Photoperiodic control of larval diapause in the yellow-spotted longicorn beetle, Psacothea hilaris: analysis by photoperiod manipulation

The photoperiodic control of diapause induction in the larvae of the yellow-spotted longicorn beetle, Psacothea hilaris (Pascoe), was investigated using a west Japan-type population collected from Ino, Kochi Prefecture, Japan. In this population, the larvae expressed a long-day photoperiodic response with a critical daylength between 13.5 and 14 h at 25 °C ; under a long daylength, the larvae pupated after the 4th or 5th instar, while the larvae entered diapause under a short daylength after 2.3 additional molts on average. When the photoperiod was changed from a short (L12:D12) to a long (L15:D9) daylength, pupation occurred in most of the individuals irrespective of the time of the change. When the photoperiod was changed from long to short at 1 or 2 weeks after hatching, all of the larvae entered diapause, whereas when the photoperiod was changed at 5 weeks after hatching or later, most of the larvae pupated. The 2 weeks exposures to a long daylength against a 'background' of a short daylength at various times revealed that the larvae of this insect are most sensitive to the photoperiod from 4 to 6 weeks after hatching.     

Interactive effects of photoperiod and temperature on diapause induction and termination in the yellow-spotted longicorn beetle, Psacothea hilaris

Abstract. The interactive effects of temperature (20 °C or 25 °C) and photoperiod (LD 12 : 12 h or LD 15 : 9 h) on diapause induction and termination are investigated in the west‐Japan type yellow‐spotted longicorn beetle, Psacothea hilaris (Pascoe) (Coleoptera: Cerambycidae). Larval diapause of P. hilaris is induced under three diapause‐inducing conditions (20 °C–SD, 20 °C–LD and 25 °C–SD), and the diapause larvae are transferred to one of four conditions (20 °C–SD, 20 °C–LD, 25 °C–SD or 25 °C–LD) for observation of pupation, which indicates termination of diapause. The intensity of diapause induced under the three conditions increases in the order 20 °C–SD < 25 °C–SD < 20 °C–LD, when assessed by the time course of pupation after the transfer. On the other hand, the effectiveness of the temperature–photoperiod combinations to terminate diapause is in the order 25 °C–SD (ineffective) < < 20 °C–LD < 25 °C–LD < 20 °C–SD. Among the temperatures (5, 10, 15 and 20 °C) examined, 15 °C is the most effective in terminating diapause under the short day; diapause in most larvae appears to have been completed in 15 days.     

Diapause avoidance induced by low temperature in the yellow-spotted longicorn beetle, Psacothea hilaris

The diapause-averting effect of low temperature on pre-diapause larvae was examined in the yellow-spotted longicorn beetle, Psacothea hilaris. Larvae that had been reared under diapause-inducing conditions (25 °C , L12:D12) were temporarily exposed to 10 °C for various periods, and returned to the initial condition. Diapause was not averted by chilling for 15 days irrespective of the age of the larvae at chilling. After a 30-day chilling treatment, all of the 40- and 60-day-old larvae averted diapause, while diapause was averted in only one-third of the 10- and 20-day-old larvae. None of the pre-diapause larvae chilled for 60 days entered diapause irrespective of the age at chilling. With diapause avoidance, larvae that overwintered in earlier instars can start growing in earliest spring without any arrest; this phenomenon probably subserves the synchronization of larval development in a population.     

Endocrine changes associated with metamorphosis and diapause induction in the yellow-spotted longicorn beetle, Psacothea hilaris

At 25 degrees C and under a long-day photoperiod, all 5th instar Psacothea hilaris larvae pupate at the next molt. Under a short-day photoperiod, in contrast, they undergo one or two additional larval molts and enter diapause; the 7th instar larvae enter diapause without further molt. The changes in hemolymph juvenile hormone (JH III) titers, JH esterase activity, and ecdysteroid titers in pupation-destined, pre-diapause, and diapause-destined larvae were examined. JH titers of the 5th instar pupation-destined larvae decreased continuously from 1.3 ng/ml and became virtually undetectable on day 13, when JH esterase activity peaked. Ecdysteroids exhibited a small peak on day 8, 1 day before gut purge, and a large peak on day 11, 2 days before the larvae became pre-pupae. The two ecdysteroid peaks are suggested to be associated with pupal commitment and pupation, respectively. JH titers of the 5th instar pre-diapause larvae were maintained at approximately 1.5 ng/ml for 5 days and then increased to form a peak (3.3 ng/ml) on day 11. JH esterase activity remained at a low level throughout. Ecdysteroid levels exhibited a large peak of 40 ng/ml on day 18, coincident with the larval molt to the 6th instar. JH titers of the 7th instar diapause-destined larvae peaked at 1.9 ng/ml on day 3, and a level of approximately 1.1 ng/ml was maintained even 30-60 days into the instar, when they were in diapause. Ecdysteroid titers remained approximately 0.02 ng/ml. Diapause induction in this species was suggested to be a consequence of high JH and low ecdysteroid titers.     

Evidence for the presence of a threshold weight for entering diapause in the yellow-spotted longicorn beetle, Psacothea hilaris

Under long-day conditions larvae of Psacothea hilaris (Coleoptera: Cerambycidae) pupate after the 4th or 5th instar, while under short-day conditions they undergo 2-4 nonstationary supernumerary molts and eventually enter diapause. To explore the possibility of a threshold weight for entering diapause, P. hilaris larvae were deprived of food on days 0 (day of ecdysis), 4 or 8 of the 4th, 5th and 6th instars under short-day conditions. Within the first 40 days of starvation, 60% of the larvae starved starting on day 0 of the 4th instar died, but all the larvae starved at later stages survived. The incidence of diapause in these survivors was determined by the occurrence of pupation after a temporary chilling at 15 degrees C for 15 days. Diapause incidence increased as the onset of starvation was delayed; from 11% in the larvae starved on day 0 of the 5th instar to 100% in the larvae starved on day 4 and day 8 of the 6th instar. Analysis of the relationship between the initial weight of a respective larva at the onset of starvation and its pupation success revealed that none of the larvae weighing 690 mg did. This finding suggests the presence of a threshold weight (about 600 mg), below which larvae are incapable of entering diapause. We discuss these findings with reference to the life history of P. hilaris.     

Diet and carbohydrate digestion in the yellow-spotted longicorn beetle Psacothea hilaris

Larval and adult Psacothea hilaris feed on mulberry wood and leaves, respectively. High levels of endogenous activity against the major dietary carbohydrates, cellulose, hemicellulose, starch and soluble sugars were secreted in the gut of larvae and adults. Activity against pectin was also high and multiple polygalacturonase (EC 3.2.1.15) components were secreted in the gut of larvae. One glycanase component, beta-EG1, which was primarily an endo-beta-1,4-glucanase (EC 3.2.1.4) and another, beta-EG2, which was mostly an endo-beta-1,4-xylanase (EC 3.2.1.8), were also secreted, while at least four additional components hydrolysed laminarin, lichenin and crystalline cellulose. The beta-glycosidase component beta-GD1 was associated with most of the beta-mannosidase (EC 3.2.1.25) and beta-xylosidase (EC 3.2.1.37) activity secreted in the gut of larvae, while another, beta-GD2, was a beta-glucosidase (EC 3.2.1.21), the activity of which was directed against cellobiose and other beta-linked disaccharides, and a beta-fucosidase (EC 3.2.1.38). A beta-galactosidase (EC 3.2.1.23), which did not hydrolyse lactose, was also secreted, as were distinct beta-N-acetylhexosaminidase (EC 3.2.1.52), trehalase (EC 3.2.1.28), alpha-L-arabinosidase (EC 3.2.1.55), alpha-galactosidase (EC 3.2.1.22) and a minimum of four alpha-glucosidase (EC 3.2.1.20) components, one of which was also likely to be associated with a peak of alpha-mannosidase (EC 3.2.1.24) activity. The alpha-glucosidase components varied in their specificity for alpha-linked disaccharides, but none was active against sucrose, which was hydrolysed by a beta-fructofuranosidase (EC 3.2.1.26) component. Overall average levels of activity in larvae were twice those of adults, but the secretion of individual carbohydrases in both was not regulated in response to the relative abundance of particular carbohydrate components in their respective diets.     

Relationship between rapid cold-hardening and cold acclimation in the eggs of the yellow-spotted longicorn beetle, Psacothea hilaris

Rapid cold-hardening (RCH) and cold acclimation (ACC) were examined in eggs of the yellow-spotted longicorn beetle, Psacothea hilaris (Pascoe) (Coleoptera: Cerambycidae). When eggs incubated at 25 degrees C were transferred directly to conditions of -22 degrees C for 2h, less than 30% survived, whereas exposure to 0 degrees C for 4h prior to transfer to -22 degrees C increased survival to nearly 60%. The rapidly enhanced cold tolerance (RCH) was transient and lost rapidly after 1h at 25 degrees C. Incubation at 15.5 degrees C for 9 days (ACC) also enhanced cold tolerance. Comparison of the cold tolerance of non-treated eggs and eggs pre-treated to give RCH, ACC, or ACC+RCH allowed the relationship between the two hardening processes to be determined. At a mild subzero temperature (-10 degrees C) an RCH effect was not detected, whereas only RCH is effective at the severest subzero temperature just above the SCP (-26 degrees C). At intermediate temperatures (-16, -22 and -25 degrees C), ACC and RCH enhanced survival in combination. Therefore, the two hardening processes have different physiological bases but operate concomitantly over a wide temperature range.     

Feeding glucose or sucrose, but not trehalose, suppresses the starvation-induced premature pupation in the yellow-spotted longicorn beetle, Psacothea hilaris

Under 25 degrees C and a long-day photoperiod, starvation induces premature pupation in 4th instar Psacothea hilaris larvae exceeding a threshold weight of 180 mg, resulting in the formation of small but morphologically normal adults. To investigate possible mechanisms underlying this phenomenon, we first measured the hemolymph trehalose and glucose levels of starved larvae. When larvae were starved after 4 days of feeding (attaining the threshold weight), glucose levels decreased 4-fold within the next 24 h, while trehalose levels, after a temporary slight decrease, increased remarkably to reach a peak just before the prepupa stage. The effects of ingesting various nutrients on the developmental fate and the hemolymph sugar titers of starving larvae were then examined. Feeding on agar blocks containing sucrose or glucose totally suppressed the occurrence of premature pupation, while trehalose, fructose, casein and starch were ineffective. Feeding on glucose or trehalose resulted in a 6-fold decrease in hemolymph glucose levels and remarkably elevated trehalose levels. Since feeding on glucose and trehalose induced similar changes in hemolymph sugar titers but trehalose was not effective in suppressing premature pupation, glucose may have exhibited its effects via gustatory mechanisms.     

Effects of photoperiod and temperature on diapause induction and termination in the swallowtail,Sericinus montelus

Abstract Sericinus montelus overwinters as diapausing pupae. In the present study, the effects of photoperiod and temperature on diapause induction and termination of diapause are investigated. The results obtained demonstrate that high temperature can reverse the effect of short day‐lengths on diapause induction. Under an LD 12 : 12 h photoperiod, all pupae enter diapause at 15, 20 and 25 °C, whereas all pupae develop without diapause at 35 °C. No pupae enter diapause under an LD 14 : 10 h photoperiod when the temperature is above 20 °C. Photoperiodic response curves obtained at 25 and 30 °C indicate that S. montelus is a long‐day species and the critical day‐length is approximately 13 h at 25 °C. At 25 °C, the duration of diapause is shortest when the diapausing pupae are maintained under an LD 16 : 8 h photoperiod and increases under LD 14 : 10 h and LD 12 : 12 h photoperiods. Under an LD 16 : 8 h photoperiod, the duration of diapause is shortest when the diapausing pupae are maintained at 25 °C, followed by 20 and 30 °C, and then at 15 °C. These results suggest that a moderate temperature favours diapause development under a diapause‐averting photoperiod in this species. The duration of diapause induced by an LD 12 : 12 h photoperiod is significantly longer at 25 °C than those at 15, 20 and 30 °C, and is shortest at 15 °C. At 25 °C, the duration of diapause induced by LD 6 : 18, LD 12 : 12 and LD 13 : 11 h photoperiods is similar and longer than 90 days. Thus, the diapause‐inducing conditions may affect diapause intensity and a photoperiod close to the critical day‐length has significant influence on diapause intensity in S. montelus.     

Change in significance of feeding during larval development in the yellow-spotted longicorn beetle, Psacothea hilaris

Larvae of the west-Japan type yellow-spotted longicorn beetle, Psacothea hilaris (Coleoptera: Cerambycidae), show a long-day type photoperiodic response at 25 degrees C; under long-day conditions, larvae pupate after the fourth or fifth instar, while under short-day conditions, they undergo a few nonstationary supernumerary molts and eventually enter diapause. In the present study, the effect of food on the development and photoperiodic response of the larvae was examined with special reference to molting and pupation. Although the pupal body size was greatly affected by the food quality and the length of feeding, the critical day length for induction of metamorphosis at 25 degrees C was always between 13.5 and 14 h. Exposure to starvation of larvae reared on the standard diet revealed that the capability to pupate is acquired after a few days of feeding in the fourth instar. In the larvae that had acquired the capability to pupate, premature pupation was induced by exposure to starvation, indicating that feeding becomes dispensable long before it is normally terminated.     

Geographic variation in cold hardiness of eggs and neonate larvae of the yellow-spotted longicorn beetle Psacothea hilaris

Cold hardiness of eggs and neonate larvae of the yellow-spotted longicorn beetle, Psacothea hilaris (Pascoe) was examined using six geographical populations in Japan. Particular attention was paid to cold hardiness of eggs and neonate larvae of the subtropical population (Ishigaki), because the east Japan populations are considered to have been introduced from a subtropical area, and the overwintering stage in the east Japan populations is incidentally shifted from the original mature larval stage to the egg or neonate larval stages. When the eggs were exposed to low temperatures for 1 h, the decrease in hatchability became significant at –12°C in the southernmost two populations (Ishigaki and Naze), and at –16°C in the northern populations. After 1 h exposure to –20°C, few eggs could hatch in the Ishigaki population, whereas 27–55% of the eggs survived in the northern populations. Pre-chilling of the eggs at 10°C for 10 days enhanced the cold hardiness in all populations. This effect was particularly distinct in the subtropical population; the eggs of the Ishigaki population became as cold hardy as those of the northern populations after acclimation. These results suggest that the subtropical population is capable of establishing itself in east Japan, where the winter is cold.     

Exploring the range expansion of the yellow-spotted longhorn beetle Psacothea hilaris hilaris in northern Italy

1. Psacothea hilaris hilaris is an exotic cerambycid detected in Lombardy (northern Italy) in 2005 and now established and undergoing an expansion phase. The species constitutes a serious pest for Ficus spp. and Morus spp.
2. The aim of this work is to update the distribution area of the species in Italy, investigating its expansion over 16 years. Data were acquired through the authors' direct field records and community science, by data mining on national and international web platforms and social networks. Reports were analysed and GPS points plotted in ArcMap 10.8.1 to obtain distribution maps.
3. Of the records collected, 34% were acquired during field surveys, 22% via e-mail and 44% through on-line forums and social networks.
4. Adults of P. h. hilaris showed a long period of activity, from April to the beginning of December, with a peak of presence from June to September. The presence of the pest has been ascertained over six provinces in Lombardy covering an area estimated at more than 1750 km² in 2021. New detections occurred within 2 km of a known infestation from the previous year in 41.6% of cases, and between 2 and 4 km away in 37.7% of cases. The mean rate of range expansion estimated from reported records was 3.17 ± 0.33 km/year from 2010 to 2021. After an initial phase of settlement, the insect is now spreading southward.
5. Given the relevant phytosanitary interest that the species may have for fig plants in Italy and in the Mediterranean Basin, it is fundamental to deepen the knowledge about its flight biology, dynamics and active dispersal capabilities in order to estimate the future range expansion.

     

Maternal temperature has different effects on the photoperiodic response and duration of larval diapause in blow fly ( Calliphora vicina ) strains collected at two latitudes

Abstract .The blow fly Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae) has a wide distribution across northern and temperate Europe. It has a facultative, maternally-induced larval diapause in response to short days. The photoperiodic response, measured at 15 and 20°C, of two populations was compared. A southern population (originating at 51° N) was sensitive to temperature at all daylengths; the incidence of diapause was greatly reduced at 20°C compared with 15°C. The photoperiodic response of a northern population (from 65° N) was sensitive to temperature only in long days; in short days (< 14 h of light) the response of this strain was identical at each temperature.
Variation in parental photoperiod and temperature were found to affect the duration of larval diapause, indicating a role for maternal effects in diapause intensity as well as incidence. However, the between-strain variation was greater than that within strains, indicating qualitative differences in diapause response. These differences may arise from the ecological conditions at the points of origin of the two strains. The northern strain from the harsher climate has a more intense diapause that follows a relatively temperature-insensitive photoperiodic response. In contrast, the southern strain has a shallow diapause and its photoperiodic response may be overridden by the experience of concurrent high temperature.     

Effects of photoperiod and temperature on the development and diapause of the bark beetle Ips typographus

Abstract:  Diapause was induced in a Central European population of Ips typographus grown at 20°C when the day length decreased below 16 h [50% diapause incidence occurred in the 14.7:9.3 h L:D (light:dark) regime]. The non-diapausing adults fed on days 2–6 and 10–14 after the ecdysis and swarmed after the second feeding bout with chorionated eggs in the ovaries and sperm in the spermiducts. Neither gonads nor the flight muscles matured and no swarming occurred in the diapausing adults. The development from egg to adult took about 34 days in both 18:6 h (no diapause) and 12:12 h L:D (diapause) regimes, but it was extended by up to 30% without diapause induction when only larvae or pupae were exposed to L:D 12:12 h. Diapause was induced in insects reared at L:D 12:12 h through the last larval and the pupal instars and/or in the adult stage. Temperature ≥ 23°C prevented diapause induction at L:D 12:12 h but diapause occurred at L:D 14:10 h associated with 26:6°C thermoperiod. The effect of thermoperiods on the developmental rate requires further research. Exposure of the non-diapausing adults to 5°C for several days blocked feeding and evoked a diapause-like state, whereas diapausing adults fed and their gonads slowly developed at this temperature. Diapausing adults exposed in forest to low night temperatures and transferred in October to 20°C readily reproduced at 18:6, but not 12:12 h L:D photoperiods. After 2-months at 5°C and darkness, they became insensitive to the photoperiod, matured and most of them also swarmed at 20°C in the 12:12 h L:D regime. In a Scandinavian population, diapause occurred at 18:6 h L:D and was terminated either by exposure to 5°C or by very long photoperiod (L:D 20:4 h) combined with high temperature (23°C).     

Effect of different artificial diets on diapause induction under controlled temperature and photoperiod in the silkworm, Bombyx mori L.

ABSTRACT. Embryonic diapause of the silkworm, Bombyx mori , is generally induced by temperature and photoperiod during the egg stage of the previous generation and not in the larval stage. However, when silkworm larvae are reared on an artificial diet instead of mulberry leaves, their diapause is strongly affected by temperature and photoperiod experienced in the larval stage, with a distinct long-day response for diapause induction. Moreover when larvae which have been reared on artificial diet under long-day condition are fed mulberry leaves even for a short period of time, most of the resultant female adults lay diapause eggs. These results suggest that the photoperiodic response of larvae for diapause induction may be strongly suppressed by some components in mulberry leaves.     

Purification, characterization, cDNA cloning and nucleotide sequencing of a cellulase from the yellow-spotted longicorn beetle, Psacothea hilaris.

A cellulase (endo-beta-1,4-glucanase, EC 3.2.1.4) was purified from the gut of larvae of the yellow-spotted longicorn beetle Psacothea hilaris by acetone precipitation and elution from gels after native PAGE and SDS/PAGE with activity staining. The purified protein formed a single band, and the molecular mass was estimated to be 47 kDa. The purified cellulase degraded carboxymethylcellulose (CMC), insoluble cello-oligosaccharide (average degree of polymerization 34) and soluble cello-oligosaccharides longer than cellotriose, but not crystalline cellulose or cellobiose. The specific activity of the cellulase against CMC was 150 micro mol.min-1.(mg protein)-1. TLC analysis showed that the cellulase produces cellotriose and cellobiose from insoluble cello-oligosaccharides. However, a glucose assay linked with glucose oxidase detected a small amount of glucose, with a productivity of 0.072 micro mol.min-1.(mg protein)-1. The optimal pH of P. hilaris cellulase was 5.5, close to the pH in the midgut of P. hilaris larvae. The N-terminal amino-acid sequence of the purified P. hilaris cellulase was determined and a degenerate primer designed, which enabled a 975-bp cDNA clone containing a typical polyadenylation signal to be obtained by PCR and sequencing. The deduced amino-acid sequence of P. hilaris cellulase showed high homology to members of glycosyl hydrolase family 5 subfamily 2, and, in addition, a signature sequence for family 5 was found. Thus, this is the first report of a family 5 cellulase from arthropods.     

Effect of photoperiod on winter and summer diapause of the soybean pod borer Leguminivora glycinivorella (Lepidoptera: Tortricidae)

To understand the geographical differences between diapause systems and synchronization of adult occurrence in the soybean pod borer Leguminivora glycinivorella (Lepidoptera: Tortricidae), we examined the timing of winter diapause termination and intensity of summer diapause using univoltine and potentially bivoltine individuals in Iwate, Japan. In laboratory rearing experiments of mature larvae maintained at constant temperature (20 °C), winter diapause intensity weakened by January without photoperiodic responses. Meanwhile, summer diapause was maintained by the long day length and presumably terminated with the photoperiodic transition from long to short day length. The intensity of summer diapause was stronger for cocoons that transitioned from a 16 h light to 8 h dark (LD 16:8) to a LD 15:9 photoperiod than for those that transitioned from LD 15:9 to LD 14:10. These results suggest that populations distributed in relatively low-latitude areas, with partly or potentially bivoltine individuals, would have a weaker summer diapause or none at all. Moreover, sexual differences in the number of days to emergence were not detected when individuals experienced a photoperiodic transition from long to short day length, suggesting that the summer diapause system may function to synchronize the emergence of males and females in the population examined.     

低温和光周期对绿盲蝽越冬卵滞育解除和发育历期的影响

为了探讨温度和光周期对绿盲蝽Apolygus lucorum Meyer-Dür越冬卵滞育解除和发育历期的影响, 系统调查了绿盲蝽越冬卵在不同温度和不同光照组合下的孵化率和孵化时间, 结果显示：绿盲蝽的越冬卵均为滞育卵,低温和光周期对绿盲蝽越冬卵的滞育解除均有影响。2℃的低温处理能够显著促进其滞育解除,在0~65 d范围内,随着低温处理时间增长,其滞育解除时间缩短,未经低温处理的越冬卵T₅₀为68.5 d,低温处理65 d的T₅₀为12.25 d,绿盲蝽越冬卵在2℃低温处理65 d后完全解除滞育;在0~40 d范围内,低温处理时间越长,绿盲蝽越冬卵的孵化率越高,在25℃、全光照的条件下不经低温处理的孵化率为68.65%,低温处理40 d后在25℃的条件下的孵化率达到99.46%。在20~26℃范围内,绿盲蝽越冬卵的滞育后发育历期随着温度的上升而缩短, 随着光周期的延长而缩短。结果说明低温处理能够提高绿盲蝽越冬卵滞育解除率,但不是其滞育解除的必要条件,低温处理与自然变温对绿盲蝽滞育解除的作用相似;高温和长光照能够促进绿盲蝽的滞育解除,缩短发育历期。     

Ecdysteroids and adult diapause in the Colorado potato beetle, Leptinotarsa decemlineata

ABSTRACT. The difference in ecdysteroid activity in short-day (10-h photo-phase) and long-day (16-h photophase) Leptinotarsa decemlineata Say adults was characterized by high performance liquid chromatography, which revealed that for short-day beetles ecdysterone and ecdysone are the major constituents of the free ecdysteroids, whereas in long-day beetles products co-migrating with 2-deoxyecdysone and 2-deoxyecdysterone are just as abundant. Ecdysteroids were detected (15 ng/ml) only in the haemolymph of females. Ligation experiments showed that the induction of the diapause syndrome by inactivation of the corpora allata or the brain not only resulted in the formation of the typical diapause proteins but also in an increase of the ecdysteroid activity. It is suggested that adult diapause in L.decemlineata is regulated by a multifactorial system.