Diapause detection and monitoring in the Mediterranean corn stalk borer

Abstract. Sesamia nonagrioides Lefebvre diapauses as mature larvae. Once diapause is induced, larvae continue to grow and moult with no apparent changes, so distinction between non-diapausing and diapausing larvae is difficult. In the present work two physiological markers of diapause induction are obtained and their efficacy in detecting diapause onset and in monitoring diapause development in field populations is evaluated. The first marker is based on the differential capacity of two photoregimes, LD 0:24 h and LD 16:8 h, to reverse diapause induction. When larvae of the first and second stadia were subjected to the diapause-inducing LD 12:12 h light cycle and then transferred to LD 0:24 h, larvae pupated after a development duration significantly higher than that of the larvae transferred to LD 168 h. This criterion was used to monitor diapause development in overwintering larvae until field-collected larvae submitted to LD 0:24 h pupated synchronously with those submitted to LD 16:8 h. This occurred between late January and early March. The second marker refers to development of imaginal wing discs. A relative index of the tracheal mass area divided by the total disc area diminishes steadily during development in non-diapausing sixth-instar larvae until it becomes zero just before pupation, whereas it remains constant in diapausing larvae. Though wing disc development was blocked in diapausing sixth-instar larvae, the disc continued to grow and, consequently, the total area of the disc was not a suitable marker of diapause induction. When the constancy of the relative area index was used to monitor diapause development and termination in overwintering larvae, the results agreed with those obtained using the first criterion and with previous field observations.     

Characterization and distribution of digestive proteases of the stalk corn borer,Sesamia nonagrioides Lef. (Lepidoptera: Noctuidae)

Larval midgut extracts from the noctuid Sesamia nonagrioides Lef. were assayed for protease activity. Total proteolytic activity, as measured by azocasein hydrolysis, showed a pH optimum in the range 10.0 to 11.5, suggesting a digestive system based largely on serine-like proteases. The ability of midgut extracts to hydrolyze specific synthetic substrates, the elucidation of the pH at which maximal hydrolysis occurs, and their sensitivity to protease inhibitors confirmed the presence of the serine endoproteases: trypsin, chymotrypsin, and elastase; and the exopeptidases: carboxypeptidase A, carboxypeptidase B, and leucine aminopeptidase. The distribution of these digestive proteases along the gut sections and among the different midgut regions was examined. All types of endoproteases and exopeptidases were mainly located in the midgut, with less than 5% of the activity in the foregut and hindgut. When the two halves of the midgut were compared, all proteolytic activities were higher in the anterior portion of the midgut. Trypsin, chymotrypsin, elastase, and carboxypeptidase B activities were mainly located in the endoperitrophic space of the midgut, with some activity in the ectoperitrophic space, whereas aminopeptidase and carboxypeptidase A activities were preferentially located in the midgut epithelium. © 1996 Wiley-Liss, Inc.     

Factors affecting the natural duration of diapause and post-diapause development in the Mediterranean corn borer Sesamia nonagrioides (Lepidoptera: Noctuidae)

Sesamia nonagrioides responds to photoperiod for diapause termination at high temperature with a Type II response curve, and only photoperiods longer than 12:12 terminate diapause. However, these photoperiods never occur in the field when diapausing larvae are competent to terminate diapause. Under a temperature similar to the natural field temperatures diapause terminates spontaneously in approximately 4 months, which ensures that the larvae reach the middle of winter without pupation. S. nonagrioides larvae pupate after going through a specific number of light-dark cycles or days: the required day number (RDN) for diapause completion. This RDN could be modulated by temperature but more research is necessary to clarify this point. In post-diapause development, when a suitable temperature threshold is considered, the absolute accumulation of heat is more important than whether the temperature received is fluctuating or constant. The temperature threshold for diapause and post-diapause development was lower than the temperature threshold of larvae in continuous development. This is important for adjusting phenological models in S. nonagrioides and in other species, and may explain why in many cases adults appear in the field when the supposed temperature threshold for development has not been attained.     

Dispersal capacity in the Mediterranean corn borer, Sesamia nonagrioides

Corn (Zea mays L.) borers are the primary target of Bacillus thuringiensis Berliner (Bt) transgenic maize. Management of corn borer resistance to Bt requires information on larval and adult dispersal capacities, a feature that is particularly unknown in Sesamia nonagrioides Lefèbvre (Lepidoptera: Noctuidae), the most damaging corn borer in Spain. Larval dispersal was studied over a 3 year period by infesting plants with egg masses and dissecting the neighbouring plants 7, 14, and 32 days later to measure larval dispersal at several ages. The number and age of larvae were recorded in the dissected plants. Only mature larvae dispersed in significant numbers; they moved at least to rows adjacent to those containing the infested plant, and down the row five plants. The percentage of larvae that dispersed from the infested plant was density‐dependent. Adult dispersal was studied with directional light and pheromone uni‐traps over 5 and 3 year periods, respectively. Directional light traps were placed in the margins between Bt and non‐Bt maize fields, half oriented towards each of the two kinds of maize field. Pheromone traps were placed in the Bt and non‐Bt fields at increasing distances (0–100 m) from the border. The numbers of males and females caught in directional light traps were not different in traps oriented towards Bt or non‐Bt fields, but the number of males caught in the third flight in Bt fields was lower than in non‐Bt fields. These results suggest that males from adjacent Bt and non‐Bt fields mate indiscriminately with females emerging in any of the two kinds of maize fields. However, male movement in the third flight may not be sufficient to randomly distribute males between the two fields.     

Genetics of resistance to the pink stem borer (Sesamia nonagrioides) in maize (Zea mays)

Our knowledge of the genetics of resistance to the pink stem borer ( Sesamia nonagrioides ) in maize ( Zea mays ) is restricted to a few crosses among maize inbreds. The objectives of this study were to enlarge our understanding of the genetics of traits related to damage by pink stem borer and yield under infestation and to use generation means analyses to compare per se and testcross performance for detecting epistatic effects. All generations, either per se or crossed to testers, were evaluated in a 10 × 10 triple lattice design under artificial infestation with S. nonagrioides in 2005 and 2006. Most traits fit an additive–dominance model; but evidence for epistasis for resistance and yield under infestation was shown. Epistasis, in general, did not appear to play an important role in the inheritance of yield under pink stem borer infestation. However, the epistasis contribution to maize yield performance could be important in some outstanding crosses such as EP42 × A637. Testcross generation means revealed epistatic effects undetected by the generation means analysis, but neither method was able to eliminate dominance effects that could prevail over epistatic effects.     

Juvenile hormone and diapause in the Mediterranean corn borer,Sesamia nonagrioides

Juvenile hormone content of Sesamia nonagrioides larvae reared under different environmental conditions that induce diapause was studied using a bioassay on newly-emerged Tribolium confusum pupae. Two analytical methods were also used to measure JHIII content. Extracts from larvae that developed under different conditions of photoperiod and temperature caused different effects in T. confusum pupae. This can be related to different diapause intensities. Extracts from diapausing larvae produced a higher juvenilizing effect than the expected, considering their JHIII titer calculated by the chromatographic analysis. This indicates that hormones other than JHIII must be present in extracts of diapausing larvae of S. nonagrioides. The analytical procedures confirm that diapausing larvae have a higher JHIII titer in the haemolymph than non-diapausing larvae. This shows that JH is involved in the maintenance of diapause in this species.     

Photoperiodic and temperature effects on the intensity of larval diapause in Sesamia nonagrioides

Abstract. The intensity of larval diapause in Sesamia nonagrioides Lef (Lepidoptera: Noctuidae) was investigated under laboratory conditions. Newly hatched larvae were exposed to different stationary photoperiods (from LD 7 : 17 h to LD 14 : 10 h), at a constant temperature of 25 °C. Diapause incidence was higher when larvae were exposed to daylengths shorter than the critical value (LD 12 : 12 h), whereas the within‐treatment variation in the larval period appeared to be significantly correlated with the photoperiod applied. The incidences of diapause and the duration of larval development were also measured after exposing larvae to short photoperiods (LD 8 : 16 h, LD 10 : 14 h or LD 12 : 12 h) in combination with various temperatures (20, 22.5 or 25 °C). Although an increase in the incidence of diapause appeared with the lowering of the temperature, no statistical differences were observed in the time needed for pupation within the photoperiodic treatments at the temperatures of 20 and 22.5 °C. Furthermore, when diapausing larvae were transferred to the long photoperiod of LD 16 : 8 h, they immediately proceeded to pupation, regardless of the photoperiod or the temperature to which they had been previously exposed, indicating that there were no differences in the intensity of diapause. Photoperiodic changes from LD 10 : 14 h to LD 12 : 12 h or to LD 14 : 10 h at different larval ages reduced the intensity of diapause with (a) early age of transfer and (b) increase of daylength. By contrast, when larvae were transferred from the long photoperiod of LD 14 : 10 h to shorter, such as LD 10 : 14 h or LD 12 : 12 h, a small increase in the intensity of diapause with the shortening of the daylength was apparent. These results support the hypothesis that insects may compare the duration of the photoperiod and could classify them as either longer or shorter in relation to the critical value.     

Effects of selection for resistance to Sesamia nonagrioides on maize yield,performance and stability under infestation with Sesamia nonagrioides and Ostrinia nubilalis in Spain

A maize synthetic population was improved for resistance to the Mediterranean corn borer (MCB, Sesamia nonagrioides) while maintaining yield. The objectives of this research were to investigate whether yield and yield stability of the maize synthetic population named EPS12 were affected by selection for MCB resistance; also to determine which genotypic and environmental covariates could explain the genotype (G), environment (E) and genotype × environment (GE) effects for yield under corn borer infestation. Plants from three cycles of selection and their testcrosses to three inbred testers (A639, B93 and EP42) were evaluated at two locations in 2 years, under MCB and European corn borer infestations. After selection EPS12 was a more stable genotype. Hybrids derived from crosses between B93 and inbreds obtained from the initial cycles of selection could be recommended for cultivation in northern Spain. The yield of crosses between cycles of selection and testers increased when there were fewer days with temperatures >25°C and higher mean maximum temperatures. Differences in yield among these genotypes were mostly explained by resistance to corn borer attack. In general, among EPS12-derived materials, genetic characteristics that contribute to increased grain yield were also responsible for increased abiotic stress tolerance.     

Electroantennogram, wind tunnel and field responses of male Mediterranean corn borer, Sesamia nonagrioides, to several blends of its sex pheromone components

The electrophysiological and behavioural responses of male Sesamia nonagrioides Lefèbvre (Lepidoptera: Noctuidae) to several synthetic blends of its sex pheromone components were investigated. Electroantennogram (EAG) recordings showed the greatest response to a 77:8:10:5 blend of the four compounds found in the female glands: (Z)-11-hexadecen-1-yl acetate, (Z)-11-hexadecen-1-ol, (Z)-11-hexadecenal and dodecyl acetate. A logistic equation was fitted to the dose-response data and differences in relative EAG responses were analysed by deriving five parameters of this equation. Flight responses of males were also evaluated in a wind tunnel where the proposed blend (77:8:10:5) also elicited the best response. Field tests confirmed the laboratory findings as the 77:8:10:5 blend captured more S. nonagrioides males than other pheromone mixtures and showed more selectivity for male S. nonagrioides over other noctuid species, Discestra trifolii Rottenberg and Lacanobia oleracea L., but not for Mythimna unipuncta Haworth, a species with a sex pheromone composition almost identical to that of S. nonagrioides.     

Cold hardiness and overwintering strategy of the pink maize stalk borer,Sesamia nonagrioides Lef (lepidoptera,noctuidae)

The cold-hardening capacity of larvae of the pink maize stalk borer,Sesamia nonagrioides Lef., was examined. Supercooling points (SCPs) of field collected diapausing larvae from south-east and south-west France and non-diapausing and diapausing laboratory-reared larvae did not differ and ranged between –5 and –8°C. Thus, this insect possesses sufficient supercooling ability to avoid freezing over its normal environmental temperature ranges. Despite this, we found thatSesamia presents paradoxical cold reactions. Mortality of cold acclimated diapausing larvae after short-term exposure to temperatures above the SCP is high, supporting the view thatSesamia is cold-sensitive. On the other hand,Sesamia could survive freezing for at least 24 h to temperatures close to the SCP. This ability does not seem to be related to haemolymph trehalose, the sole cold-accumulated compound detected by gas chromatography and mass spectrometry. Despite the mildness of the winter 1990–1991, only 5% of the field population survive and pupate in April. The main part of the population died from November to January, the period during which larvae were mainly located in the part of the corn stem above the ground and experienced air temperatures. After January, all surviving larvae were excuusively located in the root, 10 cm below the soil, where they experienced milder temperatures than air. They exhibited a constant low rate of mortality, possibly independent of the cold. In their current distribution area, survival of overwintering larvae ofS. nonagrioides is only related to the microclimate of the overwintering site and freezing tolerance capacity seems to be irrelevant. This study allows us to propose a non-pollutant pest control method based on the behavioral strategy of this insect.     

Functional response of Telenomus busseolae (Hym.: Scelionidae) an egg parasitoid of the sugarcane stem borer,Sesamia nonagrioides (Lep.: Noctuidae) at different temperatures

Telenomus busseolae Gahan is the most important egg parasitoid of sugarcane stem borers in Iran. Laboratory experiments were conducted to evaluate the functional response of T. busseolae to egg densities of Sesamia nonagrioides (Lefebvre) under four different temperatures (20, 25, 30 and 35°C). Results of logistic regression revealed a type III functional response for all temperatures tested and type of functional response was not affected by temperature. Roger's random parasite equation was fitted to observed data at 35°C. In the other data sets (collected data under 20, 25 and 30 °C) Roger's equation could not provide meaningful parameters, therefore Holling's disk equation was used to fit the data. The attack rate for T. busseolae varying from 0.001784 to 0.10704, 0.0026 to 0.152, 0.0024 to 0.144 and 0.000866 to 0.05196 per h and estimated handling times were 0.59, 0.50, 0.53 and 0.29 h at 20, 25, 30 and 35°C, respectively. Based on asymptotic 95% confidence intervals, functional response parameters did not differ significantly from 20 to 30°C, which indicates that handling time and attack rate are least affected by the temperature changes. The results suggest a similar performance of T. busseolae against S. nonagrioides eggs at temperatures of 20–30°C. These results show that T. busseolae is well adapted to temperature changes during the sugarcane growing season.     

Molecular characterization of an ecdysteroid inducible carboxylesterase with GQSCG motif in the corn borer, Sesamia nonagrioides

We obtained a full-length cDNA encoding a carboxylesterase in Sesamia nonagrioides. The complete cDNA sequence is comprised of 1838 bp with an open reading frame encoding 576 amino acid residues with predicted molecular mass of 64.24 kDa. The deduced amino acid sequence showed high identity to JHE-Related of Trichoplusia ni (65% amino acid identity) and 49-46% amino acid identity to JHEs of other lepidopterans and contained all five functional motifs of insect JHEs. The gene has been termed as SnJHE-Related (SnJHER) to denote its similarity to other insect JHE genes and the occurrence of an unusual cysteine residue immediately adjacent to the catalytic serine, instead of the conventional alanine residue. Phylogenetic analyses localised SnJHER together with TnJHER in a branch of the lepidopteran's JHEs group, with other carboxylesterases (COEs) occuring in separated groups. The JH analog methoprene did not affect the expression of SnJHER in contrast to other insect JHEs. Additionally, ecdysteroid analogs induced SnJHER gene expression. The SnJHER mRNA levels were higher in long-day non-diapausing larvae than in short-day diapausing ones. In the fifth instar of non-diapausing and ninth instar of diapausing larvae, the SnJHER mRNAs reached higher expression levels on the days close to each larval molt. In the last (sixth) non-diapausing larval instar, SnJHER mRNA levels peaked in the intermolt period but were lower than during the fifth instar.     

Differential expression of two small Hsps during diapause in the corn stalk borer Sesamia nonagrioides (Lef.)

We isolated and characterized two members of the α-crystallin/sHsp family, SnoHsp19.5 and SnoHsp20.8 from Sesamia nonagrioides (Lepidoptera: Noctuidae). The cDNAs encoded proteins of 174 and 185 amino acids, with calculated molecular weights of 19.5 and 20.8 kDa, respectively. The deduced amino acid sequences of SnoHsp19.5 and SnoHsp20.8 showed highest homology to Hsp19.7 of Mamestra brassicae and to Bombyx mori Hsp20.4, respectively. Expression patterns of SnoHsp19.5 and SnoHsp20.8 in non-diapausing individuals under different environmental conditions (heat or cold) showed different accumulation profiles for the two genes after heat and cold treatment. SnoHsp19.5 was consistently expressed, while SnoHsp20.8 gene was down-regulated in deep diapause and was up-regulated at the termination of diapause. Our results suggest that these two genes play distinctive roles in the regulation of diapause.     

Reproductive responses to photoperiod and temperature by diapausing and nondiapausing populations of Sesamia nonagrioides Lef. (Lepidoptera − Noctuidae)

Abstract.  The influence of long- and short-day cycles on ovipostion and egg hatch of the corn stalk borer, Sesamia nonagrioides were investigated at a range of temperatures. Oviposition was suppressed when insects are exposed to long days through their immature stages and then transferred to short days after mating. Moreover, mean oviposition and egg hatch increased from 15 to 27.5 °C, whereas oviposition declined significantly at 30 °C at both photoperiodic regimes. Females derived from a diapausing population exposed to long days after mating showed a significantly higher egg production compared to females derived from a nondiapausing population. However, when females from a diapausing population were left to oviposit under short days, fewer eggs were produced compared to those exposed to long days after mating. Thus, photoperiod appears to affect reproductive traits of this species in a quantitative manner. Larval diapause duration is positively correlated with fecundity of the adults. There is also a positive correlation between pupal weight of individuals derived from a diapausing population and the postdiapause fecundity of adults.     

Isolation and characterization of two digestive trypsin-like proteinases from larvae of the stalk corn borer, Sesamia nonagrioides

Two digestive trypsin-like proteinases from Sesamia nonagrioides Lef. (Lepidoptera: Noctuidae) larvae were purified by benzamidine-Sepharose affinity chromatography. The purified enzymes showed molecular size of 27 (trypsin-I) and 24 KDa (trypsin-II). Amino acid analysis and N-terminal sequencing confirmed their relationship with other trypsins from lepidopteran larvae. However, trypsin-I presented one lysine at position 11, being the first report of this amino acid in the sequence of a lepidopteran digestive trypsin. Trypsin-I had an isoelectric point of 6.0, and a Km of 2.2 x 10(-4) M and 3.9 x 10(-5) M for BApNa and BAEE, respectively. Trypsin-II presented an isoelectric point of 8.7, and Km values of 1.7 x 10(-4) M (BApNa) and 3.8 x 10(-5) M (BAEE). Both enzymes were differentially inhibited by some proteinase inhibitors. In particular, trypsin-I was inhibited by E-64 (ID50 = 6 microM) but not by lima bean trypsin inhibitor (LBI), whereas trypsin-II was inhibited by LBI (ID50 = 1 microM) and poorly by E-64 (ID50 = 85 microM). Changes in the susceptibility of the trypsin-like activity of midgut extracts from different larval instars to these inhibitors suggest that the relative proportion of these two enzymes varied through larval development, being predominant in early instars trypsin-I and in late instars trypsin-II.     

Antixenosis and larval and adult dispersal in the Mediterranean corn borer,Sesamia nonagrioides,in relation to Bt maize

Sesamia nonagrioides Lefèbvre (Lepidoptera: Noctuidae) is a key pest of maize [Zea mays L. (Poaceae)] and a main target of Bt maize in the Mediterranean area. To choose the most suitable non‐Bt refuge strategy for preventing or delaying resistance development in this maize borer, we examined its biology and behaviour. No antixenotic effects were found on numbers of eggs and egg batches per plant in choice (Bt vs. non‐Bt plants) and no‐choice assays. However, a greater ratio of young larvae dispersed from Bt than from non‐Bt plants. In addition, larvae that hatched on Bt plants tended to disperse more than those that hatched on non‐Bt plants, particularly during young growth stages. Many adults, especially females, could fly at least up to 400 m, as was found in a dispersal study with rubidium‐marked adults. The stimulation of larval dispersal by the Bt trait and the dispersal capacity of adults might compromise the efficacy of seed mixtures as an insecticide resistance management strategy.     

Environmental and genetic control of the larval diapause of the Southwestern corn borer, Diatraea grandiosella

ABSTRACT. The interaction of photoperiod and temperature in the regulation of the induction and termination of the larval diapause of the Southwestern corn borer, Diatraea grandiosella Dyar (Lepidoptera), was examined. A population originating from south-eastern Missouri had critical daylengths for diapause induction of about 15h 5min (ecological threshold) and llh (physiological threshold). The ecological threshold was more stable than was the physiological threshold at temperatures lower than 25°C. Above 25°C the diapause response was suppressed. The insect appears to measure photoperiods in a stationary manner since a stepwise increase or decrease in photoperiod did not affect the incidence of diapause. In the critical region of the photoperiodic response curve, a higher incidence of diapause was found among females than among males. Females entered diapause later than did males, but resumed active development earlier than males. The rate of diapause development was more temperature dependent than was the rate of diapause induction, yet it was also clearly under photoperiodic control. The temperature coefficient (Q₁₀) for this process was about 4. Several other factors including sex-linkage, age, and geographic adaptations are involved in controlling the rate of diapause development, even more so than they are in controlling diapause induction. In the laboratory, the intensity of diapause declined gradually without larvae being exposed to non-diapause inducing conditions. Incubation of field-collected larvae revealed that their sensitivity to diapause maintaining photoperiods had ended by January. Three generations of selection of a Mississippi population of D. grandiosella at 30°C and LD 12:12 led to the production of an essentially diapause-free strain and a diapause strain.     

Circadian clock regulates photoperiodic responses governed by distinct output pathways in the bean bug,Riptortus pedestris

Effects of RNA interference (RNAi) targeted against circadian clock genes on two distinct types of photoperiodic responses – ovarian development and lipid accumulation – were investigated in a bean bug Riptortus pedestris, to explore which physiological process in the photoperiodic response involved the circadian clock. Ovarian development and lipid accumulation are known to be regulated by distinct output pathways. Control insects showed clear photoperiodic responses; i.e. induction of ovarian development and suppression of lipid accumulation under long-day conditions, whereas opposite characteristics under short-day conditions. We found that RNAi directed against period, a negative element of the circadian clock, produced a long-day effect for both the ovarian development and lipid accumulation, while RNAi directed against Clock, a positive element of the circadian clock, produced a short-day effect for both, irrespective of photoperiod. These results indicate that the circadian clock comprised of these genes regulates a process governing both distinct photoperiodic responses.