Microbial community of predatory bugs of the genus Macrolophus(Hemiptera: Miridae)

Background

The predatory mirids of the genus Macrolophus are key natural enemies of various economically important agricultural pests. Both M. caliginosus and M. pygmaeus are commercially available for the augmentative biological control of arthropod pests in European greenhouses. The latter species is known to be infected with Wolbachia -inducing cytoplasmic incompatibility in its host- but the presence of other endosymbionts has not been demonstrated. In the present study, the microbial diversity was examined in various populations of M. caliginosus and M. pygmaeus by 16S rRNA sequencing and denaturing gradient gel electrophoresis.

Results

Besides Wolbachia, a co-infection of 2 Rickettsia species was detected in all M. pygmaeus populations. Based on a concatenated alignment of the 16S rRNA gene, the gltA gene and the coxA gene, the first is phylogenetically related to Rickettsia bellii, whereas the other is closely related to Rickettsia limoniae. All M. caliginosus populations were infected with the same Wolbachia and limoniae-like Rickettsia strain as M. pygmaeus, but did not harbour the bellii-like Rickettsia strain. Interestingly, individuals with a single infection were not found. A PCR assay on the ovaries of M. pygmaeus and M. caliginosus indicated that all endosymbionts are vertically transmitted. The presence of Wolbachia and Rickettsia in oocytes was confirmed by a fluorescence in situ hybridisation. A bio-assay comparing an infected and an uninfected M. pygmaeus population suggested that the endosymbionts had minor effects on nymphal development of their insect host and did not influence its fecundity.

Conclusion

Two species of the palaearctic mirid genus Macrolophus are infected with multiple endosymbionts, including Wolbachia and Rickettsia. Independent of the origin, all tested populations of both M. pygmaeus and M. caliginosus were infected with three and two endosymbionts, respectively. There was no indication that infection with endosymbiotic bacteria had a fitness cost in terms of development and fecundity of the predators.

     

Sampling strategies for square and boll-feeding plant bugs (Hemiptera: Miridae) occurring on cotton

Sampling methods for square and boll-feeding plant bugs (Hemiptera: Miridae) occurring on cotton, Gossypium hirsutum L., were compared with the intent to assess if one approach was viable for two species occurring from early-season squaring to late bloom in 25 fields located along the coastal cotton growing region of south Texas. Cotton fleaphopper, Pseudatomoscelis seriatus (Reuter), damages squares early-season and dominated collections using five sampling methods (approximately 99% of insects collected). A major species composition shift occurred beginning at peak bloom in coastal fields, when verde plant bug, Creontiades signatus Distant, represented 55-65% of collections. Significantly more cotton fleahoppers were captured by experienced samplers with the beat bucket and sweep net than with the other methods (30-100% more). There were more than twice as many verde plant bugs captured by experienced and inexperienced samplers with the beat bucket and sweep net than captured with the KISS and visual methods. Using a beat bucket or sweep net reduced sampling time compared with the visual method for the experienced samplers. For both species, comparing regressions of beat bucket-based counts to counts from the traditional visual method across nine cultivar and water regime combinations resulted in only one combination differing from the rest, suggesting broad applicability and ability to translate established visual-based economic thresholds to beat bucket-based thresholds. In a first look at sample size considerations, 40 plants (four 10-plant samples) per field site was no more variable than variation associated with larger sample sizes. Overall, the beat bucket is much more effective in sampling for cotton fleahopper and verde plant bug than the traditional visual method, it is more suited to cotton fleahopper sampling early-season when plants are small, it transitions well to sample for verde plant bug during bloom, and it performs well under a variety of soil moisture conditions and cultivar selections.     

Ecology and economic impact of two plant bugs (Hemiptera: Miridae) in commercial vineyards

The current study investigated the seasonal phenology, spatial distribution, feeding damage and economic impact of two plant bugs, Lygocoris inconspicuous Knight and Taedia scrupeus Say, in commercial vineyards. For both plant bugs, densities of nymphs were higher on vines located near the edge of woodlots rather than in the interior of vineyards, which may be attributed to the presence of wild vines and other alternate host-plants in wooded areas. Nymphs of both species fed on apical leaves and developing fruit clusters of vine shoots, initiating development after swelling of buds in the spring and reaching the adult stage when vines were in bloom. Confining high densities of L. inconspicuous (10 nymphs) on individual shoots early in the season resulted in significant reduction of the number of fruit clusters per shoot, even when feeding was restricted to short (7 d) duration; the average weight of fruit clusters, in contrast, was not affected to a large extent by feeding activity of nymphs. An experiment evaluating the impact of low density of L. inconspicuous (0-0.3 nymphs per shoot) indicated a marginally significant negative relationship between density of nymphs and average weight of fruit clusters. Control measures may be economically justified when population density exceeds a combined threshold of one nymph of either L. inconspicuous or T. scrupeus per 10 shoots of vines.     

Host plant use for oviposition by Trigonotylus caelestialium (Hemiptera: Miridae) and Stenotus rubrovittatus (Hemiptera: Miridae)

The main hosts and sites of oviposition for the two bugs, Trigonotylus caelestialium (Kirkaldy) (Hemiptera: Miridae) and Stenotus rubrovittatus (Matsumura) (Hemiptera: Miridae), that cause pecky rice were investigated in 24 poaceous plants. Nymphs of T. caelestialium emerged from both spikelets and leaf sheaths, while nymphs of S. rubrovittatus emerged almost exclusively from spikelets. Suitable plants for oviposition by T. caelestialium are Lolium multiflorum, Digitaria violascens and Hordeum murinum, while Poa annua, Anthoxanthum odoratum, Alopecurus aequalis and D. violascens were preferentially used by S. rubrovittatus. There was a greater difference in the number of nymphs emerging from different plants for S. rubrovittatus than for T. caelestialium. This difference may be because T. caelestialium can oviposit on leaf sheaths if the spikelets are not suitable for oviposition, whereas S. rubrovittatus only oviposits on spikelets. Although both bugs oviposited on spikelets, the internal oviposition sites were different. In D. ciliaris, T. caelestialium laid all eggs between the lemma of the first floret and the second floret, whereas S. rubrovittatus laid eggs almost exclusively inside the second floret. In contrast, in P. annua, T. caelestialium laid all eggs inside the florets, whereas S. rubrovittatus laid eggs both between and inside the florets.     

Host preferences of introduced and native parasites (Hymenoptera: Braconidae) of phytophagous plant bugs (Hemiptera: Miridae) in alfalfa-grass fields in the northeastern USA

During an eight-year field study at two widely-separated locations (NJ & DE, USA), seven species of phytophagous mirid plant bugs were found in alfalfa and alfalfa-grass fields grown for animal forage. Five species of parasites were reared from these mirids. Two parasite species introduced from Europe killed significantly higher proportions of nymphs of the two important mirid pests of alfalfa than did native parasites. The introduced Peristenus digoneutis Loan parasitized an average of 31% of first and second generation tarnished plant bugs, Lygus lineolaris, a native insect; and the introduced Peristenus conradi parasitized 22% of first generation alfalfa plant bugs, Adelphocoris lineolatus, an introduced species. In addition, Peristenus pallipes significantly parasitized nymphs of Trigonotylus caelestialium (43%) and Leptopterna dolabrata (37%); both mirids are immigrant species. Because the parasite P. pallipes significantly attacked only these two non-native mirids, and is present in Europe, it also may be an accidental introduction to North America. A native wasp, Leiophron uniformis, heavily parasitized (49%) the native garden fleahopper, Halticus bractatus. A third native species, P. pseudopallipes, occasionally parasitized a few Lygus lineolaris in alfalfa at one location. Two other mirids, Stenotus binotatus and Megaloceroea recticornis, both accidently-introduced grass-feeding species, were not parasitized by native or introduced species. It is noteworthy that the effective host ranges of all the parasites in alfalfa-grass fields were limited: four of the five parasite species significantly parasitized only one of the seven mirid species, and the other parasite significantly parasitized only two mirids. Activity of the four common parasites was correlated with the mirids' host plant: three species principally parasitized alfalfa-feeding mirids, and one species principally parasitized grass-feeding mirids.     

First record of the genus Adelphocorisella Miyamoto and Yasunaga (Hemiptera: Miridae) from Korea

Adelphocorisella lespedezae Miyamoto and Yasunaga, 1993 , which was previously known only from Japan, is reported from Korea for the first time and diagnosed. An image of its dorsal habitus as well as illustrations of the male genitalia are provided. The discovery of the mirid species also represents the first distributional record in Korea as well as the continental Palaearctic Region for the genus Adelphocorisella Miyamoto and Yasunaga, 1993 .     

Molecular phylogeny of the plant bugs (Heteroptera: Miridae) and the evolution of feeding habits

The first comprehensive cladistic analysis of Miridae, the plant bugs, is presented based on analysis of 3935 base pairs of mitochondrial (16S, COI) and nuclear (18S, 28SD3) DNA for 91 taxa in seven subfamilies. Data were analysed using maximum likelihood (ML), parsimony and Bayesian inference (BI) phylogenetic frameworks. The phylogenetic results are compared with previous hypotheses of higher relationships in the family using alternative hypothesis tests. A Bayesian relaxed molecular clock is used to examine divergence times, and ancestral feeding habits are reconstructed using parsimony and a Bayesian approach. Clades recovered in all analyses are as follows: Cimicomorpha, Miroidea and Miridae; Bryocorinae: Bryocorini; Stenodemini; Mirinae; Deraeocorinae (Clevinemini + Deraeocorini); Cylapinae; Isometopinae; Bryocorinae: Dicyphini; Orthotylini; Phylinae (Phylini + Pilophorini), and Phylinae as sister group to all the remaining mirid taxa. These results are largely congruent with former hypotheses based on morphological data with respect to the monophyly of various subfamilies and tribes; however, our results indicate that the subfamily Bryocorinae is not monophyletic, as the two tribes, Dicypini and Bryocorini, were separated in the phylogenetic results. Divergence time estimates indicate that the radiation of the Miridae began in the Permian; most genus‐level radiations within subfamilies began in the late Cretaceous, probably in response to the angiosperm radiation. Ancestral feeding state reconstructions based on Bayesian and parsimony inference were largely congruent and both reconstructed phytophagy as the ancestral state of the Miridae. Furthermore, the feeding habits of the common ancestors of Mirinae + Deraeocorinae, Bryocorinae + Cylapinae + Isometopinae + Orthotylinae, and the remaining taxa excluding Phylinae, were inferred as phytophagous. Therefore, at least three shifts from phytophagy or polyphagy to predation occurred within the Miridae. Additionally, based on the mirid host‐plant records, we discovered several trends, such as a strong relationship between host‐plant ranges and a facultative feeding habit. © The Willi Hennig Society 2011.     

Traumatic insemination in the plant bug genus Coridromius Signoret (Heteroptera: Miridae)

In traumatic insemination, males pierce females with hypodermic genitalia and ejaculate into the body cavity rather than into the genital tract. This has resulted in the evolution of female counter-adaptations in the form of paragenitalia to reduce the direct physical costs of mating. While rare in the animal kingdom, traumatic insemination is oddly prevalent in the true bug infraorder Cimicomorpha (Heteroptera), where it occurs in six families and is thought to have arisen twice. Here, we report the discovery of traumatic insemination and elaborate paragenital development in the plant bug genus Coridromius (Miridae), representing a third, independent emergence of traumatic insemination in this infraorder.     

Microbial community of predatory bugs of the genus <Emphasis Type="Italic">Macrolophus</Emphasis>(Hemiptera: Miridae)

BACKGROUND: The predatory mirids of the genus Macrolophus are key natural enemies of various economically important agricultural pests. Both M. caliginosus and M. pygmaeus are commercially available for the augmentative biological control of arthropod pests in European greenhouses. The latter species is known to be infected with Wolbachia -inducing cytoplasmic incompatibility in its host- but the presence of other endosymbionts has not been demonstrated. In the present study, the microbial diversity was examined in various populations of M. caliginosus and M. pygmaeus by 16S rRNA sequencing and denaturing gradient gel electrophoresis. RESULTS: Besides Wolbachia, a co-infection of 2 Rickettsia species was detected in all M. pygmaeus populations. Based on a concatenated alignment of the 16S rRNA gene, the gltA gene and the coxA gene, the first is phylogenetically related to Rickettsia bellii, whereas the other is closely related to Rickettsia limoniae. All M. caliginosus populations were infected with the same Wolbachia and limoniae-like Rickettsia strain as M. pygmaeus, but did not harbour the bellii-like Rickettsia strain. Interestingly, individuals with a single infection were not found. A PCR assay on the ovaries of M. pygmaeus and M. caliginosus indicated that all endosymbionts are vertically transmitted. The presence of Wolbachia and Rickettsia in oocytes was confirmed by a fluorescence in situ hybridisation. A bio-assay comparing an infected and an uninfected M. pygmaeus population suggested that the endosymbionts had minor effects on nymphal development of their insect host and did not influence its fecundity. CONCLUSION: Two species of the palaearctic mirid genus Macrolophus are infected with multiple endosymbionts, including Wolbachia and Rickettsia. Independent of the origin, all tested populations of both M. pygmaeus and M. caliginosus were infected with three and two endosymbionts, respectively. There was no indication that infection with endosymbiotic bacteria had a fitness cost in terms of development and fecundity of the predators.     

Evidence for aposematism in the plant bug Lopidea nigridea Uhler (Hemiptera: Miridae: Orthotylinae)

The biology of the plant bug Lopidea nigridea is described, with particular focus on whether this red and black species can be considered aposematic. Lopidea individuals were rejected after attack in 92 of 112 feeding trials involving three of their five most common arthropod predator species. Compared to another syntopic plant bug species, L. nigridea is aggregated in distribution, and individuals are sedentary in their basic daily habits. These life-history features, coupled with the ability to survive most attacks by arthropod predators in the feeding trials, suggest that L. nigridea is protected to some degree by an aposematic defence. How L. nigridea may acquire its distastefulness is discussed, with special attention to host plant relations in L. nigridea and in the genus Lopidea as a whole.     

Native braconid parasitism of the Tarnished plant bug (Hemiptera: Miridae) in Southern Ontario

Three agricultural regions in southern Ontario (London, Niagara and Guelph) were sampled weekly from May to September for Lygus spp. and their parasitoids in 1998, 1999 and only one region in 2000. Alfalfa was the primary crop sampled, including both “clean” and weedy alfalfa fields. Lygus lineolaris (Palisot de Beauvois) accounted for 99% of all Lygus spp. collected by sweepnet. Lygus lineolaris had two generations (mid-June and late July) on alfalfa with a partial third generation in early September which contributes to the over-wintering adults. In 1998 and 2000, an early first generation peak of Lygus nymphs was observed in mid-May on the weeds, chickweed and shepherd's purse. Overall rates of parasitism (from dissections) for native parasitoids were consistent from year to year and the means of the three regions in each growing season were below 11%. Both nymphs and adult Lygus were parasitized, with the highly mobile adults being a potential means of dispersing the parasitoids. In general weedy fields were more highly parasitized than fields of weed-free alfalfa. Six species of native braconid parasitoids were collected from L. lineolaris in southern Ontario (in decreasing order of occurrence): Peristenus pallipes (Curtis), Peristenus pseudopallipes (Loan), Leiophron lygivorus (Loan), L. solidaginis Loan, L. uniformis (Gahan), and Leiophron sp. near brevipetiolatus Loan. The large populations of Lygus and the low percent baseline parasitism in southern Ontario, particularly of second generation Lygus, support the need for introduction of a multivoltine parasitoid species in this region.     

Potential of using mineral oils for the control of the mosquito bugs Helopeltis theivora (Hemiptera: Miridae) in cashew plantations

This study investigated the influence of an nC24 mineral oil (MO) on some biological characteristics of a mosquito bug, Helopeltis theivora, reared on cashew under laboratory and greenhouse conditions. Aqueous emulsions of the oil were applied as sprays to eggs at four daily intervals after oviposition to assess impacts on egg hatch, and to cashew seedlings to assess adult longevity and female fecundity reared on the plants. Field applications were also carried out to assess the impacts of sprays on populations of the bug in a cashew orchard. The results of the laboratory study indicated that 1% (v/v) of the MO application reduced egg hatch when applied to eggs on cashew seedlings, in separate treatments, 1, 2, 3, and 4 days after oviposition (DAO). Egg mortality showed a negative linear correlation with egg maturity (DAO). The pre-oviposition period of adult females placed on sprayed cashew seedling, after deposits of 1% (v/v) sprays dried, was significantly prolonged. The longevity of males and females on dry oil-spray deposits was lower than on seedlings sprayed with water. The fecundity of females was also lower. Field sprays significantly reduced populations of the bug on mature cashew trees. Mixtures of MO and abamectin were more effective than oil alone. The results of the study indicate that mineral oils can be used to suppress populations of H. theivora in cashew plantations.     

Myrtlemiris,a new genus and new species of Australian plant bugs (Insecta: Heteroptera: Miridae): systematics,phylogeny and host associations

A new Australian genus of Orthotylinae, Myrtlemiris Cheng, Mututantri & Cassis gen.n. , is described, with nine included species described as new to science (M. agnew sp.n. , M. astartephila sp.n. , M. meanarra sp.n. , M. newmanensis sp.n. , M. rubrocuneatus sp.n. , M. russulatus sp.n. , M. silveirae sp.n. , M. tesselatus sp.n. , M. yalgoo sp.n. ). A phylogenetic analysis based on 39 morphological characters is presented for all Myrtlemiris species and 6 outgroup taxa. This analysis establishes Myrtlemiris as monophyletic, defined by a broad apophysis on the left paramere. A key to species is provided and diagnostic characters are illustrated. Myrtlemiris is endemic to southwest Western Australia. Host plants for Myrtlemiris species are near restricted to the myrtaceous tribe Chamelaucieae.     

Barcoding bugs: DNA-based identification of the true bugs (Insecta: Hemiptera: Heteroptera)

Background

DNA barcoding, the analysis of sequence variation in the 5′ region of the mitochondrial cytochrome c oxidase I (COI) gene, has been shown to provide an efficient method for the identification of species in a wide range of animal taxa. In order to assess the effectiveness of barcodes in the discrimination of Heteroptera, we examined 344 species belonging to 178 genera, drawn from specimens in the Canadian National Collection of Insects.

Methodology/Principal Findings

Analysis of the COI gene revealed less than 2% intra-specific divergence in 90% of the taxa examined, while minimum interspecific distances exceeded 3% in 77% of congeneric species pairs. Instances where barcodes fail to distinguish species represented clusters of morphologically similar species, except one case of barcode identity between species in different genera. Several instances of deep intraspecific divergence were detected suggesting possible cryptic species.

Conclusions/Significance

Although this analysis encompasses 0.8% of the described global fauna, our results indicate that DNA barcodes will aid the identification of Heteroptera. This advance will be useful in pest management, regulatory and environmental applications and will also reveal species that require further taxonomic research.     

The flight behaviour of cocoa-capsids (Hemiptera: Miridae)

The cocoa-capsid Distantiella theobroma flies from its canopy substrate spontaneously but when disturbed first falls 1.2 to 1.9 m before flying. Adults are capable of flight 18 to 20 hr after the final moult. Flight is directly sunwards when the sun is low, but when it is high the bugs fly upwards in spirals and zigzags. Flight is terminated by the sight of vertical objects. Mean flight speed is 3.1±0.5 m/sec at 23°. Potential range is at least 1.1 km in females, 2.3 km in males. The other cocoa-capsid, Sahlbergella singularis, differs in being markedly nocturnal in its dispersal flights. Males fly far more often than females.Other factors affecting flight are discussed and the implications of the flight potential for control methods are mentioned. No part of Ghana's cocoa is out of range of colonisation by either species.

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Résumé Le Capside du Cacao Distantiella theobroma s'envole spontanément et directement de son support, mais quand l'insecte est dérangé, il se laisse d'abord tomber et ne prend son vol qu'après une chute de 1,2 à 1,9 m. Les adultes sont capables de voler dans un délai de 18 à 20 h. après la dernière mue. Leur vol est orienté en direction du soleil quand celui-ci est bas sur l'horizon, mais s'il est haut l'insecte s'élève vers le ciel en effectuant des spirales et zig-zags. La vue de tout objet vertical provoque l'arrêt du vol et la pose sur ce support. La vitesse moyenne du vol est de 3,1±0,5 m/sec. à une température de 23°. Le rayon de vol est au moins de 1,1 km pour les femelles et de 2,3 km pour les mâles. L'autre punaise du cacao Sahlbergella singularis diffère de la précédente, ses vols de dispersion étant nettement nocturnes. Les mâles volent beaucoup plus souvent que les femelles.D'autres facteurs affectant le vol sont également discutés ainsi que les conséquences des potentialités de vol pour la mise au point de méthode de lutte. Toutes les zones du Ghana où se trouvent des plantations de cacao sont dans le périmètre de dispersion de chacune des deux espèces.

     

Comparative analysis of the reproductive biology of two Peristenus species (Hymenoptera: Braconidae), biological control agents of Lygus plant bugs (Hemiptera: Miridae)

Peristenus digoneutis Loan and Peristenus stygicus Loan, parasitoids of the European tarnished plant bug Lygus rugulipennis Poppius, are established in the United States for biological control of native North American Lygus species, and are being considered for deliberate release in Canada. High lifetime fecundity of parasitoids is considered a desirable attribute of biological control agents and therefore, an understanding of parasitoid reproductive biology is required. In the present study, the potential lifetime fecundity of both agents was compared under laboratory conditions to estimate the potential impact of Peristenus species on Lygus. Synovigenic P. digoneutis and P. stygicus females oviposited most actively in the first two weeks of their lifetime, with a maximum average daily oviposition rate after five days. The maximum number of eggs laid per day was 83 eggs for P. stygicus, and 36 eggs for P. digoneutis. P. digoneutis has an average potential lifetime fecundity of 385 ± 35 SE eggs produced over 22 ± 3 SE days. In contrast, P. stygicus females have a 50% higher mean potential lifetime fecundity reaching 782 ± 65 SE eggs over 28 ± 1 SE days. A positive correlation between lifetime fecundity and body size was found only for P. stygicus, and both species showed a significant relationship between lifetime fecundity and oviposition period. The present study demonstrates that the fecundity of P. digoneutis and P. stygicus is considerably higher than previously reported. Based on these findings, P. stygicus appears to be the most effective biological control agent for Lygus lineolaris (Palisot de Beauvois) when only fecundity is taken into consideration.