Effects of Age, Phase Variation and Pheromones on Male Sperm Storage in the Desert Locust, Schistocerca gregaria.

In general, sperm produced in the testis are moved into the seminal vesicle via the vas deferens in insects, where they are stored. How this sperm movement is controlled is less well understood in locusts or grasshoppers. In this study, the effects of age, phase variation and pheromones on male sperm storage were investigated in the desert locust, Schistocerca gregaria (Forskål). In this locust, a pair of ducts, the vasa deferentia, connect the testes to a pair of the long, slender seminal vesicles that are folded approximately thirty times, and where the sperm are stored. We found that phase variation affected the level of sperm storage in the seminal vesicle. Moreover, adult males that detected pheromones emitted by mature adult males showed enhanced sperm storage compared with males that received the pheromones emitted from nymphs: The former, adult male pheromones are known to promote sexual maturation of immature adults of both sexes, whereas the latter, nymphal pheromones delay sexual maturation. Most mature adult males had much sperm in the vasa deferentia at all times examined, suggesting daily sperm movement from the testes to the seminal vesicles via the vasa deferentia. As adult males aged, sperm were accumulated from the proximal part to the distal end of the seminal vesicle. Many sperm remained in the seminal vesicle after mating. These results suggest that young or new sperm located near the proximal part of the seminal vesicle could be used for mating, whereas old sperm not used for mating are stored in the distal part of the seminal vesicle.

Effects of photoperiod and aging on the adult spermatogenesis of Polygonia c-aureum (Lepidoptera: Nymphalidae), in relation to adult diapause.

Adult spermatogenesis of Polygonia c-aureum was compared between non-diapausing and diapausing butterflies before overwintering. This butterfly has seasonal polyphenism, i.e., summer and autumnal forms. Summer form butterflies that emerged in summer reproduce shortly after emergence, while autumnal forms that emerged in autumn mate in spring. Immatures were reared under either a long photoperiod, which produced the summer form without diapause or under a short photoperiod, which produced the autumnal form with diapause. We found almost no differences in adult spermatogenesis between the two seasonal forms, indicating that adult spermatogenesis is not related to adult diapause. Although adult diapause in the autumnal form is maintained under short photoperiods and terminated under long photoperiods, such a photoperiod did not affect the spermatogenesis of the autumnal form. Our earlier studies indicate that relatively few eupyrene and apyrene sperm are produced after overwintering. Although apyrene spermatogenesis occurred in young adults, eupyrene spermatogenesis did in a small scale before overwintering. These results suggest strongly that male autumnal form butterflies prepare the sperm until overwintering, which had been formed during the larval, pupal and young adult stages.

Field and Laboratory Studies on the Ecology, Reproduction, and Adult Diapause of the Asian Comma Butterfly, Polygonia c-aureum L. (Lepidoptera: Nymphalidae).

Adult diapause and reproduction of a nymphalid butterfly, Polygonia c-aureum L., were investigated in field and laboratory examinations. Laboratory studies showed that old virgin male butterflies of non-diapausing generations had heavy accessory glands and simplex, which were suppressed in diapausing generations. The number of eupyrene sperm bundles in the duplex increased with adult age, whereas testis size decreased with age. Field examinations indicated that reproductive development of both sexes of diapausing generations in autumn was suppressed, and developed in spring. We attempted to estimate the physiological age of wild-caught males, as adult male age can be estimated from the testis size. We also attempted to determine whether or not wild male butterflies had mated from the development of the accessory glands and simplex, as well as the number of eupyrene sperm bundles in the duplex, by comparing unmated males with mated males. Field examinations suggest that almost all females in a population of non-diapausing generations mated and showed a tendency toward polyandry, while in the diapausing generation, in spring, monoandry rather than polyandry predominated. This suggests a different mating strategy between non-diapausing and diapausing generations.

Effects of photoperiod, temperature and aging on adult diapause termination and post-diapause development in female Asian comma butterflies, Polygonia c-aureum Linnaeus (Lepidoptera: Nymphalidae).

Polygonia c-aureum females exhibit photoperiodically induced imaginal diapause, characterized by cessation of ovarian development. Females grown at a short daylength (SD) entered imaginal diapause, whereas those grown at a long daylength (LD) produced eggs rapidly after adult emergence at 21 °C. The termination of diapause was influenced by daylength: diapause ended faster at LD than SD. Complete termination of diapause took 30 days in unchilled females reared under LD at 21 °C. On the other hand, prompt, synchronized and strong diapause termination occurred at post-chilling periods. Photoperiods at post-chilling periods affected ovarian development, when the length of pre-chilling periods or the length of chilling periods was shorter, suggesting that these treatments were not enough to complete diapause development. Ovarian development proceeded earlier in chilled and subsequent warmed females than unchilled females. Wing damage was remarkable at post-chilling periods when females were reared under an adequate length of pre-chilling and chilling periods, especially comparing with females under pre-overwintering conditions without chilling, indicating that post-diapause reproductive development was weak in unchilled females. Thus, exposure to low temperatures is necessary for a strong diapause termination in this butterfly.

Effects of Irradiation Dose on Sperm Production, Insemination, and Male Mating Possible Period in the Sweetpotato Weevil (Coleoptera: Brentidae).

The sterile insect technique (SIT) has been used for the control or eradication of target insect pests. To successfully apply SIT, it is very important to clarify the effect of irradiation on male reproduction in the target pest, because their mating and spermiogenesis abilities affect the success of eradication program. The sweetpotato weevil, Cylas formicarius (Fabricius) (Coleoptera: Brentidae) is a notorious and worldwide pest of sweet potato. We investigated the effect of irradiation at five doses ranging from 0 (control) to 150 Gy on 9-d-old males. Survival rate of the control (no treatment) remained high from day 10 to 20 of adult life, whereas higher doses of irradiation reduced it, maximally by approximately 70%. Mating rates showed a similar tendency. Radiation dose neither affected sperm production nor sperm transfer at any dose, although spermiogenesis is active during the adult stage. However, radiation dose affected the lifetime total of ejaculated sperm number, likely because of fewer matings by irradiated males. These results suggest that use of a dose of 150 Gy or higher is appropriate for the final step of eradication of this weevil. At least, lower dose of irradiation may arise the inadequate sterilization, resulting in a failure of eradication program. We conclude that weekly release of sweetpotato weevil sterilized with high dose, achieving complete sterilization, could be useful for eradication program after reducing the population by male annihilation method.

Effects of pre-overwintering conditions on eupyrene and apyrene spermatogenesis after overwintering in Polygonia c-aureum (Lepidoptera: Nymphalidae).

Sperm polymorphism is widely known in invertebrates. In insects, Lepidoptera has two types of sperm: nucleated eupyrene (fertile) sperm and anucleated apyrene (unfertile) sperm. These sperm types are produced during post-embryogenesis, and eupyrene spermatogenesis precedes apyrene spermatogenesis. During overwintering, spermatogenesis stops and a portion of undifferentiated-stage spermatocytes degenerate. After overwintering, spermatogenesis restarts with unaffected spermatogonia. However, how new spermatozoa arise in the adult testes after overwintering is not known in Lepidoptera. In this study, we investigated the spermatogenesis events in the nymphalid butterfly Polygonia c-aureum after overwintering under various environmental conditions. Our results indicate that both eupyrene and apyrene spermatogenesis restart at any stopping stage and sperm of these types are regenerated in no particular order after adult insect overwintering. This suggests that the spermatogenesis occurring after overwintering proceeds without embryogenetic restrictions related to the developmental sequence.

Effects of juvenile hormone analogue (methoprene) and 20-hydroxyecdysone on reproduction in Polygonia c-aureum (Lepidoptera: Nymphalidae) in relation to adult diapause.

We investigated the effects of juvenile hormone analogue (methoprene) and 20-hydroxyecdysone on female and male reproduction in a nymphalid butterfly, Polygonia c-aureum. This butterfly has a facultative adult diapause controlled by the corpora allata and brain. Methoprene seems to terminate reproductive diapause, although transplantation experiments indicate that the activity of the corpora allata does not affect male mating behavior Endo (Dev Growth Differ 15:1-10, 1973a), suggesting that the brain may be involved in diapause. We found that exposure to methoprene promoted the development of ovaries and of the male accessory glands and simplex. On the other hand, exposure to 20-hydroxyecdysone did not promote the development of female and male reproductive organs and eupyrene sperm movement from the testis to the duplex in the adult stage. Ecdysteroid titer in both sexes was consistently low in adults. These results suggest that imaginal diapause is largely regulated by juvenile hormone in this butterfly.

Potential Host Range of the Larval Endoparasitoid Cotesia vestalis (=plutellae) (Hymenoptera: Braconidae).

Many parasitoid wasps are highly specialized in nature, attacking only one or a few species of hosts. Host range is often determined by a range of biological and ecological characteristics of the host including diet, growth potential, immunity, and phylogeny. The solitary koinobiont endoparasitoid wasp, Cotesia vestalis, mainly parasitizes diamondback moth (DBM) larvae in the field, although it has been reported that to possess a relatively wide lepidopteran host range. To better understand the biology of C vestalis as a potential biological control of hosts other than the DBM, it is necessary to determine suitability for potential hosts. In this study, the potential host range of the wasp and its developmental capacity in each host larva were examined under laboratory conditions using 27 lepidopteran species from 10 families. The wasp was able to parasitize 15 of the 27 species successfully. Some host species were not able to exclude C vestalis via their internal physiological defenses. When parasitization was unsuccessful, most hosts killed the parasitoid at the egg stage or early first-instar stage using encapsulation, but some host species disturbed the development of the parasitoid at various stages. No phylogenetic relationships were found among suitable and unsuitable hosts, revealing that host range in some endoparasitoids is not constrained by relatedness among hosts based on immunity.