018 ). However, predation pressure of spider mites increases early in winter, with predators of S. breviestosus disappearing in mid-winter (Ito 2020 ). In this situation, higher cold hardiness of eggs is important during winter. We observed that a few midge larvae and Agistemus mites fed on S. shii individuals, but their densities were much lower than in summer. The role of maternal effects in cold hardiness are unexplored in spider mites and should be explored further. Overwintering stages in spider mites are loosely constrained by phylogeny but vary even among sibling species (Veerman 1985 ; Sakagami 2002 ; Matsuda et al. 2014 ). We showed that evergreen hosts allow mites to overwinter in multiple life stages, and the overwintering style of S. shii is markedly different even from its sibling species S. brevisetosus (Matsuda et al. 2014 ), despite both being governed by photoperiodically controlled reproductive arrest (Oda and Ito 2021 ). The overwintering strategy of these species has evidently been shaped by low predation pressure in winter, strong cold hardiness and host availability. The importance of these factors should be evaluated to clarify the process of diversification in the overwintering strategies of spider mites. Declarations We have no pecuniary or other personal interest, direct or indirect, in any matter that raises or may raise a conflict with our duties. Acknowledgements This work was supported by a Cabinet Office grant-in-aid: the Advanced Next-Generation Greenhouse Horticulture by IoP (Internet of Plants), Japan (Grant No: D11). The authors thank the Field Science Center of Kochi University for allowing us to use the study site. Author contributions KN: planning, data collection and analysis, investigation, discussion; YN: planning, data collection and analysis, investigation, discussion; KI: planning, data collection and analysis, investigation, discussion, writing, funding acquisition Funding This work was supported by a Cabinet Office grant-in-aid and the Advanced Next-Generation Greenhouse Horticulture by IoP (Internet of Plants), Japan. Data availability The datasets generated and/or analysed during the current study are available from the corresponding author upon reasonable request. Competing interests The authors have no financial or non-financial interests to declare. References Carey DB (1994) Diapause and the host plant affiliations of lycaenid butterflies. Oikos 69:259–266. https://doi.org/10.2307/3546146 Crawley MJ (2005) Statistics: An introduction using R, 2nd edition. John Wiley & Sons Ltd., West Sussex, UK Danilevskii AS (1965) Photoperiodism and Seasonal Development of Insects. Oliver & Boyd Ltd., Edinburgh Danks HV (1987) Insect Dormancy: an Ecological Perspective. Biological Survey of Canada, Ottawa Danks HV (2006) Key themes in the study of seasonal adaptations in insects II. Life-cycle patterns. Appl Entomol Zool 41:1–13. https://doi.org/10.1303/aez.2006.1 Ehara S (1989) Four new species of spider mites (Acarina: Tetranychidae) from Japan. Proc Jpn Soc Syst Zool 40:28–38. https://doi.org/10.19004/pjssz.40.0_28 Ehara S, Gotoh T (2009) Colored guide to the plant mites of Japan. Zenkoku Noson Kyoiku Kyokai, Tokyo. (in Japanese) Gotoh T (1983) Life-history parameters of three species of Schizotetranychus on deciduous trees (Acarina: Tetranychidae). Appl Entomol Zool 18:122–128. https://doi.org/10.1303/aez.18.122 Gotoh T (1986a) Life-cycle variation in Panonychus akitanus Ehara (Acarina:Tetranychidae). Exp App Acarol 2:125–136. https://doi.org/10.1007/BF01213756 Gotoh T (1986b) Local variation in overwintering stages of Panonychus akitanus Ehara (Acarina: Tetranychidae). Exp Appl Acarol 2:137–151. https://doi.org/10.1007/BF01213757 Helle W, Overmeer WPJ (1985) Rearing Techniques. In: Helle W, Sabelis MW (eds) Spider Mites Their Biology, Natural Enemies and Control 1A. Elsevier, Amsterdam, pp 331–335 Hunter MD, McNeil JN (1997) Host-plant quality influences diapause and voltinism in a polyphagous insect herbivore. Ecology 78:977–986. https://doi.org/10.2307/2265851 Ito K (2003) Effect of leaf condition on diapause induction of a Kanzawa spider mite Tetranychus kanzawai Kishida (Acari: Tetranychidae) population on tea plants. Appl Entomol Zool 38:559–563. https://doi.org/10.1303/aez.2003.559 Ito K (2004) Deteriorating effects of diapause duration on post-diapause life history traits in the Kanzawa spider mite. Physiol Entomol 29:453–457. https://doi.org/10.1111/j.0307-6962.2004.00421.x Ito K (2020) Predators of the nest-making spider mite Schizotetranychus brevisetosus (Acari: Tetranychidae). J Acarol Soc Jpn 29:59–70. https://doi.org/10.2300/acari.29.59 Ito K, Fukuda T, Hayakawa H, Arakawa R, Saito Y (2013) Relationship between body colour, feeding, and reproductive arrest under short-day development in Tetranychus pueraricola (Acari: Tetranychidae). Exp Appl Acarol 60:471–477. https://doi.org/10.1007/s10493-013-9660-3 Ito K, Hamada E (2018) Adult female sensitivity to day-length conditions in terms of winter-egg production in Schizotetranychus brevisetosus Ehara (Acari: Tetranychidae). J Acarol Soc Jpn 27:69–76. https://doi.org/10.2300/acari.27.69 Ito K, Saito Y (2006) Effects of host-plant species on diapause induction of the Kanzawa spider mite, Tetranychus kanzawai . Entomol Exp Appl 121:177–184. https://doi.org/10.1111/j.1570-8703.2006.00471.x Ito K, Yamanishi N (2019) Production of winter eggs in Schizotetranychus brevisetosus (Acari: Tetranychidae) inhabiting evergreen Japanese blue oak. Exp Appl Acarol 78:521–534. https://doi.org/10.1007/s10493-019-00402-3 Japan Meteorological Agency (2024) Past weather. http://www.data.jma.go.jp/obd/stats/etrn/index.php (in Japanese) Koveos DS, Broufas GD (1999) Diapause induction and termination in eggs of the fruit tree red spider mite Panonychus ulmi in northern Greece. Exp Appl Acarol 23:669–679. https://doi.org/10.1023/A:1006232821653 Kroon A, Veenendaal RL, Bruin J, Egas M, Sabelis MW (2004) Predation risk affects diapause induction in the spider mite Tetranychus urticae . Exp Appl Acarol 34:307–314. https://doi.org/10.1023/B:APPA.0000049226.60183.29 Kroon A, Veenendaal RL, Bruin J, Egas M, Sabelis MW (2008) Sleeping with the enemy—predator-induced diapause in a mite. Naturwissenschaften 95:1195–1198. https://doi.org/10.1007/s00114-008-0442-4 Kroon A, Veenendaal RL, Egas M, Bruin J, Sabelis MW (2005) Diapause incidence in the two-spotted spider mite increases due to predator presence, not due to selective predation. Exp Appl Acarol 35:73–81. https://doi.org/10.1007/s10493-004-1980-x Kurota H, Shimada M (2002) Geographical variation in the seasonal population dynamics of Bruchidius dorsalis (Coleoptera: Bruchidae): constraints of temperature and host plant phenology. Environ Entomol 31:469–475. https://doi.org/10.1603/0046-225X-31.3.469 Lees AD (1953) Environmental factors controlling the evocation and termination of diapause in the fruit tree red spider mite Metatetranychus ulmi Koch (Acarina: Tetranychidae). Ann Appl Biol 40:449–486 Magiafoglou A, Hoffmann AA (2003) Cross-generation effects due to cold exposure in Drosophila serrata . Funct Ecol 17:664–672. https://doi.org/10.1046/j.1365-2435.2003.00774.x Matsuda T, Morishita M, Hinomoto N, Gotoh T (2014) Phylogenetic analysis of the spider mite sub-family Tetranychinae (Acari: Tetranychidae) based on the mitochondrial COI gene and the 18S and the 5' end of the 28S rRNA genes indicates that several genera are polyphyletic. PLoS ONE 9:e108672. https://doi.org/10.1371/journal.pone.0108672 Mochizuki M, Takafuji A (1996) Effect of photoperiod and temperature on the diapause termination of the Kanzawa spider mite, Tetranychus kanzawai Kishida (Acari: Tetranychidae). J Acarol Soc Jpn 5:83–88 Morimoto N, Takafuji A (1983) Comparison of diapause attributes and host preference among three populations of the citrus red mite, Panonychus citri (McGregor) occurring in the southern part of Okayama Prefecture, Japan. Jpn J Appl Entomol Zool 27:224–228 Mousseau TA, Fox CW (1998) The adaptive significance of maternal effects. Trends Ecol Evol 13:403–407. https://doi.org/10.1016/S0169-5347(98)01472-4 NAOJ (2023) Local Calendar for Kouchi (Kouchi). https://eco.mtk.nao.ac.jp/koyomi/dni/dni40.html Oda N, Ito K (2021) Photoperiodic control of reproductive arrest in the oak-inhabiting spider mite Schizotetranychus brevisetosus (Acari: Tetranychidae). Exp Appl Acarol 84:389–405. https://doi.org/10.1007/s10493-021-00630-6 Oku K, Yano S, Takafuji A (2003) Different maternal effects on diapause induction of tetranychid mites, Tetranychus urticae and T. kanzawai (Acari: Tetranychidae). Appl Entomol Zool 38:267–270. https://doi.org/10.1303/aez.2003.267 R Core Team (2022) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria, Vienna, Austria Sáez-Castillo AJ, Conde-Sánchez A, Martínez F (2022) DGLMExtPois: Advances in dealing with over and under-dispersion in a double GLM framework. R J 14:121–140. https://doi.org/10.32614/RJ-2023-002 Saito Y (2010) Plant mites and sociality: diversity and evolution. Springer-Verlag Tokyo Saito Y, Ito K, Sakagami T (2005) Imaginal induction of diapause in several adult-diapausing spider mites. Physiol Entomol 30:96–101. https://doi.org/10.1111/j.0307-6962.2005.00441.x Saito Y, Suzuki R (1987) Reexamination of several rearing methods for studying the life history of spider mites (Acari: Tetranychidae). Appl Entomol Zool 22:570–576. https://doi.org/10.1303/aez.22.570 Sakagami T (2002) Phylogenetic analysis of subfamily Tetranychinae (Acari: Tetranychidae) in Japan, based on 28S ribosomal DNA sequences: Does the celarius group belong to the genus Schizotetranychus ? Dissertation, Hokkaido University (in Japanese) Seck NKG, Ngom A, Noba K (2022) Modelling underdispersed count data: relative performance of Poisson model and its alternatives. Afr J Math Stat St 5:16–32. https://doi.org/10.52589/ajmss-1wpjqhyt Shinkaji N (1975) Hatching time of the winter eggs and termination of diapause in the common conifer spider mite, Oligonychus ununguis (Jacobi), on chestnut in relation to temperature (Acarina: Tetranychidae). Jpn J Appl Entomol Zool 19:144–148. https://doi.org/10.1303/jjaez.19.144 So P-M, Takafuji A (1991) Coexistence of Tetranychus urticae (Acarina: Tetranychidae) with different capacities for diapause: comparative life-history traits. Oecologia 87:146–151. https://doi.org/10.1007/bf00323792 Takafuji A, Kamibayashi M (1984) Life cycle of a non-diapausing population of the two-spotted spider mite, Tetranychus urticae Koch in a pear orchard. Res Popul Ecol 26:113–123. https://doi.org/10.1007/bf02515511 Takafuji A, Morishita M (2001) Overwintering ecology of two species of spider mites (Acari: Tetranychidae) on different host plants. Appl Entomol Zool 36:169–175. https://doi.org/10.1303/aez.2001.169 Takafuji A, So P-M, Tsuno N (1991) Inter- and intra-population variations in diapause attribute of the two-spotted spider mite, Tetranychus urticae Koch, in Japan. Res Popul Ecol 33:331–344. https://doi.org/10.1007/bf02513558 Tauber MJ, Tauber CA, Masaki S (1986) Seasonal adaptations of insects. Oxford University Press, New York Tsuda Y, Takafuji A, Kuno E (1997) Maintenance of diapause variability in the two-spotted spider mite, Tetranychus urticae , in a heterogeneous and stochastic environment. Res Popul Ecol 39:77–82. https://doi.org/10.1007/bf02765252 Veerman A (1985) Diapause. In: Helle W, Sabelis MW (eds) Spider Mites Their Biology, Natural Enemies and Control 1A. Elsevier, Amsterdam, pp 279–316 Wood SN (2011) Fast stable restricted maximum likelihood and marginal likelihood estimation of semiparametric generalized linear models. J R Stat Soc Ser B Stat Methodol 73:3–36. https://doi.org/10.1111/j.1467-9868.2010.00749.x Zar JH (2010) Biostatistical Analysis. 5th edition. Prentice Hall, Inc., New Jersey Zhou Z-S, Rasmann S, Li M, Guo J-Y, Chen H-S, Wan F-H (2013) Cold temperatures increase cold hardiness in the next generation Ophraella communa Beetles. PLoS ONE 8:e74760. https://doi.org/10.1371/journal.pone.0074760 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 12 Dec, 2024 Read the published version in Experimental and Applied Acarology → Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4084840","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":279786806,"identity":"e8a8c2c6-f813-4b2f-8df5-3b4d25273ce8","order_by":0,"name":"Kohei Nagata","email":"","orcid":"","institution":"Kochi University","correspondingAuthor":false,"prefix":"","firstName":"Kohei","middleName":"","lastName":"Nagata","suffix":""},{"id":279786807,"identity":"ac6f229a-3e4e-436e-a9b6-11e49dba48ae","order_by":1,"name":"Yamato Negoro","email":"","orcid":"","institution":"Kochi University","correspondingAuthor":false,"prefix":"","firstName":"Yamato","middleName":"","lastName":"Negoro","suffix":""},{"id":279786808,"identity":"95889881-d7a9-4ee8-a01b-ebce090a96b2","order_by":2,"name":"Katsura Ito","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAwUlEQVRIiWNgGAWjYBACCQYGxocNDAwJbFABZsJa2BiYDUnWwiYJ0kK8wyTn9z6rnFFhl8fHwGPA8KOGgd2ckBZpNnazmxvOJBezAbUw9hxjYLZsIKBFjo2N7ebDtgOJbfJvDBh4GxiYDQ4QoaUQrAVky19itEgDtTBuhGphJsoWybY0ZskZZ5KBWtgKDssckyDsF4nDxxg/9lTYJc5vYN748E2NTTLBEEMBQCdJJBuQpAUE7EjXMgpGwSgYBcMdAAAs5jQqMOp8kgAAAABJRU5ErkJggg==","orcid":"","institution":"Kochi University","correspondingAuthor":true,"prefix":"","firstName":"Katsura","middleName":"","lastName":"Ito","suffix":""}],"badges":[],"createdAt":"2024-03-12 14:32:11","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4084840/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4084840/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s10493-024-00978-5","type":"published","date":"2024-12-12T15:57:59+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":52792396,"identity":"c8600aa4-dd2d-4c01-863a-f284e831e169","added_by":"auto","created_at":"2024-03-15 20:17:59","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":157565,"visible":true,"origin":"","legend":"\u003cp\u003ePhotoperiodic response of ovipositional status in \u003cem\u003eSchizotetranychus shii\u003c/em\u003e at 20°C. The status was determined at 7 days after adult emergence. \u003cem\u003en\u003c/em\u003e= 33–36 for each treatment.\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-4084840/v1/a719184f98737a83590846af.jpeg"},{"id":52792368,"identity":"0af6c707-4d43-40d3-96e9-979e633f78ac","added_by":"auto","created_at":"2024-03-15 20:17:45","extension":"jpeg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":380901,"visible":true,"origin":"","legend":"\u003cp\u003eLife-stage composition of a \u003cem\u003eSchizotetranychus shii\u003c/em\u003e population in Nankoku, Kochi, Japan (2021–2022). (A) Proportion of each life stage, (B) density per infested leaf, and (C) adult sex ratio. F, females; M, males; A, adults; LN, larvae and nymphs; E, eggs; and T, total. X-axis is marked in 14-day intervals.\u003c/p\u003e","description":"","filename":"floatimage3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-4084840/v1/e3dffe61e55f551504ddb2a6.jpeg"},{"id":52792388,"identity":"75cb5aa7-4298-462b-b75b-b66efeb754ba","added_by":"auto","created_at":"2024-03-15 20:17:55","extension":"jpeg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":300618,"visible":true,"origin":"","legend":"\u003cp\u003eResponse curves of GAM analyses for life stages in 2021–22. Y-axis indicates the zero-centred log-scale response (density) to days elapsed. There were insufficient adult males for analysis. X-axis is marked in 14-day intervals.\u003c/p\u003e","description":"","filename":"floatimage5.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-4084840/v1/5fa7d6a7d954b7c656f8ad43.jpeg"},{"id":52792393,"identity":"68162954-d67e-4917-b0ec-1d3f2b74d87b","added_by":"auto","created_at":"2024-03-15 20:17:58","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":36845,"visible":true,"origin":"","legend":"\u003cp\u003eDays required for egg hatching under 15L conditions at 20°C. \u003cem\u003eField\u003c/em\u003e indicates the eggs collected from the study site in late October and late November, and \u003cem\u003eCulture\u003c/em\u003e refers to the new eggs obtained from the laboratory strain in the same season. Sample sizes are shown above boxes. The boxplot shows the median (thick horizontal line), the first and third quartiles (outer edges of box), and 1.5× the inter-quartile range (whiskers). Dots indicate individual data plots. See Table 4 for the results of the analyses.\u003c/p\u003e","description":"","filename":"floatimage7.png","url":"https://assets-eu.researchsquare.com/files/rs-4084840/v1/1461c6c91f4b47e87524af30.png"},{"id":52792395,"identity":"d81e6787-b6c9-4fc1-a303-40e514f0ec47","added_by":"auto","created_at":"2024-03-15 20:17:59","extension":"jpeg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":555797,"visible":true,"origin":"","legend":"\u003cp\u003eLowest and highest temperatures (solid line) and day length (dashed line) from October 2017 to March 2022 in Nankoku-Nissho, Kochi. Mean and SD are indicated. Data from Japan Meteorological Agency (2024).\u003c/p\u003e","description":"","filename":"floatimage8.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-4084840/v1/6670bb3379f340dd3fa4d385.jpeg"},{"id":71552485,"identity":"79400d6e-dae5-4c4a-a029-1ca441546e9b","added_by":"auto","created_at":"2024-12-16 16:06:38","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2121116,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4084840/v1/80cfb6e1-9d21-498a-88b5-062f0370c2dd.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Overwintering at multiple life stages in Schizotetranychus shii (Acari: Tetranychidae), a specialist of evergreen chinquapin","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eHost-plant phenology exerts strong selection pressure on overwintering strategies in herbivorous arthropods (Tauber et al. \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e1986\u003c/span\u003e; Danks \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e1987\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). The timing of leaf extension and defoliation affects the timing of herbivore development and reproduction. Their overwintering patterns are shaped so that life stages vulnerable to adverse conditions such as food deprivation, low temperature, or frost do not encounter such harsh environments. The importance of host availability in life-cycle formation is strongly supported by the associations between host phenology and winter life-stage composition in diverse herbivorous taxa (Gotoh \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e1983\u003c/span\u003e; Morimoto and Takafuji \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e1983\u003c/span\u003e; Carey \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1994\u003c/span\u003e; Hunter and McNeil \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e1997\u003c/span\u003e; Kurota and Shimada \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2002\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe success of development and reproduction in spider mites (Acari: Tetranychidae) depends on the spatio-temporal stability of hosts. Using mathematical models incorporating patchy habitats of different quality, Tsuda et al. (\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e1997\u003c/span\u003e) found that the existence of winter hosts favours non-diapausing females, or females with phenotypic plasticity whose phenotypes (diapausing or non-diapausing) match the habitat quality (i.e., females avert diapause in good habitats). Because the spider mites parasitize various hosts of different phenology and have diverse life-history strategies, they are suitable model organisms to evaluate the effects of host phenology on life-cycle formation. The significance of host availability on life cycles is suggested in \u003cem\u003eTetranychus\u003c/em\u003e (Morimoto and Takafuji \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e1983\u003c/span\u003e), \u003cem\u003eSchizotetranychus\u003c/em\u003e (Gotoh \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e1983\u003c/span\u003e; Ito and Yamanishi \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), \u003cem\u003eSasanychus\u003c/em\u003e (Gotoh \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e1986a\u003c/span\u003e, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003eb\u003c/span\u003e; Gotoh \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e1986b\u003c/span\u003e), \u003cem\u003eStigmaeopsis\u003c/em\u003e (Saito and Ueno 1979) and \u003cem\u003ePanonychus\u003c/em\u003e (Morimoto and Takafuji \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e1983\u003c/span\u003e; Koveos and Broufas \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e1999\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThese studies have traditionally associated overwintering patterns with diapause induction. In most species, short day length and low temperatures during development (Veerman \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e1985\u003c/span\u003e) or adulthood (Lees \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e1953\u003c/span\u003e; Saito et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2005\u003c/span\u003e) induce diapause, and host conditions (Ito \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Ito and Saito \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2006\u003c/span\u003e), maternal effects (Danilevskii \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e1965\u003c/span\u003e; Oku et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2003\u003c/span\u003e) and predation risks (Kroon et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2004\u003c/span\u003e, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2005\u003c/span\u003e, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2008\u003c/span\u003e) modify the primary response of day length. Despite the progress of these studies, however, the relationship between diapausing stages and overwintering stages realized in the field is still confusing (Saito \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2010\u003c/span\u003e); only diapausing individuals can overwinter in cold regions or on deciduous hosts, but this restriction is relaxed in warmer regions, where winter hosts are plentiful. In temperate populations of \u003cem\u003eTetranychus urticae\u003c/em\u003e and \u003cem\u003eT. kanzawai\u003c/em\u003e, all life stages occur throughout winter, even though populations retain sensitivity to adult diapause during short photoperiods (Takafuji and Kamibayashi \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e1984\u003c/span\u003e; So and Takafuji \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e1991\u003c/span\u003e; Takafuji et al. \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e1991\u003c/span\u003e; Takafuji and Morishita \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2001\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAdaptation to evergreen hosts, which provide temporally stable habitats, may relax the constraint of having a single overwintering stage (Gotoh \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e1986a\u003c/span\u003e, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003eb\u003c/span\u003e; Ito and Yamanishi \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Overwintering as adult females and their eggs are found in \u003cem\u003eYezonychus sapporensis\u003c/em\u003e (H. Yanagida, unpublished data, Saito \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2010\u003c/span\u003e) and \u003cem\u003eSasanychus akitanus\u003c/em\u003e (Gotoh \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e1986a\u003c/span\u003e, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003eb\u003c/span\u003e) on Sasa bamboo, which grow in cool regions and are covered by snow in winter. Gotoh (\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e1986a\u003c/span\u003e, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003eb\u003c/span\u003e) reports that \u003cem\u003eS. akitanus\u003c/em\u003e females continue to lay diapausing eggs until early winter. Although the author did not explicitly relate the scarcity of predators to the oviposition patterns, the inactivity of midge larvae and phytoseiid mites might underlie the production of winter eggs until late in the season. On the other hand, in \u003cem\u003eSchizotetranychus brevisetosus\u003c/em\u003e, which parasitizes \u003cem\u003eQuercus glauca\u003c/em\u003e L. (Fagaceae) (Ehara \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e1989\u003c/span\u003e; Ehara and Gotoh \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2009\u003c/span\u003e), adult females occupy the population in December, produce winter eggs until early March and die out (Ito and Hamada \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Ito and Yamanishi \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Oda and Ito \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). This style is distinct from \u003cem\u003eS. akitanus\u003c/em\u003e, whose females cease producing diapausing eggs and overwinter themselves and adults of different generations interbreed in spring (Gotoh \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e1986a\u003c/span\u003e). Predators are almost absent during winter in \u003cem\u003eS. brevisetosus\u003c/em\u003e (Ito \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), and this low predation pressure enables the overwintering of winter eggs. This inter-specific difference in overwintering styles may be partly explained by climatic conditions; \u003cem\u003eS. brevisetosus\u003c/em\u003e individuals can act even during winter. Despite these differences in the overwintering pattern, photoperiodism underlies reproductive arrest in both species (Gotoh \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e1986b\u003c/span\u003e; Oda and Ito \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cem\u003eSchizotetranychus shii\u003c/em\u003e (Acari: Tetranychidae) is the specialist of evergreen chinquapin \u003cem\u003eCastanopsis sieboldii\u003c/em\u003e (Ehara 1965). Individuals live gregariously in silk-web nests constructed in the depressions along leaf veins or edges. This species overwinters in various life stages and is therefore considered a non-diapausing species (Ehara and Gotoh \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). It has been assigned to the same clade as \u003cem\u003eS. brevisetosus\u003c/em\u003e in phylogenetic trees based on 18S and 28S rDNA gene sequences, whereas \u003cem\u003eS. akitanus\u003c/em\u003e is in a separate clade (Matsuda et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2014\u003c/span\u003e, Figs.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e4\u003c/span\u003e and \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e5\u003c/span\u003e). This makes \u003cem\u003eS. shii\u003c/em\u003e a suitable species for studying evolutionary changes in overwintering patterns through comparison with those in \u003cem\u003eS. brevisetosus\u003c/em\u003e. Here, we focused on the temporal changes in life-stage composition during winter in \u003cem\u003eS. shii\u003c/em\u003e. We tested the hypothesis that photoperiodic response for reproductive arrest is responsible for the formation of overwintering patterns. Based on the results, we discuss the evolution of overwintering patterns in spider mites on evergreen hosts.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003ePhotoperiodic response\u003c/h2\u003e \u003cp\u003eA laboratory strain of \u003cem\u003eS. shii\u003c/em\u003e was established from 50 individuals at various life stages collected from \u003cem\u003eC. sieboldii\u003c/em\u003e trees in Monobe Campus, Kochi University, Nankoku, Kochi Prefecture, Japan (33\u0026deg; 33' 05\"N, 133\u0026deg; 40' 30\" E, WGS84, 8 m altitude) on 8 October 2021. The strain was maintained on detached \u003cem\u003eC. sieboldii\u003c/em\u003e leaves using the leaf-culture method (Helle and Overmeer \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e1985\u003c/span\u003e). Briefly, a detached leaf was pressed on water-soaked cotton pads piled on a polyurethane mat in an insect breeding dish (100(d) \u0026times; 40(h) mm, SPL Life Sciences, Gyeonggi-do, Korea). The leaf edge was surrounded by Kimwipe\u0026reg; strips to prevent mite escape and leaf curling. Each dish was covered with a mesh lid to mitigate a temperature decrease on the leaf surface due to evaporation (Saito and Suzuki \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e1987\u003c/span\u003e). The culture was maintained under 25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5\u0026deg;C (mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD) and a photoperiod of 15 h light and 9 h dark (15L) conditions in a MIR-154 chamber (SANYO, Osaka, Japan) for two months. Relative humidity in the chamber was not controlled, but data loggers (RTR-503, T\u0026amp;D Corporation, Nagano, Japan) showed that it typically ranged between 60 and 80%.\u003c/p\u003e \u003cp\u003eWe established photoperiodic treatments from 10L to 13L at one-hour intervals in temperature gradient incubator chambers (TG-300WLED-5LE, NK System, Osaka, Japan). We also included a 10.5L treatment, which approximated the day length excluding civil twilight in mid-November (NAOJ \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), when the life-stage compositions of a nearby \u003cem\u003eS. brevisetosus\u003c/em\u003e population underwent significant change (Ito and Yamanishi \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Temperatures in the chambers were maintained at 20\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C according to the TR-51i, 52i and RTR-503 data loggers (T\u0026amp;D Corporation).\u003c/p\u003e \u003cp\u003eTwo leaf cultures were prepared per day length treatment. Ten adult females collected from the rearing culture were introduced onto each culture using a fine brush and allowed to oviposit for 3 days at 25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5\u0026deg;C under a photoperiod of 15L in the MIR-154 chamber. Subsequently, 37\u0026ndash;76 eggs were obtained per leaf. The females were removed and the dishes were maintained at their respective treatment conditions. Emerged females were separated onto \u003cem\u003eC. sieboldii\u003c/em\u003e leaf squares (1.5 cm \u0026times; 1.5 cm) daily, maintained under the same photoperiodic treatment, and the onset of oviposition from emergence was monitored (n\u0026thinsp;=\u0026thinsp;33\u0026ndash;36 per treatment). The oviposition status of each female was assessed on the seventh day from isolation, considering that oviposition occurred within 7 days at 13L (see Results), which was a long day length at the end of August (NAOJ \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2023\u003c/span\u003e) when mites are highly reproductive. Females that died within seven days were excluded from the analysis. The effects of day length on the proportions of non-ovipositing females were analysed using generalized linear models (GLM), assuming a binomial distribution with a logit link function. The \u003cem\u003eglm\u003c/em\u003e command in the software R version 4.2.2 (R Core Team \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) was used for this purpose. The critical day length (CDL), at which 50% of females oviposited, was estimated as \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(-a/b\\)\u003c/span\u003e\u003c/span\u003e, where \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(a\\)\u003c/span\u003e\u003c/span\u003e and \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(b\\)\u003c/span\u003e\u003c/span\u003e represent the intercept and the slope of the fitted model, respectively (Oda and Ito \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). The model assumptions, especially the distribution and deviance of residuals, were assessed visually.\u003c/p\u003e \u003cp\u003eTo assess the depth of reproductive arrest, we also counted the number of females starting oviposition within 30 days of isolation in the 10L, 10.5L and 11L treatments. This threshold was selected based on previous studies on the duration of diapause termination in temperate regions (Mochizuki and Takafuji \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e1996\u003c/span\u003e; Ito \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Ito et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). The proportions of ovipositing females among the day length treatments were compared using Chi-square tests.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eField survey of life stages\u003c/h2\u003e \u003cp\u003eApproximately 30 leaves (ranging from 12 to 32) with web nests were collected from the Monobe Campus using a 10\u0026times; magnifying grass. These leaves were immediately observed using a stereomicroscope (SZ-61 and SZX-7, Olympus, Tokyo, Japan), and the life stages of \u003cem\u003eS. shii\u003c/em\u003e were divided into four categories: \u003cem\u003eegg\u003c/em\u003e, \u003cem\u003eimmature\u003c/em\u003e (larvae and nymphs), \u003cem\u003eadult female\u003c/em\u003e and \u003cem\u003emale\u003c/em\u003e. Sampling was conducted weekly from 8 October 2021 to 28 January 2022 and then biweekly until 22 April 2022, when colonies were disturbed by predators. The periodic changes in stage composition were tested via Chi-square tests with Yates\u0026rsquo; correction using the \u003cem\u003echisq.test\u003c/em\u003e function in R. Adult sexes were pooled because males were rare.\u003c/p\u003e \u003cp\u003eThe association between the observation date and the density of each life stage was analysed by a generalised additive model (GAM) using the \u003cem\u003egam\u003c/em\u003e function in the \u003cem\u003emgcv\u003c/em\u003e package version 1.9.0 (Wood \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2011\u003c/span\u003e) in R. Parameter estimation followed the procedure outlined by Crawley (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). The statistical model was as follows:\u003cdiv id=\"Equ1\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equ1\" name=\"EquationSource\"\u003e\n$$\\text{log }\\left(y\\right)={\\beta }_{0}+s\\left(x\\right)+\\text{l}\\text{o}\\text{g}\\left(n\\right)$$\u003c/div\u003e\u003cdiv class=\"EquationNumber\"\u003e1\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003ewhere \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(y\\)\u003c/span\u003e\u003c/span\u003e is the number of individuals at each life stage, \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\({\\beta }_{0}\\)\u003c/span\u003e\u003c/span\u003e is the model intercept, \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(s\\left(x\\right)\\)\u003c/span\u003e\u003c/span\u003e is the smooth function against the day \u003cem\u003ex\u003c/em\u003e and \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\text{l}\\text{o}\\text{g}\\left(n\\right)\\)\u003c/span\u003e\u003c/span\u003e is the offset term expressed as the logarithm of leaf number. The best spline function was selected using the residual maximum likelihood (REML) method (Wood \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). For each life stage, the GAM model and a simple linear model were compared by likelihood ratio (LR) tests using the \u003cem\u003eanova\u003c/em\u003e function in R.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eDays required for hatching in the field (transplant experiment)\u003c/h2\u003e \u003cp\u003eA new laboratory strain was established from 15 adult females from the Monobe Campus on 13 September 2022 and maintained under 15L conditions at 25\u0026deg;C for a month. Three sets of experiments commenced on 15 October, 19 November and 24 December 2022. In each set, 10 (October) or 15 (November and December) adult females from the laboratory strain were placed on each of two leaves (November) or one leaf (December) prepared as above (the leaf culture), and allowed to oviposit for 3 days. Following the removal of females, the leaf cultures were enclosed in a mesh cage (internal dimension: 30 cm \u0026times; 30 cm \u0026times; 30 cm, BugDorm, Taichung, Taiwan) to prevent predator intrusion and exposure to rain. The cage was fixed to a plastic stand (10 cm in height) set on the forest floor using vinyl ropes. The leaf cultures were checked for egg hatching on-site using a 10\u0026times; magnifying grass every 3 days, and subsequently daily after hatching began. Deformed or discoloured eggs were regarded as dead. For cases where precise counting was difficult, the leaves were assessed in the laboratory with a stereomicroscope.\u003c/p\u003e \u003cp\u003eThe average days required for hatching among groups were compared via Welch\u0026rsquo;s ANOVA using the \u003cem\u003eoneway.test\u003c/em\u003e function in R. The survival rates of eggs were compared via Chi-square tests with Yates\u0026rsquo; correction, and pairwise comparisons were conducted via Tukey\u0026rsquo;s multiple test for proportions (Zar \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2010\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eHatchability of autumnal eggs in the field\u003c/h2\u003e \u003cp\u003eEggs were collected from the Monobe Campus on (I) 24 October and (II) 20\u0026ndash;26 November 2023 to measure the days required for hatching described as below. Each month, several field-collected leaves with eggs were arbitrarily excised with a razor blade, and these pieces were placed on wet cotton pads in a breeding dish. The leaf pieces were maintained under 15L conditions at 20\u0026deg;C, and hatching was assessed daily, with larvae being removed each time they were observed.\u003c/p\u003e \u003cp\u003eAs a control, 20 females from the new laboratory strain (derived from 70 individuals at various life stages from the Monobe Campus on 23 July 2023 and maintained at 25\u0026deg;C/16L) were allowed to oviposit for 2 days on a detached leaf. Eggs that were laid on the first day were removed with a fine brush to adjust the age of eggs, and the occurrence of hatching was recorded as above. In other words, newly laid eggs were obtained for this strain.\u003c/p\u003e \u003cp\u003eThe data were structured as a two-way design (months \u0026times; strain) and were analysed by GLM. However, the data were heavily underdispersed from Poisson distribution due to the mixing of data obtained in different environments (see Results), and established analytical methods are not applicable to such cases (S\u0026aacute;ez-Castillo et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Seck et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Accordingly, we adopted a GLM with a two-way design (months \u0026times; groups), assuming a COM-Poisson distribution with a log link function and a constant dispersion parameter, using the \u003cem\u003eglm.CMP\u003c/em\u003e in the DGLMExtPoison package version 0.2.3 (S\u0026aacute;ez-Castillo et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The significance of each term was assessed by comparing it with the full model using likelihood ratio (LR) tests by \u003cem\u003elrtest\u003c/em\u003e in R.\u003c/p\u003e \u003c/div\u003e"},{"header":"RESULTS","content":"\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003ePhotoperiodic response\u003c/h2\u003e \u003cp\u003eThe developmental period of adult females under 13L was 5.4\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2 days, with a range of 4 to 8 days, and a 97% oviposition rate within 7 days (n\u0026thinsp;=\u0026thinsp;33). Accordingly, females ovipositing within 7 days were defined as reproducing. Figure\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e1\u003c/span\u003e shows the response curve of oviposition rates against photoperiod. All females under 10L, 10.5L and 11L never oviposited within seven days (0%, n\u0026thinsp;=\u0026thinsp;34\u0026ndash;36), but most females laid eggs under 12L (80.6%, n\u0026thinsp;=\u0026thinsp;36) and 13L. The logistic analysis estimated a positive intercept and a negative slope defined by \u003cem\u003ey\u003c/em\u003e\u0026thinsp;=\u0026thinsp;57.73\u0026ndash;4.91\u003cem\u003ex\u003c/em\u003e, with residual deviance\u0026thinsp;=\u0026thinsp;5.56 and degrees of freedom (df)\u0026thinsp;=\u0026thinsp;3. The CDL was estimated as 11.8L. Though there were no apparent outliers, residual analysis suggested that the 12L datapoint is slightly deviated from the prediction line of the normal Q\u0026ndash;Q plot and had a higher leverage.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eEven when kept under short-day conditions (10\u0026ndash;11L), 64\u0026ndash;86% of the non-ovipositing females survived 30 days after emergence, and 90\u0026ndash;96% of them oviposited within that period (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Except for survival rates, no differences were found between 10L and the other two daylength treatments.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe proportions of surviving females 30 days after emergence and their ovipositing rate (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;36 for all treatments). The results of Chi-square test and, if significant, Tukey\u0026rsquo;s multiple comparisons for proportions (Zar \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2010\u003c/span\u003e) are indicated by different lower-case letters.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDaylength\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e10L\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e10.5L\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e11L\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003eχ\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003edf\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSurvival rate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.64\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.86\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.86\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e7.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.029\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOviposition rate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.730\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eField survey of life stages\u003c/h2\u003e \u003cp\u003eThe proportion of life stages changed significantly during the field survey (Fig.\u0026nbsp;2A, \u003cem\u003eχ\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;566.27, df\u0026thinsp;=\u0026thinsp;42, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001). In October, the proportion of life stages did not significantly vary (\u003cem\u003eχ\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;5.06, df\u0026thinsp;=\u0026thinsp;6, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.537) and the proportions of eggs remained moderate (45.1\u0026ndash;64.0%). The proportion of eggs gradually decreased to 11.8% by 3 December (\u003cem\u003eχ\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;22.86, df\u0026thinsp;=\u0026thinsp;7, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.002). Conversely, the proportion of immature stages increased from 23.1% on 8 October to 52.4% on 26 November (\u003cem\u003eχ\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;23.56, df\u0026thinsp;=\u0026thinsp;7, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.001). The proportion of adults increased from 15.4% on 8 October to 41.2% on 3 December (\u003cem\u003eχ\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;16.00, df\u0026thinsp;=\u0026thinsp;8, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.042), slowly decreased to 1.2% on 7 April (1.2%) (\u003cem\u003eχ\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;109.77, df\u0026thinsp;=\u0026thinsp;11, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001) with a concurrent increase in the proportion of eggs and rose again to 20.4% on 22 April (\u003cem\u003eχ\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;62.70, df\u0026thinsp;=\u0026thinsp;1, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001). The proportion of eggs rose from 3 December to 25 February (\u003cem\u003eχ\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;122.89, df\u0026thinsp;=\u0026thinsp;9, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001), whereas the proportions of immature stages, including larvae (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e), declined during this period (0.7\u0026ndash;47.1%, \u003cem\u003eχ\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;93.78, df\u0026thinsp;=\u0026thinsp;9, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe number of larvae (L) and nymphs (N) of \u003cem\u003eSchizotetranychus shii\u003c/em\u003e between 2021\u0026ndash;2022. The number of teleiochrysalis females (Tf) is shown separately.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"16\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c12\" colnum=\"12\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c13\" colnum=\"13\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c14\" colnum=\"14\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c15\" colnum=\"15\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c16\" colnum=\"16\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDate\u003c/p\u003e \u003cp\u003eMonth\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e12\u003c/p\u003e \u003cp\u003eN\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e19\u003c/p\u003e \u003cp\u003eN\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e26\u003c/p\u003e \u003cp\u003eN\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3\u003c/p\u003e \u003cp\u003eD\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e10\u003c/p\u003e \u003cp\u003eD\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e17\u003c/p\u003e \u003cp\u003eD\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003e24\u003c/p\u003e \u003cp\u003eD\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003e7\u003c/p\u003e \u003cp\u003eJ\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003e14\u003c/p\u003e \u003cp\u003eJ\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c11\"\u003e \u003cp\u003e21\u003c/p\u003e \u003cp\u003eJ\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c12\"\u003e \u003cp\u003e28\u003c/p\u003e \u003cp\u003eJ\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c13\"\u003e \u003cp\u003e11\u003c/p\u003e \u003cp\u003eF\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c14\"\u003e \u003cp\u003e25\u003c/p\u003e \u003cp\u003eF\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c15\"\u003e \u003cp\u003e11\u003c/p\u003e \u003cp\u003eM\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c16\"\u003e \u003cp\u003e25\u003c/p\u003e \u003cp\u003eM\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eL\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c13\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c14\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c15\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c16\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eN\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c12\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c13\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c14\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c15\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c16\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e(Tf)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c13\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c14\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c15\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe mean density of each life stage per sampled leaf is shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e2\u003c/span\u003eB. The total density remained below 3.0 until 7 January but then increased to approximately 15.5 on 22 April, with an increase in egg density after December (representing 70.8\u0026ndash;96.4% of the population). The density of immature stages peaked on 26 November (1.1) and gradually decreased toward 25 February (0.0). The immature stages reappeared on 11 March (0.3) and increased towards 7 April (7.6). The adults of the next generation emerged on 22 April (3.2), and some were observed moving onto newly extended leaves. The density of adults did not show a clear seasonal trend. Sex ratios fluctuated between 8\u0026ndash;50% until 28 January (except for 0% on 24 December), and males were absent between 11 February to 7 April (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e2\u003c/span\u003eC). However, the change in sex ratios over the survey period was not significant due to the small number of males (\u003cem\u003eχ\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;26.05, df\u0026thinsp;=\u0026thinsp;21, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.205).\u003c/p\u003e \u003cp\u003eThe results of GAM models are summarised in Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e3\u003c/span\u003e and Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. The response curve in adult females showed no conspicuous trend until March, although density increased in late April (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e3\u003c/span\u003eA). The curves for immature stages decreased toward late February and began to increase thereafter (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e3\u003c/span\u003eB). The curve for eggs fluctuated until November and gradually increased after December (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e3\u003c/span\u003eC). The smooth term for adult females was not significant (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.576, Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e), which suggests that spline fitting is unnecessary. However, the smooth terms were significant for immature stages and eggs (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001). Also, LR tests revealed that spline fitting was no better than a simple linear model for females (df\u0026thinsp;=\u0026thinsp;2.32, deviance\u0026thinsp;=\u0026thinsp;3.47, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.220), but improved fit for immature stages (df\u0026thinsp;=\u0026thinsp;3.72, deviance\u0026thinsp;=\u0026thinsp;29.84, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001) and eggs (df\u0026thinsp;=\u0026thinsp;8.03, deviance\u0026thinsp;=\u0026thinsp;180.74, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe summary of GAM analyses for each life stage in 2021\u0026ndash;22 (n\u0026thinsp;=\u0026thinsp;22). F: adult females, LN: larvae and nymphs, and E: eggs. The data of adult males were not analysed because of insufficient sample size. The results of LR tests based on REML methods are shown.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStage\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eEstimated df\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eχ\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eDeviance (%)\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;REML\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eF\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.576\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e11.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e53.94\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLN\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e54.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e74.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e56.74\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eE\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e7.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e231.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e81.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e97.18\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"6\"\u003e\u003csup\u003e*\u003c/sup\u003eDeviance explained by the smooth term \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(s\\left(x\\right)\\)\u003c/span\u003e\u003c/span\u003e in Eq.\u0026nbsp;(\u003cspan refid=\"Equ1\" class=\"InternalRef\"\u003e1\u003c/span\u003e)\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eDays required for hatching in the field (transplant experiment)\u003c/h2\u003e \u003cp\u003eThe hatchability of eggs placed in the field on October 15 (n\u0026thinsp;=\u0026thinsp;21) and November 19 (n\u0026thinsp;=\u0026thinsp;85) were 100.0% and 51.8%, respectively, but all eggs set on 24 December died during development. The survival rates of the eggs were significantly differed among months (\u003cem\u003eχ\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;70.353, df\u0026thinsp;=\u0026thinsp;2, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001), and pairwise comparisons revealed differences between December and the other two months (Tukey\u0026rsquo;s multiple test for proportions, \u003cem\u003eα\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.05). Days required for hatching were 13.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.8 (n\u0026thinsp;=\u0026thinsp;21) and 20.1\u0026thinsp;\u0026plusmn;\u0026thinsp;7.9 days (n\u0026thinsp;=\u0026thinsp;44) for the eggs set in October and November, respectively (Welch\u0026rsquo;s ANOVA, \u003cem\u003eF\u003c/em\u003e\u0026thinsp;=\u0026thinsp;24.446, \u003cem\u003edf\u003c/em\u003e\u003csub\u003e\u003cem\u003e1\u003c/em\u003e\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;1, \u003cem\u003edf\u003c/em\u003e\u003csub\u003e\u003cem\u003e2\u003c/em\u003e\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;51.689, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001). The last eggs to hatch did so on 2 November and 5 January, respectively.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eHatchability of autumnal eggs in the field\u003c/h2\u003e \u003cp\u003eFigure \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e4\u003c/span\u003e shows the days for field and laboratory eggs to hatch under 15L conditions at 20\u0026deg;C. The hatch times in the field were highly variable, especially in November, and one egg needed more than 10 days to hatch. A COM-Poisson GLM showed underdispersion (i.e., higher average with lower variance, Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e4\u003c/span\u003e) as suggested by the dispersion parameter of 1.31 (\u0026gt;\u0026thinsp;1). The hatch times in November were significantly longer than those in October (Table\u0026nbsp;4A, Month), and the hatch times in culture were also longer than those in the field (Treatment). The interaction between treatments was highly significant (Month:Treatment). The LR tests revealed significance in all terms (Table\u0026nbsp;4B).\u003c/p\u003e \u003c/div\u003e\u003cp\u003e\u003cstrong\u003eTable 4.\u003c/strong\u003e \u0026nbsp;(A) Estimated coefficients of a COM-Poisson GLM on the days required for egg hatching (Fig. 4). (B) LR tests (compared with the full model; df = 57). \u003c/p\u003e\n\u003cp\u003e(A)\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"640\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"41.58878504672897%\" colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003eSource\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.576323987538942%\" valign=\"top\"\u003e\n \u003cp\u003eEstimate\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.370716510903426%\" valign=\"top\"\u003e\n \u003cp\u003eSE\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.576323987538942%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cem\u003ez\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.576323987538942%\" valign=\"top\"\u003e\n \u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.3115264797507788%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"41.58878504672897%\" colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003eFixed effects\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.576323987538942%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.370716510903426%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.576323987538942%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.576323987538942%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.3115264797507788%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"12.1875%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.53125%\" valign=\"top\"\u003e\n \u003cp\u003eIntercept\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" valign=\"top\"\u003e\n \u003cp\u003e1.04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.40625%\" valign=\"top\"\u003e\n \u003cp\u003e0.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" valign=\"top\"\u003e\n \u003cp\u003e10.42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e\u0026lt; 0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"12.1875%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.53125%\" valign=\"top\"\u003e\n \u003cp\u003eMonth (Nov)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" valign=\"top\"\u003e\n \u003cp\u003e0.78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.40625%\" valign=\"top\"\u003e\n \u003cp\u003e0.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" valign=\"top\"\u003e\n \u003cp\u003e6.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e\u0026lt; 0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"12.1875%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.53125%\" valign=\"top\"\u003e\n \u003cp\u003eTreatment (Culture)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" valign=\"top\"\u003e\n \u003cp\u003e1.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.40625%\" valign=\"top\"\u003e\n \u003cp\u003e0.11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" valign=\"top\"\u003e\n \u003cp\u003e13.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e\u0026lt; 0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"12.1875%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.53125%\" valign=\"top\"\u003e\n \u003cp\u003eMonth:Treatment\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026ndash;0.79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.40625%\" valign=\"top\"\u003e\n \u003cp\u003e0.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026ndash;5.66\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e\u0026lt; 0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"12.1875%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.53125%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.40625%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"12.1875%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.53125%\" valign=\"top\"\u003e\n \u003cp\u003edf\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" valign=\"top\"\u003e\n \u003cp\u003e57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.40625%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"12.1875%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.53125%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.40625%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"41.71875%\" colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003eDispersion coefficient \u0026nu; (logit link)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.40625%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"12.1875%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"29.53125%\" valign=\"top\"\u003e\n \u003cp\u003eIntercept\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" valign=\"top\"\u003e\n \u003cp\u003e1.31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.40625%\" valign=\"top\"\u003e\n \u003cp\u003e0.18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" valign=\"top\"\u003e\n \u003cp\u003e7.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.625%\" colspan=\"2\" valign=\"top\"\u003e\n \u003cp\u003e\u0026lt; 0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e(B)\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"38.25%\" valign=\"top\"\u003e\n \u003cp\u003eNull model\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.75%\" valign=\"top\"\u003e\n \u003cp\u003edf\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003eStatistic*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"38.25%\" valign=\"top\"\u003e\n \u003cp\u003eTreatment\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.75%\" valign=\"top\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e29.81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026lt; 0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"38.25%\" valign=\"top\"\u003e\n \u003cp\u003eMonth\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.75%\" valign=\"top\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e112.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026lt; 0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"38.25%\" valign=\"top\"\u003e\n \u003cp\u003eTreatment + Month\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"11.75%\" valign=\"top\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e26.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026lt; 0.001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e*\u0026ndash;2(logL\u003csub\u003e0\u003c/sub\u003e/L\u003csub\u003e1\u003c/sub\u003e), where L\u003csub\u003e0\u003c/sub\u003e and L\u003csub\u003e1\u003c/sub\u003e are the maximum likelihoods of the null (reduced) model and the full model, respectively.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eWe demonstrated that various stages of \u003cem\u003eS. shii\u003c/em\u003e coexist during winter and that stage compositions changed with time. Although this pattern appears to be consistent with the traditional view that \u003cem\u003eS. shii\u003c/em\u003e is a non-diapausing species (Ehara and Gotoh \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2009\u003c/span\u003e), with overwintering patterns being regulated by photoperiod like \u003cem\u003eS. brevisetosus\u003c/em\u003e or common diapausing species (Ito and Yamanishi \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), our findings indicate that short photoperiod strongly induced reproductive arrest in adult females, and that the photoperiodic response followed a sigmoid curve ranging from 0\u0026ndash;100% of the population (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e1\u003c/span\u003e). This study represents the first to reveal a tight relationship between population dynamics of species overwintering in multiple life stages and photoperiodism.\u003c/p\u003e \u003cp\u003eThe females of \u003cem\u003eS. shii\u003c/em\u003e arrested reproduction in response to short photoperiods, as reported in \u003cem\u003eS. brevisetosus\u003c/em\u003e on evergreen oak (Oda and Ito \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Females completely arrested oviposition under 10\u0026ndash;11L conditions, with a CDL of 11.8L (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e1\u003c/span\u003e), which corresponds with the day length in Kochi city at the beginning of October (NAOJ \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Consequently, immature stages that developed in late September (12L) or early October (11.5L) may cease reproduction, which explains the observed reduction of the egg proportion after late October (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e2\u003c/span\u003eA, B, Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e3\u003c/span\u003eC). Although this CDL is slightly shorter than the consensus value of 10 geographic populations of \u003cem\u003eS. brevisetosus\u003c/em\u003e (12.2L, Ito and Yamanishi \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), it is close to CDL of a population at a nearby site (Noichi). Therefore, the season of oviposition arrest is likely to be similar between the two species, but the overwintering patterns of the two species differ significantly. In \u003cem\u003eS. brevisetosus\u003c/em\u003e, adult females that emerge after early October do not oviposit, and the proportion of eggs steeply declined from October to November (Ito and Yamanishi \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Most of these females produce winter eggs from December to March, and only females and eggs are found throughout winter (Ito and Yamanishi \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Oda and Ito \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). In contrast, in \u003cem\u003eS. shii\u003c/em\u003e, the season for the two life stages occur much later in February and is short-lived (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e2\u003c/span\u003eA).\u003c/p\u003e \u003cp\u003eThis interspecific difference may be caused by differences in the time that it takes for females to resume activity after reproductive arrest, and earlier reactivation in \u003cem\u003eS. shii\u003c/em\u003e may complicate its overwintering pattern. More than 90% of non-reproductive \u003cem\u003eS. shii\u003c/em\u003e females oviposit within 30 days at 20\u0026deg;C without chilling treatment (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), whereas \u003cem\u003eS. brevisetosus\u003c/em\u003e females that had developed under 10L and 11L conditions take 49.5\u0026thinsp;\u0026plusmn;\u0026thinsp;6.9 (n\u0026thinsp;=\u0026thinsp;5) and 45.5\u0026thinsp;\u0026plusmn;\u0026thinsp;6.5 days (n\u0026thinsp;=\u0026thinsp;16), respectively, to oviposit under the same experimental conditions (N. Yamanishi and K. Ito, unpubl. data). Although statistical tests between the two species are not conducted due to the difference in data structure, these results predict that \u003cem\u003eS. shii\u003c/em\u003e females would resume oviposition sooner than \u003cem\u003eS. brevisetosus\u003c/em\u003e under short-day conditions. Indeed, eggs collected in the field in October all consisted of old eggs, which were soon to hatch, but eggs in November included a higher proportion of new eggs, which needed a longer time to hatch (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e4\u003c/span\u003e, Table\u0026nbsp;4). In addition, all eggs successfully hatched in both months, and the hatching time was short. Diapausing eggs of spider mites usually need a long period of chilling to hatch (Lees \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e1953\u003c/span\u003e; Shinkaji \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e1975\u003c/span\u003e; Koveos and Broufas \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e1999\u003c/span\u003e), such as in \u003cem\u003eS. sapporensis\u003c/em\u003e eggs on evergreen bamboos, which require more than 40 days at 5\u0026deg;C (Gotoh \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e1986b\u003c/span\u003e); given this, the \u003cem\u003eS. shii\u003c/em\u003e eggs studied here may not be in diapause. Furthermore, the transplant experiments of laboratory eggs to the field conditions suggest that eggs in mid-October and November successfully hatched within the year (13.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.8 and 20.1\u0026thinsp;\u0026plusmn;\u0026thinsp;7.9 days, respectively; see Results). Taken together, females emerging in October stop oviposition but soon begin to lay non-diapausing eggs in November, when immature stages of the preceding generation are still growing. These eggs would hatch within the year and constitute the source of immature stages after December (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e2\u003c/span\u003eA, B, Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e3\u003c/span\u003eB).\u003c/p\u003e \u003cp\u003eThe field survey demonstrated that immature stages (likely derived from new eggs above) develop throughout winter. The proportion of immature stages reached 46.7\u0026ndash;52.4% from 19 November to 3 December, caused by of a gradual decrease in the egg density by hatching and increase in the nymph density (Figs.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e2\u003c/span\u003eB, \u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e3\u003c/span\u003eB). Although the period (degree-days) from hatching to adult emergence is unknown in \u003cem\u003eS. shii\u003c/em\u003e, it takes approximately 20 days at 25\u0026deg;C/15L in the closely related \u003cem\u003eS. brevisetosus\u003c/em\u003e (M. Muto-Yamawaki, unpubl. data) and appears to be similar from the present daylength experiments (data not shown). If so, immature stages emerged early in November may mature within or across the year, because the daily highest temperatures in November ranged from 16.4 to 22.3\u0026deg;C (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e5\u003c/span\u003e) (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Furthermore, immature stages that emerge in the later season have a slow developmental rate in January and February. The mean temperature of these months drops to 6.0\u0026deg;C and 7.1\u0026deg;C on average, respectively (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e4\u003c/span\u003e, Japan Meteorological Agency \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2024\u003c/span\u003e), but the highest temperature rises to approximately 15\u0026deg;C in both months, which permits development. Furthermore, the development of immature stages during winter is supported by the fact that immature stages of various ages occur in December, but only nymphs, including teleiochrysalis stages, are present from January to March (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Furthermore, adult males were found until the end of January (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e1\u003c/span\u003eC). These males may be composed of new adults or survivors from previous years, and their ability to survive through the coldest season suggests that temperatures in the study site are nonlethal.\u003c/p\u003e \u003cp\u003eThe arrest of oviposition despite the availability of winter habitats by evergreen hosts of \u003cem\u003eS. shii\u003c/em\u003e may be attributed to a physiological shift that confers cold hardiness to their eggs, which is known as trans-generational effects in insects (Mousseau and Fox \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e1998\u003c/span\u003e; Magiafoglou and Hoffmann \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Zhou et al. \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Sub-zero temperatures over several months are stressful for spider mites in the summer form (Veerman \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e1985\u003c/span\u003e). Indeed, in the transplant experiment, all eggs that were collected from constant 25\u0026deg;C/15L conditions and placed in the field in mid-December did not survive (see Results). Still, this result does not agree with the pattern of wild eggs, which increased after December, the coldest season (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e1\u003c/span\u003eB). That is, cold hardiness in wild eggs in December is evidently higher than in laboratory-derived eggs (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e2\u003c/span\u003eA, B). Living on evergreen hosts provides the reproductive advantage of earlier initiation of reproduction than on deciduous hosts, especially where predation pressure in winter is minimal (Ito and Hamada \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). However, predation pressure of spider mites increases early in winter, with predators of \u003cem\u003eS. breviestosus\u003c/em\u003e disappearing in mid-winter (Ito \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). In this situation, higher cold hardiness of eggs is important during winter. We observed that a few midge larvae and \u003cem\u003eAgistemus\u003c/em\u003e mites fed on \u003cem\u003eS. shii\u003c/em\u003e individuals, but their densities were much lower than in summer. The role of maternal effects in cold hardiness are unexplored in spider mites and should be explored further.\u003c/p\u003e \u003cp\u003eOverwintering stages in spider mites are loosely constrained by phylogeny but vary even among sibling species (Veerman \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e1985\u003c/span\u003e; Sakagami \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Matsuda et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). We showed that evergreen hosts allow mites to overwinter in multiple life stages, and the overwintering style of \u003cem\u003eS. shii\u003c/em\u003e is markedly different even from its sibling species \u003cem\u003eS. brevisetosus\u003c/em\u003e (Matsuda et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2014\u003c/span\u003e), despite both being governed by photoperiodically controlled reproductive arrest (Oda and Ito \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). The overwintering strategy of these species has evidently been shaped by low predation pressure in winter, strong cold hardiness and host availability. The importance of these factors should be evaluated to clarify the process of diversification in the overwintering strategies of spider mites.\u003c/p\u003e"},{"header":"Declarations","content":"\n\u003cp\u003eWe have no pecuniary or other personal interest, direct or indirect, in any matter that raises or may raise a conflict with our duties.\u003c/p\u003e\u003ch2\u003eAcknowledgements\u003c/h2\u003e\n\u003cp\u003eThis work was supported by a Cabinet Office grant-in-aid: the Advanced Next-Generation Greenhouse Horticulture by IoP (Internet of Plants), Japan (Grant No: D11).\u003c/p\u003e\n\u003cp\u003eThe authors thank the Field Science Center of Kochi University for allowing us to use the study site.\u003c/p\u003e\n\u003ch2\u003eAuthor contributions\u003c/h2\u003e\n\u003cp\u003eKN:\u0026nbsp;planning, data collection and analysis, investigation, discussion;\u0026nbsp;YN:\u0026nbsp;planning, data collection and analysis, investigation, discussion; KI: planning, data collection and analysis, investigation, discussion, writing, funding acquisition\u003c/p\u003e\n\u003ch2\u003eFunding\u003c/h2\u003e\n\u003cp\u003eThis work was supported by a Cabinet Office grant-in-aid and the Advanced Next-Generation Greenhouse Horticulture by IoP (Internet of Plants), Japan.\u003c/p\u003e\n\u003ch2\u003eData availability\u003c/h2\u003e\n\u003cp\u003eThe datasets generated and/or analysed during the current study are available from the corresponding author upon reasonable request.\u003c/p\u003e\n\n\u003ch2\u003eCompeting interests\u003c/h2\u003e\n\u003cp\u003e\u0026nbsp;The authors have no financial or non-financial interests to declare.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eCarey DB (1994) Diapause and the host plant affiliations of lycaenid butterflies. Oikos 69:259\u0026ndash;266. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2307/3546146\u003c/span\u003e\u003cspan address=\"10.2307/3546146\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCrawley MJ (2005) Statistics: An introduction using R, 2nd edition. John Wiley \u0026amp; Sons Ltd., West Sussex, UK\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDanilevskii AS (1965) Photoperiodism and Seasonal Development of Insects. Oliver \u0026amp; Boyd Ltd., Edinburgh\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDanks HV (1987) Insect Dormancy: an Ecological Perspective. Biological Survey of Canada, Ottawa\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDanks HV (2006) Key themes in the study of seasonal adaptations in insects II. Life-cycle patterns. Appl Entomol Zool 41:1\u0026ndash;13. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1303/aez.2006.1\u003c/span\u003e\u003cspan address=\"10.1303/aez.2006.1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEhara S (1989) Four new species of spider mites (Acarina: Tetranychidae) from Japan. Proc Jpn Soc Syst Zool 40:28\u0026ndash;38. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.19004/pjssz.40.0_28\u003c/span\u003e\u003cspan address=\"10.19004/pjssz.40.0_28\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEhara S, Gotoh T (2009) Colored guide to the plant mites of Japan. Zenkoku Noson Kyoiku Kyokai, Tokyo. (in Japanese)\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGotoh T (1983) Life-history parameters of three species of \u003cem\u003eSchizotetranychus\u003c/em\u003e on deciduous trees (Acarina: Tetranychidae). Appl Entomol Zool 18:122\u0026ndash;128. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1303/aez.18.122\u003c/span\u003e\u003cspan address=\"10.1303/aez.18.122\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGotoh T (1986a) Life-cycle variation in \u003cem\u003ePanonychus akitanus\u003c/em\u003e Ehara (Acarina:Tetranychidae). Exp App Acarol 2:125\u0026ndash;136. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/BF01213756\u003c/span\u003e\u003cspan address=\"10.1007/BF01213756\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGotoh T (1986b) Local variation in overwintering stages of \u003cem\u003ePanonychus akitanus\u003c/em\u003e Ehara (Acarina: Tetranychidae). Exp Appl Acarol 2:137\u0026ndash;151. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/BF01213757\u003c/span\u003e\u003cspan address=\"10.1007/BF01213757\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHelle W, Overmeer WPJ (1985) Rearing Techniques. In: Helle W, Sabelis MW (eds) Spider Mites Their Biology, Natural Enemies and Control 1A. Elsevier, Amsterdam, pp 331\u0026ndash;335\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHunter MD, McNeil JN (1997) Host-plant quality influences diapause and voltinism in a polyphagous insect herbivore. Ecology 78:977\u0026ndash;986. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2307/2265851\u003c/span\u003e\u003cspan address=\"10.2307/2265851\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIto K (2003) Effect of leaf condition on diapause induction of a Kanzawa spider mite \u003cem\u003eTetranychus kanzawai\u003c/em\u003e Kishida (Acari: Tetranychidae) population on tea plants. Appl Entomol Zool 38:559\u0026ndash;563. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1303/aez.2003.559\u003c/span\u003e\u003cspan address=\"10.1303/aez.2003.559\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIto K (2004) Deteriorating effects of diapause duration on post-diapause life history traits in the Kanzawa spider mite. Physiol Entomol 29:453\u0026ndash;457. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/j.0307-6962.2004.00421.x\u003c/span\u003e\u003cspan address=\"10.1111/j.0307-6962.2004.00421.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIto K (2020) Predators of the nest-making spider mite \u003cem\u003eSchizotetranychus brevisetosus\u003c/em\u003e (Acari: Tetranychidae). J Acarol Soc Jpn 29:59\u0026ndash;70. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2300/acari.29.59\u003c/span\u003e\u003cspan address=\"10.2300/acari.29.59\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIto K, Fukuda T, Hayakawa H, Arakawa R, Saito Y (2013) Relationship between body colour, feeding, and reproductive arrest under short-day development in \u003cem\u003eTetranychus pueraricola\u003c/em\u003e (Acari: Tetranychidae). Exp Appl Acarol 60:471\u0026ndash;477. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10493-013-9660-3\u003c/span\u003e\u003cspan address=\"10.1007/s10493-013-9660-3\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIto K, Hamada E (2018) Adult female sensitivity to day-length conditions in terms of winter-egg production in \u003cem\u003eSchizotetranychus brevisetosus\u003c/em\u003e Ehara (Acari: Tetranychidae). J Acarol Soc Jpn 27:69\u0026ndash;76. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2300/acari.27.69\u003c/span\u003e\u003cspan address=\"10.2300/acari.27.69\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIto K, Saito Y (2006) Effects of host-plant species on diapause induction of the Kanzawa spider mite, \u003cem\u003eTetranychus kanzawai\u003c/em\u003e. Entomol Exp Appl 121:177\u0026ndash;184. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/j.1570-8703.2006.00471.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1570-8703.2006.00471.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIto K, Yamanishi N (2019) Production of winter eggs in \u003cem\u003eSchizotetranychus brevisetosus\u003c/em\u003e (Acari: Tetranychidae) inhabiting evergreen Japanese blue oak. Exp Appl Acarol 78:521\u0026ndash;534. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10493-019-00402-3\u003c/span\u003e\u003cspan address=\"10.1007/s10493-019-00402-3\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJapan Meteorological Agency (2024) Past weather. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://www.data.jma.go.jp/obd/stats/etrn/index.php\u003c/span\u003e\u003cspan address=\"http://www.data.jma.go.jp/obd/stats/etrn/index.php\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (in Japanese)\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKoveos DS, Broufas GD (1999) Diapause induction and termination in eggs of the fruit tree red spider mite \u003cem\u003ePanonychus ulmi\u003c/em\u003e in northern Greece. Exp Appl Acarol 23:669\u0026ndash;679. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1023/A:1006232821653\u003c/span\u003e\u003cspan address=\"10.1023/A:1006232821653\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKroon A, Veenendaal RL, Bruin J, Egas M, Sabelis MW (2004) Predation risk affects diapause induction in the spider mite \u003cem\u003eTetranychus urticae\u003c/em\u003e. Exp Appl Acarol 34:307\u0026ndash;314. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1023/B:APPA.0000049226.60183.29\u003c/span\u003e\u003cspan address=\"10.1023/B:APPA.0000049226.60183.29\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKroon A, Veenendaal RL, Bruin J, Egas M, Sabelis MW (2008) Sleeping with the enemy\u0026mdash;predator-induced diapause in a mite. Naturwissenschaften 95:1195\u0026ndash;1198. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s00114-008-0442-4\u003c/span\u003e\u003cspan address=\"10.1007/s00114-008-0442-4\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKroon A, Veenendaal RL, Egas M, Bruin J, Sabelis MW (2005) Diapause incidence in the two-spotted spider mite increases due to predator presence, not due to selective predation. Exp Appl Acarol 35:73\u0026ndash;81. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10493-004-1980-x\u003c/span\u003e\u003cspan address=\"10.1007/s10493-004-1980-x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKurota H, Shimada M (2002) Geographical variation in the seasonal population dynamics of \u003cem\u003eBruchidius dorsalis\u003c/em\u003e (Coleoptera: Bruchidae): constraints of temperature and host plant phenology. Environ Entomol 31:469\u0026ndash;475. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1603/0046-225X-31.3.469\u003c/span\u003e\u003cspan address=\"10.1603/0046-225X-31.3.469\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLees AD (1953) Environmental factors controlling the evocation and termination of diapause in the fruit tree red spider mite \u003cem\u003eMetatetranychus ulmi\u003c/em\u003e Koch (Acarina: Tetranychidae). Ann Appl Biol 40:449\u0026ndash;486\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMagiafoglou A, Hoffmann AA (2003) Cross-generation effects due to cold exposure in \u003cem\u003eDrosophila serrata\u003c/em\u003e. Funct Ecol 17:664\u0026ndash;672. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1046/j.1365-2435.2003.00774.x\u003c/span\u003e\u003cspan address=\"10.1046/j.1365-2435.2003.00774.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMatsuda T, Morishita M, Hinomoto N, Gotoh T (2014) Phylogenetic analysis of the spider mite sub-family Tetranychinae (Acari: Tetranychidae) based on the mitochondrial COI gene and the 18S and the 5' end of the 28S rRNA genes indicates that several genera are polyphyletic. PLoS ONE 9:e108672. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1371/journal.pone.0108672\u003c/span\u003e\u003cspan address=\"10.1371/journal.pone.0108672\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMochizuki M, Takafuji A (1996) Effect of photoperiod and temperature on the diapause termination of the Kanzawa spider mite, \u003cem\u003eTetranychus kanzawai\u003c/em\u003e Kishida (Acari: Tetranychidae). J Acarol Soc Jpn 5:83\u0026ndash;88\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMorimoto N, Takafuji A (1983) Comparison of diapause attributes and host preference among three populations of the citrus red mite, \u003cem\u003ePanonychus citri\u003c/em\u003e (McGregor) occurring in the southern part of Okayama Prefecture, Japan. Jpn J Appl Entomol Zool 27:224\u0026ndash;228\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMousseau TA, Fox CW (1998) The adaptive significance of maternal effects. Trends Ecol Evol 13:403\u0026ndash;407. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/S0169-5347(98)01472-4\u003c/span\u003e\u003cspan address=\"10.1016/S0169-5347(98)01472-4\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNAOJ (2023) Local Calendar for Kouchi (Kouchi). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://eco.mtk.nao.ac.jp/koyomi/dni/dni40.html\u003c/span\u003e\u003cspan address=\"https://eco.mtk.nao.ac.jp/koyomi/dni/dni40.html\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOda N, Ito K (2021) Photoperiodic control of reproductive arrest in the oak-inhabiting spider mite \u003cem\u003eSchizotetranychus brevisetosus\u003c/em\u003e (Acari: Tetranychidae). Exp Appl Acarol 84:389\u0026ndash;405. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10493-021-00630-6\u003c/span\u003e\u003cspan address=\"10.1007/s10493-021-00630-6\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOku K, Yano S, Takafuji A (2003) Different maternal effects on diapause induction of tetranychid mites, \u003cem\u003eTetranychus urticae\u003c/em\u003e and \u003cem\u003eT. kanzawai\u003c/em\u003e (Acari: Tetranychidae). Appl Entomol Zool 38:267\u0026ndash;270. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1303/aez.2003.267\u003c/span\u003e\u003cspan address=\"10.1303/aez.2003.267\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eR Core Team (2022) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria, Vienna, Austria\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eS\u0026aacute;ez-Castillo AJ, Conde-S\u0026aacute;nchez A, Mart\u0026iacute;nez F (2022) DGLMExtPois: Advances in dealing with over and under-dispersion in a double GLM framework. R J 14:121\u0026ndash;140. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.32614/RJ-2023-002\u003c/span\u003e\u003cspan address=\"10.32614/RJ-2023-002\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSaito Y (2010) Plant mites and sociality: diversity and evolution. Springer-Verlag Tokyo\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSaito Y, Ito K, Sakagami T (2005) Imaginal induction of diapause in several adult-diapausing spider mites. Physiol Entomol 30:96\u0026ndash;101. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/j.0307-6962.2005.00441.x\u003c/span\u003e\u003cspan address=\"10.1111/j.0307-6962.2005.00441.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSaito Y, Suzuki R (1987) Reexamination of several rearing methods for studying the life history of spider mites (Acari: Tetranychidae). Appl Entomol Zool 22:570\u0026ndash;576. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1303/aez.22.570\u003c/span\u003e\u003cspan address=\"10.1303/aez.22.570\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSakagami T (2002) Phylogenetic analysis of subfamily Tetranychinae (Acari: Tetranychidae) in Japan, based on 28S ribosomal DNA sequences: Does the \u003cem\u003ecelarius\u003c/em\u003e group belong to the genus \u003cem\u003eSchizotetranychus\u003c/em\u003e? Dissertation, Hokkaido University (in Japanese)\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSeck NKG, Ngom A, Noba K (2022) Modelling underdispersed count data: relative performance of Poisson model and its alternatives. Afr J Math Stat St 5:16\u0026ndash;32. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.52589/ajmss-1wpjqhyt\u003c/span\u003e\u003cspan address=\"10.52589/ajmss-1wpjqhyt\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShinkaji N (1975) Hatching time of the winter eggs and termination of diapause in the common conifer spider mite, \u003cem\u003eOligonychus ununguis\u003c/em\u003e (Jacobi), on chestnut in relation to temperature (Acarina: Tetranychidae). Jpn J Appl Entomol Zool 19:144\u0026ndash;148. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1303/jjaez.19.144\u003c/span\u003e\u003cspan address=\"10.1303/jjaez.19.144\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSo P-M, Takafuji A (1991) Coexistence of \u003cem\u003eTetranychus urticae\u003c/em\u003e (Acarina: Tetranychidae) with different capacities for diapause: comparative life-history traits. Oecologia 87:146\u0026ndash;151. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/bf00323792\u003c/span\u003e\u003cspan address=\"10.1007/bf00323792\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTakafuji A, Kamibayashi M (1984) Life cycle of a non-diapausing population of the two-spotted spider mite, \u003cem\u003eTetranychus urticae\u003c/em\u003e Koch in a pear orchard. Res Popul Ecol 26:113\u0026ndash;123. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/bf02515511\u003c/span\u003e\u003cspan address=\"10.1007/bf02515511\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTakafuji A, Morishita M (2001) Overwintering ecology of two species of spider mites (Acari: Tetranychidae) on different host plants. Appl Entomol Zool 36:169\u0026ndash;175. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1303/aez.2001.169\u003c/span\u003e\u003cspan address=\"10.1303/aez.2001.169\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTakafuji A, So P-M, Tsuno N (1991) Inter- and intra-population variations in diapause attribute of the two-spotted spider mite, \u003cem\u003eTetranychus urticae\u003c/em\u003e Koch, in Japan. Res Popul Ecol 33:331\u0026ndash;344. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/bf02513558\u003c/span\u003e\u003cspan address=\"10.1007/bf02513558\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTauber MJ, Tauber CA, Masaki S (1986) Seasonal adaptations of insects. Oxford University Press, New York\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTsuda Y, Takafuji A, Kuno E (1997) Maintenance of diapause variability in the two-spotted spider mite, \u003cem\u003eTetranychus urticae\u003c/em\u003e, in a heterogeneous and stochastic environment. Res Popul Ecol 39:77\u0026ndash;82. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/bf02765252\u003c/span\u003e\u003cspan address=\"10.1007/bf02765252\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVeerman A (1985) Diapause. In: Helle W, Sabelis MW (eds) Spider Mites Their Biology, Natural Enemies and Control 1A. Elsevier, Amsterdam, pp 279\u0026ndash;316\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWood SN (2011) Fast stable restricted maximum likelihood and marginal likelihood estimation of semiparametric generalized linear models. J R Stat Soc Ser B Stat Methodol 73:3\u0026ndash;36. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/j.1467-9868.2010.00749.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1467-9868.2010.00749.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZar JH (2010) Biostatistical Analysis. 5th edition. Prentice Hall, Inc., New Jersey\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhou Z-S, Rasmann S, Li M, Guo J-Y, Chen H-S, Wan F-H (2013) Cold temperatures increase cold hardiness in the next generation \u003cem\u003eOphraella communa\u003c/em\u003e Beetles. PLoS ONE 8:e74760. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1371/journal.pone.0074760\u003c/span\u003e\u003cspan address=\"10.1371/journal.pone.0074760\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Castanopsis sieboldii (Fagaceae), critical day length (CDL), diapause, evergreen host plants, photoperiodic response, reproductive arrest","lastPublishedDoi":"10.21203/rs.3.rs-4084840/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4084840/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eHost availability in winter affects the overwintering strategies of herbivorous arthropods. Spider mites (Acari: Tetranychidae) overwinter as eggs or adult females, but some do so as multiple life stages on evergreen hosts. For example, in \u003cem\u003eSchizotetranychus brevisetosus\u003c/em\u003e, adult females and their eggs stay on host leaves in mid-winter. However, few studies have focused on proximate factors generating such overwintering stages. Here, we investigated photoperiodic responses and life-stage compositions in winter in a population of \u003cem\u003eSchizotetranychus shii\u003c/em\u003e, a specialist of Japanese chinquapin (Fagaceae). The proportion of non-ovipositing females at 20\u0026deg;C followed a sigmoid curve with increasing photoperiod, and the critical day length (CDL) was estimated as 11.8L, which corresponds to the environments from late September to early October. Although females grown under 10\u0026ndash;11L conditions never oviposited within 7 days, 90\u0026ndash;96% of them started oviposition within only 30 days without chilling (n\u0026thinsp;=\u0026thinsp;23\u0026ndash;31). In the field, all life stages were observed to occur throughout winter, but their proportions varied drastically. The proportion of eggs declined from early October (62%) to early December (12%), as predicted by CDL, but steeply increased toward late February (96%), during which only adult females and eggs remained. In summary, a short photoperiod in October arrests oviposition in emerging females, but they soon commence oviposition in November while immature stages are still growing, and individuals at all life stages (including a new generation) coexist until all immature stages mature in February. This novel pattern suggests that evergreen hosts allow spider mites to evolve overwintering strategies with little phylogenetic constraint.\u003c/p\u003e","manuscriptTitle":"Overwintering at multiple life stages in Schizotetranychus shii (Acari: Tetranychidae), a specialist of evergreen chinquapin","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-03-15 20:16:20","doi":"10.21203/rs.3.rs-4084840/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"89d4f724-7590-4115-8d18-dd5501dede5b","owner":[],"postedDate":"March 15th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-12-16T16:03:07+00:00","versionOfRecord":{"articleIdentity":"rs-4084840","link":"https://doi.org/10.1007/s10493-024-00978-5","journal":{"identity":"experimental-and-applied-acarology","isVorOnly":false,"title":"Experimental and Applied Acarology"},"publishedOn":"2024-12-12 15:57:59","publishedOnDateReadable":"December 12th, 2024"},"versionCreatedAt":"2024-03-15 20:16:20","video":"","vorDoi":"10.1007/s10493-024-00978-5","vorDoiUrl":"https://doi.org/10.1007/s10493-024-00978-5","workflowStages":[]},"version":"v1","identity":"rs-4084840","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4084840","identity":"rs-4084840","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}