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A Chromosome-Level Genome Assembly of the Potato Leafhopper Empoasca fabae (Hemiptera: Cicadellidae).

The potato leafhopper, Empoasca fabae (Harris, 1841), is a highly polyphagous, migratory insect pest of eastern North America that feeds on more than 200 herbaceous and woody plant species, causing substantial losses to forage and field crops. Despite its agricultural and ecological importance, no genome has been available for this species. Here, we present the first chromosome-level genome assembly of E. fabae, generated from Oxford Nanopore long reads, Illumina short reads, and Omni-C proximity-ligation data. The final assembly spans 908 Mb across 132 scaffolds, with 99.8% of the assembly captured in ten chromosome-length scaffolds (nine autosomes and an X chromosome) with a scaffold N50 of 96.2 Mb. The assembly is highly complete, recovering 92.9% of conserved hemipteran single-copy orthologs from protein annotations, and is composed of 47.6% repetitive sequence, dominated by long terminal repeat retrotransposons and unclassified elements. Read-depth comparison between male and female individuals supports assignment of a single sex-linked chromosome, consistent with an XO sex determination system. BRAKER3 gene annotation predicted 31,406 protein-coding genes after retaining the longest isoform per locus. Comparative genome analysis of the two closest related Typhlocybinae species with genomes available, Matsumurasca onukii and Hebata decipiens, revealed extensive chromosome-scale collinearity while defining a shared core gene repertoire. This reference genome provides a foundation for comparative and population genomic studies and for investigating genetic traits in this economically important crop pest species.

Animals↗

Glandular trichomes of Solanum berthaultii and its hybrids with Solanum tuberosum affect nymphal emergence, development, and survival of Empoasca fabae (Homoptera: Cicadellidae).

The influence of foliar glandular trichomes on developmental biology of the potato leafhopper, Empoasca fabae (Harris) (Homoptera: Cicadellidae), was examined by selective removal of trichomes on resistant and susceptible potato germplasm comprising two glandular trichome-bearing genotypes (PI 473331 and PI 473334) of wild potato, Solanum berthaultii (Hawkes), two interspecific hybrids of S. berthaultii with Solanum tuberosum L. (Q174-2 and NY123), and two commercial S. tuberosum cultivars ('Elba' and 'Allegany'). Adult mortality was affected, whereas nymphal emergence was unaffected by removal of glandular trichome exudates. No nymphs were observed emerging from leaves of PI 473331 or PI 473334. Regardless of the trichome removal treatment, no nymphs completed development on PI 473331, and only a small percentage of nymphs survived to adulthood on NY123 and PI 473334. PI 473331 had the greatest frequency of adults and nymphs with trichome exudate deposition on their bodies, followed by PI 473334. Nymphs, but not adults, had exudate accumulation when caged on foliage of Q174-2. Removal of type A trichome heads and type B droplets eliminated exudate deposition. The results of these studies provide evidence that phenolic oxidation chemistry and the physical barrier imposed by trichomes of S. berthaultii are responsible for much of the observed resistance but that other factors possibly linked to the presence of trichomes aid in the expression on resistance.

Animals↗

Estimating the potato leafhopper Empoasca fabae (Homoptera: Cicadellidae) overwintering range and spring premigrant development by using geographic information system.

The potato leafhopper, Empoasca fabae (Harris), is a circular migratory pest of many crops in the United States that overwinters in the southern states. Northward migrant population arrival to the northern states occurs earlier in the north central states compared with northeastern states. Migrant leafhopper arrival to the north varies from year to year depending on factors influencing the development of spring migrants in the overwintering areas and on timing of weather systems capable of transporting the migrants northward. An estimate of the potato leafhopper minimum temperature survival, the geographic limits of the potato leafhopper overwintering range, leafhopper spring development in the overwintering areas, and the identification of the spring migration initiation northwards can help to predict the leafhopper arrival time in the northern states. In the current study, geographic information system (GIS) was used to estimate the potato leafhopper minimum temperature survival and premigrant development. The minimum winter temperature was estimated by overlaying minimum temperature isolines with potato leafhopper collection data taken during the winter, The geographic limits of the overwintering range were estimated using the minimum temperature survival to create a condition-based model by using ArcMap-GIS 8.2. The estimated overwintering range was larger and covered areas further north than previously estimated and included Missouri, Kansas, Kentucky, Virginia, and Maryland. The use of degree-day accumulation to estimate days of first adult emergence in the overwintering areas resulted in earliest adult emergence in the south central region. First adult emergence in south central and southeastern areas occurred before the detection of potato leafhoppers in the north central United States. These data suggested that the difference in population arrival between the north central states and the northeastern states was more dependent on factors affecting the migration and weather conditions encountered along the migration pathway.

Animal Migration↗

Glandular trichome extracts from Medicago sativa deter settling by the potato leafhopper Empoasca fabae.

Extracts of glandular trichomes from Medicago sativa were tested for their ability to disrupt the settling behavior of the potato leafhopper, Empoasca fabae. Erect and procumbent glandular trichomes were mechanically isolated from stem sections of resistant genotype "G98A," and nonglandular trichomes were collected from susceptible cultivar "Ranger." Isolated trichomes were extracted with chloroform, acetone, and ethanol, and the resulting crude extracts were applied to the surface of a sachet containing an artificial diet. Leafhoppers were offered a two-way choice between crude trichome extracts from G98A and Ranger. All three of G98A solvent extracts caused various degrees of diet rejection, resulting in the crude Ranger trichome extracts being preferred over G98A extracts. Overall, the fewest leafhoppers settled on the ethanolic extracts. Additional bioassays documented a dose response associated with G98A ethanolic extracts when compared with Ranger trichome extracts and a solvent control. No difference in preference behavior was detected between Ranger trichome extracts and a solvent control. Gas chromatography-mass spectrometry analysis revealed a homologous series of nonvolatile fatty acid amides C(n)H(2n+1)NO (n = 19-23) unique to G98A glandular trichome extracts.

Amides↗

Subcellular effects and localization of binding sites of phytohemagglutinin in the potato leafhopper, Empoasca fabae (Insecta: Homoptera: Cicadellidae).

To identify the means by which phytohemagglutinin (PHA) exerts its toxicity on the potato leafhopper, four different methods (thick and semi-thin sectioning combined with immunofluorescent staining, in vitro receptor autoradiography, and immunoelectron microscopy) were used to elucidate the PHA target tissue, binding site, and its effects on this tissue. Sixteen 1- or 2-day-old female potato leafhoppers were fed for 36 h on each of three treatments: a control, diet or a diet containing either the PHA-E subunit or the PHA-L subunit. The PHA-E subunit, but not PHA-L, had previously been shown to be lethal. The insects were then prepared for both light and confocal microscopy. Analysis of images showed that PHA bound only to the surface of midgut epithelial cells of the potato leafhopper. PHA-E caused severe disruption, disorganization, and elongation of the brush border microvilli, and swelling of the epithelial cells into the lumen of the gut, leading to complete closure of the lumen. Furthermore, PHA-E stimulated the division of midgut epithelial cell nuclei, leading to two nuclei in each cell. Nuclei later elongated and degraded. In contrast, PHA-L had little effect on the epithelial cells of the midgut. It did not strongly bind to the surface of epithelial cells and caused much less disruption of brush-border microvilli, less disorganization of the cells and less elongation of nuclei. Strong binding of PHA occurred solely on the cell membrane of the brush border microvilli of epithelial cells. In contrast, the controls (i.e., midgut tissue, blocking agent, PHA, and antibodies) showed that midgut tissue was not autofluorescent and showed no fluorescent binding signal. Analysis of both bright- and dark-field images obtained by autoradiography and immunoelectron microscopy confirmed these findings.

Animals↗

Within-field manipulation of potato leafhopper (Homoptera: Cicadellidae) and insect predator populations using an uncut alfalfa strip.

The effects of a 3-m wide uncut alfalfa strip on the within field distribution of Empoasca fabae (Harris) (Homoptera: Cicadellidae) and several species of insect predators were examined for 3 yr (1998-2000). The objectives were to determine whether this uncut strip would serve as a trap crop for E. fabae and a refuge for insect predators at first harvest. Empoasca fabae and insect predators in the families Coccinellidae, Nabidae, Anthocoridae, Chrysopidae, and Hemerobiidae were collected weekly using sweep samples and sticky traps from the uncut strips and up to 40 m into the alfalfa regrowth. For 2 yr, both small- (0.34 ha) and large-scale (approximately 11.3 ha) field trials showed higher numbers of E. fabae in 73% of the uncut strips for 2-3 wk after harvest. Similarly, the number of insect predators found within <50% of the uncut strips was also higher during the first or second week after harvest. In 1999, however, we did not observe higher numbers of E. fabae in the uncut strips. Differences may be because of higher E. fabae population numbers in 1999 compared with 1998 and 2000. This research provides alfalfa growers a potential cultural management technique for E. fabae while conserving predatory insects.

Animals↗

RAPD-PCR for identification of Zoophthora radicans isolates in biological control of the potato leafhopper.

Biological control studies require the ability to distinguish released pathogens from locally occurring isolates of the same species. We have developed a technique that differentiates genotypes using random amplified polymorphic DNA (RAPD) for the apomictic species Zoophthora radicans (Zygomycota: Entomophthorales), a pathogen of the potato leafhopper, Empoasca fabae (Homoptera: Cicadellidae). RAPD analysis was performed on Z. radicans isolates released in test plots in 1990 and 1991 for leafhopper control; isolates later recovered from the same plots and diverse other isolates were included in the analysis. RAPD fragment profiles of five recovered isolates proved very similar to those of the released isolates and different from all other isolates tested; they are probable descendants of the released isolates. One of the recovered isolates had RAPD profiles similar to isolates derived from aphids and probably represents a population endemic at the release site. In addition to verifying the successful establishment of our experimental releases, RAPD analysis revealed clear relationships among isolates derived from the same host taxon. We propose that this simple and relatively inexpensive method will be valuable in determining the establishment and spread of organisms released in biological control studies.

Animals↗

Mass spectral characterization of fatty acid amides from alfalfa trichomes and their deterrence against the potato leafhopper.

A homologous series of N-(3-methylbutyl)amides of normal saturated C14, C15, C16, C17 and C18 fatty acids were identified as major components of glandular trichome extracts from Medicago sativa G98A, an alfalfa genotype resistant to the potato leafhopper, Empoasca fabae. A second homologous series of N-(2-methylpropyl)amides of C14 through C18 normal fatty acids were minor components. Saturated free fatty acids C12, C13, C14, C15, C16, C17 and C18 were present in trace amounts, as was the N-(3-methylbutyl)amide of linoleic acid (C18:2). N-(3-methylbutyl)amides and N-(2-methylpropyl)amides of C14 through C18 fatty acids, along with the N-(3-methylbutyl)amide of linoleic acid, were synthesized and bioassayed for leafhopper deterrence by applying the compounds to the surface of a sachet containing an artificial diet. Leafhoppers were then offered a two-way choice between diet surfaces treated with the synthetic amides or an untreated control. N-(3-methylbutyl)amides and N-(2-methylpropyl)amides of C14 through C18 fatty acids did not deter leafhopper settling in a dose-dependent fashion. In contrast, when tested singly, N-(3-methylbutyl)amide of linoleic acid exhibited dose-dependent deterrence against leafhopper settling. Fatty acid amides localized in alfalfa glandular trichomes likely contribute to leafhopper resistance.

Amides↗

The arthropod fauna of Quebec vineyards with particular reference to phytophagous arthropods.

A 3-yr study using different sampling and trapping techniques showed that the arthropod pest fauna in two commercial vineyards in southwestern Quebec was qualitatively and quantitatively different than that of Ontario, Canada, and New York state. We hypothesize that a colder winter climate in addition to the agronomic activity of earthing up around the vines in autumn to protect the roots from freezing in winter contributed to low numbers of pests, such as the grape berry moth, Endopiza viteana Clemens (Lepidoptera, Tortricidae). Once in 3 yr, the density of this pest approached, in one of the vineyards, the action threshold recommended for New York. Therefore, it should be monitored on an annual basis. Another phytophagous arthropod that has the potential to cause sporadic economic damage is the potato leafhopper, Empoasca fabae (Harris). The Asiatic garden beetle, Maladera (= Autoserica) castanea (Arrow), was reported for the first time in Canada. The tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), was also captured by sampling. However, its status as a pest has yet to be clarified.

Animals↗

Host effects of glandular-haired alfalfa on alfalfa weevil (Coleoptera: Curculionidae) and potato leafhopper (Homoptera: Cicadellidae) populations in Virginia.

Cultivars of glandular-haired alfalfa, Medicago sativa L., such as '54H69', are currently available and marketed as being resistant to potato leafhopper, Empoasca fabae (Harris). Between 2000 and 2002, studies were conducted to compare the effects of '54H69' and a standard, nonglandular-haired alfalfa cultivar, 'Choice', on alfalfa weevil, Hypera postica (Gyllenhal), and potato leafhopper populations at Campbell and Montgomery counties, Virginia. '54H69' had no effect on alfalfa weevil populations. At each location, densities of alfalfa weevil in '54H69' and 'Choice' were similar, but pest pressure was higher at Campbell Co. than at Montgomery Co. and always exceeded the economic threshold before insecticide was applied. Densities of potato leafhopper also did not differ between '54H69' and 'Choice' in any year at the two locations. Insecticide treatment effectively reduced potato leafhopper densities in the two cultivars, although populations were below the economic threshold at both locations when the insecticides were applied. Overall, postinsecticide treatment comparisons showed that the densities of alfalfa weevil and potato leafhoppers were similar or higher in untreated '54H69' compared with insecticide-treated 'Choice'. In addition, there were no differences in seasonal dry yields between '54H69' and 'Choice' in any year at either location. Our results indicate that the glandular-haired alfalfa '54H69' does not provide acceptable resistance to potato leafhopper and also does not offer a yield advantage to growers in Virginia.

Animals↗

Aspects of potato leafhopper (Homoptera: Cicadellidae) biology on Solanum berthaultii and other potato genotypes.

Survival of nymphs and adult males and females of potato leafhopper, Empoasca fabae (Harris), on six potato (Solanum spp.) genotypes with varied levels of resistance was evaluated and related to type, density, and droplet/head size of foliar glandular trichomes. Survival was greater on Solanum tuberosum cultivars ('Allegany' and 'Elba') compared with accessions of the wild S. berthaultii (PI 473331 and PI 473334). The barrier provided by glandular trichomes may affect survival of E.fabae by restricting nymph and adult feeding. PI 473331 was the most pubescent host, bearing type A and type B trichomes and the most unsuitable host for E. fabae. Allegany possesses leaflets with low trichome densities and small gland diameter and was the most suitable host for adult E. fabae survival. Females generally had higher survival rates than males. Nymphal and adult leafhoppers differed greatly in their survival rates, particularly on S. tuberosum x S. berthaultii hybrids (Q174-2 and NY123). Interestingly, nymphs had longer (190 d) mean lethal time (LT50) than adults (31 d) when held on Q174-2, and a lower LT50 (19 d) when held on NY123 compared with adults (104 d). Starvation, dehydration, or both were likely associated with mortality of nymphs confined on PI 473331 and PI 473334. Nymphs on these accessions died at a comparable rate to those confined on a starvation treatment and more rapidly than those supplied with water only, sucrose solution or other potato genotypes. Darkened trichome exudates were observed on insects held on PI 473331 and PI 473334, consistent with previous reports that glandular trichomes of S. berthaultii contain phenolic oxidation chemistry. This study aids in the identification of the probable mechanisms of resistance to E. fabae used by S. berthaulti and its hybrids with S. tuberosum.

Animals↗

Potato leafhopper (Homoptera: Cicadellidae) injury disrupts basal transport of 14C-labelled photoassimilates in alfalfa.

The potato leafhopper, Empoasca fabae (Harris), is a key pest of alfalfa, Medicago sativa L., in part because of the leafhopper's ability to disrupt upward translocation within phloem tissues. To determine if leafhopper injury also disrupts basal translocation necessary for regrowth and perenniality of alfalfa, we used radiolabeled 14CO2 to measure the basal transport of photoassimilates in injured and healthy plants. In one experiment, less 14C was transported to lower stem tissue of leafhopper-injured plants in comparison to the same tissue of healthy plants in early vegetative and early reproductive stages of alfalfa development. In a second experiment, less 14C was transported to lower stem, crown, and root tissues of injured plants in comparison to the same tissues of healthy, early reproductive plants. The disruption of basal transport caused by potato leafhopper may impact carbon storage and mobilization subsequent to defoliation, winter survival, and nitrogen fixation.

Animals↗

Role of the glandular trichomes in resistance of perennial alfalfa to the potato leafhopper (Homoptera: Cicadellidae).

Experiments were performed to elucidate resistance of glandular-haired alfalfa, Medicago sativa L., to the potato leafhopper, Empoasca fabae (Harris). The primary objective was to examine the potential role of the glandular trichomes and stem lignification for imparting resistance to this pest. During free-choice foliar discoloration experiments, the resistant alfalfa clone FGplh13 expressed lower levels of injury than the susceptible P5373 after 13 d. No-choice tests performed using nymphal potato leafhoppers showed higher levels of mortality associated with the resistant glandular-haired clone FGplh13, and no nymphs survived after 48 h. Additional no-choice experiments using FGplh13 and P5373 alfalfa with the glandular and nonglandular trichomes intact or removed show that mortality of nymphs and adults decreased after removal of the glandular trichomes from FGplh13. Nymphal and adult mortality and nymphal development time did not differ on FGplh13 with the trichomes removed and P5373 with the trichomes intact or removed. Firstinstar, and possibly second-instar, potato leafhoppers were entrapped by an exudate produced by the glandular trichomes on the FGplh13 alfalfa. Stem lignification and the number of vascular bundles did not differ between FGplh13 and P5373 alfalfas. The glandular trichomes on FGplh13 alfalfa appeared to provide the major host resistance factor, with resistance to adults being chemically based and resistance to nymphs being chemically and mechanically based. To maintain levels of potato leafhopper resistance, breeders would appear to benefit by continuing to select for the expression of the glandular trichome phenotype.

Animals↗

Influence of herbicide tolerant soybean production systems on insect pest populations and pest-induced crop damage.

Conventional soybean weed management and transgenic herbicide-tolerant management were examined to assess their effects on soybean insect pest populations in south Georgia in 1997 and 1998. Soybean variety had very little impact on the insect species observed, except that maturity group effects were observed for stink bug, primarily Nezara viridula (L.), population densities on some sampling dates. Stink bugs were more abundant on the early maturing varieties in mid-season. Velvetbean caterpillar, Anticarsia gemmatalis Hübner, potato leafhopper, Empoasca fabae (Harris), and grasshoppers Melanoplus spp. were more numerous on either conventional or herbicide-tolerant varieties on certain dates, although these differences were not consistent throughout the season. Soybean looper, Pseudoplusia includens (Walker), threecornered alfalfa hopper, Spissistilus festinus (Say), and whitefringed beetles, Graphognathus spp , demonstrated no varietal preference in this study. Few weed treatment differences were observed, but if present on certain sampling dates, then pest numbers were higher in plots where weeds were reduced (either postemergence herbicides or preplant herbicide plus postemergence herbicide). The exception to this weed treatment effect was grasshoppers, which were more numerous in weedy plots when differences were present. In post emergence herbicide plots, there were no differences in insect pest densities between the conventional herbicides (e.g., Classic, Select, Cobra, and Storm) compared with specific gene-inserted herbicide-tolerant materials (i.e., Roundup and Liberty). Defoliation, primarily by velvetbean caterpillar, was different between soybean varieties at some test sites but not different between herbicide treatments. We did not observe differences in seasonal abundance of arthropod pests between conventional and transgenic herbicide-tolerant soybean.

Animals↗

Influence of plant ontogeny and abiotic factors on resistance of glandular-haired alfalfa to potato leafhopper (Hemiptera: Cicadellidae).

Laboratory experiments were conducted to characterize the trichome-based defense of glandular-haired alfalfa, Medicago sativa L., against the potato leafhopper, Empoasca fabae (Harris). Within-plant variability in leafhopper resistance was examined by caging adult leafhoppers to either basal or apical stem internodes of the leafhopper-resistant, glandular-haired M. sativa genotype G98A or the susceptible, nonglandular-haired M. sativa 'Ranger'. Young, actively secreting glandular trichomes are located on apical internodes of G98A, whereas senesced gland heads are found on older, basal internodes of G98A. After 96 h, the highest cumulative leafhopper mortality and lowest number of excretory droplets were associated with apical internodes of G98A. No difference was detected in mortality and feeding levels among insects caged to basal internodes of G98A and basal and apical internodes of Ranger. The influence of abiotic factors on leafhopper resistance was evaluated by caging adult leafhoppers to either G98A or Ranger under four combinations of low and high light (250 and 1,000 micromol s(-1) m(-2)) and temperature regimes (17 and 30 degrees C). After 96 h, the highest cumulative mortality was associated with leafhoppers confined to G98A under high light and high temperature conditions. Temperature level and plant type also had an effect on the production of excretory droplets, resulting in the highest number of excretory droplets being associated with Ranger under the high temperature regime. These results indicate that certain regions of M. sativa G98A are better protected against the potato leafhopper than others and that temperature influences resistance levels of glandular-haired alfalfa.

Animals↗

Yield and forage quality of glandular-haired alfalfa under alfalfa weevil (Coleoptera: Curculionidae) and potato leafhopper (Hemiptera: Cicadellidae) pest pressure in Virginia.

Cultivars of glandular-haired alfalfa, Medicago sativa L., such as '54H69', are currently available and marketed as being resistant to potato leafhopper, Empoasca fabae (Harris) (Hemiptera: Cicadellidae). 54H69 and a standard, nonglandular-haired alfalfa 'Choice' were evaluated at two locations in Virginia over a 3-yr period. Dry matter yields and concentrations of crude protein and acid detergent fiber were compared at the first, second, and third harvests. Overall, the two cultivars produced similar dry matter yields of comparable forage quality in the absence of insecticides at both locations in each year. Untreated 54H69 did not produce greater dry matter yields than untreated Choice under either light or heavier potato leafhopper pest pressure. Concentrations of crude protein did not vary between the two cultivars at any harvest. Some differences in concentrations of acid detergent fiber were detected between cultivars, but these differences were not consistent among years, harvests, or between locations. Further comparisons between untreated 54H69 and treated Choice were made, but few significant differences were detected in dry matter yields or forage quality. An economic analysis for the study indicated that a grower planting 54H69 would realize less net revenue than a grower planting Choice, largely because of the seed premium for the glandular-haired cultivar and the evident need to treat 54H69 with insecticide for control of alfalfa weevil, Hypera postica (Gyllenhal) (Coleoptera: Curculionidae), and potato leafhopper.

Animals↗

Host-plant-mediated competition via induced resistance: interactions between pest herbivores on potatoes.

Plant-mediated competition among insect herbivores occurs when one species induces changes in plant chemistry, nutrition, or morphology that render plants resistant to attack by others. We explored plant-mediated interspecific interactions between the potato leafhopper (Empoasca fabae) and the Colorado potato beetle (Leptinotarsa decemlineata), two important pests on potatoes. Leafhoppers colonize fields in advance of beetles, and thus the possibility exists that previous feeding by leafhoppers induces changes in potato plants that have adverse consequences for beetles. The consequences of leafhopper-induced resistance for beetle performance were studied in the greenhouse, field cages, and in large open-field plots. Potato plants were exposed to four densities of leafhoppers (none, low, moderate, and high), and visible feeding symptoms were measured as percentage leaf curling, chlorosis, and necrosis. The oviposition preference, performance, and survivorship of Colorado potato beetles were then measured on the four categories of induced plants in field-cage and greenhouse settings. In open field plots, survival on the four categories of induced plants was determined by placing cohorts of beetle adults onto plants and measuring the densities of resulting eggs, larvae, and emerging Fl adults. Leafhopper-induced symptoms on potato plants were density dependent, with the percentage of curled, chlorotic, and necrotic leaves increasing with leafhopper density. Previous feeding by leafhoppers adversely affected oviposition and larval performance of beetles. Fewer egg masses were deposited on plants that incurred high levels of leafhopper feeding. Similarly, larval development was delayed and emerging adult beetles weighed less when fed induced foliage from the high leafhopper-density treatment. Beetles survived less well in the field on plants experiencing moderate and high levels of leafhopper feeding as evidenced by lower densities of eggs, larvae, and emerging F1 adults. Overall, leafhoppers and beetles competed through feeding-induced changes in plant quality. Notably, the asymmetric interaction took place at a large spatial scale in open field plots and had negative consequences that persisted to the next beetle generation. Ultimately, to establish an effective management strategy for crop pests such as leafhoppers, it is essential to consider the positive indirect effects of induced resistance along with the negative direct effects on crop yield.

Animals↗