Male-killing virus in a noctuid moth Spodoptera litura.

A female-biased sex ratio is considered advantageous for the cytoplasmic elements that inhabit sexually reproducing organisms. There are numerous examples of bacterial symbionts in the arthropod cytoplasm that bias the host sex ratio toward females through various means, including feminization and male killing. Recently, maternally inherited RNA viruses belonging to the family Partitiviridae were found to cause male killing in moths and flies, but it was unknown whether male-killing viruses were restricted to Partitiviridae or could be found in other taxa. Here, we provide compelling evidence that a maternally inherited RNA virus, Spodoptera litura male-killing virus (SlMKV), selectively kills male embryos of the tobacco caterpillar Spodoptera litura, resulting in all-female broods. SlMKV injected into uninfected S. litura can also be inherited maternally and causes male killing. SlMKV has five genomic segments encoding seven open reading frames, has no homolog of known male-killing genes, and belongs to an unclassified group of arthropod-specific viruses closely related to Tolivirales. When transinfected into larvae, both male and female recipients allow SlMKV to proliferate, but only males die at the pupal stage. The viral RNA levels in embryonic and pupal male killing suggest that the mechanism of male killing involves the constitutive expression of viral products that are specifically lethal to males, rather than the male-specific expression of viral products. Our results, together with recent findings on male-killing partiti-like viruses, suggest that diverse viruses in arthropods tend to acquire male killing independently and that such viruses may be important components of intragenomic conflict in arthropods.

Diapause in Univoltine and Semivoltine Life Cycles.

Although it is generally more adaptive for insects to produce additional generations than to have longer life cycles, some insects produce one or fewer generations per year (univoltine or semivoltine life cycles, respectively). Some insects with the potential to produce multiple generations per year produce a univoltine life cycle in response to environmental conditions. Obligatory univoltine insects have a single long diapause or multiple diapauses in different seasons. Semivoltine insects have multiple diapauses in different years, a prolonged diapause for more than a year, or diapause controlled by a circannual rhythm. Diapause in these insects greatly varies among species both in the physiological mechanism and in the evolutionary background, and there is no general rule defining it. In this review, we survey the physiological control of univoltine and semivoltine insects' diapause and discuss the adaptive significance of the long life cycles. Although constraints such as slow development are sometimes responsible for these life cycles, the benefits of these life cycles can be explained by bet-hedging in many cases. We also discuss the effect of climate warming on these life cycles as a future area of research.

Geographic Variation in Photoperiodic Response for Induction of Pseudopupal Diapause in Epicauta gorhami (Coleoptera: Meloidae).

Geographical variations in the threshold of environmental cues for diapause induction are important in understanding the life history strategy of insects. Larvae of the bean blister beetle, Epicauta gorhami (Coleoptera: Meloidae), feed on grasshopper eggs and undergo hypermetamorphosis. They normally enter diapause as a pseudopupa (fifth instar). However, when the larvae are reared under long-day and high-temperature conditions, they do not enter pseudopupal diapause but pupate directly from the fourth instar. In addition, this insect is known to modify its photoperiodic response for induction of pseudopupal diapause depending on food availability for larvae. In this study, the larval photoperiodic responses for diapause induction were examined for three populations of E. gorhami collected from the northernmost (Morioka, 39.7°N), central (Kazo, 36.1°N), and southernmost (Takanabe, 32.1°N) parts of its distribution range, and the responses were examined under both fully fed and food-deprived (FD) conditions. Diapause incidence decreased in FD larvae for all populations in long-day conditions, and the critical day-lengths for the diapause incidence were calculated as 14.81, 13.97, and 13.99 h in the Morioka, Kazo, and Takanabe populations, respectively. The results indicate the presence of a geographical variation in larval photoperiodic response, in which diapause is induced earlier in higher latitudinal areas. From these results, together with data for developmental threshold temperature and thermal constant, the Morioka and Kazo populations were suggested to have a partial bivoltine life cycle under the conditions of extremely early oviposition season, a sufficient summer temperature, and poor larval food availability.

Microhabitat Variation in Egg Diapause Incidence in Summer Within a Local Population: An Adaptation to Decline in Host-Plant Suitability in Trigonotylus caelestialium (Hemiptera: Miridae).

Diapause induction in multivoltine insects is an ecophysiological event that is generally triggered by seasonal cues such as photoperiod and temperature. The rice leaf bug, Trigonotylus caelestialium (Kirkaldy), feeds on various Poaceae grasses and produces several generations a year. Previous studies have shown that adults produce diapause and nondiapause eggs under short-day and long-day conditions, respectively. However, there is a distinct mid-summer peak in diapause incidence before an autumnal increase in diapause incidence in the field, which cannot be explained by the laboratory results. The present study was performed to examine the environmental factors affecting the diapause incidence in mid-summer and the adaptive significance of this phenomenon. Seasonal trends in diapause incidence differed significantly among three sites located 150-400 m apart from each other and with different host plants. The suitability of host plants differs depending on species and seasonally. Therefore, the microhabitat difference in diapause trend is believed to be due to the difference in host plants. When field-collected female adults laying diapause eggs in late June were fed a seasonally deteriorating host (the orange foxtail, Alopecurus aequalis Sobol. [Poales: Poaceae]), they kept laying diapause eggs, whereas when fed a suitable host (the wheat, Triticum aestivum L. [Poales: Poaceae]) for 5 d, they changed oviposition mode to lay nondiapause eggs. These results indicate that host-plant suitability affects the oviposition mode of T. caelestialium. Diapause-egg oviposition in mid-summer in T. caelestialium has adaptive significance as a bet-hedging strategy against unpredictable dietary conditions.

Life-History Traits of a Subtropical Cerambycid Beetle, Ropica honesta (Coleoptera: Cerambycidae).

Ropica honesta Pascoe is a small-sized cerambycid that has been recorded in tropical to subtropical Asia. In this study, life-history traits were examined for a local population collected from Iriomote Island (24.3°N, 123.8°E), Okinawa, Japan, by rearing insects on artificial diet as larval food. The egg period was 5.9 ± 0.3 d at 25°C. There was no significant difference in the duration of the larval, pupal, and adult preoviposition periods between long-day (14:10 [L:D]) and short-day (12:12 [L:D]) photoperiods at both 20 and 25°C. These periods at 25°C (14:10 [L:D]) were 28.5 ± 1.4, 8.4 ± 0.5, and 9.6 ± 1.9 d (mean ± SD), respectively. The relationship between the developmental rate and temperature followed the law of total effective temperature; thus, the developmental threshold temperature and thermal constant were estimated based on these data. Together with the finding that R. honesta may not have diapause in the egg stage, it is suggested that this beetle does not have diapause in the life cycle. Furthermore, when larvae were reared on natural food (dead twigs of hardwoods) adults emerged from the twigs 47.6 ± 2.9 d after oviposition, and this value was close to the total duration of the egg to pupal periods. Together with the data for annual temperature of the habitat and the fact that food resources for the species (dead twigs and leaves of hardwoods) are available throughout the year, we conclude that R. honesta develops and reproduces all year round, with five generations at maximum.

Rearing Theretra oldenlandiae (Lepidoptera: Sphingidae) Larvae on an Artificial Diet.

The hawk moth Theretra oldenlandiae (Fabricius) is an important insect pest because in the larval stage it feeds on agricultural crops and ornamental plants such as the eddoe and garden balsam. In this study, we established methods for rearing T. oldenlandiae in the laboratory using an artificial diet containing dry powder of a wild grass Cayratia japonica (Thunb.) Gagnep. Several artificial diets were tested with different ratios of a commercial diet, Insecta LFM, and the dry leaf powder, and including different antibiotics, and the composition of the standard diet on which larvae performed best was determined. The standard diet contains 20 g of Insecta LFM, 4 g of leaf powder, 100 ml of water, 75 mg of chloramphenicol, and 200 µl of propionic acid. Larvae reared on the standard diet became larger pupae than those reared on C. japonica leaves. This result suggests that the larvae have growth potential that is masked on C. japonica leaves, and that C. japonica may not be the most suitable host species for T. oldenlandiae larvae in terms of nutrient level.

Northward expansion of the bivoltine life cycle of the cricket over the last four decades.

Recent climate warming has affected some life-history traits of insects, including voltinism and body size. The magnitude of changes in these traits may differ latitudinally within a species because of the differing lengths of season available for growth. The present study aims to estimate the change in voltinism of the lawn ground cricket, Polionemobius mikado (Shiraki) (Orthoptera: Trigonidiidae), over the last four decades by comparing the body size between adults collected from a wide range of latitudes in Japan in recent years (2015-2017) and those collected four decades ago (1969-1976). The body size of adults collected in recent years showed a latitudinal saw-tooth cline, in the same way as body size did four decades ago, and the cline shifted northward over the last four decades: In 2015-2017, the body size decreased slightly with increasing latitude from 31°N to 36°N, and then increased to 40°N, and again decreased from 40°N to 44°N. Comparison of the body size between recent years and four decades ago revealed that the body size has decreased significantly at the middle latitudes (36-40°N), suggesting that the proportion of smaller bivoltine individuals there has increased over the last four decades. The sum of effective temperatures for postdiapause embryonic development at around 36°N in recent years was comparable to that at 31-35°N four decades ago, at which P. mikado populations were bivoltine. Taken together, these findings suggested that the latitudinal range suitable for the bivoltine life cycle of P. mikado has expanded northward over the last four decades because of climate warming. This is the first report that shows that a decrease in body size can be caused by climate warming via an increase in voltinism.

Adaptive significance of precocious pupation in the bean blister beetle, Epicauta gorhami (Coleoptera: Meloidae), a hypermetamorphic insect.

Larvae of the bean blister beetle, Epicauta gorhami Marseul (Coleoptera: Meloidae), feed on grasshopper eggs in soil and undergo hypermetamorphosis. They normally enter diapause as a pseudopupa at the fifth instar, a form characteristic of hypermetamorphosis for meloid beetles. However, fourth-instar larvae exposed to long days and high temperature avoid pseudopupal diapause and pupate directly from the fourth instar. Fourth-instar larvae also tend to pupate precociously with a smaller body size if they are deprived of food. In these larvae, the critical day-length controlling induction of pseudopupal diapause becomes shorter than that for fully-fed larvae. In this study, we examined how the reaction norm of food-deprived E. gorhami larvae functions in nature by rearing insects from the egg stage outdoors in different seasons with manipulation of the food supply. The results indicated that most fully-fed larvae entered pseudopupal diapause, whereas food-deprived larvae tended to pupate precociously without entering diapause, especially early in the season. The resulting smaller adults reproduced early in the autumn and their progeny attained the pseudopupal stage before winter, indicating that the reaction norm may have an adaptive role in controlling seasonal development in the face of food shortages, producing a bivoltine life cycle.

Effect of increased male and female age at mating on the reproductive performance of Cnaphalocrocis medinalis (Crambidae: Lepidoptera).

The success of mating disruption using synthetic sex pheromones depends not only on preventing mating, but also on delayed mating in the target insect. Using the rice leaffolder moth, Cnaphalocrocis medinalis (Guenée), we determined the effect of increased age at mating imposed on males only (male treatment), females only (female treatment),or on both sexes simultaneously (both sexes treatment). These increased age treatments had a negative effect on the percentage of mating, the total number of eggs, and the hatchability. The female reproductive performance in C. medinalis was decreased with increased moths' age. The both sexes treatment had the most potent negative effect on reproductive performance. Longevity of mated moths and duration of the preoviposition period in C. medinalis were not significantly different among these increased age treatments. The underlying mechanisms causing a decline in female reproductive performance of C. medinalis when increased age was imposed on males versus females and the potential of using mating disruption strategies to control the populations in paddy fields are discussed.

Effects of temperature and photoperiod on termination of pseudopupal diapause in the bean blister beetle, Epicauta gorhami.

Larvae of the bean blister beetle, Epicauta gorhami (Coleoptera: Meloidae), feed on grasshopper eggs in soil and undergo hypermetamorphosis. This beetle undergoes larval diapause in the fifth instar as a pseudopupa, a form characteristic of hypermetamorphosis in meloid beetles. This pseudopupal diapause is maintained for a long period if the larvae are kept under continuous diapause-inducing short-day conditions. In the present study, the effects of temperature and photoperiod on termination of pseudopupal diapause in E. gorhami were examined using pseudopupae obtained under 12L-12D at 25°C. Diapause was terminated by exposure to low or high temperature, but not by transfer to long-day conditions. The pseudopupal stage comprises diapause and post-diapause phases despite its unchanging appearance. The optimum low temperature for diapause termination was 10-15°C, and a higher or lower temperature was less effective. There was an individual variation in the low temperature requirement for diapause termination and post-diapause development may not occur until this requirement is satisfied. Although under natural conditions pseudopupae may encounter low temperatures that are effective for diapause termination at different ages, our results show that pseudopupae at various ages respond similarly to low temperature exposure. This response may ensure that resumption of development synchronizes in a population.

Quantitative short-day photoperiodic response in larval development and its adaptive significance in an adult-overwintering cerambycid beetle, Phytoecia rufiventris.

The chrysanthemum longicorn beetle, Phytoecia rufiventris, overwinters in the adult stage and reproduces in spring. Larvae of this beetle develop during summer inside a host stem or root. In the present study, photoperiodic control of larval development and its adaptive significance were examined in this beetle using an artificial diet. Larvae showed a short-day photoperiodic response at 25°C with a critical day length of around 14 h; larvae reared under short-day conditions pupated, whereas those reared under long-day conditions entered summer diapause with some supernumerary molts and did not pupate. A similar response was found at 30°C, but with a shorter critical day length. Below the critical day length, a shorter day length corresponded to a shorter larval period. Larvae transferred from long-day conditions to various photoperiods showed a similar quantitative response. Field rearing of larvae starting at various times of year showed that pupation occurs within a relatively short period in early autumn. Field rearing of pupae and adults at various times indicated that only pupation in early autumn results in a high survival rate until winter. Earlier or later pupation led to a low survival rate due to death before overwintering in the adult and pupal stages, respectively. Thus, in P. rufiventris, timing of pupation regulated by the quantitative short-day photoperiodic response is vital for survival. Relatively lower developmental threshold in the pupal stage supports this hypothesis.

Photoperiodic response of larvae of the yellow-spotted longicorn beetle Psacothea hilaris after removal of the stemmata.

The role of the stemmata in photoperiodism has been examined in holometabolic insects, but the only reliable results in Coleoptera have been obtained in Leptocarabus kumagaii (Carabidae), the larvae of which do not respond to photoperiod without stemmata. In the present study, photoperiodism was examined in another coleopteran, Psacothea hilaris (Pascoe) (Cerambycidae), after surgical removal of the stemmata. Larvae reared under short-day conditions and transferred to long-day conditions on day 2 of the 5th instar pupated without further larval molts, whereas those continuously reared under short-day conditions underwent supernumerary molts and did not pupate. When the stemmata were removed on day 2 of the 5th instar, the larvae pupated under long-day conditions but did not do so under short-day conditions. However, under long-day conditions some underwent supernumerary molts before pupation. Larvae from which the sensilla trichodeum were removed showed a similar response to that of stemmata-deficient larvae, and larvae from which stemmata were removed at a younger stage (day 2 of the 4th instar) responded to photoperiod similarly to intact larvae. Thus, supernumerary molts under long-day conditions after removal of the stemmata were attributed to injury due to surgery, rather than a change in photoperiodic photoreception. Therefore, we conclude that larvae of P. hilaris show a photoperiodic response after removal of stemmata, in contrast to larvae of L. kumagaii.

Different photoreceptor organs are used for photoperiodism in the larval and adult stages of the carabid beetle, Leptocarabus kumagaii.

The role of the two distinct retinal photoreceptor organs in photoreception for photoperiodism was examined in the carabid beetle, Leptocarabus kumagaii, by surgical removal. This beetle shows long-day and short-day photoperiodic responses in the larval and adult stages, respectively. Larval diapause in the final instar is induced under short-day conditions whereas pupation occurs without diapause under long-day conditions. Adult reproductive diapause is terminated under short-day conditions but maintained under long-day conditions. The stemmata of the larvae and compound eyes of the adults were removed and the responses of the animals to photoperiod were compared to those of intact beetles. When all the stemmata were removed, larvae pupated without entering diapause under both long-day and short-day conditions, indicating that the larvae lacking stemmata were incapable of photoreception for photoperiodism. As in other holometabolous insects, the stemmata migrated into the brain during metamorphosis and remained rudimentarily in the optic lobe of the adult brain. However, these stemmata-derived organs were found to be no longer necessary for photoperiodism, because adults lacking the stemmata-derived organs responded to photoperiod normally. By contrast, removal of the compound eyes in adults resulted in the termination of reproductive diapause under both long-day and short-day conditions, indicating that photoreception for photoperiodism in the adult stage is performed by the compound eyes. Therefore, the site of photoperiodic photoreception in L. kumagaii appear to change from the stemmata to the compound eyes during metamorphosis.

Artificial selection for responsiveness to photoperiodic change alters the response to stationary photoperiods in maternal induction of egg diapause in the rice leaf bug Trigonotylus caelestialium.

Female adults of the rice leaf bug Trigonotylus caelestialium (Kirkaldy) (Heteroptera: Miridae) produce non-diapause eggs under long-day conditions, whereas they produce diapause eggs under short-day conditions. These egg-production modes change following a photoperiodic change from long-day to short-day conditions or vice versa, with individual variations in responsiveness shown in the time from the photoperiodic change to the mode change. Strains of this insect with higher or lower responsiveness to photoperiodic change were established after several generations of selection, indicating that the individual variation has a genetic basis. The selected strains that were more responsive and less responsive to one photoperiodic change were found to be less responsive and more responsive to the opposite photoperiodic change, respectively, indicating a significant negative correlation between responsiveness to reciprocal photoperiodic changes. The selected strains also had a significantly different incidence of diapause-egg producers in stationary photoperiods compared to a non-selected strain, showing that selections for responsiveness to photoperiodic change were essentially the same as selections for a higher or lower incidence of diapause-egg producers. These results indicate that responsiveness to photoperiodic change is one aspect of the tendency to produce diapause or non-diapause eggs.

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.

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.

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.