G.M. Chippendale

Juvenile hormone regulation of the larval diapause of the Southwestern corn borer, Diatraea grandiosella☆

Abstract The hormonal mechanism which controls the larval diapause of the southwestern corn borer was examined. The onset of this facultative mature larval diapause is marked by a transition from a spotted to an immaculate larval form, and during diapause individuals may undergo one or more stationary larval ecdyses. Experiments were designed to uncover the nature of the humoral mechanism regulating this diapause state. The finding that injecting diapause larvae with 20-hydroxyecdysone only brought about a stationary larval ecdysis suggests that diapause was not maintained by the lack of ecdysone. Neck ligations performed on larvae which had just entered diapause resulted in a premature termination of diapause, and larval-pupal ecdysis occurred in the thoraco-abdominal section, suggesting that a cephalic factor was necessary for the maintenance of diapause. This finding was further supported by the discovery that injecting 20-hydroxyecdysone into the thoraco-abdominal section of previously ligated diapause larvae also resulted in a premature termination of diapause and larval-pupal ecdysis, indicating that ecdysone only initiated the pupal moulting cycle when the cephalic factor was absent. Further experiments led to the conclusion that the juvenile hormone is the cephalic factor. Topical treatment with a juvenile hormone mimic caused non-diapause mature larvae to become immaculate and enter diapause. Periodical topical application of this mimic to diapause larvae prolonged diapause and increased the number of stationary larval ecdyses. These findings suggest that the initiation and maintenance of diapause are regulated by juvenile hormone titre. Results indicate that larvae retain a high titre of juvenile hormone until the last stages of diapause. Injection of 20-hydroxyecdysone into early or middiapause larvae only caused stationary larval ecdyses, while the same injection into larvae in the late stages of diapause caused some of them to pupate. Histological studies of the neurosecretory cells, corpus cardiacum-allatum complex, and prothoracic glands showed that the endocrine system was not inactive during diapause. A new hypothesis is therefore proposed which recognizes the existence of hormonal activity during larval diapause and emphasizes the principal regulatory role of juvenile hormone.

Migration of the monarch butterfly, Danaus plexippus: Energy sources

Abstract The origin and nature of the metabolic reserves of the monarch butterfly during its autumn migration through Missouri were examined. Information was obtained about the butterflys protein, glycogen, lipid, and lipid-soluble pigment composition. The principal results showed that lipids, which made up ca. 45 per cent of the insects dry weight, were the major reserve. Analyses of the lipid classes showed that triglycerides were the dominant class in each sex. Separate analyses of the thorax and abdomen showed that >90 per cent of the lipids were present in the massive fat body which is restricted to the abdomen. The adult female rapidly incorporated U-14C- d -glucose into abdominal glycerides. The role of larval reserves and nectar consumption in migration and ovarial maturation was discussed.

Diapause of the southwestern corn borer, Diatraea grandiosella: Further evidence showing juvenile hormone to be the regulator

Abstract Further evidence is presented to demonstrate the involvement of juvenile hormone (JH) in regulating diapause in the final larval stage of the southwestern corn borer. Diatraea grandiosella. JH titres in the haemolymph were measured throughout the entire diapause period. Additional results showed that actively secreting corpora allata are necessary to maintain diapause because allatectomized larvae terminated diapause prematurely. A topical application of JH mimic 2 days after the allatectomy prevented this premature termination of diapause. Intact nervous connections between the brain and the corpora allata were necessary for the maintenance of JH secretion. Other surgical work showed that the brains of nondiapausing larvae exhibited a higher ecdysiotropic activity than those of pre-, early-or mid-diapausing larvae. A single application of a JH mimic was more effective in maintaining a diapause-like state in nondiapausing larvae than were repeated topical applications of C18-JH or an implantation of active corpora allata, suggesting that JH was more rapidly metabolized than was the JH mimic. The oxygen consumption of diapausing larvae which had received repeated topical applications of JH mimic was not significantly elevated over that of the controls indicating that treated larvae maintained a low metabolic rate even though they reverted to the spotted morph. A single application of 0.03 μg JH mimic/larva was sufficient to prolong diapause, thereby confirming that JH is necessary for diapause maintenance.

Photoperiodic and thermoperiodic interactions in the regulation of the larval diapause of Diatraea grandiosella

Abstract The effect of various combinations of photoperiod and temperature on the induction and termination of the mature larval diapause of a Missouri strain of the southwestern corn borer. Diatraea grandiosella, was examined. Larval exposure to regimes in which the low phase of a 30°:23°C thermoperiod coincided with a scotophase of 10 to 14 hr duration led to high incidence of diapause. Larval exposure to 30°:24°C, 33°:21°C, and 36°:18°C thermoperiods with half cycles of 12 hr in continuous darkness yielded a diapause incidence of 16%, 22%, and 59%, respectively, whereas exposure to a 30°:24°C thermoperiod in continuous illumination yielded a completely nondiapause generation. Larval exposure to one of a series of 36°:18°C thermoperiods in which the duration of the high phase was increased in 2 hr increments from 0 to 24 hr in continuous darkness showed that “short-day” thermoperiods yielded a high incidence of diapause. However, no clearly defined critical thermoperiod was observed. An examination of photoperiodic and thermoperiodic effects on diapause development showed that, in general, those combinations of temperature and light cycles which were diapause inductive also retarded diapause development. The relationship between seasonal photoperiods and thermoperiods in southeastern Missouri was examined.

Metamorphic changes in fat body proteins of the southwestern corn borer, Diatraea grandiosella☆

Abstract Developmental changes in the fat body proteins of the southwestern corn borer, Diatraea grandiosella, were studied from the beginning of the fifth instar to the newly ecdysed pupal stage. The dry weight of the perivisceral fat body was found to increase markedly from a mean of 1·15 mg/insect at 10 days of age to 7·9 mg in the newly ecdysed pupa (16 days). This rise was partially attributed to the increase in fat body protein content which took place during the same period. The fat body protein content rose from about 0·25 mg/insect at 10 days to a maximum of about 1·7 mg/insect in the newly ecdysed pupa. The rate of protein biosynthesis during this period was estimated using leucine-14C as a tracer. Results showed that 14-day-old larval fat body, unlike that of newly ecdysed pupae, had a high rate of protein biosynthesis. The chronology of synthesis and accumulation of the major proteins was determined by subjecting the soluble fat body proteins to disk electrophoresis. A changing electrophoretic pattern was detected between 10 and 15 days, which coincided with the period of active protein biosynthesis. Analyses of 15-day-old mature larval fat body revealed the presence of three major soluble proteins whose titre greatly increased after pupal ecdysis. The relationship between fat body and haemolymph proteins and the role of the major fat body proteins in adult differentiation are discussed.

Temperature and photoperiodic regulation of diapause of the southwestern corn borer, Diatraea grandiosella

Abstract The effect of temperature and photoperiod on the onset and termination of the mature larval diapause of the southwestern corn borer, Diatraea grandiosella , was examined. The results showed that diapause induction was an extremely temperature-dependent process. Larvae reared at 23°C under short days all entered diapause whereas 90 per cent of those reared at 27°C developed continuously. A photoperiodic response was only demonstrable at 25°C, when diapause was instituted following larval exposure to daily photophases ranging from 8 to 14 hr. An examination of the sensitivity of immature larvae to low temperatures revealed that all instars showed some susceptibility to low temperature induction of diapause. No intermediate instar was found in which the developmental programme could be switched entirely from a diapause to a non-diapause one, or vice versa. Diapause development was also found to be primarily temperature regulated. The rate of termination of diapause at 30°C was significantly higher than that at 25 or 23°C. Continuous exposure to light rather than a daily photoperiod produced the highest rate of diapause development. It was also shown that a period of chilling (5°C) did not accelerate diapause development. The significance of the results in relation to the evolution and geographical distribution of the species is discussed.

Nature and fate of the nutrient reserves of the periodical (17 year) cicada

Abstract The metabolic reserves of Magicicada cassini , a periodical cicada with a 17 year life cycle, were examined. Analysis of males and females which were collected shortly after emergence and just before death provided information about the nature and fate of the reserves which had been accumulated during the attenuated larval period. Results showed that young females had a mean weight of 565 mg compared with only 351 mg for equivalent males. Lipids and soluble proteins were the principal reserve materials. Glycogen was a relatively minor reserve of both sexes. While young females contained about twice as much soluble protein (110 mg) as the males (51 mg), they contained three times as much total lipid material (58–19 mg). Lipids were fractionated into neutral and phospholipids and the absolute amounts of each class were determined; triglycerides were the main component. Further analyses of old females provided information about the utilization of reserves for oogenesis. The data showed that even though the adults feed, lipids and soluble proteins stored during the larval stages are essential for egg maturation. The results are discussed in relation to the special requirements of an insect which has a protracted immature stage, yet a fleeting adult existence.

Free amino acids of the haemolymph of the southwestern corn borer and the European corn borer in relation to their diapause

The titres of free amino acids present in the haemolymph of diapausing larvae of the southwestern corn borer, Diatraea grandiosella and the European corn borer, Ostrinia nubilalis, were examined. High titres of serine were found in the haemolymph of both species. Serine may serve as a storage form of compounds that are required for the synthesis of uric acid and other purines. The high titres of proline found in the haemolymph of O. nubilalis during the fall and winter may contribute to the freezing tolerance of this species. Alanine accumulated in the haemolymph of both species during the winter.

Juvenile hormone and the induction of larval polymorphism and diapause of the southwestern corn borer, Diatraea grandiosella

Abstract The southwestern corn borer, Diatraea grandiosella , enters diapause as an immaculate mature larva which is a polymorphic variant of the spotted non-diapause larva. Because dormant immaculate larvae could be obtained by treating last stage non-diapause larvae with a juvenile hormone mimic (JHM), experiments were conducted to determine whether these hormonally induced immaculate larvae (HIL) were physiologically comparable to the normal environmentally induced immaculate diapause larvae (EIL). Comparative data obtained about pupation rates, response to JHM, metabolic reserves, oxygen consumption, and the state of spermatogenesis of HIL and EIL led to the conclusion that the HIL were physiologically similar to the EIL. The results demonstrate that the developmental programme of non-diapause larvae could be switched and ‘diapause’ induced solely by the topical application of JHM. We believe that the data further support our hypothesis that the larval diapause of D. grandiosella is initiated and maintained by the juvenile hormone.

Larval diapause of the European corn borer, Ostrinia nubilalis: Further experiments examining its hormonal control

Abstract The possible involvement of juvenile hormone (JH) in controlling the mature larval diapause of the European corn borer, Ostrinia nubilalis , was examined using biological and chemical assays for JH titres, topical applications of JH mimic, and injections of 20-hydroxy-ecdysone. Bioassays of extracts of larval haemolymph showed that (1) 4th instar pre-diapausing larvae had a higher JH titre ( ca. 1450 Galleria Units (GU)/ml) than equivalent non-diapausing larvae ( ca. 340 GU/ml), and that (2) 5th instar pre-diapausing larvae contained a JH titre of ca. 320 GU/ml, which declined to ca. 90 GU/ml in newly-diapaused larvae. Chemical assasys carried out on extracts of whole larvae showed that early diapausing larvae contained an extremely low titre of JH. In addition, the application of JH mimic or 20-hydroxy-ecdysone or both agents to diapausing larvae failed to reveal the presence of a functional JH titre during diapause. The application of JH mimic to early 5th instar non-diapausing larvae produced moribund larval-pupal intermediates rather than supernumerary larvae. Our results, therefore, suggest that although JH may control some phases of diapause induction, it is not involved in maintaining diapause.