Oosorption in the stink bug Plautia stali: role of Juvenile hormone in the induction of oosorption

To clarify the role of Juvenile hormone (JH) in the induction of oosorption in females of the stink bug Plautia stali Scott (Heteroptera: Pentatomidae), the effects of extirpation and implantation of the corpus cardiac‐corpus allatum complex (CC‐CA) are examined in fed and starved adults, respectively. Removal of CC‐CA induces oosorption in fed females, whereas CC‐CA implantation stimulates ovary development in starved females. Transection of the nervous connections between the brain and CC‐CA also exerts a stimulatory effect on ovary development. Uptake of yolk protein by the oocytes, assessed in terms of incorporation of a fluorescence dye, occurs on the day after food deprivation but ceases within 1 day after allatectomy. When females are deprived of food, beginning on day 3 of adult life, and treated with JH III skipped bisepoxide (JHSB₃) on the same day, their ovaries develop in a dose‐dependent fashion. Approximately half of the starved females that received JHSB₃ application on day 5 undergo oosorption in the terminal oocytes. This indicates that the critical starvation period for oosorption induction is approximately 2 days, and the earlier half of this period may reflect the time required for the brain to detect poor nutritional condition. During the latter half, in response to food deprivation, the brain inhibits JH biosynthesis by the corpus allatum through nervous connections, resulting in a low JH titre, which in turn induces oosorption.     

Localization of the photoreceptor for photoperiodism in the stink bug, Plautia crossota stali

The brown-winged green bug Plautia crossota stali Scott (Heteroptera: Pentatomidae) shows a long-day photoperiodic response with respect to the control of adult diapause. The location of the photoreceptor for this response was examined by surgical removal of putative photoreceptor organs. Even after both ocelli were removed, the insects responded normally to the photoperiod. After bilateral removal of the compound eyes, the insects developed reproductive organs and the volume of the corpus allatum increased regardless of photoperiod. Therefore, the compound eyes play a major role in the reception of photic information for photoperiodism in P. c. stali. However, because removal of the bilateral compound eyes did not completely prevent the response to photoperiod, photoreceptors other than the compound eyes can also receive photic information for photoperiodism.     

Symbiont of the stink bug Plautia stali synthesizes rough-type lipopolysaccharide

The structures and biosynthesis of lipopolysaccharide (LPS), a major component of the outer membrane of Gram-negative bacteria, have been studied extensively in cultured bacteria such as Escherichia coli. In contrast, little is known about the structures and biosynthesis of the LPS of unculturable bacteria, including insect symbionts, many of which are Gram-negative bacteria. A brown-winged green bug, Plautia stali, is known to harbor a single species of gamma-proteobacterium in the posterior mid-gut caeca. To characterize the features of its LPS, we analyzed the genome sequence of the symbiont, and identified the putative genes involved in LPS synthesis. Genes involved in the synthesis of lipid A and the core oligosaccharide were found in the genome, but waaL, which encodes the O-antigen ligase, was not. Furthermore, we characterized the LPS of this symbiont using SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Toll-like receptor 4 (TLR4) stimulation assays. Consistent with the genomic analysis, the SDS-PAGE analysis suggested that the symbiont had rough-type LPS, which lacked the O-antigen. The TLR4 stimulation assay demonstrated that LPS purified from the symbionts activated NF-κB-dependent reporter expression, indicating the existence of a bioactive lipid A portion in the LPS. These results suggest that the P. stali symbiont produces rough-type LPS.     

Biological activities of juvenile hormone III skipped bisepoxide in last instar nymphs and adults of a stink bug, Plautia stali

Juvenile hormone III skipped bisepoxide (JHSB₃), methyl (2R,3S,10R)-2,3;10,11-bisepoxyfarnesoate was recently determined as a novel juvenile hormone (JH) in a stink bug, Plautia stali. To further confirm the biological function of JHSB₃ in this insect, its juvenilizing, reproduction-stimulating and diapause-terminating activities and the presence in the hemolymph were examined. Topical application of JHSB₃ to last instar nymphs inhibited their metamorphosis in a dose-dependent fashion. In allatectomized and diapausing adults, JHSB₃ application exerted stimulatory effects on the development of ovaries and ectadenia in females and males, respectively. JHSB₃ was detected from the hemolymph of reproductively active females by gas chromatography-mass spectrometry analysis while its titer in the hemolymph collected from diapausing adults was too low to be detected. These results demonstrated that JHSB₃ has biological function as a JH in P. stali. Topical application of JHSB₃, its stereoisomers and 10R-JH III also indicated that compounds with the 2R,3S-configuration were more potent than those with the 2S,3R-configuration and 2,3-double bond.     

A putative juvenile hormone in a stink bug,Plautia stali: The corpus allatum produces and releases a JH-active product different from any known JHs in vitro

Summary

A radiochemical method was adopted to analyze the in vitro products of the corpus allatum (CA) of Plautia stali. The radiolabel derived from ³H-methionine added to the incubation medium was incorporated and released by CA as two main radiolabelled products. They showed Rf values of about 0.3 and 0.5, respectively, in the thin layer chromatography (TLC) system used. The release of these products was shown to be CA-specific since in control incubations using the brain, midgut, aorta and flight muscle, virtually no release of these products was observed. The locations where these main products migrated on the TLC did not coincide with spots of synthetic standards of JH I-III or JHB₃, a JH found in higher Diptera. An addition of precursors of JH III, farnesoic acid or farnesol stimulated the CA to biosynthesize the products with an Rf value of 0.5 up to about 10-fold, suggesting that the product in question may have a sesquiterpenoid skeleton similar to JH III. Topical applications of the hexane extracts of the medium in which the CA had been incubated exerted a juvenilizing, metamorphosis-inhibiting effect on final instar nymphs in a dose-dependent fashion. The nymphs treated with the hexane extracts at a high dose moulted to intermediates with reduced forewings and scutellum, as well as nymphs implanted with the CA from reproductively active females. Based on this juvenilizing effect found in the hexane extracts, the JH-active fraction was determined after TLC separation. This assay indicated that the products found at an Rf value of about 0.5 was JH-active. These results suggest the presence of a new JH different from any known JHs in P. stali.     

Effects of low temperature on diapause termination and body colour change in adults of a stink bug, Plautia stali

Abstract. Diapause adults of Plautia stali Scott maintained at 20°C under short day conditions (LD 12:12 h) were exposed to four temperatures of 5–20°C to examine the effect on diapause development which was assessed in terms of oviposition. Diapause adults kept at 20°C under short day conditions changed their body colour gradually from brown to green and started egg laying after a prolonged preoviposition period. Those transferred to either 10 or 15°C also showed colour change but did not lay eggs. Bugs exposed to 5°C underwent neither body colour change nor oviposition and died more rapidly than those kept at higher temperatures. When 30-day-old diapause adults were chilled at 5, 10 or 15°C for 30 or 60 days and returned to 20°C and long day conditions (LD 16:8 h), the preoviposition period varied primarily depending on the chilling, but not on the temperature. On the other hand, when 60day-old diapause adults chilled for 30 days were observed at 20°C and long day conditions, their preoviposition period tended to be longer as the chilling temperature was lower In this case, a temperature of 10°C appeared to intensify diapause. Therefore, the effect of chilling on diapause development varied depending on the age at which insects were chilled. When chilled bugs were transferred to short day conditions at 20°C, most females failed to lay any eggs and some turned green, then after a while, some green bugs changed to brown again. These results indicate that bugs remained sensitive to short day conditions even after a 60-day chilling at 10 or 15°C.     

Prevalence of a trypanosomatid in the southern green stink bug, Nezara viridula

The southern green stink bug, Nezara viridula (L.), and certain of its host plants were examined to determine the prevalence and biological characteristics of an intestinal trypanosomatid. Promastigotes with short (< or = 17.5 microm excluding flagellum) and long forms (> or = 25.0 microm) usually infected < 50% of the bugs before August and > 50% (maximum 95%) during August-October, but prevalence was not host-density dependent. The flagellate was detected in adults and in all nymphal instars, at all sampling sites where at least 10 bugs were captured, and in bugs from all host plants sampled (soybean, red clover, vetch). Of bugs with flagellates, 27% were heavily infected (> 20 flagellates per 160X microscope field). Weights of infected and uninfected adults did not differ. Live flagellates were detected in bug feces and in one stem of red clover. When bugs were fed soybean pods, tomatoes, or snap beans in the laboratory, only once were flagellates detected in plant tissue (snap beans). The flagellate was cultured in modified Medium 199. This flagellate is prevalent in N. viridula populations in Louisiana and apparently does not cause significant pathological effects in N. viridula or its host plants, including soybean.     

Developmental response of the brown-winged green bug, Plautia stali (Heteroptera: Pentatomidae), to food shortage

To study developmental response of the brown-winged green bug, Plautia stali Scott, to food shortage we reared nymphs under restricted feeding conditions produced by shortening the feeding period after molt or withholding food from second to fifth instars. For second instars, molting rates were significantly reduced as the feeding period was shortened. Shortening the feeding periods for third to fifth instars also reduced molting rates, but less so; some nymphs were able to complete their developments even if no food was given to any of the instars. Compared with controls for which feeding was not restricted, nymphs that successfully reached the next instar had reduced postmolt body size as a result of restricted feeding for all instars except the fifth (2-day feeding for second instar, and 1-day feeding for third and fourth instars), whereas instar duration was unchanged or only slightly prolonged for all instars. These results suggest that only nymphs with nutritional accumulation over a specific threshold in each instar can progress to the next instar, and that, particularly for second to fourth instars, nymphs develop on schedule without prolonging instar durations to compensate for reduced growth under conditions of food shortage.     

Maternal age and egg weight affect offspring performance in the predatory stink bug Podisus nigrispinus

To investigate influences of maternal age and egg weight on developmental time and body weight of offspring in the Neotropical pentatomid Podisus nigrispinus, a study was carried out using large (0.36 mg) and small (0.32 mg) eggs of young (two-week- old) females and large eggs of old (seven-week-old) females. Incubation period of eggs was influenced only by egg weight, i.e. small eggs took a relatively shorter time to hatch. Both maternal age and egg weight had significant effects on total nymphal and egg-to- adult developmental times. Overall, offspring of young parents had shorter developmental times. None of the developmental periods were significantly affected by sex. No significant differences were found among various offspring groups for percentages of egg hatch, nymphal survival, and sex ratio. Variation of egg weight in relation to the number of eggs per day observed within the first month of life suggests a trade-off between weight of individual eggs and oviposition rate. It is concluded that for mass production of the predators in view of their use in augmentative biocontrol programmes, young parents (2–4 weeks old) should be preferred.     

Precocious induction of vitellogenin with JH III in the twospotted stink bug, Perillus bioculatus (Heteroptera: Pentatomidae)

The effect of juvenile hormone (JH) III on the hemolymph composition of vitellogenin was examined in Perillus bioculatus. Adult females were treated topically with JH III, and the premature presence of vitellogenin in the hemolymph was then detected using electrophoresis and Western blot analyses. JH III treatment resulted in a dose-dependent early production of vitellogenin that was detectable 48 h before vitellogenin was present in non-treated insects. Vitellogenin was not observed in the hemolymph of JH III-treated adult males. The techniques reported here may be useful for the detection, isolation and characterization of compounds with JH-like activity in P. bioculatus and other species of Heteroptera (which are thought to have JH-like substances other than the JHs with known chemical identity). These same techniques may also provide a method for researchers to investigate the interactions of JH-like compounds and other substances, such as ecdysteroid, in the regulation of vitellogenesis in Heteroptera.     

Response of the parasitoid Telenomus podisi to induced volatiles from soybean damaged by stink bug herbivory and oviposition

Abstract

Egg parasitoids have a short time frame in which their host eggs are suitable for parasitism, and in several systems these parasitoids respond to plant volatiles induced by oviposition on the plant (either in isolation or in combination with feeding damage) as a means of finding suitable hosts. It is known that the parasitoid of pentatomid eggs Telenomus podisi responds to damage done to soybeans by female Euschistus heros, its preferred host. In this study our aim was to determine the type of E. heros damage to soybean (herbivory, oviposition or a combination of both) necessary for attraction of T. podisi. In a Y-tube olfactometer the parasitoid has shown to respond to the undamaged plant over clean air and herbivory-damaged plants over undamaged plants. However, the parasitoids did not respond to the treatments where oviposition occurred, either in isolation or in combination with herbivory. Analysis of volatile blends revealed that herbivory plus oviposition damage to soybean induced a volatile blend different to those induced when herbivory or oviposition occurred separately. These results, along with other results from this system, suggest that T. podisi uses plant volatile cues associated with female E. heros damage in order to be present when E. heros lays its eggs, and thus ensure its resource is optimal for parasitism.     

Sentinel eggs underestimate rates of parasitism of the exotic brown marmorated stink bug,Halyomorpha halys

Native to eastern Asia, the brown marmorated stink bug (BMSB), Halyomorpha halys (Stål), has become a serious invasive pest in North America. Consequently, accurate assessment of parasitism rates under field conditions is critical for determining baseline parasitism rates of native egg parasitoids on BMSB, and for future evaluations of native or exotic parasitoid biological control release strategies and impacts. BMSB sentinel (laboratory-laid) egg masses have typically been used for this purpose, even though they could be providing misleading estimates of parasitoid activity. Accordingly, we compared the use of BMSB sentinel (laboratory-laid) and wild (naturally field-laid) egg masses in 2012 and 2013 to examine rates of parasitism and the parasitoid community composition of indigenous egg parasitoids in outdoor ornamental nurseries. Wild egg masses consistently had higher rates of parasitism than sentinel egg masses. In 2012, wild egg masses had a mean percent parasitism of 28.4% compared to 4.6% in sentinel egg masses, while in 2013 the difference between the two methods increased even further with a mean percent parasitism of 55.3% in wild egg masses compared to 0.8% in sentinel eggs. Furthermore, we found greater total numbers of parasitoids (889, 42) and greater species richness (seven, five), when using wild egg masses compared to sentinel egg masses, respectively. While sentinel egg masses provide a rapid and convenient way to assess the presence of natural enemies, our findings indicate that using sentinel egg masses could dramatically underestimate actual rates of parasitism and provide inaccurate estimates of parasitoid community composition. Future studies should address potential mechanisms underlying these patterns such as lack of certain host cues required by parasitoids in sentinel compared to wild egg masses.     

Insecticidal potential of Bacillus thuringiensis for the biological control of neotropical brown stink bug

Euschistus heros (Hemiptera: Pentatomidae) is considered a major insect pest in soybean production in Brazil. The indiscriminate application of pesticides to fields leads to reduction in biodiversity, selection of resistant populations, emergence of new pest outbreaks, and damage to non‐target organisms. The objective of this research was to evaluate the effects of exposure to Cry proteins present in new isolates of Bacillus thuringiensis Berliner, commercial formulations, and Bt soybean on E. heros development. The feeding preference of E. heros for Bt‐ vs. non‐Bt soybean was also evaluated. All treatments caused significant mortality to E. heros except the Bt soybean. Tests with combinations of isolated Bt‐toxins indicated that combinations had greater efficacy than other treatments (>98% mortality). The results demonstrate that E. heros is susceptible to B. thuringiensis toxins, which may contribute to the management of this insect in soybean agro‐ecosystems.     

Roles of the circadian clock and endocrine regulator in the photoperiodic response of the brown‐winged green bug Plautia stali

The physiological mechanisms underlying photoperiodism in insects have been studied extensively, although the associated molecular machinery remains largely unknown. In the present study, we investigate the roles of the circadian clock gene cycle (cyc) and the endocrine regulator gene myoinhibitory peptide (Mip) in the photoperiodic response of the brown‐winged green bug Plautia stali Scott (Hemiptera, Pentatomidae). Typically, adult females of this species develop their ovaries under long‐day conditions, whereas they suppress its development under short‐day conditions. We find that RNA interference (RNAi) directed against cyc causes malfunction of the circadian clock governing the locomotor activity rhythm and yields abnormal activity profiles not only under constant darkness, but also under light/dark conditions. RNAi directed against cyc and Mip disrupts the photoperiodic response in ovarian development. cyc RNAi suppresses the ovarian development even under long‐day conditions, whereas Mip RNAi induces it even under short‐day conditions. We propose that the core circadian clock gene cyc regulates the photoperiodic response and that Mip is the causal regulator of juvenile hormone biosynthesis in the corpus allatum. Neither photoperiod, nor cyc RNAi affect Mip mRNA levels, and therefore it remains unknown how the photoperiodic information is processed and mediated by Mip.     

Mate location in the southern green stink bug,Nezara Viridula (Heteroptera: Pentatomidae), mediated through substrate-borne signals on ivy

We observed mate location behavior in Nezara viridulaon the ivy Hedera helix.The female stays motionless on the plant stem, calling with rhythmic repetitions of her calling song. The male walks on the plant, answering with his calling and courtship songs. On encountering petioles or branching points, the male stops and straddles his legs across the fork. Only after another repetition of the female's call does the male proceed to walk again. This behavior may enable the male to compare the vibrations in the two substrates and choose the stronger one to follow. On plants where visual and airborne acoustic communication can be prevented by leaves, substrateborne vibrational signals perceived by receptors in legs would enhance the likelihood of mate location in N. viridula.     

The effect of vibratory disturbance on sexual behaviour of the southern green stink bug Nezara viridula (Heteroptera, Pentatomidae)

The effect of vibratory disturbance on sexual behaviour and substrate-borne sound communication of the southern green stink bug, Nezara viridula L. was studied. Disturbance signals do not change the time N. viridula males need to locate the source of vibratory signals, but decrease the number of males responding with the calling and courtship song to calling females. Female N. viridula proceed calling during stimulation with disturbance signals but some of them change the song rhythm by skipping one or more signal intervals or emitting the repelling signals. The number of females which change the dominant frequency of the calling song decreases proportionally with increasing differences between the dominant frequency of the disturbance signals and the emitted female calling song. Variation of the song dominant frequency probably serves females to avoid interference by increasing the signal to noise ratio. Signal duration and repetition rate do not change significantly when the female is stimulated with the disturbance signals. This indicates that frequency shift by calling females is the main strategy for reducing interference by competitive signalers in N. viridula vibrational communication.     

Localization of selected pathogens of cotton within the southern green stink bug

Southern green stink bugs, Nezara viridula (L.) (Hemiptera: Pentatomidae), are pests of cotton recently shown to ingest, retain, and introduce some pathogens of cotton into bolls. The objective of this study was to determine where pathogen colonization occurs in N. viridula after ingestion. Laboratory‐reared adult N. viridula were fed sterile green beans soaked in water (control) or beans previously soaked in a suspension of one of three opportunistic bacterial pathogens [Pantoea agglomerans (Ewing & Fife), Pantoea ananatis (Serrano) Truper & De’Clari, or Klebsiella pneumoniae (Schroeter)] or a yeast [Nematospora coryli (Peglion)]. The insect rostrum, head, and alimentary canal were subsequently processed to determine the presence of pathogenic organisms. Overall, the alimentary canal exhibited significantly more bacterial colony forming units per g tissue (mean ± SEM = 4 806.3 ± 397.2) than the head (443.4 ± 397.2) and rostrum (46.0 ± 397.2); concentrations in the head and rostrum did not differ significantly. All four pathogens were detected in the alimentary canal, but only P. agglomerans and N. coryli were detected in the rostrum and head. Although P. ananatis and K. pneumoniae were only detected in the alimentary canal, their ranges of concentrations were similar to those of P. agglomerans and N. coryli in this tissue. Non‐selective media indicated that other bacterial fauna was also detected in all study insects. Thus, N. viridula does not discriminate with regard to ingestion of pathogens. The observed distribution pattern of tested bacteria and yeast in certain tissues of N. viridula clarifies the propensity for transmission of P. agglomerans and N. coryli. Results are discussed in relation to future research avenues examining selective colonization of certain pathogens and frequency of host infection by adults harboring the pathogenic organisms.     

Prevalence,damage, management and insecticide resistance of stink bug populations (Hemiptera: Pentatomidae) in commodity crops

1. Pest management of stink bugs (Hemiptera: Pentatomidae) in soybean [Glycine max (L.) Merr.], corn (Zea mays L.) and cotton (Gossypium spp.) agroecosystems has become a major concern in several countries of the Americas.
2. In this review, we report an overview on geographical distribution, injury, damage and methods used to control (plant resistance mechanisms, biological control) the most important stink bugs in the Americas, with an emphasis on Brazil, the implications of the trend towards decreased susceptibility of stink bug populations to insecticides and the current difficulties of the management of these insect pests.
3. Currently, the Neotropical brown stink bug Euschistus heros (Fabricius) is less susceptible to organophosphate insecticides than in the past. A slight reduction in E. heros susceptibility to pyrethroids and, to a lesser extent, to neonicotinoids has also been observed. In addition, the green‐belly stink bug [Dichelops melacanthus (Dallas)] is more tolerant to the three classes of insecticides (neonicotinoids, organophosphates and pyrethroids) than E. heros.
4. Metabolic detoxification is involved in organophosphate, neonicotinoid and pyrethroid differences in susceptibility. Restricted availability of insecticides with different modes of action could favour the selection of resistant phenotypes in stink bug populations.

     

Chemical cues from Spodoptera litura larvae elicit prey-locating behavior by the predatory stink bug, Eocanthecona furcellata

Two chemical compounds eliciting prey-locating behavior of the predatory stink bug, Eocanthecona furcellata, were isolated from solvent extracts of Spodoptera litura larvae and identified. A hexane eluted fraction from silica gel chromatography of the solvent extracts of S. litura larvae was attractive to E. furcellata nymphs when assayed with a linear track olfactometer. The hexane fraction was found to contain n-pentadecane (2500 ng/larva), n-tetracosane (54 ng/larva), n-heptadecane (41 ng/larva), n-heptacosane (61 ng/larva), n-nonacosane (147 ng/larva), n-hentriacontane (200 ng/larva) and squalene (323 ng/larva). The synthetic n-pentadecane was attractive to the bugs, although its activity was slightly lower than that of the hexane fraction. A mixture of synthetic n-pentadecane and the other six hydrocarbons was also attractive, although no improved attractiveness was observed by the addition of the latter compounds. On the other hand, a 15%-ether-in- hexane eluted fraction from silica gel chromatography stimulated bugs to display a proboscis-protruding behavior. A neutral-layer fraction of the 15% fraction, which contained E-phytol (480 ng/larva), also elicited this behavior. Synthetic E-phytol had the same effect on the predators as the neutral-layer fraction.