Adam R. Zeilinger

Colonization preference of Euschistus servus and Nezara viridula in transgenic cotton varieties, peanut, and soybean

Producers of Bt cotton, Gossypium hirsutum L. (Malvaceae), in the southeastern USA face significant losses from highly polyphagous stink bug species. These problems may be exacerbated by crop rotation practices that often result in cotton, peanut, Arachis hypogaea L., and soybean, Glycine max (L.) Merrill (both Fabaceae), growing in close proximity to one another. Because all of these crops are hosts for the major pest stink bug species in the region, we experimentally examined colonization preference of these species among the crops to clarify this aspect of their population dynamics. We planted peanut, soybean, Bt cotton, and glyphosate‐tolerant (RR) non‐Bt cotton at three sites over 3 years in replicated plots ranging from 192 to 1 323 m2 and calculated odds ratios for colonization of each crop for Nezara viridula (L.) and Euschistus servus (Say) (both Hemiptera: Pentatomidae). In four of five experiments, both E. servus and N. viridula preferred soybean significantly more often than Bt cotton, non‐Bt cotton, and peanut. Neither N. viridula nor E. servus showed any preference between non‐Bt and Bt cotton in any experiment. Both species had higher numbers in Bt and non‐Bt cotton relative to peanut; this was not significant for any single experiment, but analyses across all experiments indicated that N. viridula preferred Bt and non‐Bt cotton significantly more often than peanut. Our results suggest that soybean in the landscape may function as a sink for stink bug populations relative to nearby peanut and cotton when the soybean is in the reproductive stage of development. Stink bug preference for soybean may reduce pest pressure in near‐by crops, but population increases in soybean could lead to this crop functioning as a source for later‐season pest pressure in cotton.

Competition between stink bug and heliothine caterpillar pests on cotton at within‐plant spatial scales

Outbreaks of non‐target pests associated with transgenic Bt cotton threaten the economic and ecological benefits of the technology in cotton‐producing countries. In the southeastern USA, stink bug pests, namely Nezara viridula L. and Euschistus servus Say (both Heteroptera: Pentatomidae), have recently become severe problems associated with Bt cotton, requiring continued insecticide use. However, the causes of non‐target pest outbreaks remain unclear. Release from competition with Bt‐susceptible pest species, in addition to other mechanisms, may contribute to increased stink bug populations in Bt cotton. We investigated the competitive interactions between the two stink bug species and the Bt‐susceptible pests Helicoverpa zea (Boddie) and Heliothis virescens Fabricius (both Lepidoptera: Noctuidae) on non‐Bt cotton. We tested for competition effects on stink bug growth rates in no‐choice experiments at two spatial scales: a single cotton boll and a branch with multiple developing bolls. Although caterpillars of the two species had equivalent effects on resource availability, they had distinct effects on stink bug growth rates. Fourth instar H. zea reduced growth rates of both stink bug species by 60% when caged on a single cotton boll and reduced growth rates of only E. servus by 36% when caged on a cotton branch. In contrast, H. virescens had no effect on stink bug growth rates. Resource competition was apparent in the interactions between H. zea and E. servus, but interference competition may have contributed to the interactions as well. Competitive release of stink bug populations in Bt cotton is possible, and should be more likely for E. servus than for N. viridula. Understanding the causes of non‐target pest outbreaks in Bt cotton will contribute to improved environmental risk assessments of future releases of Bt cotton and related transgenic crops.

Competitive release and outbreaks of non-target pests associated with transgenic Bt cotton.

The adoption of transgenic Bt cotton has, in some cases, led to environmental and economic benefits through reduced insecticide use. However, the distribution of these benefits and associated risks among cotton growers and cotton-growing regions has been uneven due in part to outbreaks of non-target or secondary pests, thereby requiring the continued use of synthetic insecticides. In the southeastern USA, Bt cotton adoption has resulted in increased abundance of and damage from stink bug pests, Euschistus servus and Nezara viridula (Heteroptera: Pentatomidae). While the impact of increased stink bug abundance has been well-documented, the causes have remained unclear. We hypothesize that release from competition with Bt-susceptible target pests may drive stink bug outbreaks in Bt cotton. We first examined the evidence for competitive release of stink bugs through meta-analysis of previous studies. We then experimentally tested if herbivory by Bt-susceptible Helicoverpa zea increases stink bug leaving rates and deters oviposition on non-Bt cotton. Consistent with previous studies, we found differences in leaving rates only for E servus, but we found that both species strongly avoided ovipositing on H. zea-damaged plants. Considering all available evidence, competitive release of stink bug populations in Bt cotton likely contributes to outbreaks, though the relative importance of competitive release remains an open question. Ecological risk assessments of Bt crops and other transgenic insecticidal crops would benefit from greater understanding of the ecological mechanisms underlying non-target pest outbreaks and greater attention to indirect ecological effects more broadly.

Blocking the Transmission of a Noncirculative Vector-Borne Plant Pathogenic Bacterium

The successful control of insect-borne plant pathogens is often difficult to achieve due to the ecologically complex interactions among pathogens, vectors, and host plants. Disease management often relies on pesticides and other approaches that have limited long-term sustainability. To add a new tool to control vector-borne diseases, we attempted to block the transmission of a bacterial insect-transmitted pathogen, the bacterium Xylella fastidiosa, by disrupting bacteria-insect vector interactions. X. fastidiosa is known to attach to and colonize the cuticular surface of the mouthparts of vectors; a set of recombinant peptides was generated and the chemical affinities of these peptides to chitin and related carbohydrates was assayed in vitro. Two candidates, the X. fastidiosa hypothetical protein PD1764 and an N-terminal region of the hemagglutinin-like protein B (HxfB) showed affinity for these substrates. These proteins were provided to vectors via an artificial diet system in which insects acquire X. fastidiosa, followed by an inoculation access period on plants under greenhouse conditions. Both PD1764 and HxfAD1-3 significantly blocked transmission. Furthermore, bacterial populations within insects over a 10-day period demonstrated that these peptides inhibited cell adhesion to vectors but not bacterial multiplication, indicating that the mode of action of these peptides is restricted to limiting cell adhesion to insects, likely via competition for adhesion sites. These results open a new venue in the search for sustainable disease-control strategies that are pathogen specific and may have limited nontarget effects.

Is Vector Control Sufficient to Limit Pathogen Spread in Vineyards

ABSTRACT Vector control is widely viewed as an integral part of disease management. Yet epidemiological theory suggests that the effectiveness of control programs at limiting pathogen spread depends on a variety of intrinsic and extrinsic aspects of a pathosystem. Moreover, control programs rarely evaluate whether reductions in vector density or activity translate into reduced disease prevalence. In areas of California invaded by the glassy-winged sharpshooter (Homalodisca vitripennis Germar), Pierces disease management relies heavily on chemical control of this vector, primarily via systemic conventional insecticides (i.e., imidacloprid). But, data are lacking that attribute reduced vector pressure and pathogen spread to sharpshooter control. We surveyed 34 vineyards over successive years to assess the epidemiological value of within-vineyard chemical control. The results showed that imidacloprid reduced vector pressure without clear nontarget effects or secondary pest outbreaks. Effects on disease prevalence were more nuanced. Treatment history over the preceding 5 yr affected disease prevalence, with significantly more diseased vines in untreated compared with regularly or intermittently treated vineyards. Yet, the change in disease prevalence between years was low, with no significant effects of insecticide treatment or vector abundance. Collectively, the results suggest that within-vineyard applications of imidacloprid can reduce pathogen spread, but with benefits that may take multiple seasons to become apparent. The relatively modest effect of vector control on disease prevalence in this system may be attributable in part to the currently low regional sharpshooter population densities stemming from areawide control, without which the need for within-vineyard vector control would be more pronounced.

A Likelihood-Based Biostatistical Model for Analyzing Consumer Movement in Simultaneous Choice Experiments

ABSTRACT Consumer feeding preference among resource choices has critical implications for basic ecological and evolutionary processes, and can be highly relevant to applied problems such as ecological risk assessment and invasion biology. Within consumer choice experiments, also known as feeding preference or cafeteria experiments, measures of relative consumption and measures of consumer movement can provide distinct and complementary insights into the strength, causes, and consequences of preference. Despite the distinct value of inferring preference from measures of consumer movement, rigorous and biologically relevant analytical methods are lacking. We describe a simple, likelihood-based, biostatistical model for analyzing the transient dynamics of consumer movement in a paired-choice experiment. With experimental data consisting of repeated discrete measures of consumer location, the model can be used to estimate constant consumer attraction and leaving rates for two food choices, and differences in choice-specific attraction and leaving rates can be tested using model selection. The model enables calculation of transient and equilibrial probabilities of consumer-resource association, which could be incorporated into larger scale movement models. We explore the effect of experimental design on parameter estimation through stochastic simulation and describe methods to check that data meet model assumptions. Using a dataset of modest sample size, we illustrate the use of the model to draw inferences on consumer preference as well as underlying behavioral mechanisms. Finally, we include a users guide and computer code scripts in R to facilitate use of the model by other researchers.

Do Counts of Salivary Sheath Flanges Predict Food Consumption in Herbivorous Stink Bugs (Hemiptera: Pentatomidae)?

ABSTRACT Counts of salivary sheaths and salivary flanges have been widely used in studies of feeding behavior and crop damage of pestiferous stink bugs (Hemiptera: Pentatomidae) and other sheath-feeding Hemiptera. While salivary flanges can effectively predict crop damage by stink bugs, previous studies have assumed that food consumption (e.g., ingestion) and preference can also be inferred from flange data. Yet this assumption has remained untested. We investigated the relationship between the number of stink bug salivary flanges and consumption of cotton bolls for two important agricultural pest species: Nezara viridula (L.) and Euschistus servus (Say). We inferred food consumption rates from measures of relative growth rate and excreta quantity. To measure excreta, we quantified the color intensity, or chromaticity, of excreta using digital image analysis. We found a positive relationship between growth rate and the number of flanges for fifth instars of E. servus. However, we found no relationship between growth or excretion and the number of flanges for all stages of N. viridula and for E. servus adults. Our results indicate that counts of salivary flanges should not be used to infer food consumption or preference in studies on N. viridula and E. servus adults, but can be used in studies of E. servus nymphs. Species-and stage-specific differences in the relationship between consumption and salivary flanges suggests distinct feeding strategies among species and stages; such differences may be potentially important in determining crop damage from pestiferous stink bugs.

A chitinase is required for Xylella fastidiosa colonization of its insect and plant hosts

Xylella fastidiosa colonizes the xylem network of host plant species as well as the foregut of its required insect vectors to ensure efficient propagation. Disease management strategies remain inefficient due to a limited comprehension of the mechanisms governing both insect and plant colonization. It was previously shown that X. fastidiosa has a functional chitinase (ChiA), and that chitin likely serves as a carbon source for this bacterium. We expand on that research, showing that a chiA mutant strain is unable to grow on chitin as the sole carbon source. Quantitative PCR assays allowed us to detect bacterial cells in the foregut of vectors after pathogen acquisition; populations of the wild-type and complemented mutant strain were both significantly larger than the chiA mutant strain 10 days, but not 3 days, post acquisition. These results indicate that adhesion of the chiA mutant strain to vectors may not be impaired, but that cell multiplication is limited. The mutant was also affected in its transmission by vectors to plants. In addition, the chiA mutant strain was unable to colonize host plants, suggesting that the enzyme has other substrates associated with plant colonization. Lastly, ChiA requires other X. fastidiosa protein(s) for its in vitro chitinolytic activity. The observation that the chiA mutant strain is not able to colonize plants warrants future attention to be paid to the substrates for this enzyme.

Behavioural and chemical mechanisms of plant-mediated deterrence and attraction among frugivorous insects

1. Herbivory often induces systemic plant responses that affect the host choice of subsequent herbivores, either deterring or attracting them, with implications for the performance of both herbivore and host plant. Combining measures of herbivore movement and consumption can efficiently provide insights into the induced plant responses that are most important for determining choice behaviour.

Plant Water Stress and Vector Feeding Preference Mediate Transmission Efficiency of a Plant Pathogen

Abstract Pathogen spread by arthropod vectors is the outcome of pathogen–vector–plant interactions, as well as how these interactions are impacted by abiotic and biotic factors. While plant water stress impacts each component of the Pierces disease pathosystem (Xylella fastidiosa Wells et al., insect vectors, and grapevines), the outcome of interactions in relation to pathogen spread is unknown. The objectives of this study were 1) to determine the role of plant water stress on vector acquisition and inoculation of X. fastidiosa under choice and no-choice conditions for source or recipient vines, and 2) to provide insights into the effects of vineyard irrigation regimes on spread of X. fastidiosa by using a host–vector epidemic model. Under no-choice conditions, pathogen acquisition increased as water stress increased in source plants, while inoculation was not affected by water status of recipient vines. Thus, under no-choice conditions, plant water stress increased transmission of X. fastidiosa. However, when vectors had a choice of an uninfected well-watered versus an infected water-stressed grapevine, transmission efficiency declined as water stress levels increased. While our experimental results produced wide uncertainty estimates, the epidemiological modeling suggested a non-linear relationship between water stress and pathogen spread: moderate water stress enhances pathogen spread but severe or no stress produce equivalent spread. In summary, both host plant condition and vector host preference interacted to determine transmission efficiency of X. fastidiosa.