Characterization of eight polymorphic microsatellite markers in the tarnished plant bug,Lygus lineolaris (Palisot de Beauvois)

A partial genomic library of the tarnished plant bug, Lygus lineolaris, enriched for microsatellite sequences was screened to identify marker loci. Eight polymorphic loci suitable for population genetic studies were identified by screening 192 field‐collected insects. The observed number of alleles ranged from four to 21 with an average of 12.25 (SE ± 1.94) while the effective number of alleles ranged from 1.23 to 11.05 with an average of 4.49 (SE ± 1.15). No linkage disequilibria or significant deviations from Hardy–Weinberg expectations were detected at any of the loci. Seven of the eight L. lineolaris microsatellite loci were transferable to Lygus hesperus.     

Changes in abundance of native and introduced parasites (Hymenoptera: Braconidae), and of the target and non-target plant bug species (Hemiptera: Miridae), during two classical biological control programs in alfalfa.

High numbers of tarnished plant bugs [Lygus lineolaris (Palisot)], were once common in alfalfa, as was a low level of parasitism (9%) by the native Peristenus pallipes (Curtis). After the bivoltine European parasite Peristenus digoneutis Loan became well established, average parasitism of the first and second generations increased to 64%, and tarnished plant bug numbers dropped by 65%. This reduced host density eventually caused a decline in total parasitism by both parasite species to 22%. A few P. digoneutis also attacked the alfalfa plant bug, Adelphocoris lineolatus (Goeze), but did not reduce this pest or increase its parasitism rate. At another location, where P. digoneutis is not established, parasitism of first generation alfalfa plant bugs, an adventive (accidently introduced) pest, was increased to 21% by the introduced univoltine parasite, Peristenus conradi Marsh, and a slight reduction in the pest may have resulted. P. digoneutis did not parasitize the meadow plant bug, Leptopterna dolabrata (L.), an adventive pest of forage grasses, so did not affect this mirid or its parasite. Neither introduced parasite eliminated the native parasites of the tarnished or alfalfa plant bugs. The narrow host ranges of the braconid parasites of mirid nymphs are contrasted with the broad host range of the native tachinid parasite [Phasia robertsoni (Towns.)] of adult mirids. The major changes in mirid abundance and their mortality by parasites that slowly occurred during this 19-year study demonstrate the need for long-term field research, to adequately document and understand these complex interactions.     

Efficiency of volatile baited sticky traps for the Tarnished Plant Bug (Lygus lineolaris) in strawberry fields

Insecticides have adverse effects on human health and the environment. Thus, the development of non-chemical replacement to manage insect pests is urgent. An alternative is to bait sticky traps with attractive insect sex pheromones or plant volatiles. In Quebec, the tarnished plant bug (Lygus lineolaris) is a major insect pest. We tested the efficiency of sex pheromones (mixture of hexyl butyrate (HB), (E)-2-hexenyl butyrate (E2HB) and (E)-4-oxo-2-hexenal (KA)) and sunflower (Helianthus annuus) volatiles (pinene, sabinene and phenylacetaldehyde) as sticky trap baits for the tarnished plant bug in strawberry fields of the Laurentians in Southern Québec, Canada. The pheromones decreased the number of tarnished plant bug caught in the traps compared to a control. The sunflower volatile did not have any effect on the number of individuals caught in the traps. Different Lygus species use HB, E2HB and KA in different ratios for sexual signals and alarm signals and for species recognition. GC-MS analysis of the sex pheromone bait revealed that the ratios between the three main compounds did not match the intended ratio for the L. lineolaris species. This mismatch probably explains our results. Individuals were not attracted to sunflower volatiles. Our results point to the difficulty of manufacturing and using sex pheromones as baits. Future work should assess the effect of several pheromone ratios.     

Biological control of Lygus lineolaris by Peristenus spp. in strawberry

Peristenus digoneutis Loan (Hymenoptera: Braconidae) was introduced to the US for biological control of the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois) (Hemiptera: Miridae), and has since spread through much of the northeast. The purpose of this study was to determine if P. digoneutis and a native congener, Peristenus pallipes (Curtis), parasitize L. lineolaris in strawberry (where it is a key pest), and what factors relate to parasitism levels. During 1997–1999 we monitored parasitism on 17 strawberry farms in 14 counties in eastern and western New York State. We found that in eastern NY (where P. digoneutis has been established since the early 1990s), overall mean parasitism was 19.7% (ranging from 0 to 70%), mostly by P. digoneutis. Mean parasitism was significantly lower (12.3%, ranging from 0 to 58%) in western NY (where P. digoneutis was first recorded in 1999), and was mostly by P. pallipes. P. pallipes parasitism was significantly lower in eastern than western NY, suggesting the potential for competitive interaction with P. digoneutis. The insecticide regime of a farm was an important factor influencing parasitism rate, which was 5- to 6.5-fold higher on organic or casually sprayed farms than on intensely treated farms, though pest density under these three regimes was not significantly different. L. lineolaris density, and parasitism rate in nearby alfalfa and abandoned fields were also significant factors for parasitism in strawberry.     

RNA interference‐mediated knockdown of IAP in Lygus lineolaris induces mortality in adult and pre‐adult life stages

In recent years, RNA interference (RNAi) has been validated as a viable approach for functional genetic studies in non‐model organisms. In this report we demonstrate the efficacy of RNAi in the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois) (Miridae: Hemiptera). A L. lineolaris inhibitor of apoptosis gene (LlIAP) has been identified and cloned. The translated sequence encodes a 381 amino acid protein similar to other insect IAPs and contains two conserved baculovirus inhibitor of apoptosis protein repeat (BIR) domains. Microinjection of double stranded RNA (dsRNA) corresponding to two disparate portions of the gene resulted in decreased LlIAP mRNA quantities relative to controls. Both nymphs and adult specimens injected with IAP dsRNA exhibited significantly reduced lifespan compared with those injected with non‐insect dsRNA (eGFP). Thus, RNAi‐mediated knockdown of LlIAP expression has been correlated with a lethal phenotype in adults and nymphs.     

Effects of different Beauveria bassiana isolates on field populations of Lygus lineolaris in pigweed (Amaranthus spp.)

The tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), is a pest of various fruit, vegetable, fiber, and seed crops; including cotton. Lygus spp. populations often build on alternate host plants before moving to cotton, and in the midsouthern U.S. wild host plants, such as pigweed (Amaranthus spp.), play a major role in L. lineolaris population development. Three isolates of the entomopathogenic fungus Beauveria bassiana (Balsamo) were evaluated for L. lineolaris control in redroot pigweed (Amaranthus retroflexus L.): one from L. lineolaris in Mississippi (TPB3); one from Lygus hesperus (Knight) in California (WTPB2); and one commercial isolate from Mycotrol^® (GHA). Fungal applications resulted in moderate to high mycosis in adults (33 to 80%) and moderate mycosis in nymphs (36 to 53%) that were collected from field plots at 2 days post-treatment and incubated under laboratory conditions. Although TPB3 was previously found to be more pathogenic in laboratory bioassays, there was not a consistent separation of this isolate from the other two isolates in field trials. Where differences in adult mycosis or mortality were observed, TPB3 was the most pathogenic. However, in one field trial 7 day mortality for nymphs treated with GHA was higher than those treated with TPB3 or WTPB2. Infection rates at 2, 7, and 14 days post-treatment from caged and non-caged adults suggested that movement of adults among plots occurred, which could have masked some treatment effects. Fungal treatments did not significantly reduce populations relative to controls. This may have been caused by delayed mortality rates under field conditions and/or difficulties with estimating population change under field conditions characteristic of wild host plant populations (e.g., heterogeneous populations, adult movement, and small plot size). Further work evaluating time–dose–mortality over dynamic temperatures, spring and fall field trials on this and other wild hosts, and improved methods for estimating populations on wild hosts are needed.     

Local and landscape scale drivers of Euschistus servus and Lygus lineolaris in North Carolina small grain agroecosystems

1. Crop production sequences influence arthropod populations in temporally unstable row crop systems. Winter wheat (Triticum aestivum L.) represents one of the earliest abundant crops in south-eastern United States. This study aims to understand primary source habitats driving brown stink bug, Euschistus servus (Say), and tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), population abundance in wheat.
2. To better understand these relationships, adult and nymphal densities were in wheat fields weekly from flowering through harvest in 2019 and 2020. Geospatial data were used to measure landscape composition surrounding sampled fields. We investigated the influence of landscape predictors on E. servus and L. lineolaris abundance using generalized linear mixed modelling.
3. Field size, proportion of agriculture, proportion of wheat area, and proportion of soybean Glycine max L.) area from the previous year in the surrounding landscape were associated with E. servus abundance in wheat. Similarly, L. lineolaris abundance was associated with proportion of wheat area and soybean area from the previous year.
4. These results reveal the influence of soybean area planted the previous year on insect pest densities the following spring in wheat. Further, results suggest agricultural landscapes dominated by wheat are associated with decreased pest abundance across the sampled region.

     

Impact of Nosema (Microsporidia) infection and fumagillin treatment on Lygus lineolaris (Hemiptera: Miridae)

A consistent supply of healthy tarnished plant bug, Lygus lineolaris (Palisot de Beauvois) (Hemiptera: Miridae), is necessary for the development of novel management strategies targeting this pest. After being in culture for several years, a substantial portion of a tarnished plant bug colony was found to be infected with a Nosema (Microsporidia) species. Studies were subsequently undertaken to evaluate the impact of Nosema infection on tarnished plant bug productivity and to test the efficacy of fumagillin to treat this infection. Using buffalo black stain, infections could not be reliably detected in adult tarnished plant bugs until adults were 6-8days post eclosion. Nosema infections reduced adult longevity and fecundity. Maximum fecundity was restored using a concentration of 16.8ppm fumagillin while maximum longevity for females was at a concentration of 33.6ppm fumagillin incorporated into the tarnished plant bug diet. Minimum infection scores were obtained at 67.2ppm, the highest concentration tested. A field survey of tarnished plant bugs in Mississippi found Nosema infections in 3% of wild tarnished plant bugs.     

Bioassay method for assessing the virulence of Beauveria bassiana against tarnished plant bug, Lygus lineolaris (Hem., Miridae)

A standard bioassay method for assessing the pathogenicity of Beauveria bassiana (Balsamo) Vuillemin (GHA strain) against second instar tarnished plant bug, Lygus lineolaris (Palisot de Beauvois) (Hem., Miridae) was developed. Several types of inoculation methods, assay containers and incubation times were tested. Our goal was to minimize control mortality and maximize treatment mortality. Five inoculation methods (immersing broccoli florets or bean pods, spraying broccoli florets or bean pods, and immersing insects) and four types of plastic containers (114‐, 171‐, 228‐ and 455‐ml) were tested. Immersing insects directly in a fungal suspension was the most effective inoculation method, which resulted in a treatment mortality of 70–81.3% at a concentration of 1 × 10⁷ conidia/ml. The 114‐ml plastic container was the most suitable assay container when 10 tarnished plant bug nymphs were treated together, resulting in a control mortality of only 6% 12 days after treatment. Within the first 6 days after treatment, 71.1% of the insects were killed, compared with a total mortality of 81.3% after 12 days. Nymphs infected with the test fungus changed colour from green to black. Mycelial outgrowth and sporulation on the cadavers demonstrated that most nymphs died of fungal infection. A total of 61.1 and 80.5% of the cadavers showed signs of mycelial outgrowth 9 days after death among those that were surface sterilized and those that were not, respectively.     

Identification of Lygus hesperus by DNA barcoding reveals insignificant levels of genetic structure among distant and habitat diverse populations

Background

The western tarnished plant bug Lygus hesperus is an economically important pest that belongs to a complex of morphologically similar species that makes identification problematic. The present study provides evidence for the use of DNA barcodes from populations of L. hesperus from the western United States of America for accurate identification.

Methodology/Principal Findings

This study reports DNA barcodes for 134 individuals of the western tarnished plant bug from alfalfa and strawberry agricultural fields in the western United States of America. Sequence divergence estimates of <3% reveal that morphologically variable individuals presumed to be L. hesperus were accurately identified. Paired estimates of F_st and subsequent estimates of gene flow show that geographically distinct populations of L. hesperus are genetically similar. Therefore, our results support and reinforce the relatively recent (<100 years) migration of the western tarnished plant bug into agricultural habitats across the western United States.

Conclusions/Significance

This study reveals that despite wide host plant usage and phenotypically plastic morphological traits, the commonly recognized western tarnished plant bug belongs to a single species, Lygus hesperus. In addition, no significant genetic structure was found for the geographically diverse populations of western tarnished plant bug used in this study.     

The development of sampling techniques for populations of the tarnished plant bug,Lygus lineolaris (hemiptera: Miridae)

Summary The relationship of ‘mean crowding’ to mean density showed that counts of nymphs and adults of the tarnished plant bug,Lygus lineolaris (Beauv.), on birdsfoot trefoil were aggregated and that the index of basic contagion approached zero for each developing stage. Aggregation increased gradually from the first to the fourth instar and then decreased through the adult stage. The mortality process from the first to the fifth instar was inversely density dependent. Inter-sample variance was the major source of population variance although significant variance was occasionally associated with blocks and plots owing to heterogeneity of the host stand. The most appropriate sample unit was a 3 sq. ft. area of foliage and substrate. This was divided into four quarters; application of the variance component technique indicated that in most cases one quarter drawn at random was optimal for sampling different stages. The number of samples required to attain a given level of precision varied inversely with population density. The cost function for sampling was determined for specified levels of precision.