Spatial pattern of parasitism in Diatraea saccharalis Fab. (Lep., Crambidae) populations at two different spatial scales in sugarcane fields in Brazil

Abstract:  The biological control of Diatraea saccharalis is regarded as one of the best examples of successful classical biological control in Brazil. Since the introduction of the exotic parasitoid, Cotesia flavipes , the decrease in D. saccharalis infestation in sugarcane fields has been attributed to the effectiveness of this agent. Native Tachinidae fly parasitoids ( Lydella minense and Paratheresia claripalpis ) have also been implicated in the success. Quantitative data confirming the actual contribution of these agents to the control of D. saccharalis are, however, rather scant. The purpose of this study was to investigate the spatial pattern of parasitism of these parasitoids in D. saccharalis populations at two large spatial scales (fields and zones). To investigate this subject, a large data set comprising information collected from a sugarcane mill located in the state of São Paulo, Brazil (São João sugarcane mill) was analysed. When regressions between the proportion parasitism against host density were computed, the percentage of significant regressions with either a positive or a negative slope was very small at both spatial scales for both parasitoid species. Regressing the densities of tachinid-parasitized hosts against host densities per field showed that these parasitoids presented a 'moderate aggregative' response to host densities, as 53.33% of the regressions were positively significant. Cotesia flavipes was 'weakly aggregated' on host densities at the field level, because only 33.33% of the regressions were positively significant. At the zone level, neither aggregative nor spatial proportion parasitism responses were evident for either parasitoid species due to the small percentage of significant regressions computed.     

Frequency of alleles conferring resistance to Bacillus thuringiensis maize in Louisiana populations of the southwestern corn borer

Transgenic maize [Zea mays L. (Poaceae)] expressing Bacillus thuringiensis proteins (Bt maize) has become the most important tool for managing stalk borers in maize in the USA. The current strategy for delaying the evolution of resistance in target insects for Bt maize is referred to as high dose/refuge strategy. A key requirement of the strategy is that initial resistance allele frequencies in field insect populations are low (e.g., <0.001). More than 200 iso‐line families of the southwestern corn borer, Diatraea grandiosella Dyar (Lepidoptera: Crambidae), a major target stalk borer pest of Bt maize, were developed from Louisiana populations and evaluated for Bt resistance using a modified F₂ screening method during 2005. No major resistance alleles were detected in these populations. The results showed that the expected Bt resistance allele frequency in the Louisiana populations was <0.0035 with 95% probability and a detection power of 83.9 ± 0.6%. The F₂ screen indicates that Bt resistance allele frequencies in D. grandiosella are low among the Louisiana populations and should meet the rare resistance allele requirement of the ‘high dose/refuge’ strategy.     

Identification of two components of the female sex pheromone of the sugarcane‐borer Diatraea flavipennella (Lepidoptera: Crambidae)

Analysis by gas chromatography with electroantennographic detection of extracts of pheromone glands derived from calling females of the sugarcane‐borer Diatraea flavipennella revealed two antennally active compounds. These components were identified as (Z)‐9‐hexadecenal (Z9–16:Ald) and (Z)‐11‐hexadecenal (Z11–16:Ald) by comparison of the retention times of the natural compounds and the synthetic compounds supported by two‐dimensional gas chromatography – time‐of‐flight mass spectrometric analysis and the positions of the double bounds in the chains were confirmed from the mass spectral fragmentation patterns of their dimethyldisulphide adducts. The analysis indicated that Z9–16:Ald and Z11–16:Ald were present in the sex pheromone in the proportions 25 : 75. Trace amounts of tetradecanal, hexadecanal, (Z)‐7‐hexadecenal (Z7–16:Ald), (Z)‐9‐hexadecen‐1‐ol and (Z)‐11‐hexadecen‐1‐ol were also found in the extract, but of these only Z9–16:Ald and Z11–16:Ald appeared to be antennally active. Behavioural bioassays demonstrated that a binary blend composed of Z9–16:Ald and Z11–16:Ald in the ratio of 25 : 75 induced a response in D. flavipennella virgin males similar to that elicited by live virgin females or by an hexane extract of the pheromone glands of calling females. Z9–16:Ald and Z11–16:Ald are, therefore, considered to be the major constituents of the female sex pheromone of D. flavipennella.     

Use of ablated stalks to assess field rates of parasitism of Diatraea saccharalis (Lepidoptera: Crambidae) by Cotesia flavipes (Hymenoptera: Braconidae)

We evaluated a technique that used ablated sections of corn stalks infested with larvae of Diatraea saccharalis F. to assess parasitism by Cotesia flavipes (Cameron) in a corn field. Stalkborer larvae were retrieved successfully from the artificially infested, ablated stalks for at least 96 h after deployment in the field. Levels of parasitism in ablated stalks attached to corn plants were comparable to levels of parasitism measured using whole plants that were artificially infested. Olfactometer comparisons showed preference by female C. flavipes for both whole plants and ablated stalks containing larval D. saccharalis, over uninfested stalks and plants. This technique provides a means to estimate parasitism of stalkborer larvae by C. flavipes without destructive sampling of agronomic plants.     

Inheritance of resistance to Bacillus thuringiensis Cry1Ab protein in the sugarcane borer (Lepidoptera: Crambidae)

Inheritance traits of a Cry1Ab-resistant strain of the sugarcane borer, Diatraea saccharalis (F.) were analyzed using various genetic crosses. Reciprocal parental crosses between Cry1Ab-susceptible and Cry1Ab-resistant populations, F₁ by F₁ crosses, and backcrosses of F₁ with the Cry1Ab-resistant population were successfully completed. Larval mortality of the parental and cross-populations were assayed on Cry1Ab diet and Bacillus thuringiensis (Bt)-corn leaf tissue. Maternal effects and sex linkage were examined by comparing the larval mortality between the two F₁ populations. Dominance levels of resistance were measured by comparing the larval mortality of the Cry1Ab-resistant, -susceptible, and -heterozygous populations. Number of genes associated with the resistance was evaluated by fitting the observed mortality of F₂ and backcross populations with a Mendelian monogenic inheritance model. Cry1Ab resistance in D. saccharalis was likely inherited as a single or a few tightly linked autosomal genes. The resistance was incompletely recessive on Bt corn leaf tissue, while the effective dominance levels (D_ML) of resistance increased as Cry1Ab concentrations decreased with Cry1Ab-treated diet. D_ML estimated based on larval mortality on intact Bt corn plants reported in a previous study ranged from 0.08 to 0.26. This variability in D_ML levels of Cry1Ab resistance in D. saccharalis suggests that Bt corn hybrids must express a sufficient dose of Bt proteins to make the resistance genes functionally recessive. Thus, Bt resistant heterozygous individuals can be killed as desired in the “high/dose refuge” resistance management strategy for Bt corn.     

(11Z)-hexadec-11-enal enhances the attractiveness of Diatraea saccharalis main pheromone component in wind tunnel experiments

Abstract:  GC-EAD and GC-MS analysis of pheromone gland extracts of sugarcane borer, Diatraea saccharalis , revealed two antennally active compounds, (9 Z ,11 E )-hexadeca-9,1-dienal and (11 Z )-hexadec-11-enal, in approximately 10 : 1 ratio. Various doses of identified compounds were investigated in wind tunnel experiments individually and in a 10 : 1 ratio. At all tested doses (9 Z ,11 E )-hexadeca-9,1-dienal alone elicited upwind orientation and source location only in a minority of tested males. An admixture of (11 Z )-hex-11-enal enhanced the attractiveness of (9 Z ,11 E )-hexadeca-9,11-dienal significantly. This two-component blend (100 pg) was as attractive as natural pheromone extracted from three female pheromone glands. The data suggest that (11 Z )-hexadec-11-enal is a part of the D. saccharalis sex pheromone.     

High basal defense gene expression determines sorghum resistance to the whorl‐feeding insect southwestern corn borer

Southwestern corn borer (SWCB, Diatraea grandiosella) and fall armyworm (FAW, Spodoptera frugiperda) are major pests of sorghum in the southern United States. Host plant resistance is a desirable means for reducing plant damage and yield losses from both insects. In this study, we evaluated 12 sorghum lines for whorl‐stage resistance to leaf‐feeding SWCB and FAW in greenhouse and laboratory bioassays. Differential plant responses were detected against the two insects. Among 12 lines tested, CM1821, Della and PI196583 were resistant to both insects, while BTx2752 was largely susceptible. Line R.09110 was resistant to SWCB, but susceptible to FAW, whereas Redbine‐60 was susceptible to SWCB, but not to FAW. In addition, we quantified various chemical components in the plants and determined their association with insect resistance. Tannin and chlorophyll in leaves did not show any significant correlation with resistance to either insects, but contents of soluble protein in general were negatively correlated with resistance to both insects. Endogenous soluble sugar and dhurrin were only positively correlated with resistance to SWCB, but not with FAW resistance. To gain some molecular insight into resistance mechanism of sorghum to SWCB, we performed qPCR reactions for key genes encoding enzymes involved in dhurrin and jasmonic acid (JA) biosynthesis on selected resistant or susceptible lines. Although these genes were rapidly and strongly induced by insect feeding in all lines, the observed resistance is likely explained by higher constitutive dhurrin contents in some resistant lines and higher basal JA biosynthesis in others. Our results suggest that sorghum utilizes multiple strategies to defend itself against SWCB.     

Effects of entomopathogenic bacterium Photorhabdus temperata infection on the intestinal microbiota of the sugarcane stalk borer Diatraea saccharalis (Lepidoptera: Crambidae)

Photorhabdus temperata is an entomopathogenic bacterium that is associated with nematodes of the Heterorhabditidae family in a symbiotic relationship. This study investigated the effects of P. temperata infection on the intestinal microbiota of the sugarcane stalk borer Diatraea saccharalis. Histopathology of the infection was also investigated using scanning electron microscopy. Groups of 20 larvae were infected by injection of approximately 50 bacterial cells directly into the hemocoel. After different periods of infection, larvae were dissected and different tissues were used for bacterial cell quantification. P. temperata was highly virulent with an LD₅₀ of 16.2 bacterial cells at 48 h post-infection. Infected larvae started dying as soon as 30 h post-infection with a LT₅₀ value of 33.8 h (confidence limits 32.2–35.6) and an LT₉₀ value of 44.8 h (CL 40.8–51.4). Following death of the larvae, bacteria from the midgut did not invade the hemocoel. In the midgut epithelium, P. temperata occupied the space underneath the basal lamina. The cultivable intestinal bacterial populations decreased as soon as 1 h post-infection and at 48 h post-infection, 90% of the gut microbiota had died. The role of P. temperata in control of the midgut microbiota was discussed.     

Effects of soybean proteinase inhibitors on development of the soil mite Scheloribates praeincisus (Acari: Oribatida)

Proteinase inhibitors (PI) are present in plant tissues, especially in seeds, and act as a defense mechanism against herbivores and pathogens. Serine PI from soybean such as Bowman–Birk (BBPI) and Kunitz have been used to enhance resistance of sugarcane varieties to the sugarcane borer Diatraea saccharalis (Fabricius) (Lepidoptera: Crambidae), the major pest of this crop. The use of these genetically-modified plants (GM) expressing PI requires knowledge of its sustainability and environmental safety, determining the stability of the introduced characteristic and its effects on non-target organisms. The objective of this study was to evaluate direct effects of ingestion of semi-purified and purified soybean PI and GM sugarcane plants on the soil-dwelling mite Scheloribates praeincisus (Berlese) (Acari: Oribatida). This mite is abundant in agricultural soils and participates in the process of organic matter decomposition; for this reason it will be exposed to PI by feeding on GM plant debris. Eggs of S. praeincisus were isolated and after larvae emerged, immatures were fed milled sugarcane leaves added to semi-purified or purified PI (Kunitz and BBPI) or immatures were fed GM sugarcane varieties expressing Kunitz and BBPI type PI or the untransformed near isogenic parental line variety as a control. Developmental time (larva-adult) and survival of S. praeincisus was evaluated. Neither Kunitz nor BBPI affected S. praeincisus survival. On the other hand, ingestion of semi-purified and purified Kunitz inhibitor diminished duration of S. praeincisus immature stages. Ingestion of GM senescent leaves did not have an effect on S. praeincisus immature developmental time and survival, compared to ingestion of leaves from the isogenic parental plants. These results indicate that cultivation of these transgenic sugarcane plants is safe for the non-target species S. praeincisus.     

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

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