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.     

Oviposition and development of the tarnished plant bug (Heteroptera: Miridae) on field maize

Reduced insecticide use in cotton, Gossypium hirsutum L., as a consequence of the Boll Weevil Eradication Program and the broad adoption of Bt cotton, have helped make the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), a consistent pest of cotton each year in the mid-south. Maize, Zea mays L., has been implicated as having a role in the season-long dynamics of tarnished plant bug infestations in cotton. To date, no published information exists describing the quality of maize as a host for tarnished plant bug. No-choice field studies indicated that adult tarnished plant bug females oviposited into maize leaves, tassels, and ears. Laboratory studies showed that first-instar tarnished plant bugs could successfully develop to the adult stage when fed maize silks at the R1 growth stage, tassels before (VT) and during (R1) pollen shed, and milk stage (R3) kernels from the tip and base of the ear. The proportion of nymphs surviving to the adult stage on these tissues was often similar to that of broccoli, Brassica oleracea L. Nymphs did not develop to adults when fed V5 or R1 maize leaves. However, survival of first instars to the adult stage was improved when nymphs fed on tassels with pollen for 6 d and then moved to silks or leaves. Another field study showed that tarnished plant bugs reproduced in maize mainly during the tassel (VE and VT) and the R1-R3 ear growth stages, and a single new generation was produced in maize during these stages. The highest population recorded during the study (24 June 2005) consisted mostly of nymphs and was estimated to be 29,600/ha (12,000/acre). These studies showed that maize is a suitable host for tarnished plant bug reproduction and development, and its production plays a significant role in the population dynamics of the tarnished plant bug in the mid-south.     

Injury to cotton by adult Lygus hesperus (Hemiptera: Miridae) of different gender and reproductive states

Lygus hesperus Knight (Hemiptera: Miridae) is a key pest of cotton (Gossypium spp.) in the western United States that injures floral buds (squares) and developing fruit (bolls). However, no clear relationship between Lygus population level and plant injury has been established. Age-dependent feeding activity by L. hesperus is a possible source of variation that has not been examined for its influence in studies of the impact of Lygus on cotton. Recent video-based laboratory studies indicated that feeding behaviors and trivial movement varied among L. hesperus adults of different gender and reproductive states (prereproductive; reproductive and unmated; and reproductive and mated). We compared within-plant distributions and accumulations of feeding injury to intact cotton plants corresponding to adult L. hesperus of different gender and reproductive states. Adult females, regardless of reproductive state, were observed on squares and axillary buds more often than were males. Additionally, prereproductive adults were observed on squares and axillary buds more often than were mated or unmated reproductive adults, regardless of gender. Plants that were exposed to prereproductive adults exhibited more abscised squares and more squares with injured anthers compared with plants exposed to reproductive adults. However, feeding injury did not differ by insect mating status or gender. These results are consistent with results of our previous video-based assays, and indicate adult reproductive state represents a source of variation that should be controlled in studies to evaluate Lygus-induced injury to cotton and other crop plants.     

Comparison of direct and indirect sampling methods for tarnished plant bug (Hemiptera: Miridae) in flowering cotton

A complex of hemipterans, especially the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois) (Hemiptera: Miridae), has become a major target of insecticides in flowering cotton, Gossypium hirsutum L., in the mid-southern United States. Sampling protocols for this complex during this period of cotton development are poorly established, resulting in uncertainty about when infestations warrant treatment. Nine direct and indirect sampling methods were evaluated for bias, precision, and efficiency in cotton throughout the Mid-South during 2005 and 2006. The tarnished plant bug represented 94% of the bug complex in both years. Sweep-net and black drop-cloth methods were more efficient than other direct sampling methods, but they were biased toward adults and nymphs, respectively. Sampling dirty blooms was the most efficient indirect sampling method. The sweep-net, whole-plant, and dirty-bloom methods were more accurate than the other sampling methods evaluated based on correlations with other sampling methods. Variability attributed to the person collecting the sample was significant for all sampling methods, but least significant for the dirty-square method. Further research is needed to establish thresholds based on sweep-net, drop-cloth, dirty-square, and dirty-bloom sampling methods as these methods provide the best combinations of accuracy and efficiency for sampling tarnished plant bugs in cotton.     

State-dependent sampling bias in insects: implications for monitoring western tarnished plant bugs

Insect populations vary in the proportion of individuals exhibiting a particular ‘state’ (e.g., developmental stage, sex, egg load, or nutritional status). Because an insect's developmental state often determines both its size and its behavior, it is likely that this will also affect the probability of being sampled. We propose that a comprehensive approach to pest management must consider the interaction between the structure of a pest population (i.e., the relative number of individuals in each state) and any state‐dependent sampling bias. To illustrate the usefulness of this method we sampled populations of the western tarnished pest bug, Lygus hesperus Knight (Heteroptera: Miridae), in cotton fields. Our sampling technique utilized large cages to measure the absolute densities of each L. hesperus stage and adult sex within a population. This technique allowed us to document a wide range of absolute stage structures and sex ratios across 10 L. hesperus populations in California. Using a combination of cage samples and sweep net samples, we quantified the state‐dependent sampling bias by calculating the efficiency of sweep sampling as a function of L. hesperus developmental stage and sex. We found that the efficiency of sweep nets increased steadily with each successive developmental stage (i.e., nymphal instar) of L. hesperus. We also found that sweep nets are slightly more efficient in capturing male vs. female L. hesperus adults. Since other studies have documented that the stage and sex of L. hesperus can affect feeding impact on cotton flower buds, our results suggest that accurate predictions of Lygus damage will need to incorporate stage and sex‐dependent sampling biases.     

Control of western tarnished plant bug Lygus hesperus Knight (Hemiptera: Miridae) in California organic strawberries using alfalfa trap crops and tractor-mounted vacuums

A key economic pest of strawberries in California is the western tarnished plant bug, Lygus hesperus Knight (Hemiptera:Miridae). Alfalfa (Medicago sativa L.) is a highly attractive plant host to western tarnished plant bug, and we hypothesized that it can be successfully managed as a trap crop for pest suppression in strawberries. Completely randomized design trap cropping experiments were established on an organic strawberry farm from 2002 to 2004. Western tarnished plant bug adults and nymphs were significantly more abundant in alfalfa trap crops than in comparable edge strawberry rows. Over 3 experimental yr, twice-weekly summer vacuuming of alfalfa trap crops with a tractor-mounted vacuuming device reduced adult and nymph abundance by 72 and 90%, respectively, in trap crops. This summer vacuuming of alfalfa trap crops also significantly reduced damage caused by western tarnished plant bug in associated unvacuumed organic strawberries (June and July 2002, June 2003, and June and July 2004) compared with either an untreated control (2003) or the organic strawberry grower's standard whole field vacuuming treatment. Vacuuming of alfalfa trap crops reduces an organic grower's costs (tractor, tractor fuel, and driver time) by 78% compared with current whole field vacuuming practices. An economic analysis of a whole hectare model indicates that a positive return from the use of vacuumed trap crops could be realized in 2004. The overall potential positive net return for the 3 mo of vacuumed alfalfa trap crop treatments in 2004 was calculated at +$1,829/ha.     

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.     

Temperature dependent development of Lygus hesperus (Hemiptera: Miridae) nymphs

Lygus hesperus Knight (Hemiptera: Miridae) is a key pest of fruit and vegetable crops, forages, and cotton (Gossypium spp.) in the western United States. Accurate models describing relationships between temperature and L. hesperus development are critical to the study of seasonal L. hesperus population dynamics. Development of L. hesperus nymphs was assessed at nine constant temperatures from 10 to 37.8 degrees C. The relationships between temperature and development for each L. hesperus instar, and for the entire nymphal stage, were best described by six-parameter biophysical models indicating both low- and high-temperature inhibition of development. Development rates asymptotically approached zero with decreasing temperature in the lower thermal range, and decreased with increasing temperatures above 32.2 degrees C. Nymphs did not survive from egg hatch to adulthood at either 10 or 37.8 degrees C, and nymph mortality was > 90% at both 12.8 and 35.0 degrees C. The fifth instar exhibited the longest stadium, whereas the shortest stadia were associated with the second and third instars. Development rates of males and females did not differ, and the ratio of males to females was not different from 1:1 at any temperature. Our temperature-dependent development rate models for L. hesperus nymphs will facilitate control of insect physiological age in controlled laboratory experiments, and should be useful in planning and interpreting field studies on L. hesperus population dynamics.     

Host plants of the tarnished plant bug (Heteroptera: Miridae) in Central Texas

The tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), has taken on added importance as a pest of cotton in the Cotton Belt after successful eradication efforts for the boll weevil (Anthonomus grandis grandis Boheman). Because the Southern Blacklands region of Central Texas is in advanced stages of boll weevil eradication, blooming weeds and selected row crops were sampled during a 3-yr study to determine lygus species composition and associated temporal host plants. L. lineolaris was the sole lygus species in the region. Thirteen previously unreported host plants were identified for L. lineolaris, of which 69% supported reproduction. Rapistrum rugosum L. Allioni and Ratibida columnifera (Nuttall) Wooton and Standley were primary weed hosts during the early season (17 March to 31 May). Conyza canadensis L. Cronquist variety canadensis and Ambrosia trifida L. were primary weed hosts during the midseason (1 June to 14 August) and late-season (15 August to 30 November), respectively. Sisymbrium irio L. and Lamium amplexicaule L. sustained L. lineolaris populations during the overwintering period (1 December to 16 March). The proportion of females and numbers of nymphs found in R. rugosum, C. canadensis, A. trifida, and S. irio suggests these weeds supported reproductive adults during the early, mid-, and late season and overwintering period, respectively. Medicago sativa L. was the leading crop host for L. lineolaris; Glycine max L. Merrill did not yield L. lineolaris. Few L. lineolaris were collected in Gossypium hirsutum L. These results provide a more comprehensive assessment of host plants contributing to L. lineolaris populations in central Texas.     

Lygus hesperus （Heteroptera： Miridae） tolerates high concentrations of dietary nickel

Nickel hyperaccumulator plants contain unusually elevated levels of Ni （〉 1 000 μg Ni/g）. Some insect herbivores, including Lygus hesperus （Western tarnished plant bug）, have been observed feeding on the California Ni hyperaccumulator Streptanthus polygaloides. This bug may be able to utilize S. polygaloides as a host either through its feeding behavior or by physiological tolerance of Ni. This experiment determined the Ni tolerance of L hesperus by offering insects artificial diet amended with 0, 0.4, 1, 2, 4.5, 10, 20 and 40 mmol Ni/L and recording survival. Survival varied due to Ni concentration, with diets containing 10 mmol Ni/L and greater resulting in significantly lower survival compared to the control （0 mmol Ni/L） treatment. Insects tolerated diet containing as much as 4.5 mmol Ni/L, a relatively elevated Ni concentration. I conclude that L hesperus can feed on S. polygaloides because it is Ni-tolerant, probably due to physiological mechanisms that provide it with resistance to plant chemical defenses including elemental defenses such as hyperaccumulated Ni.     

Physiological consequences of laboratory rearing of Lygus hesperus (Hemiptera: Miridae)

Several aspects of the basic biology of the western tarnished plant bug, Lygus hesperus Knight, are poorly known despite the economic importance of this species. Among these are the factors regulating the adult diapause. Reports of recent studies questioned the validity of earlier reports of diapause in L. hesperus, in part because of the demonstrated loss of diapause response in insects obtained from long-standing laboratory colonies. However, use of laboratory reared insects would facilitate additional diapause research, so long as those insects exhibit a diapause response similar to that of the field population. L. hesperus, originating as eggs from field-collected insects, were reared in the laboratory for four generations to examine corresponding changes in selected biological characteristics. Over the course of the four generations, incidence of diapause in both L. hesperus genders decreased whereas the frequency of oviposition by virgin females increased. Measurable changes were not observed in frequency of occurrence of a specific fat body type (glass bead fat) or nymphal development time. These results suggest L. hesperus used in diapause research should be as close to the field population as possible, but no further removed than three generations. Results further demonstrate variability among different biological characteristics in their responses to selection from laboratory rearing. Collectively, these findings demonstrate the importance of understanding the influences of rearing on specific biological characteristics under study, and the need to verify the similarity of laboratory-reared insects to their native counterparts in studies used to draw inferences regarding the field population.     

Olfactory Responses of <Emphasis Type="Italic">Anaphes iole</Emphasis> (Hymenoptera: Mymaridae) to Volatile Signals Derived from Host Habitats

Anaphes iole Girault is a frequent parasitoid of Lygus spp. eggs in the United States, and has potential as a biological control agent against Lygus hesperus Knight in different crops. Feeding and oviposition by L. hesperus induce emission of plant volatiles, but studies to date do not address the role of plant volatiles in the host-searching behavior of A. iole. In this study, a four-arm olfactometer was used to test the responses of female parasitoids to odors emanating from cotton (Gossypium hirsutum L., Malvaceae) plants damaged by L. hesperus females, L. hesperus males, larvae of the nonhost Spodoptera exigua Hubner, or mechanically, or to odors from L. hesperus females alone. In addition, various plants damaged by L. hesperus females were evaluated in the olfactometer: cotton, alfalfa (Medicago sativa L., Fabaceae), common groundsel (Senecio vulgaris L., Asteraceae), annual ragweed (Ambrosia artemisifolia L., Asteraceae), and redroot pigweed (Amaranthus retroflexus L., Amaranthaceae). In all olfactometry bioassays, treatment odors were compared against three controls (humidified air). Results showed that A. iole females were consistently attracted to odors derived from different plant–L. hesperus complexes, while odors from plants subjected to nonhost (S. exigua) or mechanical damage and L. hesperus females alone were not attractive or only variably attractive. These findings suggest that while searching for hosts A. iole females use specific volatiles induced by L. hesperus feeding and oviposition to locate hosts inhabiting a wide variety of plants, including annual and perennial species from four plant families. It was suggested that future research should seek to identify the chemical elicitors involved in the release of plant volatiles attractive to A. iole females.     

Evaluation of peracid activated organophosphates in studies of insecticide resistance conferred by insensitive acetylcholinesterases

Peracid activated organophosphorus (OP) insecticides were evaluated against electric eel and a range of insect acetylcholinesterases (AChE). Incubation of chlorpyrifos and sulprofos with meta-chloroperoxybenzoic acid (MCPBA) produced active metabolites capable of distinguishing between AChE enzymes conferring OP susceptibility and resistance in populations of the tobacco whitefly, Bemisia tabaci Gennadius, and the beet armyworm, Spodoptera exigua Hubner. These products were also active against enzymes present in the lygus bug Lygus hesperus Knight and the greenhouse whitefly, Trialeurodes vaporariorum Westwood. Although profenofos was active against electric eel AChE, incubation of the OP with MCPBA did not enhance its anti-AChE properties toward the eel or insect AChEs.     

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.     

Cotton plants expressing a hemipteran-active Bacillus thuringiensis crystal protein impact the development and survival of Lygus hesperus (Hemiptera: Miridae) nymphs

The plant bugs Lygus hesperus Knight (Hemiptera: Miridae) and L. lineolaris (Palisot de Beauvois) have emerged as economic pests of cotton in the United States. These hemipteran species are refractory to the insect control traits found in genetically modified commercial varieties of cotton. In this article, we report the isolation and characterization of a 35 kDa crystal protein from Bacillus thuringiensis, designated TIC807, which causes reduced mass gain and mortality of L. hesperus and L. lineolaris nymphs when presented in an artificial diet feeding assay. Cotton plants expressing the TIC807 protein were observed to impact the survival and development of L. hesperus nymphs in a concentration-dependent manner. These results, demonstrating in planta activity of a Lygus insecticidal protein, represent an important milestone in the development of cotton varieties protected from Lygus feeding damage.     

Development and population growth of Lygus hesperus on selected weed hosts, artificial diet and cotton in the laboratory

Laboratory studies were conducted to obtain basic biological information of Lygus hesperus Knight (Hemiptera: Miridae) on alfalfa (Medicago sativa L.), pigweed (Amaranthus palmeri S. Wats.), Russian thistle (Salsola iberica Sennen and Pau), green bean (Phaseolus vulgaris L.), artificial diet, and cotton (Gossypium hirsutum L.) square and cotton boll at 27 degrees C. The nymphal developmental duration was 28.8 d on cotton bolls, nearly twice longer, compared with approximately 15 d in other hosts. The nymphal survivorship was significantly lowest on the cotton boll (18%), whereas the survivorship in other hosts ranged from 33% (cotton square) to 93% (green bean). The average total nymphal survivorship was approximately 60% and the most nymphal mortalities occurred on the first three instars. The life-table parameters were obtained only in cotton square, artificial diet, green bean, and alfalfa. The average adult longevity in artificial diet was the shortest (18.7 d),with the longest longevity observed (36.1 d) on green beans. The intrinsic rates of population increase (r(m)) were much higher in artificial diet (0.0771) and green beans (0.0671) than in alfalfa (0.0327) and cotton square (0.0317). Although statistically significant, the difference in r(m) values between alfalfa and cotton square was very small, indicating the similarity in reproductive suitability of cotton square and alfalfain a no-choice test. Thus, even though cotton is not a preferred host, when blooming alfalfa and roadside weeds are mowed in the Texas High Plains during June-July, it is the most likely time that Lygus bugs may move from alfalfa and other host plants into adjacent cotton fields and become effective pest of actively fruiting cotton.     

Characterization of digestive proteolytic activity in Lygus hesperus Knight (Hemiptera: Miridae)

The tarnished plant bug, Lygus hesperus Knight, is a pest that causes considerable economic losses to vegetables, cotton, canola, and alfalfa. Detailed knowledge of its digestive physiology will provide new opportunities for a sustainable pest management approach to control this insect. Little is known about the different protease class contributions to the overall digestion of a specific protein. To this end, the proteolytic activities in female adult L. hesperus salivary gland and midgut homogenates were quantified over a range of pH's and time points, and the contribution of different classes of proteases to the degradation of FITC-casein was determined. In the salivary gland, serine proteases were the predominant class responsible for caseinolytic activity, with the rate of activity increasing with increasing pH. In contrast, both aspartic and serine proteases contributed to caseinolytic activity in the midgut. Aspartic protease activity predominated at pH 5.0 and occurred immediately after incubation, whereas serine protease activity predominated at pH 7.5 after a 9h delay and was resistant to aprotinin. The salivary serine proteases were distinctly different from midgut serine proteases, based on the tissue-specific differential susceptibility to aprotinin and differing pH optima. Collectively, the caseinolytic activities complement one another, expanding the location and pH range over which digestion can occur.     

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.     

Field evaluation of efficacy and persistence of an insect vacuum device against the tarnished plant bug (Hemiptera: Miridae) in a day-neutral strawberry field

The efficacy of a vacuuming device to reduce Lygus lineolaris (Palisot de Beauvois) populations was evaluated over 3 yr in a day-neutral strawberry field. Immediately after treatments, a significant reduction of population estimates (by tapping of flower clusters) was observed for adults (75% of the time) and nymphs (50% of the time). Under the same conditions, control (i.e., vacuum turbine off) treatments significantly reduced adult population estimates 25% of the time. Lygus lineolaris tapping samples taken from strawberry plants adjacent to the treated zone did not show significant variations over sampling time, suggesting that no escape behavior occurred. This was supported by adult catches on sticky traps located in zones adjacent to the treated one. Our results suggest that vacuuming had inconsistent effect on tarnished plant bug populations and that L. lineolaris mobility plays a marginal role in the use of this control method.