Performance of crosses among flint maize populations under infestation by Sesamia nonagrioides (Lepidoptera: Noctuidae)

Flint maize, Zea mays L., varieties provide some interesting agronomic characteristics and kernels that possess a better ability than other kernels for developing high-quality flour. The pink stem borer, Sesamia nonagrioides Lefebvre, is an important constraint for the maize crop in Mediterranean regions. The objective of this work was to identify a "flint x flint" heterotic pattern that would perform well under artificial infestation by S. nonagrioides. A 10-population diallel was evaluated under infestation by S. nonagrioides in 2 yr. Variety effects were the only significant effects involved in stem and ear resistance to S. nonagrioides attack. Variety effects and average heterosis effects were the only significant effects for grain yield under artificial infestation conditions. Considering variety effects and cross-performance, the heterotic pattern Basto/Enano levantixo x Longfellow (BA/EL x LO) would be recommended for obtaining flint maize hybrids tolerant to S. nonagrioides attack because BA/EL had the most favorable variety effects for stem resistance, LO exhibited the most positive variety effects for grain yield, and the cross BA/EL x LO yielded significantly more than the remaining crosses.     

Antibiosis of the pith maize to Sesamia nonagrioides (Lepidoptera: Noctuidae)

Thirteen inbred lines of maize (Zea mays L.) with different levels of stem resistance to the stem borer Sesamia nonagrioides Lefevbre were evaluated in the field and the laboratory to determine the antibiotic resistance to this pest. Inbreds CM151, CO125, and EP39 had antibiotic pith as well as stem resistance, so the pith could play a role in stem resistance. Inbreds A509, F473, and PB130 did not have antibiotic pith but had stem resistance; therefore, other mechanisms could confer stem resistance. Finally, the inbred MS1334 had antibiotic pith and did not show stem resistance; thus, other factors could compensate the effect of the pith. Therefore, although pith antibiotic compound seems to play a role in the defense against S. nonagrioides attack, it is not the only possible mechanism of defense.     

Egg laying preference of Sesamia nonagrioides (Lepidoptera: Noctuidae) among primary and secondary hosts

Sesamia nonagrioides Lefébvre (Lepidoptera: Noctuidae) has a fairly wide range of host plants. The present study tests the egg-laying preference of S. nonagrioides on four potential host plants (maize, sweet and fiber sorghum and johnsongrass) with respect to several life history traits. No-choice and two-choice tests in all possible combinations were conducted. Concerning no-choice tests, significantly higher number of eggs were laid on sweet sorghum (282.5 eggs/female) in relation to the other tested plants. Moreover, viability of eggs that were laid on sweet sorghum was significantly higher in relation to those that were laid on johnsongrass. Longevity of females reared on maize plants was significantly greater compared with the other test plants except johnsongrass. Furthermore, pre-oviposition period of females reared on maize was significantly longer (2.4-fold) compared with that on sweet sorghum and johnsongrass. Similarly post-oviposition period of females reared on maize was significantly longer compared with that on fiber sorghum. On two choice tests significantly fewer eggs were laid on sweet sorghum than on maize. On the other hand, significantly more eggs were laid on sweet sorghum than on fiber sorghum and johnsongrass. We discuss the potential of secondary hosts to be used as trap plants.     

Diversity of trypsins in the Mediterranean corn borer Sesamia nonagrioides (Lepidoptera: Noctuidae), revealed by nucleic acid sequences and enzyme purification

The existence of a diverse trypsin gene family with a main role in the proteolytic digestion process has been proved in vertebrate and invertebrate organisms. In lepidopteran insects, a diversity of trypsin-like genes expressed in midgut has also been identified. Genomic DNA and cDNA trypsin-like sequences expressed in the Mediterranean corn Borer (MCB), Sesamia nonagrioides, midgut are reported in this paper. A phylogenetic analysis revealed that at least three types of trypsin-like enzymes putatively involved in digestion are conserved in MCB and other lepidopteran species. As expected, a diversity of sequences has been found, including four type-I (two subtypes), four type-II (two subtypes) and one type-III. In parallel, four different trypsins have been purified from midgut lumen of late instar MCB larvae. N-terminal sequencing and mass spectrometric analyses of purified trypsins have been performed in order to identify cDNAs coding for major trypsins among the diversity of trypsin-like sequences obtained. Thus, it is revealed that the four purified trypsins in MCB belong to the three well-defined phylogenetic groups of trypsin-like sequences detected in Lepidoptera. Major active trypsins present in late instar MCB lumen guts are trypsin-I (type-I), trypsin-IIA and trypsin-IIB (type-II), and trypsin-III (type-III). Trypsin-I, trypsin-IIA and trypsin-III showed preference for Arg over Lys, but responded differently to proteinaceous or synthetic inhibitors. As full-length cDNA clones coding for the purified trypsins were available, three-dimensional protein models were built in order to study the implication of specific residues on their response to inhibitors. Thus, it is predicted that Arg73, conserved in type-I lepidopteran trypsins, may favour reversible inhibition by the E-64. Indeed, the substitution of Val213Cys, unique for type-II lepidopteran trypsins, may be responsible for their specific inhibition by HgCl2. The implication of these results on the optimisation of the use of protease inhibitors for pest control, and on the identification of endoprotease-mediated resistance to Bacillus thuringiensis Cry-toxins is discussed.     

Susceptibility to the Cry1F toxin of field populations of Sesamia nonagrioides (Lepidoptera: Noctuidae) in Mediterranean maize cultivation regions

Maize hybrids expressing the Cry1F toxin provide efficient control of lepidopteran pests. The Mediterranean corn borer, Sesamia nonagrioides (Lefèvre), is one of the most damaging pests of maize in the Mediterranean basin. In this work we firstly determined the efficacy of maize hybrids expressing the Cry1F toxin (event TC1507) to control neonates of S. nonagrioides. Leaf tissue feeding bioassays revealed that TC1507 maize is highly effective against this pest, and the percentage mortality obtained was comparable to that obtained with a Cry1Ab-expressing maize hybrid (Compa CB, event 176), which is known to be highly efficacious against S. nonagrioides. Secondly, interpopulation variation in the susceptibility to the Cry1F insecticidal protein was established for nine field-collected populations of S. nonagrioides (three Spanish, two French, two Italian, one Greek, and one Turkish). Estimates of the susceptibility of larvae to the Cry1F toxin showed low variability in lethal concentrations and growth inhibition concentrations among field populations. Moreover, no significant differences were found when they were grouped by geographical areas [Western Mediterranean (Spain and France) versus Eastern Mediterranean (Italy, Greece and Turkey)] or by history of exposure to Bt plants (Spanish vs. other populations). Therefore, the minor differences found in field populations can be attributed to natural variation in sensitivity to Cry1F. The importance of establishing baselines of susceptibility for resistance detection is discussed. Future changes in susceptibility of S. nonagrioides populations to Cry1F could be documented based on this baseline data.     

Genetic differentiation of Mediterranean horse mackerel (Trachurus mediterraneus) populations as revealed by mtDNA PCR-RFLP analysis

The genetic population structure of Mediterranean horse mackerel, Trachurus mediterraneus , from seven locations throughout the Black, Marmara, Aegean and eastern Mediterranean seas was investigated using restriction fragment length polymorphism (RFLP) analysis of the mtDNA 16S rDNA region. An approximately 2000-bp segment was screened in 280 individuals using six restriction enzymes, resulting in 10 composite haplotypes. The most common haplotype was present in 56.42% individuals; the next most frequent haplotype was present in 22.85% individuals. Average haplotype diversity within samples was moderate (0.38), and nucleotide diversity was low (0.00435). Mean nucleotide divergence for the seven sampling sites was 0.0028. Nucleotide divergence among samples was moderate, with the highest value detected between the Aegean Sea (Izmir) and the eastern Black Sea (Trabzon) populations (0.007055), and the lowest (−0.000043) between the Marmara Sea (Adalar) and the western Black Sea (Sile) populations. In Monte Carlo pairwise comparisons of haplotype frequencies, the Sinop from the middle Black Sea, Trabzon from the eastern Black Sea, and Iskenderun Bay from the north-eastern Mediterranean Sea exhibited highly significant (P   <   0.001) geographical differentiation from each other and from all other populations. Mantel's test indicated that the nucleotide divergence among populations of T. mediterraneus was not significantly associated with their geographical isolation ( r  = −0.2963; P   >   0.05). Consequently, the mtDNA 16S rDNA region provided evidence for the existence of three distinct T. mediterraneus populations (Sinop, Trabzon and Iskenderun Bay) in the Black and north-eastern Mediterranean seas.     

Inheritance of resistance to ear damage caused by Sesamia nonagrioides (Lepidoptera: Noctuidae) in maize

Sesamia nonagrioides Lefebvre is a major insect pest of maize (Zea mays L.) in northwestern Spain. The inheritance of ear resistance in field corn to this pest has not been studied. This work aims to determine the importance of genetic (additive, dominance, and epistatic effects) and environmental effects in the inheritance of ear resistance to S. nonagrioides. Three field corn inbreds (CM109, EP31, and EP42) were used as parents and two crosses (CM109 x EP31 and CM109 x EP42) were made. These inbreds show different ear resistance levels to S. nonagrioides, with CM109 more resistant than EP31 and EP42. For each cross, parents (P1, P2), F1, F2, and backcrosses to each parent (BC1 and BC2) were evaluated. Correlations among ear damage traits showed that general appearance of the ear should be useful indicator of ear resistance. Ear resistance was dominant to susceptibility and was transmitted from inbreds to their hybrids. The additive-dominance model fit the generation mean analysis for both crosses and the degree of genetic control varied depending on the cross and trait. For both crosses, additive and dominance effects were significant for most ear damage traits. Epistatic gene effects were significant for husk and shank damage, and gene effects for number and length of tunnels were not significant. Because ear resistance involved additive and dominance effects for this set of inbreds, breeding procedures based on both types of gene action should be effective.     

Resistance of early maturing sweet corn varieties to damage caused by Sesamia nonagrioides (Lepidoptera: Noctuidae)

In Mediterranean countries, the principal pest of maize, Zea mays L., is Sesamia nonagrioides Lefebvre. The objective of this work was to study the resistance of the four early maturing varieties of sweet corn, Baby Orchard, Orchard Baby, Dorinny Sweet, and Golden Early Market, and to check the relationship among resistance to S. nonagrioides and infestation dates. In a previous study, these varieties had showed a confusing behavior, being the most resistant varieties to S. nonagrioides attack in some environments and the most susceptible in others. These varieties were again evaluated along with three medium maturing varieties, used as testers because they were more stable in both environments, by using two infestation dates. Plants were more damaged in the late infestation, but the effect of the infestation depends on the variety studied. Orchard Baby and Baby Orchard were the most resistant varieties under two infestation dates. These two varieties are closely related. Therefore, we can use either one as a source of precocity and resistance for modern sweet corn. Although many studies showed that early maturing inbreds had greater damage caused by S. nonagrioides and European corn borer than late maturing inbreds and hybrids, our results showed that the earliest maturing varieties were the most resistant.     

Mean generation analysis of the damage caused by Sesamia nonagrioides (Lepidoptera: Noctuidae) and Ostrinia nubilalis (Lepidoptera: Crambidae) in sweet corn ears

Sesamia nonagrioides (Lefebvre) and Ostrinia nubilalis (Hübner) are the main maize (Zea mays L.) pests in Mediterranean countries. To develop insect-resistant cultivars, it is helpful to know the genetic control of the resistance. Our objective was to determine the genetic control of the resistance to both borers. For each of two crosses (EP59 x P51 and 15125 x EP61), six generations were evaluated (P1, P2, F1, F2, BC1, and BC2). Genetic effects x environment interactions were not significant. For the O. nubilalis resistance traits; both crosses fitted an additive-dominance model. EP59 x P51 had important dominance and additive effects, whereas for 15125 x EP61 we did not detect significant genetic effects, but significant year effects were detected. For S. nonagrioides infestation, both crosses fitted to an additive-dominance model. There were additive effects for most traits in EP59 x P51. The cross I5125 x EP61 showed significant dominance effects for several traits. Resistance to both corn borers fit an additive-dominance model, but genetic effects depend on the cross evaluated. In the resistance to S. nonagrioides, additive effects were important for shank resistance, which is a useful trait for avoiding S. nonagrioides damage on the ear. Early sowings are recommended to make good use of the resistance to both corn borers. In the late sowings, damage is so high that resistant plants are not able to control the pest.     

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.     

Susceptibility of Spanish populations of the corn borers Sesamia nonagrioides (Lepidoptera: Noctuidae) and Ostrinia nubilalis (Lepidoptera: Crambidae) to a Bacillus thuringiensis endotoxin

Baseline susceptibility to the Cry1Ab delta-endotoxin from Bacillus thuringiensis (Berliner) was determined for four populations of Sesamia nonagrioides (Lefebvre) and two populations of Ostrinia nubilalis (Hübner) from Spain. This study shows that S. nonagrioides is at least as susceptible as O. nubilalis to B. thuringiensis Cry1Ab protein. We found small differences in susceptibility among the Spanish populations of S. nonagrioides that can be attributed to natural variation, because there are no records of B. thuringiensis products being used on corn crops in Spain. There were no differences in susceptibility to Cry1Ab toxin between the two populations of O. nubilalis.     

Fine structure of the female sex pheromone-producing glands in Sesamia nonagrioides lef. (Lepidoptera : Noctuidae)

The morphology and cytology of the female sex pheromone-producing glands of Sesamia nonagrioides (Lepidoptera : Noctuidae) consist of modified intersegmental membranes; they are unusual compared with those described in other moths. There are 2 different glandular areas (a ventral area, GI, around the ostium bursae and a ring-shaped area, GII, around the ovipositor), instead of a single area (between segments 8 and 9 + 10) as in most other species. Ultrastructural features, such as developed basal invaginations and dilated intercellular spaces indicate transport of material from the hemolymph to the glandular cells, while the presence of abundant mitochondria, numerous free ribosomes, rough endoplasmic reticulum, and inclusions reveal high cellular activity. The smooth endoplasmic reticulum is absent in S. nonagrioides, in contrast to its extensive development in glands producing similar compounds in other moths. The cuticle, abnormally thick (7–8 times thicker than the glandular epithelium), is traversed by dilated pore canals forming cuticular gaps, connected with epicuticular canals. This system allows the storage and the passage of pheromone in and through the cuticle.     

Free phenols in maize pith and their relationship with resistance to Sesamia nonagrioides (Lepidoptera: Noctuidae) attack

The stem borer Sesamia nonagrioides (Lefèbvre) is the most important insect pest of maize, Zea mays L., in northwestern Spain. Among the metabolites present in maize, phenolic compounds could play an important role in resistance. The objective of this work was to determine whether a relationship between phenols and the amount of resistance exists. Amounts of free phenolic compounds in the pith of 13 inbred maize lines that differ in resistance were measured. The phenolic compounds identified were p-coumaric acid, cafeic acid, ferulic acid, vanillic acid, syringic acid, chorogenic acid, sinapic acid, p-hydroxybenzoic acid, and vanillin. The amount of free p-coumaric acid was correlated with the resistance level. Higher quantities of p-coumaric in the pith could contribute to general resistance to stem borer attack. Jointly with ferulic acid, p-coumaric could provide resistance mechanisms through cell wall fortification and lignification. The other compounds showed no or an unclear relationship with resistance. The vanillic acid showed a decreased tendency after silking, when maize is most attractive for S. nonagrioides, suggesting this acid could act as a chemoattractant for S. nonagrioides larvae or adults. Future studies that focus on these phenolic compounds could be useful in understanding S. nonagrioides resistance.     

Chemical stimuli from corn plants affect host selection and oviposition behavior of Sesamia nonagrioides (Lepidoptera: Noctuidae)

The role of chemical stimuli in host selection and oviposition of Sesamia nonagrioides (Lef) was studied in the laboratory using eight corn hybrids. Female oviposition preference was evaluated on plants of the eight hybrids, their extracts in pentane and methanol, and volatiles collected by steam distillation in ether. Results from two-choice bioassays indicate various degrees of female preference to the hybrids. Four hybrids received more eggs than control plants, whereas the remainder received fewer eggs. Relatively similar results were obtained when filter paper sticks baited with pentane and steam distillation extracts were evaluated. Methanol extracts from all eight hybrids received fewer eggs than the methanol controls. Results showed that pentane and steam distillation extracts contain chemicals that elicit female oviposition response, while methanol extracts may contain chemicals that deter or repel females to oviposition.     

RFLP analysis of genetic variation in North American populations of the fall armyworm moth Spodoptera frugiperda (Lepidoptera: Noctuidae)

A high level of genetic variability was detected in North American fall armyworm (FA W), Spodoptera frugiperda (J. E. Smith), populations by restriction fragment length polymorphism (RFLP) analysis of genomic DNA. In nearly all cases individual larvae could be differentiated using 22 probe-enzyme combinations. Laboratory colonies formed distinct groups based on RFLP analysis. Individuals from a colony of the previously determined ‘rice strain’ formed a group showing obvious deviation in RFLP patterns from the other five populations, which included a population from a ‘com strain’ colony. The results were consistent with previous studies using allozymes which indicated that there are two genetically differentiated forms of the fall armyworm. Diagnostic markers were also found that distinguish these two groups.     

Genetic structure of natural populations ofDryas iulia (Lepidoptera: Nymphalidae) revealed by enzyme polymorphism and mitochondrial DNA (mtDNA) restriction fragment length polymorphism (RFLP)

Dryas iulia appears to have undergone a mode of evolution different from that of other members of its subfamily (Heliconiinae). While other species constitute highly subdivided and inbred populations, those ofD. iulia are thought to be large and uniform. Analyzing six samples from Southern Brazil (state of Rio Grande do Sul) in relation to three enzyme systems (EST, LAP, and PGM) and their mtDNA RFLP patterns, we found that they are very similar at the molecular level. TheF statistics for enzyme polymorphism data revealed that inbreeding makes a great contribution to the population homozygosity, sinceF _IS equals 0.1322 andF _ST equals 0.0023. Since the chi-square test showed thatF _ST is not significant, we conclude that all localities belong to the same population. The mtDNA differentiation was about 12 times greater than for nuclear genes;F _ST was equivalent to 0.0265. We suggest that this difference is due to a higher dispersal of males, in relation to females.     

Genetic variability and demographic history of Heliothis virescens (Lepidoptera: Noctuidae) populations from Brazil inferred by mtDNA sequences

Intra- and inter-population genetic variability and the demographic history of Heliothis virescens (F.) populations were evaluated by using mtDNA markers (coxI, coxII and nad6) with samples from the major cotton- and soybean-producing regions in Brazil in the growing seasons 2007/08, 2008/09 and 2009/10. AMOVA indicated low and non-significant genetic structure, regardless of geographical scale, growing season or crop, with most of genetic variation occurring within populations. Clustering analyzes also indicated low genetic differentiation. The haplotype network obtained with combined datasets resulted in 35 haplotypes, with 28 exclusive occurrences, four of them sampled only from soybean fields. The minimum spanning network showed star-shaped structures typical of populations that underwent a recent demographic expansion. The recent expansion was supported by other demographic analyzes, such as the Bayesian skyline plot, the unimodal distribution of paired differences among mitochondrial sequences, and negative and significant values of neutrality tests for the Tajima's D and Fu's F(S) parameters. In addition, high values of haplotype diversity (?) and low values of nucleotide diversity (π), combined with a high number of low frequency haplotypes and values of θ(π)<θ(W), suggested a recent demographic expansion of H. virescens populations in Brazil. This demographic event could be responsible for the low genetic structure currently found; however, haplotypes present uniquely at the same geographic regions and from one specific host plant suggest an initial differentiation among H. virescens populations within Brazil.     

Evaluation of the European Union Maize Landrace Core Collection for resistance to Sesamia nonagrioides (Lepidoptera: Noctuidae) and Ostrinia nubilalis (Lepidoptera: Crambidae)

Two corn borer species are the principal maize insect pests in Europe, the European corn borer, Ostrinia nubilalis (Hübner), and the pink stem borer, Sesamia nonagrioides (Lefebvre). Hence, it would be advisable to evaluate the European maize germplasm for corn borer resistance to generate European varieties resistant to corn borer attack. The creation of the European Union Maize Landrace Core Collection (EUMLCC) allowed the screening of most of the variability for European corn borer resistance present among European maize local populations from France, Germany, Greece, Italy, Portugal, and Spain, testing a representative sample. The objective of this study was the evaluation of stem and ear resistance of the EUMLCC to European corn borer and pink stem borer attack. Trials were made at two Spanish locations that represent two very different maize-growing areas. Populations that performed relatively well under corn borer infestation for stem and ear damage were 'PRT0010008' and'GRC0010085', among very early landraces; 'PRT00100120' and 'PRT00100186', among early landraces; 'GRC0010174', among midseason landraces; and 'ESP0070441', among late landraces. Either the selection that could have happen under high insect pressure or the singular origin of determined maize populations would be possible explanations for the higher corn borer resistance of some landraces. Landraces 'PRT0010008', 'FRA0410090', 'PRT00100186', and 'ESP0090214' would be selected to constitute a composite population resistant to corn borers and adapted to short season, whereas populations 'ESP0090033', 'PRT00100530', 'GRC0010174', and 'ITA0370005' would be used to make a resistant composite adapted to longer season.     

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.