Ontogeny of the circadian system governing ecdysial rhythms in a holometabolous insect, Diatraea grandiosella (Pyralidae)

ABSTRACT. Three eclosion rhythms of Diatraea grandiosella Dyar (Lepidoptera, Pyralidae) showed different phase relationships to the LD cycle, different degrees of synchronization and different sensitivity to a brief light perturbation. Egg hatching occurred mainly at dawn, whereas pupation and adult emergence occurred mainly at dusk. The egg hatching was the most synchronized rhythm of the three, pupation the least. The egg hatching rhythm was more sensitive to a brief light perturbation than was the adult emergence rhythm. A light pulse in the middle of the night caused greater disturbance in the egg hatching rhythm than did a pulse placed in the early night. The three rhythms persisted in both DD and LL. LL free-running rhythm gradually became desynchronized and broke down at the third cycle. The phase relationships reveal difficulties in applying the 'external coincidence model' to this species' photoperiodic response.     

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

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 rôle of juvenile hormone.     

Influence of fertilizer on southwestern corn borer, Diatraea grandiosella, infestation and damage to field corn

Nitrogen (N) and phosphorus (P) fertilizers have differing effects on southwestern corn borer (SWCB). Diatraea grandiosella Dyar, densities and damage to field corn, Zea mays L. Nitrogen fertilization of corn is associated with an increase in number of stalks infested and girdled by corn borers, while addition of P has the opposite effect. When combinations of N and P were applied, there were no significant differences in SWCB infestation or damage from plots receiving no fertilizer. The effect of combinations of N and P on SWCB were intermediate to either element alone.

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Résumé Nous avons étudié les corrélations entre l'utilisation d'engrais azotés et phosphatés d'une part, l'infestation et l'alimentation de D. grandiosella sur maïs en champs d'autre part. Les formules d'engrais comparées étaient: pas d'engrais, 90 et 179 kg N/ha sans P, 45 et 90 kg P/ha sans N, et les combinaisons de ces deux teneurs en N et P.La contamination était plus élevée dans les lots ayant reçu de l'azote que dans ceux traités sans N, mais la présence d'azote n'a pas réduit le décapage des tiges et la verse dus à l'alimentatin larvaire. A l'inverse, les lots n'ayant reçu que du phosphore avaient des pourcentages plus faibles de tiges contaminées par D. grandiosella et ainsi décapées et versées, que ceux qui n'avaient pas reçu P. Il n'a pas eu de différences significatives dans les contaminations ou les dégats entre les lots ayant reçu différentes combinaisons de N et P, et les effets de ces combinaisons sur l'insecte étaient intermédiaires à ceux de N et P seuls.

     

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.     

Haemolymph ecdysteroid titres in diapause and nondiapause larvae of the European corn borer, Ostrinia nubilalis

The ecdysteroid titres of last-instar prediapausing, diapausing and nondiapausing larvae of Ostrinia nubilalis were determined by radioimmunoassay. In the nondiapause larvae a major peak of ecdysteroid activity preceded pupation by 24 h and continued through the pupal ecdysis. This peak was correlated with head and thorax critical periods as well as with changes in behaviour and physiology marking the transition from feeding larva to prepupa. Nondiapause larvae also displayed a rise in ecdysteroid titre during the feeding phase of development. This rise was approx one tenth that of the major peak and lasted 32 h. It was not correlated with any overt changes in larval physiology or behaviour. The diapause ecdysteroid profile was distinctive in that the levels measured were all lower than the lowest of the nondiapause curve. No peaks were observed in the diapause titres. Prepupal changes such as spinning and the cessation of feeding were not correlated with any increase in ecdysteroid levels. During diapause termination, under a long-day photoperiod, no increases in ecdysteroid titre were observed for the first 10 days. After 12 days individuals began to show ecdysteroids above the diapause levels. Pupation started after 16 long days.     

Relationship between the fatty acids of fat body triacylglycerol and lipophorin diacylglycerol of non-diapause and diapause larvae of the southwestern corn borer,Diatraea grandiosella

The fatty acids of the triacylglycerol reserves in the fat body and of the diacylglycerol of lipophorin in the hemolymph of non-diapause and diapause larvae of D. grandiosella were compared. For both non-diapause and diapause larvae palmitate, palmitoleate, oleate, and linoleate were the predominant fatty acids present in fatty body triacylglycerol, and palmitate, oleate, and linoleate were the predominant fatty acids present in lipophorin diacylglycerol. However, differences were detected in the relative amounts of oleate and linoleate present in lipophorin diacylglycerol of non-diapause and diapause larvae. The relative amount of linoleate in lipophorin diacylglycerol declined during diapause, whereas that of oleate remained relatively high during diapause. The fatty acid profile of lipophorin diacylglycerol from non-diapause larvae treated with a juvenile hormone analog to induce a diapause-like state more closely matched that of diapause larvae than that of non-diapause larvae. The differences detected in the fatty acid composition of lipophorin diacylglycerol in non-diapause and diapause larvae appear to be due mainly to the different physiological states rather than to the different rearing temperatures employed. The results are discussed in relation to the essential role fatty acids, especially oleate, play in the survival of diapause larvae.     

Cold hardiness of larvae of the southwestern corn borer, Diatraea grandiosella

The cold-hardening capacity of field-collected larvae from southeast Missouri and laboratory-reared larvae of the southwestern corn borer, Diatraea grandiosella Dyar, was examined. Supercooling points of non-diapause and diapause larvae collected from maize plants grown in Missouri (36°30 N lat.) were ca.-7.0°C. The hemolymph melting points of diapause field larvae (-0.8°C) were significantly lower than those of non-diapause larvae collected in July (-0.5°C). The supercooling points of hemolymph from non-diapause and diapause field larvae ranged randomly from-10° to-18°C. Supercooling points of non-diapause laboratory larvae increased from-13° to-10°C prior to pupation, whereas those of diapause larvae increased similarly before the onset of diapause, but then decreased during diapause to ca.-17°C. No change in supercooling points or capacity to survive in the presence of ice was observed in diapause laboratory larvae acclimated at 4°C for 63 days. Laboratory and field larvae began to freeze at ca.-1.5°C in the presence of ice, but survived to several degrees below their melting points. The high supercooling points of field larvae appeared to be due to the presence of an environmental ice-nucleator. Although data for laboratory larvae indicate sufficiently low supercooling points to permit winter survival in southeastern Missouri, considerable larval mortality occurs in the field due to inoculative freezing and the presence of an ice-nucleator.     

Juvenile hormone and a protein associated with the larval diapause of the southwestern corn borer,Diatraea grandiosella

Pre-diapausing larvae of the southwestern corn borer, Diatraea grandiosella, were found to store an electrophoretically discrete protein fraction which makes up about 20% of the soluble proteins of the fat body of newly diapaused individuals. This fraction appears to be restricted to the larval fat body and haemolymph. It accumulates in the fat body of pre-diapausing larvae and reaches a maximum titre in newly diapaused individuals. The protein is utilized gradually during diapause so that only traces of it remain after diapause has terminated.The protein fraction was found to accumulate in the fat body of non-diapausing larvae which had been treated with a juvenile hormone (JH) mimic. This treatment induces a diapause-like state. The involvement of JH in the accumulation and utilization of the protein was investigated by examining the proteins of the fat body after larvae had been allatectomized, or had received a JH mimic application, or had received both treatments. The results showed that non-diapausing larvae began to accumulate the protein fraction within 8 days of receiving a single application of JH mimic. Allatectomy of pre-diapausing larvae was shown to suppress the accumulation of the protein. This suppression was partially reversed by a topical application of JH mimic. JH mimic treatments and allatectomy of diapausing larvae were also shown to cause a decrease in the titre of most of the proteins of the fat body including the diapause-associated protein.The diapause-associated protein has a molecular weight of about 41,000, and contains 15–17 different amino acid residues, of which aspartate, leucine, lysine, and glutamate predominate, comprising about 43, 37, 35 and 32 residues per molecule, respectively. We suggest that diapauselin is a suitable name for this protein. Our findings provide the necessary framework for undertaking a more detailed analysis of the hormonal regulation of diapauselin synthesis and utilization.     

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

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.     

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

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.     

The role of juvenile hormone in the larval diapause of the European corn borer,Ostrinia nubilalis

The possible role of juvenile hormone (JH) in the induction and termination of larval diapause in the European corn borer, Ostrinia nubilalis, was investigated using topical applications of both JH I and a JH mimic as well as by monitoring JH titers with the Galleria bioassay. Neither JH nor the JH mimic ZR515 was capable of influencing diapause termination when administered topically. The Galleria bioassay revealed little or no JH in the hemolymph of mid diapause (>30 days) insects, indicating no demonstrable role for JH in diapause maintenance. When ZR515 was administered to nondiapause, newly ecdysed fifth instar larvae the pupal molting cycle was delayed. By use of photoperiodic regimes we were able to show that the molting delay was not equivalent to diapause induction. The Galleria bioassay showed differences in JH titer profiles between diapause and nondiapause animals during the final larval stadium. The nondiapause insects showed titers that decline rapidly to trace amounts following the molt to fifth instar then rose prior to pupation. The diapause insects had generally higher titers and exhibited a more gradual decline after the molt. No evidence was obtained to support the hypothesis that JH plays a key role in the induction, maintenance, or termination of larval diapause.     

In vitro release of lipophorin from the fat body of nondiapause and diapause larvae of the Southwestern corn borer,Diatraea grandiosella

The release of lipophorin and total protein was examined from the fat body of nondiapause and diapause larvae of the southwestern corn borer, Diatraea grandiosella, incubated in vitro in Grace's medium. The characteristics of the released lipophorin were compared to those of the high-density lipophorin present in the hemolymph of nondiapause and diapause larvae. Over a 4 h incubation period, the fat body of nondiapause larvae released about 1.5 times more total protein and 2 times more lipophorin per mg dry weight than did that of diapause larvae. Lipophorin isolated from the medium in which fat bodies of nondiapause and diapause larvae had been incubated and from the plasma of nondiapause and diapause larvae had similar mean densities of 1.115, 1.112, 1.117 and 1.119 g/ml, respectively. Although the lipid classes detected in lipophorin isolated from the fat body incubation medium and hemolymph were identical, more polar lipids and less diacylglycerol were associated with lipophorin isolated from fat body incubation medium then were associated with lipophorin isolated from the hemolymph. Sterols accounted for about 11% of the total lipids of lipophorin isolated from the fat body incubation medium, whereas they accounted for about 20% of the total lipids of lipophorin from hemolymph. We conclude that the fat body of feeding nondiapause larvae and nonfeeding diapause larvae releases high-density lipophorin.     

Water involvement in diapause and the resumption of morphogenesis of the southwestern corn borer, Diatraea grandiosella

The effect of water on diapause and the resumption of morphogenesis of the southwestern corn borer was examined. Diapause larvae were held under dry conditions or with access to water in various environmental regimes known to permit different rates of diapause development. Results showed that diapause development occurred at a higher rate in larvae held dry than in those continuously supplied with contact water. Larvae did not, therefore, require an exogenous water source to complete diapause development. Although the availability of water promoted the post-diapause pupal moulting cycle of larvae which had completed diapause development, those which maintained a tissue water content > ca. 55% did not require contact water to complete this cycle. The loss of water in diapause larvae held under dry conditions or with access to water along with the quantity imbibed following various periods of dehydration was estimated. The physiological significance of the results in relation to other similar data is discussed.

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Zusammenfassung Es wurde der Einfluss von Wasser auf Diapause und Wiederaufnahme der Morphogenesis bei Diatraea grandiosella untersucht. Reife Diapauselarven wurden aus Laborzuchten erhalten oder im Freiland in Südost-Missouri gesammelt. Sie wurden trocken gehalten oder hatten Zugang zu Wasser und zwar in verschiedenen Umgebungsbedingungen, bei denen bekanntermassen die Diapauseentwicklung mit verschiedener Geschwindigkeit erfolgt. Die Ergebnisse zeigten, dass die Diapauseentwicklung rascher ablief bei Larven, die trocken gehalten wurden als bei solchen mit dauerndem Kontaktwasser. Die Larven brauchen demnach keine exogene Wasserquelle zur vollständigen Diapauseentwicklung. Zwar begünstigt die Verfügbarkeit von Wasser nach der Diapause das Einsetzen der Verpuppungsphase; dennoch brauchten diejenigen Larven, deren Gewebswassergehalt bei über ca. 55% lag, überhaupt kein Kontaktwasser, um diese Phase, d.h. die Verpuppung, abzuschliessen. Der Wasserverlust von Diapauselarven bei trockenen und nassen Bedingungen wurde bestimmt und ebenso die aufgenommene Wassermenge nach Trockenperioden. Die physiologische Bedeutung der Ergebnisse in Beziehung zu anderen, ähnlichen Daten wird diskutiert.

     

Brain neurosecretory cell and ecdysiotropin activity of the non-diapausing,pre-diapausing and diapausing southwestern corn borer, Diatraea grandiosella Dyar

The location and number of brain neurosecretory cells were studied in the larval southwestern corn borer. One posterior, two median and two lateral groups of paraldehyde-fuchsin positive cells were found in each cerebral hemisphere.Implantation of brain parts containing different groups of neurosecretory cells revealed that the median neurosecretory cells contained higher ecdysiotropic activity than the other cell groups. In vitro culture of ecdysial gland with brain or brain-parts extract showed also that the median neurosecretory cells contained much higher ecdysiotropic activity than other neurosecretory cells. To estimate the ecdysiotropic activity of pre-diapausing 6th instar larvae, their brain or brain extract was incubated in culture medium containing an ecdysial gland from a day-4 last-instar non-diapausing larva. Data showed that the ecdysiotropic activity in the pre-diapausing larvae was far lower than in non-diapausing and diapausing larvae.     

Significance of water contact as a factor terminating larval diapause in a stem borer, Busseola fusca

Previous studies revealed that diapause larvae of Busseola fusca (Lepidoptera: Noctuidae) began to respond to water and pupated in artificial wet conditions only in late diapause. Experiments are designed to clarify how water is involved in the termination of larval diapause. Neither feeding the larvae on fresh sorghum stems (6 weeks old) nor allowing them to drink water stimulated a break in the larval diapause. Diapause larvae transferred to artificial wet conditions pupated without increase in fresh weight. It is thus concluded that water contact is more significant as a factor terminating diapause in B. fusca than water uptake.

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Résumé Des études antérieures avaient montré que la diapause des chenilles de B. fusca (Lep., Noctuidae) commence par une réaction à l'eau et qu'elles ne se nymphosent dans des conditions artificiellement humides qu'en fin de diapause. Des expériences ont été conçues pour préciser l'intervention de l'eau dans la terminaison de la diapause. Ni l'alimentation des chenilles sur des tiges fraiches de jeune sorgho (de 6 semaines), ni la possibilité de boire de l'eau n'ont stimulé une rupture de la diapause larvaire. Des chenilles en diapause transférées dans des conditions artificiellement humides se sont nymphosées sans augmentation du poids frais. On en a conclu que le contact avec de l'eau était plus important que l'absorption d'eau comme facteur de fin de diapause chez B. fusca.

     

Effect of artificial wetting and rainfall on the larval diapause of a stem borer, Busseola fusca in Western Kenya

Clarification of factors enabling diapause termination was carried out in a stem borer, Busseola fusca. Diapausing larvae very rarely pupated under dry conditions. Water apparently played a significant role in the termination of late diapause. During the late diapause period, larvae in the field were exposed to rains of the short rainy season. This precipitation, however, terminated diapause in only a few larvae, which indicate that termination requires sustained exposure to wet conditions that are not usually met during the short rains.

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Résumé Les causes de la fin de la diapause ont été précisées sur B. fusca. Les chenilles diapausantes se nymphosent très rarement en conditions sèches. L'eau joue apparement un rôle important dans la fin de la dernière phase de la diapause. Dans la nature, les chenilles sont exposées pendant cette période aux plues de la brève saison de pluies. Cependant, ces précipitations n'interrompent la diapause que d'un nombre limité de chenilles, ce qui montre que la fin de la diapause exige une exposition prolongée à des conditions humides que l'on ne rencontre pas généralement pendant la brève saison des pluies.

     

Seasonal adaptations of populations of the southwestern corn borer, Diatraea grandiosella, from tropical and temperate regions

Seasonal adaptations of populations of the southwestern corn borer, Diatraea grandiosella, obtained from south-central Mexico (19°N latitude) and southeast Missouri (37°N latitude) were compared. Day length and temperature were found to serve as environmental cues to programme the larval diapause of both populations, but different critical values were observed. The critical day length for diapause induction was about 13 hr light/day for Mexican larvae and about 15 hr light/day for Missouri larvae, and was relatively stable at 20 to 30°C. Mexican larvae displayed a less-intense diapause than did Missouri larvae. Some diapausing Mexican larvae maintained at 25 or 30°C pupated in about 15 days, regardless of the day length to which they were exposed. The rate of diapause development of Mexican larvae was high at day lengths between 14 hr and 16 hr, whereas that of Missouri larvae was accelerated at day lengths of 16 hr at 25 and 30°C. Diapause development of Mexican larvae was virtually unaffected by chilling at 10°C, whereas that of Missouri larvae continued at a low rate at 10°C. Selection of Mexican larvae for diapause showed that only four generations were needed to significantly increase the incidence of diapause.     

Population consequences of diapause in a model system: the European corn borer

The diapause biology of the European corn borer (ECB), Ostrinia nubilalis (Hübn.), is described based on natural and controlled environment studies of feral and lab-reared ECB's in North Carolina (NC). The diapause response is described as a function of photophase (h of light/day) as well as a function of larval age (instar) at onset of diapause-inducing conditions. A critical photophase of 14.4 h and a critical mean larval instar of 3.3 is found in the lab studies and supported by three years of insectary studies. Seven years of black light trapping of ECB moths in Goldsboro, NC, revealed the likelihood of up to four moth flights/year.Information about the diapause biology of this insect is used to explain both the number of flights and the relative magnitude of the final moth flights. On average, the majority of ECB lineages pass through three generations/year with early maturing ECB's producing a significant and predictable fourth generation. The timing and magnitude of the fourth flight can be partly explained on the basis of the critical photophase and the timing and age structure of previous ECB generations. In most years, the fourth flight is smaller than the third due to the majority of the fourth generation's predisposition towards diapause. However, in at least one case (1977), the fourth flight was unusually large and could be predicted by slight temporal shifts in the previous three flights resulting in the majority of the fourth generation larvae averting diapause. The value of the ECB-diapause interaction as a model system for the explanation and prediction of dynamic phenological events is discussed.

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Résumé La diapause d'O. nubilalis Hubn. est décrite d'après des études en conditions naturelles et programmées de souches sauvages ou élevées au laboratoire en Caroline du Nord. La diapause est décrite comme une fonction de la photophase (heures de jour/24 heures) et du stade larvaire au début des conditions inductrices de la diapause. Une photophase critique de 14,4 h et un stade critique larvaire de 3,3 ont été établis au laboratoire après 3 ans d'études en insectarium. 7 ans de piégeage à la lumière noire à Goldsboro, ont montré la vraisemblance de l'existence de 4 vols par an. Les données sur la diapause de cet insecte sont utilisées pour expliquer tant le nombre de vols que l'importance relative des derniers vols. En moyenne, la majorité des lignées ont 3 générations par an, O. nubilalis précoces produisant une quatrième génération conséquente et prédictible. La data et l'importance du 4ème vol peuvent être partiellement expliquées d'après la photophase critique, et la data et la structure en âge des générations précédentes. La plupart des années, le 4ème vol est moins important que le 3ème par suite de la prédisposition à la diapause de la majorité de la 4ème génération. Cependant, dans un cas au moins, en 1977, le 4ème était anormalement important et pouvait être prédit par de faibles changements temporels dans les 3 précédents vols, détournant de la diapause la majorité des chenilles de 4ème génération. La discussion porte sur la valeur du modèle fourni par la diapause de O. nubilalis pour expliquer et prédire la dynamique des évéments phénologiques.

     

Lipids and fatty acids of the southwestern corn borer,Diatraea grandiosella

The lipids of the adults and of several immature stages of the southwestern corn borer, Diatraea grandiosella, were studied after they were fed natural corn stalks or artificial diets. Linoleic acid (18:2) was the major fatty acid of the neutral lipids in both the natural and the artificial diets, but aleic acid (18:1) was the principal neutral lipid in all insect stages. Also, linoleic acid and oleic acid were the principal acids in the insect phospholipids of all stages. The content of linoleic acid in the natural diet was also high, but that in the artificial diet appeared to be much too low for insect requirements. Phosphatidyl choline (PC) and phosphatidyl ethanolamine (PE) were the major phospholipids in all growth stages. Thus, in larvae diapausing in the field, the unsaturated fatty acid content of PC was 59·3 per cent, primarily 16:1 and 18:1, and PE was 87·4 per cent, primarily 18:1, 18:2, and 18:3, and the fatty acids in the number 1- and 2-positions of PC were 53·6 and 97·2 per cent unsaturated, respectively. The haemolymph of diapausing southwestern corn borer larvae contained primarily glycerides but also had some PC and PE. Fat body from diapausing larvae contained primarily 16:0, 16:1, and 18:1 in a ratio of 1 : 1 : 2. Thus lipids of the southwestern corn borer do not reflect dietary lipids as closely as do other insects studied.