Neuroendocrine roles of the brain in the regulation of 20-hydroxyecdysone responsiveness in two types of diapause pupae of the cabbage armyworm, Mamestra brassicae

The cabbage armyworm, Mamestra brassicae, has winter-and aestival- diapause pupae (WD- and AD-pupae) showing differences in the strength of diapause. We tried to quantify diapause-strength by measuring the doses of 20-hydroxyecdysone (20-E) required to induce adult development in WD-, AD- and decerebrated non-diapause pupae (ND-pupae). The role of the brain in the regulation of diapause-strength was studied through the decerebration and brain-reimplantation of WD-and AD-pupae. The 20-E doses required for adult development were small within the first 2 days of pupation, and increased thereafter to reach a constant level about 10 days after pupation in AD- and decerebrated ND-pupae. The required 20-E doses in WD-pupae increased for more than 40 days after pupation. When 0-day-old WD- and 0-day-old AD-pupae were decerebrated, required 20-E doses increased after pupation and reached a constant about 10 days later. The required 20-E dose reached a constant level in decerebrated WD-pupae that was smaller than that observed for decerebrated ND- and WD-pupae. Furthermore, the required doses increased when 0-day-old WD-pupal brains were reimplanted into decerebrated WD- and decerebrated ND-pupae. In WD-, AD- and decerebrated ND-pupae, diapause-strength can be represented as the 20-E dose required for adult development. Diapause-strength is weak after pupation, increases thereafter, and reaches a constant about 10 days later in AD- and decerebrated ND-pupae. In WD-pupae, diapause-strength increases for more than 40 days after pupation and reaches a level that is twice that estimated for AD-pupae. Brains of diapausing WD-pupae may secrete a factor that suppresses the 20-E responsiveness of pupal organs, for the purpose of maintaining winter-diapause.     

Hourglass and oscillator expressions of photoperiodic diapause response in the cabbage moth Mamestra brassicae

Abstract. Both oscillator and hourglass features are found in the photoperiodic response that controls the pupal winter diapause of Mamestra brassicae. The expression of oscillatory response to extended long-night cycles is temperature dependent, i.e. circadian resonance appears at 23 and 25^oC but not at 20 and 28^oC. At 20^oC, scanning of extended scotophases by a short light pulse does not reveal any clear circadian rhythmicity. However, a circadian feature of the photoperiodic response is indicated even at 20^oC by a bistability phenomenon, i.e. either one of the two dark periods in symmetrical skeleton photoperiods determines the diapause response depending on the phase angle with the preceding (entraining) light-dark cycles. At 20 and 25^oC, the incidence of diapause increases as a function of the number of light–dark cycles regardless of the cycle length (T) , if T is 24 h or 2 X 24h (with a 12 h light period). A non-diel cycle (r=36h) is less effective, suggesting that disturbance of the circadian organization partly impairs the diapause-inducing function. The inductive effect of a long night is largely affected by temperature, and becomes saturated with eight cycles at 20^oC and 14 cycles at 25^oC. Presumably, an hourglass mechanism measures the dark time, and a circadian component involved in some later sequence of the photoperiodic response may or may not be expressed depending on the mode of interaction between them.     

Sex pheromone components of the cabbage armyworm, Mamestra brassicae: Isolation, identification and field experiments

Glass capillary gaschromatography coupled to a mass spectrometer has shown that in addition to Z-11-hexadecenyl acetate, hexadecyl acetate and Z-11-heptadecenyl acetate are present in pheromone glands of calling virgin females of Mamestra brassicae L. (Lepidoptera: Noctuidae, Hadeninae). Electroantennography revealed positive responses for Z 11–17: Ac and 16: Ac, while behavioural tests were only positive for Z 11–17: Ac.

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Résumé Il a été procédé à l'identification des phéromones par chromatographie en phase gazeuse et par spectrographie de masse. Outre l'acétoxy-1 hexadécène-11Z, on identifié l'acétoxy-heptadécène-11Z et l'acétoxy-1-hexadécane dans les glandes des femelles vierges de M. brassicae. L'antennographie a montré des réactions positives à l'acétoxy-hepta-décène-11Z et à l'acétoxy-1-hexadécane, tandis que seul l'acétoxyhepta-décène-11Z provoque un comportement sexuel des mâles.

     

Control of ommochrome synthesis by both juvenile hormone and melanization hormone in the cabbage armyworm,Mamestra brassicae

Summary Pigmentation of last instar larvae of the cabbage armyworm,Mamestra brassicae is of two types: melanin in the cuticle and ommochrome in the epidermis. The latter was found to be primarily xanthommatin. When allatectomy was performed 8 h before head capsule slippage (HCS) in the last larval molt, later ommochrome synthesis was inhibited. Application of juvenile hormone (JH) up to 12 h after HCS (9 h before ecdysis) (activity: methopreneJH I>JH II>JH III) restored ommochrome synthesis. After that time it has less and less effect.Removal of the suboesophageal ganglion from the larvae 8 h before HCS prevented both later ommochrome synthesis and melanization. Melanization of isolated abdomens was restored by implantation of 3 suboesophageal ganglia or injection of melanization and reddish coloration hormone (MRCH) 18 h after HCS. Restoration of ommochrome synthesis required exogenous JH in addition to melanization hormone from suboesophageal ganglion or MRCH. Therefore, melanization appears to be critical for the later onset of ommochrome synthesis even in a larva which has been exposed to JH during the critical period.Abbreviations CC·CA corpora cardiaca-corpora allata complex - JH juvenile hormone - MRCH melanization and reddish coloration hormone - HCS head capsule slippage     

Mortality of eggs, larvae and pupae and larval dispersal of the cabbage moth, Mamestra brassicae, in white cabbage in south-eastern Norway

The mortality of eggs, larvae and pupae and larval dispersal of the cabbage moth, Mamestra brassicae (L.) (Lepidoptera: Noctuidae) was investigated in a series of small-scale field experiments in white cabbage, Brassicae oleracea var. capitata (L.), and in the laboratory during 1990–1992 in south–eastern Norway. The highest mortality was found in young larvae and in hibernating pupae. In 1990, larval mortality in the first instar was 80% (range 9–97% for the individual cohorts). Most larvae died within the first 1–3 days after hatching. The dispersal activity during these days was high, and failure to establish feeding sites and predation were probably the main mortality causes. Pupal mortality during winter was 90% on average for 1990–1993 (range 81–100%). The main mortality factor was probably unfavourable weather conditions, and indications of cold stress were found. The impact from parasitoids and diseases was generally low. Trichogramma semblidis (Aurivillius) (Trichogrammatidae) was reared from M. brassicae eggs in very low numbers in 1991. Larval parasitism increased from < 1% in 1990 to almost 24% in 1992, and was almost totally caused by the braconids Microplitis mediator (Haliday) and Aleiodes (Aleiodes) sp. Predation of frozen larvae on the soil surface was 75% on average (range 63–96%) during 1990–1992 in first instar larvae and decreased gradually with larval age. The consumption rates of Philonthus atratus (Gravenhorst) (Staphylinidae) and the carabids Bembidion tetracolum (Say), Pterostichus melanarius (Illiger) and Harpalus rufipes (Degeer) on M. brassicae eggs and larvae were investigated in non-choice experiments in the laboratory. A preliminary survival model based on estimates of the mortality factors identified in this study is presented.     

Aestival dormancy in the cabbage moth Mamestra brassicae L. (Lepidoptera: Noctuidae)

Summary In a geographically wide distribution the life cycles of different populations of the cabbage moth Mamestra brossicae are adapted to a remarkable diversity of climatic conditions. This is undoubtedly a proof of its success in adaptation. Some populations living in regions characterized by a drought period interrupting the growth season are capable of distinguishing between one critical day length signalling the onset of the drought period and another signalling the end of the growth season. This study, therefore, is primarily concerned with the geographical patterns in the variability of the adaptional responses of populations exposed to environmental conditions requiring different strategies and tactics in, synchronizing individual, life cycles. It is also a contribution to our understanding of evolutionary mechanisms maintaining median responses to photoperiodically inductive day lengths in geographically different populations. The populations investigated originated from regions differing in predictability of the incidence, onset and duration of a drought period: Freiburg (48.0°N, Southern Germany), Avignon (44.0°N, Southern France), and Argelès (42.5°N, Southern France). Geographical variation with respect to both onset and duration of a drought period consequently results in clinal variation of the variability of innate day length thresholds triggering aestival dormancy and of innate duration of aestivation. In this paper we considered the influence of geographically changing temperatures on aestival dormancy induction. Even in southern populations of M. brassicae a temperature dependent switch off-mechanism exists which prevents aestival dormancy under certain environmental conditions. The effective temperatures vary geographically, too. What the geographical patterns in adaptive responses really are, is discussed.This research was supported by the Deutsche Forschungsgemeinschaft (Sa 259/3-1)     

Aestival dormancy in the cabbage moth Mamestra brassicae L. (Lepidoptera: Noctuidae)

Summary A highly specific recognition system, capable of foreseeing and distinguishing between two critical points in time, exists in Mamestra brassicae (Lepidoptera: Noctuidae). Both points in time, the onset of a drought period and the end of the growth season, require different growth patterns of the pupae. In order to minimize the likelihood of weather-induced mortality and to maximize fitness, individuals of M. brassicae must enter aestival dormancy or hibernal diapause, respectively, before the onset of drought or frost. This study is primarily concerned with aestival dormancy. Normally, the pupal period of dormancy-free developing individuals amounts to approximately 20 to 30 days. A modified pupal period of approximately 35 to 80 days is defined as aestival dormancy. The onset of aestival dormancy is triggered by day lengths exceeding an innate individaul-specific threshold. The results reported in this paper indicate that the photoperiodic response curve represents largely the genetic variability within a population with respect to the thresholds triggering aestival dormancy. This variability in thresholds is considered to reflect the frequency of correlation of a distinct day length with a certain onset of drought period in the past. Furthermore, the innate thresholds are characterized by a temperature dependent norm of reaction. Our results also indicate, that a strong genetical component is involved in variability of duration of the pupal period. This variability in duration of aestivation reflects the frequency of drought periods of a certain length in the past. The adaptive significance of both the variation in day length thresholds and duration of aestival dormancy is discussed with respect to the number of generations per season, and the synchronization of the individual life cycles with the seasonal changing environmental conditions.This study is dedicated to Prof. H.J. Müller, Jena, for his 75th anniversaryThis research was supported by the Deutsche Forschungsgemeinschaft (Sa 259/3-1)     

Plant volatiles activating specific olfactory receptor neurons of the cabbage moth Mamestra brassicae L. (Lepidoptera, Noctuidae)

Herbivore insects are suitable model organisms for studying how plant odor information is encoded in olfactory receptor neurons (RNs). By the use of gas chromatography linked to electrophysiological recordings from single RNs, screening for sensitivity to naturally produced plant odorants is possible in order to determine the molecular receptive ranges of the neurons. Using this method, we have in this study of the cabbage moth, Mamestra brassicae, classified 21 types of olfactory RNs according to their responses to odorants present in the host plants of Brassicae, in the related species of Arabidopsis, as well as in essential oils of nonhost plants like ylang-ylang. Most of the RNs were tuned to one or a few structurally similar compounds, showing minimal overlap of their molecular receptive ranges. Whereas some RNs displayed a novel tuning, others were tuned to the same compounds as neurons in other insect species. We also found colocation in the same sensillum of 3 RN types with the same response characteristics and tuning as 3 colocated types described in heliothine moths living on different host plants. The presence of similar RN types across different insect species implies conservation or reappearance of the RN types, independent of the evolution of host plant ranges.     

Ecdysteroid levels during post-diapause development and 20-hydroxyecdysone-induced development in male pupae of Mamestra configurata Wlk.

Post-diapause development in male pupae of Mamestra configurata Wlk. was characterized by the appearance of large, transitory peaks of ecdysone (2.8 μg/g live wt) at day 8 and 20-hydroxyecdysone (2.2 μg/g) at day 12 which declined to low levels prior to adult eclosion at day 28.Treatment of diapausing pupae with 20-hydroxyecdysone elicited a progression of dose-dependent physiological and pathological effects, including termination of diapause, development, accelerated development, and accelerated development leading to malformation and death. At a dose of 7.5 μg 20-hydroxyecdysone/g, all treated pupae terminated diapause, developed with little mortality and produced a high proportion of morphologically perfect adults. However, there were no large peaks of ecdysone or 20-hydroxyecdysone in treated pupae, possibly due to feedback inhibition by 20-hydroxyecdysone.At doses greater than 7.5 μg/g, development was accelerated markedly, survival decreased precipitously (0% at 15 μg/g) and the proportion of malformed adults increased sharply. Pupae that received a lethal dose of 20-hydroxyecdysone died almost synchronously after undergoing accelerated development for 18–20 days, indicating that they encounter a common, hormone-induced developmental block. Pupae receiving 15 μg/g also showed no edcysone or 20-hydroxyecdysone peaks, but had a prolonged period of hyperecdysonism which likely caused their accelerated development and death.     

Molecular cloning and functional expression of chitinase-encoding cDNA from the cabbage moth, Mamestra brassicae

Chitinase is a rate-limiting and endo-splitting enzyme involved in the bio-degradation of chitin, an important component of the cuticular exoskeleton and peritrophic matrix in insects. We isolated a cDNA-encoding chitinase from the last larval integument of the cabbage moth, Mamestra brassicae (Lepidoptera; Noctuidae), cloned the ORF cDNA into E. coli to confirm its functionality, and analyzed the deduced amino acid sequence in comparison with previously described lepidopteran chitinases. M. brassicae chitinase expressed in the transformed E. coli cells with the chitinase-encoding cDNA enhanced cell proliferation to about 1.6 times of the untransformed wild type strain in a colloidal chitin-including medium with only a very limited amount of other nutrients. Compared with the wild type strain, the intracellular levels of chitin degradation derivatives, glucosamine and N-acetylglucosamine were about 7.2 and 2.3 times higher, respectively, while the extracellular chitinase activity was about 2.2 times higher in the transformed strain. The ORF of M. brassicae chitinaseencoding cDNA consisted of 1686 nucleotides (562 amino acid residues) except for the stop codon, and its deduced amino acid composition revealed a calculated molecular weight of 62.7 and theoretical pI of 5.3. The ORF was composed of N-terminal leading signal peptide (AA 1-20), catalytic domain (AA 21-392), linker region (AA 393-498), and C-terminal chitin-binding domain (AA 499-562) showing its characteristic structure as a molting fluid chitinase. In phylogenetic analysis, the enzymes from 6 noctuid species were grouped together, separately from a group of 3 bombycid and 1 tortricid enzymes, corresponding to their taxonomic relationships at both the family and genus levels.     

First evidence of pupal summer diapause in Pieris brassicae L.: the evolution of local adaptedness

Pieris brassicae L. is adapted to a wide variety of local climates throughout the Palaeartic by means of geographic variation in daylengths and temperatures triggering winter diapause. We report for the first time a population from Sotogrande in southern Spain (strain S) that additionally exhibits a pronounced summer diapause in the pupal stage as a response to long daylengths. As well as having diapause triggered by long daylengths, this population develops directly at short, ecologically relevant daylengths. This unusual short-day type response is expressed at temperatures of 21 degrees C and above. The absence of cryptic summer diapause in a population from southern France indicates that this summer diapause evolved locally in Spain or Northern Africa in a population of P. brassicae which lacked this trait. The PPRs of strains S, H (Denmark) and B (France) and crosses between them suggest that summer diapause is inherited as a recessive allele which has a clinal decrease of frequency in populations northwards. The population from southern France shows that this simple pattern of inheritance is further modified by an unknown genetic mechanism controlling the expression of summer diapause near the border between aestivating and non-aestivating populations. On the basis of the photoperiodic response and climatic data we predict P. brassicae to have five or more generations each year in southern Spain.     

Interaction of cyclic nucleotides and ecdysterone in breaking the pupal diapause of the bertha armyworm, Mamestra configurata WLK.

Cyclic GMP and cyclic AMP have distinct and opposite effects upon the action of ecdysterone in diapausing pupae of the Bertha armyworm, $\text{Mamestra}$$\text{configurata}$. Cyclic GMP enhanced the effectiveness of suboptimal doses of ecdysterone in breaking diapause; the amount of cyclic GMP required to lower the ED50 of ecdysterone by half was 80 μg/g. Dibutyryl cyclic GMP had no apparent effect on the action of ecdysterone over a wide dose range (0.07 – 70 μg/g). On the other hand, cyclic AMP and dibutyryl cyclic AMP effectively blocked the diapause-breaking action of ecdysterone when administered simultaneously with the steroid hormone. The amount of cyclic AMP required to reduce the incidence of diapause termination from 100% to 50% was 60 μg/g; for dibutyryl cyclic AMP the amount required was only 14 μg/g. No cyclic nucleotide tested in the study could by itself break the pupal diapause of $\text{M.}$$\text{configurata}$. The concept that cyclic GMP and cyclic AMP provide at least different if not opposing regulatory influences in certain insect systems is discussed briefly in the light of these observations.     

Discrimination between enantiomers of linalool by olfactory receptor neurons in the cabbage moth Mamestra brassicae (L.)

Plants emit complex blends of volatiles, including chiral compounds that might be detected by vertebrates and invertebrates. Insects are ideal model organisms for studying the underlying receptor neuron mechanisms involved in olfactory discrimination of enantiomers. In the present study, we have employed two-column gas chromatography linked to recordings from single olfactory receptor neurons of Mamestra brassicae, in which separation of volatiles in a polar and a chiral column was performed. We here present the response properties of olfactory receptor neurons tuned to linalool. The narrow tuning of these receptor neurons was demonstrated by their strong responses to (R)-(-)-linalool, the weaker responses to the (+)-enantiomer as well as a few structurally related compounds, and no responses to the other numerous plant released volatiles. The enantioselectivity was verified by parallel dose-response curves, that of (R)-(-)-linalool shifted 1 log unit to the left of the (S)-(+)-linalool curve. A complete overlap of the temporal response pattern was found when comparing the responses of the same strength. Analysis of the spike amplitude and waveform indicated that the responses to the two enantiomers originated from the same neuron.     

Identification of a new hobo element in the cabbage moth, Mamestra brassicae (Lepidoptera)

A complete hobo-like element, called Mbhobo, was identified in the cabbage moth, Mamestra brassicae. This element has a high sequence similarity to the HFL1 hobo element of Drosophila melanogaster. Amplification of Mbhobo termini indicated that transposition occurred into a 5'-GTGGGTAC-3' target sequence that was duplicated upon insertion. This target site conforms to the consensus sequence established for the insertion sites of insect hAT elements. Mbhobo has a single 1935 bp long ORF with significant homology to the D. melanogaster HFL1 hobo transposase. FISH experiments evidenced Mbhobo clusters located in heterochromatic regions of Z and W sex chromosomes and in heterochromatic areas of chromosome pair 10.     

Dynamics of energy metabolism in ontogenesis of striped shield bug (Graphosoma lineatum L.) and cabbage moth (Mamestra brassicae L.)

Dynamics of growth and oxygen consumption during ontogenesis of insects with direct (striped shield bug Graphosoma lineatum L.) and indirect (cabbage moth Mamestra brassicae L.) development have been compared. The correlation between a character of energy metabolism alteration and peculiarities of development of the insects has been shown. Cyclic decrease of oxygen consumption during molt and sharp dropping during metamorphosis have been observed in insects with indirect development. The decrease of oxygen consumption has been observed in insects with direct development only during molts. The coefficient a of allometric dependence of oxygen consumption on body weight of imago for cabbage moth was two times higher than that for striped shield bug.     

Inheritance of the photoperiodic response controlling imaginal summer diapause in the cabbage beetle, Colaphellus bowringi

Adults of the cabbage beetle Colaphellus bowringi display a summer diapause in response to the exposure of their larvae to long photoperiods. In the present study, the inheritance of the photoperiodic response controlling summer diapause in C. bowringi by crossing a high diapause strain (D strain) with a laboratory selected nondiapause strain (N strain) was investigated under different photoperiods at 22, 25 and 28 °C. The beetles in both reciprocal crosses and backcrosses showed a clear short-day response for the induction of diapause at all temperatures, similar to that of the D strain, suggesting that photoperiodic response of this beetle is heritable. The diapause incidences in the progeny from all the crosses under LD 15:9 or LD 12:12 at 25 °C suggest that genetic and genetic-environmental interactions are involved in diapause induction. The incidence of diapauses in F₁ progeny was significantly lower than that in the D × D strain but significantly higher than that in the N × N strain, indicating that the diapause capability is inherited in an incomplete dominant manner. The incidence of diapause was greater among the offspring of hybrid females when those females had a D strain mother or grandmother rather than a N strain mother or grandmother, indicating that maternal effects on diapause induction are stronger than paternal effects. The laboratory selected nondiapause strain also showed a short-day photoperiodic response at a low temperature of 22 °C, indicating that the photoperiodic photoreceptor and photoperiodic clock still function in the nondiapause strain.