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.     

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.     

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.     

Transformation of maize by 2,4-dihydroxy-7-methoxy-2H-1,4-benzo- xazin-3(4H)-one resistant Agrobacterium strains

One of the important factors responsible for recalcitrance of maize tissue towards Agrobacterium-mediated transformation is the presence of 2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (DIMBOA), an inhibitory metabolite found in maize cells. DIMBOA-resistant strains of Agrobacterium tumefaciens were used to transfer genes coding for GUS (-glucuronidase) and NPTII (neomycine phosphotransferase II) in maize shoot apical meristems derived from 20 day-old seedlings and immature embryos. GUS expression was higher (21–34%) in the apical meristem and was dependent on the type of infecting strain and explant-age. The PCR analysis of selected tissues confirmed the presence of GUS gene in the transformed cells.     

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.     

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.     

Decomposition of 2,4-Dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one in Aqueous Solutions

Cyclic hydroxamic acids present in some species of Gramineae have been reported to be important in resistance of these plants to fungi and insects. Since the nonglucosylated forms of these acids are unstable in aqueous solution, in vitro methods for the measurement of their antibiotic properties have been difficult. Kinetics of the decomposition of 2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (DIMBOA), the major hydroxamate in corn (Zea mays L.) extracts, were studied in buffered aqueous solutions from pH 5 to 7.5 at temperatures from 20 to 80 C. Kinetics were apparently first order under all conditions tested; energies of activation (24 to 28 kcal/mol) were nearly pH-independent. DIMBOA decomposed rapidly (half-life = 5.3 hours at 28 C, pH 6.75) relative to the time required for many procedures which have been used to demonstrate the biological activity of DIMBOA. The rate of disappearance of inhibitory activity of DIMBOA toward Erwinia carotovora was indistinguishable from the rate of decomposition of DIMBOA. Contrary to reports, yields of 6-methoxy-2-benzoxazolinone (MBOA) were not quantitative. Gas-liquid chromatography analytical procedures were developed for quantitation of trimethylsilyl and acetyl derivatives of MBOA. As measured by ultraviolet spectroscopy and/or gas-liquid chromatography, conversion of DIMBOA to MBOA ranged from 40 to 75% of theoretical in aqueous buffers, bacterial growth medium, and ethyl acetate extracts of corn tissue resuspended in buffer. Yields varied with temperature, pH, and constituents in the medium.     

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.     

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.     

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.     

Degradation of diazinon by 2,4-dihydroxy-7-methoxy-2h-1, 4-benzoxazin-3(4h)-one in maize

A cyclic hydroxamate, 2,4-dihydroxy-7-methoxy-2H- 1,4-benzoxazin-3(4H)-one (DIMBOA), was isolated and identified from shoots of 6-day-old corn seedlings grown in the dark. From 100 g of plant tissue 100 mg of DIMBOA were isolated. This hydroxamate was very effective in catalysing the hydrolysis of the pyrimidinyl organophosphate insecticide, diazinon (O, O-diethyl- O-[6- methyl-2-(1-methylethyl)-4-pyrimidinyl] phosphorothioate) to 6- methyl-2-(1-methylethyl)-4-hydroxypyrimidine and diethyl phosphorothioic acid. The optimum pH for hydrolytic activity was 5 and at pH values equal to or higher than the pK_a of the hydroxamic group (6.95) most of the activity was lost.     

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.     

Fluconazole analogues containing 2H-1,4-benzothiazin-3(4H)-one or 2H-1,4-benzoxazin-3(4H)-one moieties,a novel class of anti-Candida agents

As a part of our program to develop new antifungal agents, a series of fluconazole analogues was designed and synthesized wherein one of the triazole moieties in fluconazole was replaced with 2H-1,4-benzothiazin-3(4H)-one or 2H-1,4-benzoxazin-3(4H)-one moiety. The new chemical entities thus synthesized were screened against various fungi and it was observed that the compounds 4a and 4i are potent inhibitors of Candida strains. The structure–activity relationship for these compounds is discussed.     

Factors That Influence the Activity of 2,4-Dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one on Erwinia Species in Growth Assays

Factors affecting the inhibitory activity of 2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (DIMBOA) against Erwinia carotovora, a nonpathogen of Zea mays L., and against a maize pathovar of Erwinia chrysanthemi (ECZ) were examined. Most experiments were performed with DIMBOA dissolved in a bacterial growth medium containing 10 g/liter of sucrose, inorganic salts, and 1 g/liter of casamino acids at pH 6.75. When temperature and pH were held constant, inhibition of E. carotovora varied linearly with the logarithm of the initial cell population. By altering temperatures, assays with constant pH and initial cell populations were performed under conditions of varying DIMBOA stability. When E. carotovora was grown at 24, 28, 32, and 36 C in the presence of 0.1 to 0.5 mm DIMBOA, the inhibition of bacterial growth was maintained long after DIMBOA had decomposed in the medium to levels which, if added initially, would not have been inhibitory. When assays were performed at pH 5.5, the pH of aqueous maize extracts, E. carotovora was more inhibited than at pH 6.75; however, ECZ was substantially less inhibited at the lower pH.     

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.     

2,4-Dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one, an Inhibitor from Zea mays with Differential Activity against Soft Rotting Erwinia Species

[2,4-Dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one] DIMBOA was extracted with ethyl acetate from acidified water homogenates of corn (Zea mays L.) seedlings. Pure DIMBOA or ethyl acetate extracts of corn tissue were added to bacterial growth medium at five concentrations (measured as hydroxamates). DIMBOA and corn extracts were more inhibitory to soft rot bacteria (Erwinia spp.) that are nonpathogenic to corn than to soft rot bacteria that are corn pathogens. The inhibitory activity of DIMBOA was similar to that of the ethyl acetate extracts. Both corn extracts and DIMBOA prolonged the lag phase of bacterial growth without significantly changing log phase growth rates. At various concentrations of the inhibitor, 50 to 100% of the activity of corn extracts inhibitory to different bacterial isolates was attributable to DIMBOA. Extracts of DIMBOA-deficient plants (genotype bxbx) were not inhibitory to Erwinia spp. It was concluded that DIMBOA is the major active component in those corn extracts which are inhibitory to soft rot Erwinia species.     

The aglycone specificity-determining sites are different in 2, 4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA)-glucosidase (Maize beta -glucosidase) and dhurrinase (Sorghum beta -glucosidase)

The maize beta-glucosidase isozyme Glu1 hydrolyzes a broad spectrum of substrates in addition to its natural substrate DIMBOAGlc (2-O-beta-d-glucopyranosyl-4-hydroxy-7-methoxy-1,4-benzoxazin-3-on e), whereas the sorghum beta-glucosidase isozyme Dhr1 hydrolyzes exclusively its natural substrate dhurrin (p-hydroxy-(S)-mandelonitrile-beta-d-glucose). To study the mechanism of substrate specificity further, eight chimeric beta-glucosidases were constructed by replacing peptide sequences within the C-terminal region of Glu1 with the homologous peptide sequences of Dhr1 or vice versa, where the two enzymes differ by 4 to 22 amino acid substitutions, depending on the length of the swapped regions. Five Glu1/Dhr1 chimeras hydrolyzed substrates that are hydrolyzed by both parental enzymes, including dhurrin, which is not hydrolyzed by Glu1. In contrast, three Dhr1/Glu1 chimeras hydrolyzed only dhurrin but with lower catalytic efficiency than Dhr1. Additional domain-swapping within the C-terminal domain of Glu1 showed that replacing the peptide (466)FAGFTERY(473) of Glu1 with the homologous peptide (462)SSGYTERF(469) of Dhr1 or replacing the peptide (481)NNNCTRYMKE(490) in Glu1 with the homologous peptide (477)ENGCERTMKR(486) of Dhr1 was sufficient to confer to Glu1 the ability to hydrolyze dhurrin. Data from various reciprocal chimeras, sequence comparisons, and homology modeling suggest that the Dhr1-specific Ser-462-Ser-463 and Phe-469 play a key role in dhurrin hydrolysis. Similar data suggest that DIMBOAGlc hydrolysis determinants are not located within the extreme 47-amino acid-long C-terminal domain of Glu1.     

Leucine Transport Is Affected by Bacillus thuringiensis Cry1 Toxins in Brush Border Membrane Vesicles from Ostrinia nubilalis Hb (Lepidoptera: Pyralidae) and Sesamia nonagrioides Lefebvre (Lepidoptera: Noctuidae) Midgut

The pore-forming activity of Cry1Ab, Cry1Fa and Cry1Ca toxins and their interaction with leucine transport mediated by the K⁺/leucine cotransporter were studied in brush border membrane vesicles (BBMVs) isolated from the midgut of Ostrinia nubilalis and Sesamia nonagrioides. In both species, as in other Lepidoptera, leucine uptake by BBMVs can take place in the absence of cations, but it can also be driven by a K⁺ gradient. Experiments with the voltage-sensitive fluorescent dye 3,3′-diethylthiacarbocyanine iodide proved that Cry1Ab, a Bacillus thuringiensis toxin active in vivo, enhanced the membrane permeability to potassium in O. nubilalis BBMVs. This result is in agreement with similar effects observed in S. nonagrioides BBMV incubated with various Cry1 toxins active in vivo. The effect of the above toxins was tested on the initial rate of 0.1 mM leucine influx. Instead of an increase in leucine influx, a reduction was observed with the Cry1 toxins active in vivo. Cry1Ab and Cry1Fa, but not the inactive toxin Cry1Da, inhibited in a dose-dependent manner leucine uptake both in the absence and in the presence of a K⁺ gradient, a clear indication that their effect is independent of the channel formed by the toxins and that this effect is exerted directly on the amino acid transport system.     

Evaluation of damage caused by stalk borers,Sesamia spp. (Lepidoptera: Noctuidae), on sugarcane quality in Iran

The noctuid stalk borers, Sesamia spp. (S. cretica Led. and S. nonagrioides Lef.), damage a considerable percentage of sugarcane internodes annually in the province of Khuzestan, Iran. A study was conducted to quantify the relationship between stalk borer damage and sugar quality to estimate the monetary loss incurred at these levels of damage at harvest time. Three commercial varieties of sugarcane (SP70‐1143, CP69‐1062 and CP48‐103) were studied to determine the effects of damage on sugar quality. Stalks were separated into seven groups: control (without damage); one to five bored internodes per stalk; and more than five bored internodes per stalk. The percentage of bored internodes and several parameters, such as pol and brix, were measured. Sugar in the control group was 123.78, 123.72 and 104.84 kg/ton for SP70‐1143, CP69‐1062 and CP48‐103, respectively. Sugar per ton, juice purity, pol and brix were all significantly inversely correlated to the percentage of bored internodes. Estimated sugar losses for every 1% bored internode were 0.17, 0.39 and 0.23% (equal to 210, 482 and 241 kg sugar) in SP70‐1143, CP69‐1062 and CP48–103, respectively. These varieties could tolerate damage of up to 18.1, 11.7 and 18.9% bored internodes, respectively, compared to the control. Regression slopes for CP69‐1062 were slightly higher than those of the other two varieties. Therefore, based on quality damage, CP69‐1062 is susceptible to stalk borer damage.