Biochemical systematics of Japanese fireflies of the subfamily Luciolinae and their flash communication systems

Japanese fireflies of the subfamily Luciolinae are biochemically analyzed using 13 allozymes, and the phylogenetic relationships obtained from this analysis are compared with their flash communication systems. As a result, the Japanese Luciolinae can be divided into three groups.Hotaria parvula andH. tsushimana together withLuciola yayeyamana andL. kuroiwae from the first group, and they use the same communication system.L. lateralis, Curtos okinawana, andC. costipennis make up the second group, and their communication systems are also the same.L. cruciata makes up the last one, and its communication system is different from the other fireflies of Luciolinae. Therefore, their taxonomical arrangement and communication systems are not congruent. However, the genetic similarity deduced by allozymic analysis of the members of the Japanese Luciolinae is highly consistent with their flash communication systems.     

A comparative study of spectral sensitivity curves in three diurnal and eight nocturnal species of Japanese fireflies

The spectral properties of the eyes of 3 species of diurnal and 8 species of nocturnal Japanese fireflies, in many cases males and females, were determined by an electroretinographic method. With the exception of Hotaria parvula males, which had a λ_max of 580 nm, almost all species studied possessed a maximum around 500–540 nm. The eyes of diurnal and nocturnal species did not differ significantly in their sensitivity maxima. As in North American species of fireflies (Lall, 1981a,b) congruency existed between visual sensitivity peaks and light emission maxima in Luciola cruciata, L. lateralis and Hotaria parvula. In agreement with Seliger et al. (1982a,b) we conclude that an adaptation of the visual sensitivity to the light produced need not have occurred and that evolutionary adaptation of light emission to an existing ancestral green-sensitivity of the eye is the more likely course of events.     

Variation in flash speed of Japanese firefly,Luciola cruciata (Coleoptera: Lampyridae), identifies distinct southern “quick‐flash” population on Goto Islands,Japan

Luciola cruciata, a bioluminescent firefly endemic to Japan, communicates with species‐specific flashing patterns. Previous studies reported that flying males possess three distinct flashing frequencies: slow (4 s), intermediate (3 s) and fast (2 s). Our study focused on geographical variations in flashing patterns in the northwestern part of the Kyushu region of southern Japan (including its isolated islands) and their relationship to the mitochondrial COII region of firefly DNA. We found that “quick‐flash (1 s)” fireflies occur in the Goto Islands, at the western tip of Kyushu, and that, although they possess a unique haplotype, they do not differ much genetically from “fast‐flash” fireflies inhabiting mainland Kyushu. In contrast, although fireflies inhabiting the Tsushima and Iki Islands also possess a unique haplotype, their flashing patterns were the same (fast‐flash) as fireflies inhabiting mainland Kyushu. We determined that flash patterns of L. cruciata in Japan should be classified according to four flashing patterns: slow, intermediate, fast and quick.     

CCD imaging of basal bioluminescence in larval fireflies: clues on the anatomic origin and evolution of bioluminescence.

The anatomic and biochemical origin of beetle bioluminescence is still poorly understood. Through CCD imaging we report that larvae and pupae of the Brazilian fireflies Aspisoma lineatum and Cratomorphus sp emit a continuous weak glow throughout the entire body during all stages. This luminescence is especially developed after feeding, ecdysis and in the pupal stage, gradually disappearing as the cuticle becomes sclerotized and the adult emerges. This weak glow arises from the fat body, which consists of small lobes spread all over the body cavity. According to their pigmentation, these lobes can be divided in whitish and pinkish, and display different luciferase isozymes. Morphological studies suggest that the jelly-like ventral lanterns in the 8th abdominal segment evolved from these white lobes, providing a rationale for the widespread location of lanterns in larvae of different bioluminescent beetles. The biological and biochemical function of this weak diffuse bioluminescence is discussed in the context of the larval life-history.     

Adult firefly abundance is linked to weather during the larval stage in the previous year

1. Much is known about the brief adult phase of fireflies. However, fireflies spend a relatively long developmental period under the soil surface. Climatic and soil conditions may directly affect the eggs, larvae, and pupae, and also affect them indirectly through predators, competitors, and prey items. Climatic conditions during the early life stages of this iconic species are therefore relevant to their hypothesised decline within the context of global warming. 2. Data on the abundance of fireflies were extracted from the publicly available citizen dataset across North America over a period of 9 years. The effects of weather in the 24 months before the observations of firefly abundance were documented based on 6761 observations. 3. Climatic conditions during both the larval and adult phases have a non‐linear effect on adult firefly abundance. Maximum winter and spring temperatures and mean precipitation in the 20‐month period before the observations had the greatest impact on the abundance of firefly adults. Low maximum soil moisture during the 5–19 months preceding the observations affected the adult abundance negatively, and high maximum soil moisture affected it positively. 4. After correcting the firefly abundance for these weather effects, it is estimated that the abundance of fireflies increased over the time period of this study. 5. This study suggests that early life climatic conditions have a small but significant impact on adult firefly abundance with a total R² of 0.017.     

Parasitism and oviposition of Melittobia acasta on Bombus terrestris: Impact of larval overwintering and host status on pupation and adult emergence

Melittobia acasta (Walker) are microhymenopteran ectoparasitoids of the pupae and prepupae of the commercially‐used pollinator bumblebee species Bombus terrestris L. The female parasitoids puncture the host cuticle with their sting and feed oozing hemolymph. This study shows that M. acasta parasitize 100% pupae and 84% prepupae of B. terrestris but are ineffective on the larvae of the bees. The female parasitoids lay a significantly higher number of eggs on pupae (67.7 ± 16.2 female^?1) compared to prepupae (20.5 ± 14.5 female^?1). The parasitoids differ in their choice for oviposition sites and fecundity on different locations of B. terrestris pupae, and they show most preference for oviposition (32%) as well as fecundity (34.9 ± 15.1 female^?1) on the petiole of the host. Larvae of the parasitoids overwinter at low temperatures but larval overwintering duration and post‐diapause rearing on original or new hosts do not affect their pupation and adult emergence. Larvae have a higher percentage of pupation (88.0–94.4%) and adult emergence (84.4–92.9%) both on the original and the new host, thus indicate that the parasitoids are highly capable of reproduction in B. terrestris colonies.     

Morphological features,development and reproduction of Melittobia acasta on Bombus terrestris

Morphological features, development and reproduction behavior of the parasite Melittobia acasta (Walker) were studied when reared on the pupae of the bumblebee Bombus terrestris L. in the laboratory under 23°C, 50% relative humidity and 12 h light : 12 h dark conditions. The parasites laid transparent white and elongated eggs. Newly hatched larval size and shape were very similar to eggs but they were identified by their body segments. Larvae increased their body size through moulting and transformed into a vermiform shape. Male pupae were shiny brown with dots. The female pupae were distinguished by their black shiny color, shorter size and the presence of compound eyes. Adult male pupae were dark brown and dwarf‐winged, whereas female pupae were macropterous and brachypterous. Reproduction took place by fertilization and also parthenogenetically. Mean fecundity within 5 days by mated (47.9 ± 30.5 female^?1) and virgin (7.4 ± 6.8 female^?1) females were statistically different. Mated females laid fertilized eggs that produced adult males or females, whereas virgin females laid unfertilized eggs that produced males. Development durations of the virgin female originated eggs, larvae, pupae and adults were statistically identical with those of mated females. The parasites were female‐biased and foundress number did not affect offspring sex ratio. This study shows that both mated and virgin females of M. acasta can produce many offspring on B. terrestris pupae within a short period, indicating that they are dangerous parasites of the bumblebee in a mass rearing system.     

CRISPR/Cas9-based functional analysis of yellow gene in the diamondback moth,Plutella xylostella

The diamondback moth, Plutella xylostella (L.), is an economically important pest of cruciferous crops worldwide. This pest is notorious for rapid evolution of the resistance to different classes of insecticides, making it increasingly difficult to control. Genetics-based control approaches, through manipulation of target genes, have been reported as promising supplements or alternatives to traditional methods of pest management. Here we identified a gene of pigmentation (yellow) in P. xylostella, Pxyellow, which encodes 1674 bp complementary DNA sequence with four exons and three introns. Using the clustered regularly interspersed palindromic repeats (CRISPR)/CRISPR-associated protein 9 system, we knocked out Pxyellow, targeting two sites in Exon III, to generate 272 chimeric mutants (57% of the CRISPR-treated individuals) with color-changed phenotypes of the 1st to 3rd instar larvae, pupae, and adults, indicating that Pxyellow plays an essential role in the body pigmentation of P. xylostella. Fitness analysis revealed no significant difference in the oviposition of adults, the hatchability of eggs, and the weight of pupae between homozygous mutants and wildtypes, suggesting that Pxyellow is not directly involved in regulation of growth, development, or reproduction. This work advances our understanding of the genetic and insect science molecular basis for body pigmentation of P. xylostella, and opens a wide avenue for development of the genetically based pest control techniques using Pxyellow as a screening marker.     

Fly parasitoid Megaselia opacicornis uses defensive secretions of the leaf beetle Chrysomela lapponica to locate its host

Larvae of the leaf beetle Chrysomela lapponica derive a defensive secretion from salicyl glucosides found in the host plant Salix borealis. This secretion protects beetle larvae from some natural enemies, but does not appear to repel parasitoids. We tested the hypothesis that the fly parasitoid Megaselia opacicornis (Diptera, Phoridae) uses the larval defensive secretion of Ch. lapponica in its search for prey. In the field, nearly 30 times more M. opacicornis individuals were caught on leaves coated with sticky resin next to a source of secretion than on control leaves. In the laboratory, M. opacicornis females laid six times more eggs next to a cotton ball soaked in secretion than next to one soaked in water. Fly females also lay more eggs on prey rich in larval secretion than on secretion-poor prey. In the field, removal of defensive secretion from beetle prepupae resulted in a 7.5-fold reduction of oviposition by fly females. Parasitoids were nearly twice as likely to lay eggs on prepupae, rich in secretion, as on pupae, which contain little secretion. Fly offspring reared from beetle prepupae reached a 21% larger body mass than those reared from pupae. Finally, M. opacicornis females avoided host prepupae already parasitized by the tachinid fly Cleonice nitidiuscula, which possess little secretion. These experiments indicate that host plant-derived defensive secretions are used by this parasitoid for host location. Adaptation of parasitoids to use defensive secretions of hosts may selectively favor an increase in diet breadth in specialist herbivores.     

Studies on Tyrosinase in Housefly

Occurrence of protyrosinase in the prepupae of housefly, Musca vicina Maquart, and its activation were described. The prepupae possessing no appreciable tyrosinase activity could be separated from the other aged pupae by putting them into water. Homogenate prepared from the prepupae contains protyrosinase and has no activation system for the proenzyme. As has been reported by Ohnishi²⁾ a certain activator occurred naturally in the aged pupae apparently activates the protyrosinase in vitro. However, contrary to Ohnishi’s results³⁾ it was found that this protyrosinase can be activated by the treatment with sodium dodecyl sulfate in vitro.     

Differences in pupal cold hardiness and larval food consumption between overwintering and non‐overwintering generations of the common yellow swallowtail,Papilio machaon (Lepidoptera: Papilionidae), from the Osaka population

To clarify differences in pupal cold hardiness and larval food consumption between overwintering and non‐overwintering generations of the common yellow swallowtail, Papilio machaon, we reared larvae from the Osaka population under photoperiods of 16 h light : 8 h dark (LD 16:8) (long day) or LD 12:12 (short day) at 20°C. We examined the relationship between food consumption and weight during the final larval stadium and pupae, and measured the pupal supercooling point (SCP). Although the ratio of assimilation to consumption did not differ significantly between photoperiods, the ratio of assimilation to pupal weight differed significantly between individuals reared under long and short days. All diapausing pupae were brown, whereas 56% of non‐diapausing pupae were green with the remainder brown. The mean pupal body length (L), dorsal width (W₁) and lateral width (W₂) were larger in non‐diapausing than in diapausing pupae, and the W₁/L and W₁/W₂ ratios differed significantly between non‐diapausing and diapausing pupae. SCP was approximately –20°C and did not differ among pupae 5, 15 and 30 days after pupation under long‐day conditions. However, under short‐day conditions, mean SCP gradually decreased, stabilizing at approximately –24 to –25°C by 30 days after pupation. After freezing, some diapausing pupae emerged as adults, whereas all non‐diapausing pupae died. Both egestion and assimilation were greater under long‐day conditions. The results revealed that pupae of this papilionid exhibit seasonal polyphenism in physiological and morphological traits. Energy from food appears to be expended on increasing cold hardiness in the overwintering generation and on reproduction in the non‐overwintering generation.     

Kinetic properties of highly purified luciferase from fireflies Luciola mingrelica.]

Luciferase of the fireflies Luciola mingrelica was isolated from dried lanterns of fireflies and purified by chromatography on DEAE-Sephadex. The homogeneity of the preparation was determined by polyacrylamide gel disc electrophoresis. The molecular weight of the enzyme equal to 45000 was determined by disc electrophoresis in the presence of sodium dodecyl sulfate. The kinetic properties of the enzyme (V and Km for luciferin and ATP) within the pH-range of 7,0--8,5 were studied. The kinetic curves of the pH-dependences of log V and log Km for both substrates are bell-shaped, with a slope equal to 2. At pH optimum (7,7--7,9) the Km values for luciferin and ATP are 6,6 mkM and 0,3 mM, respectively. The properties of luciferase L. m. were compared to those of luciferase from fireflies Phophinus pyralis previously described in literature.     

TiLIA: a software package for image analysis of firefly flash patterns

As flash signaling patterns of fireflies are species specific, signal‐pattern analysis is important for understanding this system of communication. Here, we present time‐lapse image analysis (TiLIA), a free open‐source software package for signal and flight pattern analyses of fireflies that uses video‐recorded image data. TiLIA enables flight path tracing of individual fireflies and provides frame‐by‐frame coordinates and light intensity data. As an example of TiLIA capabilities, we demonstrate flash pattern analysis of the fireflies Luciola cruciata and L. lateralis during courtship behavior.     

A comparative study of cannibalism and predation in seven species of flour beetle

Abstract 1. The present study quantified egg and pupal cannibalism, and interspecific predation on eggs and pupae, by larvae and adults of seven species of flour beetle (Tribolium spp.) under laboratory conditions: T. anaphe, T. brevicornis, T. castaneum, T. confusum, T. destructor, T. freemani, and T. madens. 2. Variation among species in cannibalism and predation propensities did not reflect taxonomic affinities within the genus, indicating that these behaviours were shaped by ecology at species level. 3. Within species, larvae and adults displayed different propensities for cannibalism and predation, leading to the conclusion that these behaviours evolve independently in the two life stages. 4. All species behaved as intraguild predators to some degree, especially in the adult stage. 5. Three general patterns of cannibalism and predation were described by principal component mapping and cluster analysis. 6. The first group comprised three cosmopolitan pest species that were more voracious as adults than as larvae: T. castaneum, T. confusum, and T. destructor. It is proposed that stored product environments select for high adult voracity because the costs associated with emigration from such rare, but resource‐rich, habitats intensifies interference competition among adults. 7. The second group consisted of species that inhabit natural environments and that were more voracious as larvae: T. anaphe, T. freemani, and T. madens. Habitats for these species are probably numerous, but generally poor in quality, a situation that intensifies larval competition, while favouring earlier adult emigration. 8. The largest species, T. brevicornis, demonstrated inconsistent voracity between life stages and was the only species with chemically defended pupae. 9. It is proposed that consumption of eggs provides primarily nutritional benefits, whereas consumption of pupae has a more important role in interference competition.     

The ecological impact of an introduced population on a native population in the firefly Luciola cruciata (Coleoptera: Lampyridae)

In Japan, Tatsuno Town has been famous for many Luciola cruciata fireflies emerging every summer at least since the 1920s. However, in the 1960s, L. cruciata fireflies were intentionally introduced from the Lake Biwa area into Matsuo-kyo, the most famous habitat of fireflies in that town. In this study, I examined ambient temperatures and flash rates of L. cruciata at four sites including Matsuo-kyo in the Tatsuno area and two sites in the Lake Biwa area. The linear regression of flash rates on temperatures indicated that the Matsuo-kyo population was distinct from the other three populations native to the Tatsuno area, but similar to the two populations native to the Lake Biwa area in terms of flash rates. These results were also supported by a recent molecular biological study, suggesting that the introduced fireflies had a strong ecological impact on the native ones at Matsuo-kyo. The present study emphasizes that we should not transport and release L. cruciata fireflies without careful consideration.     

The effect of temperature on the development and life cycle regulation of the pine weevil Hylobius abietis and the potential impacts of climate change

• 1 The pine weevil Hylobius abietis is widely distributed in the Palaearctic region where it is a major pest. Although predominantly semi‐voltine, with a 2‐year life cycle, the generation time across its range can vary from 1 to 4 years. The duration of the life cycle and the seasonal timing of weevil activity affect the economic impact and management of this pest, all of which are likely to change in a warming climate.
• 2 To determine the effect of temperature and tree species on weevil growth and development, laboratory experiments were performed with eggs, larvae, prepupae, pupae and adults, using, as appropriate, the host species Scots pine Pinus sylvestris L. and Sitka spruce Picea sitchensis (Bong.) Carr. under constant or alternating temperatures.
• 3 The development rate was linearly related to temperature, with developmental thresholds for eggs, larvae and pupae of 8, 4.5 and 7.3 °C, respectively. Day‐degrees were estimated for each life stage. Larval development was affected by tree species, being slower on Sitka spruce than on Scots pine, and was faster under alternating than constant temperatures.
• 4 The development time for prepupae was highly variable, with an apparent facultative prepupal diapause initiated by temperature. The temperature range 20–17.5 °C marked the transition between median prepupal development times of approximately 25 and 90 days. The prepupal stage may serve to minimize the risk of overwintering mortality in the pupal stage and help to synchronize the life cycle.
• 5 Larval and adult mass was positively related to developmental temperature, demonstrating an inverse temperature size rule, and weevils were heavier when developing on Scots pine than Sitka spruce. Development in alternating temperatures reduced weevil mass on Scots pine. The influence of temperature on weevil mass is likely to have a positive effect on fecundity and overwintering survival. The effects of climate change on development, voltinism and weevil mass are discussed.

     

不同林分白蜡树云斑白条天牛种群空间格局地统计学分析

云斑白条天牛是我国重要林业害虫,在黄河三角洲地区对白蜡树造成严重危害。利用地统计学方法,通过调查城市行道绿化林、庭院绿化林、公路林和片林4种不同林分类型的白蜡树林地,以产卵刻槽、排粪孔和羽化孔为调查指标,对危害白蜡云斑白条天牛种群卵、幼虫、成虫(蛹)的空间格局进行了研究。结果表明:除片林样点成虫(蛹)的半变异函数模型为指数模型外,危害白蜡云斑白条天牛种群卵、幼虫、成虫(蛹)在不同林分类型下的半变异函数模型均为球形模型,其指明空间分布均为聚集分布,种群的聚集分布习性与林分类型相关性不强。但不同林分类型云斑白条天牛种群的空间相关性地统计学参数存在差异,片林调查样点的卵、幼虫和成虫(蛹)的变程a、块金值C0和基台值C0+C均小于其他3种林分类型的统计值,片林样点成虫(蛹)的半变异函数模型不同于其他林分类型为指数模型。     

Intraguild predation among Scolothrips longicornis (Thysanoptera: Thripidae), Neoseiulus californicus and Typhlodromus bagdasarjani (Acari: Phytoseiidae) under laboratory conditions

This study was carried out on the ability of predatory thrips Scolothrips longicornis Priesner to feed on 2 phytoseiid species and vice versa. Also the effect of predation of Neoseiulus californicus (McGregor) on Typhlodromus bagdasarjani Wainstein and Arutunjan and vice versa was evaluated. The larvae, prepupae, and pupae of thrips and the eggs, larvae, and protonymphs of phytoseiids were selected as intraguild prey. The intraguild predation (IGP) among S. longicornis and 2 phytoseiid species was unidirectional and in favor of phytoseiids, i.e., S. longicornis was not able to feed on larval stages of 2 phytoseiids. However, N. californicus and T. bagdasarjani fed on the 1st instar larvae (1.39 and 0.80 per day), 2nd instar larvae (0.87 and 0.55 per day), prepupae (0.51 and 0.48 per day), and pupae of thrips (0.51 and 0.49 per day, respectively). Both phytoseiids fed on eggs, larvae, and protonymphal stages of each other. Females of N. californicus consumed more phytoseiid larvae (2.49 per day) than T. bagdasarjani, which consumed 1.08 N. californicus larvae per day. When Tetranychus urticae was presented as an extraguild prey, intensity of IGP between 2 species of phytoseiids and on larval stages of S. longicornis reduced significantly. Therefore, it is concluded that (i) IGP existed among the 3 examined species and lack of feeding of S. longicornis on 2 phytoseiid species can be justified by its feeding type (monophagy), (ii) N. californicus was much more prone to IGP than was T. bagdasarjani.