Hybridizing transgenic Bt cotton with non-Bt cotton counters resistance in pink bollworm.

Extensive cultivation of crops genetically engineered to produce insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) has suppressed some major pests, reduced insecticide sprays, enhanced pest control by natural enemies, and increased grower profits. However, these benefits are being eroded by evolution of resistance in pests. We report a strategy for combating resistance by crossing transgenic Bt plants with conventional non-Bt plants and then crossing the resulting first-generation (F1) hybrid progeny and sowing the second-generation (F2) seeds. This strategy yields a random mixture within fields of three-quarters of plants that produce Bt toxin and one-quarter that does not. We hypothesized that the non-Bt plants in this mixture promote survival of susceptible insects, thereby delaying evolution of resistance. To test this hypothesis, we compared predictions from computer modeling with data monitoring pink bollworm (Pectinophora gossypiella) resistance to Bt toxin Cry1Ac produced by transgenic cotton in an 11-y study at 17 field sites in six provinces of China. The frequency of resistant individuals in the field increased before this strategy was widely deployed and then declined after its widespread adoption boosted the percentage of non-Bt cotton plants in the region. The correspondence between the predicted and observed outcomes implies that this strategy countered evolution of resistance. Despite the increased percentage of non-Bt cotton, suppression of pink bollworm was sustained. Unlike other resistance management tactics that require regulatory intervention, growers adopted this strategy voluntarily, apparently because of advantages that may include better performance as well as lower costs for seeds and insecticides.

Enhanced stability in host-parasitoid interactions with autoparasitism and parasitoid migration.

Previous studies based on simple non-spatial model have suggested that autoparasitism, in which females develop as primary endoparasitoids of hosts while males develop at the expense of primary parasitoids, stabilizes host-parasitoid steady state. To date, however, how the stabilizing role of autoparasitism would be affected by more complex spatial factors has not been adequately investigated. To address the issue, here we analyzed a spatially extended two-patch host-parasitoid model and compared it with the corresponding non-spatial model. Results showed that in the non-spatial model and the case of autoparasitoid, the host-parasitoid steady states can be unstable if the host׳s intrinsic rate of growth and/or carrying capacity is sufficiently large. However, in the spatially extended two-patch model with parasitoid migration, the unstable host-parasitoid steady states in each local patch may become stable, provided there is certain spatial unevenness in host growth and/or carrying capacity. Therefore, the migration of parasitoid together with spatial unevenness in host growth and/or carrying capacity stabilizes the host-parasitoid interactions. The stabilizing effects are stronger with the host density-dependent migration of parasitoid than with the random migration of parasitoid. In the case of primary parasitoid, the model demonstrated similar stabilizing effects associated with the migration of parasitoid. However, the parameter conditions for stability are much more stringent than in the case of autoparasitoid. We concluded that the stabilizing effects of parasitoid migration and autoparasitism can add to each other, leading to more stable host-parasitoid interactions.

Population impacts of Wolbachia on Aedes albopictus.

Prior studies have demonstrated that Wolbachia, a commonly occurring bacterium capable of manipulating host reproduction, can affect life history traits in insect hosts, which in turn can have population-level effects. Effects on hosts at the individual level are predicted to impact population dynamics, but the latter has not been examined empirically. Here, we describe a biological model system based on Aedes albopictus (Asian tiger mosquito) that allows for measurement of population dynamics, which has not been accomplished in prior field trials or laboratory designs. The results demonstrate the studied populations to be robust and allow for persistent, closed populations with overlapping generations, which are regulated solely through density-dependent, intraspecific competition for limited resources. Using a novel experimental design, we compare populations that are either uninfected or infected with Wolbachia. The results show differences that include population size, eclosion rates, adult survivorship, and fecundity. The aposymbiotic populations were generally larger and adults longer lived relative to the infected populations. The outcome is discussed in context with naturally occurring Wolbachia invasions, proposed autocidal strategies, and the utility of the developed system as a biological platform for hypothesis testing and improved parameterization.

Mutation in the Cadherin Gene Is a Key Factor for Pink Bollworm Resistance to Bt Cotton in China.

Transgenic crops producing Bacillus thuringiensis (Bt) toxins are widely planted for insect control, but their efficacy may decrease as insects evolve resistance. Understanding the genetic basis of insect resistance is essential for developing an integrated strategy of resistance management. To understand the genetic basis of resistance in pink bollworm (Pectinophora gossypiella) to Bt cotton in the Yangtze River Valley of China, we conducted an F2 screening for alleles associated with resistance to the Bt (Cry1Ac) protein for the first time. A total of 145 valid single-paired lines were screened, among which seven lines were found to carry resistance alleles. All field parents in those seven lines carried recessive resistance alleles at the cadherin locus, including three known alleles, r1, r13 and r15, and two novel alleles, r19 and r20. The overall frequency of resistance alleles in 145 lines was 0.0241 (95% CI: 0.0106-0.0512). These results demonstrated that resistance was rare and that recessive mutation in the cadherin gene was the primary mechanism of pink bollworm resistance to Bt cotton in the Yangtze River Valley of China, which will provide a scientific basis for implementing targeted resistance management statics of pink bollworm in this region.

Wolbachia infections that reduce immature insect survival: predicted impacts on population replacement.

BACKGROUND: The evolutionary success of Wolbachia bacteria, infections of which are widespread in invertebrates, is largely attributed to an ability to manipulate host reproduction without imposing substantial fitness costs. Here, we describe a stage-structured model with deterministic immature lifestages and a stochastic adult female lifestage. Simulations were conducted to better understand Wolbachia invasions into uninfected host populations. The model includes conventional Wolbachia parameters (the level of cytoplasmic incompatibility, maternal inheritance, the relative fecundity of infected females, and the initial Wolbachia infection frequency) and a new parameter termed relative larval viability (RLV), which is the survival of infected larvae relative to uninfected larvae. RESULTS: The results predict the RLV parameter to be the most important determinant for Wolbachia invasion and establishment. Specifically, the fitness of infected immature hosts must be close to equal to that of uninfected hosts before population replacement can occur. Furthermore, minute decreases in RLV inhibit the invasion of Wolbachia despite high levels of cytoplasmic incompatibility, maternal inheritance, and low adult fitness costs. CONCLUSIONS: The model described here takes a novel approach to understanding the spread of Wolbachia through a population with explicit dynamics. By combining a stochastic female adult lifestage and deterministic immature/adult male lifestages, the model predicts that even those Wolbachia infections that cause minor decreases in immature survival are unlikely to invade and spread within the host population. The results are discussed in relation to recent theoretical and empirical studies of natural population replacement events and proposed applied research, which would use Wolbachia as a tool to manipulate insect populations.

Seasonal Migration Patterns of Ostrinia furnacalis (Lepidoptera: Crambidae) Across the Bohai Strait in Northern China.

The Asian corn borer (ACB), Ostrinia furnacalis (Guenée), is a notorious polyphagous insect pest in China and other regions of East Asia. The long-distance flight behavior of the ACB moths, however, is not well understood, especially in the field conditions. In this study, nightly monitoring data for multiple years (2003-2017) on the long-distance flight of adults crossing the Bohai Strait in northern China, showed that a large number of the adults frequently flew across the Bohai Strait from May to September with the peak migrations usually occurred, followed in order by September, June, July, and May, and the number of southward migrants was larger than that of northward migrants. From May to September in 2010, 2011, and 2017, a subsample of trapped ACB females was dissected (879 individuals in 2010, 197 individuals in 2011, and 247 individuals in 2017), and the results showed that the sex ratio of the trapped ACB moths was unbiased each month from May to September. While the proportion of mated females in northward populations (May to July = 92.85 ± 4.86%) was significantly higher than that in southward population (August and September = 74.53 ± 8.55%) . The seasonal pattern in the proportion of sexually mature females was similar to the above proportion of mated females. We conclude that the ACBs have a strong propensity and ability for long-distance migration, although local dispersal is generally considered to be the primary movement of the ACB. These findings may be helpful to improve the forecasting systems and the pest management schemes for the ACBs.

Density-dependent intraspecific competition in the larval stage of Aedes aegypti (Diptera: Culicidae): revisiting the current paradigm.

Density-dependent intraspecific competition has been considered an important determinant of the dynamics of larval stages of Aedes aegypti. A model was published in 1984 providing a mathematical description of this density dependence, based on field data, that has since been widely used. This description, however, is based on the strong assumption that all mortality is density-dependent. We re-examine the data without this premise and find a reduced importance of density dependence, as well as a different functional form. Based on these discrepancies, we emphasize that the characterization of density dependence in the larval stages of Ae. aegypti should be based on a more complete dataset, and we use artificially generated data to explore how such additional information could help developing a better description of this density dependence. We review other empirical studies on larval competition, discuss the need for further dedicated studies, and provide a few simple guidelines for the design of such studies.

A killer-rescue system for self-limiting gene drive of anti-pathogen constructs.

A number of genetic mechanisms have been suggested for driving anti-pathogen genes into natural populations. Each of these mechanisms requires complex genetic engineering, and most are theoretically expected to permanently spread throughout the target species' geographical range. In the near term, risk issues and technical limits of molecular methods could delay the development and use of these mechanisms. We propose a gene-drive mechanism that can be self-limiting over time and space, and is simpler to build. This mechanism involves one gene that codes for toxicity (killer) and a second that confers immunity to the toxic effects (rescue). We use population-genetic models to explore cases with one or two independent insertions of the killer gene and one insertion of the rescue gene. We vary the dominance and penetrance of gene action, as well as the magnitude of fitness costs. Even with the fitness costs of 10 per cent for each gene, the proportion of mosquitoes expected to transmit the pathogen decreases below 5 per cent for over 40 generations after one 2:1 release (engineered:wild) or after four 1:2 releases. Both the killer and rescue genes will be lost from the population over time, if the rescue construct has any associated fitness cost. Molecular approaches for constructing strains are discussed.

Seasonal Migration Pattern of Nilaparvata lugens (Hemiptera: Delphacidae) Over the Bohai Sea in Northern China.

The brown planthopper Nilaparvata lugens (Stål) (Hemiptera: Delphacidae) is a major, regionally migratory pest of rice crops in Asia. Despite intensive studies, the seasonal pattern of migration in this species remains largely unknown, especially in northern China. Analysis of monitoring data of light trapping at Beihuang island in northern China showed that brown planthopper migrants could be found at the island in any month from July to October. However, the daily number of brown planthopper migrants varied considerably from day to day, month to month, and year to year. Most of migrants were caught from July to September, with fewer in October. Simulation of backward trajectories showed that there was temporal variation in the source areas of brown planthopper migrants trapped at Beihuang. A majority of migrants trapped at Beihuang in July came from south of Beihuang. In contrast, migrants caught in August and September could be from any direction around the island. Results suggested that the brown planthopper migrants likely traveled northward in July, and Multidirectionally in August and September in northern China. Some of brown planthopper in northeastern China could escape the 'Pied Piper effect' and migrate southward in September.

Introducing desirable transgenes into insect populations using Y-linked meiotic drive - a theoretical assessment.

The use of genetic drive mechanisms to replace native mosquito genotypes with individuals bearing antipathogen transgenes is a potential strategy for repressing insect transmission of human diseases such as malaria and dengue. Antipathogen transgenes have been developed and tested, but efficient gene drive mechanisms are lacking. Here we theoretically assess the feasibility of introducing antipathogen genes into wild Aedes aegypti populations by using a naturally occurring meiotic drive system. We consider the release of males having both a Y-linked meiotic drive gene and an X-linked drive-insensitive response allele to which an antipathogen gene is linked. We use mathematical models and computer simulations to determine how the post-introduction dynamics of the antipathogen gene are affected by specific genetic characteristics of the system. The results show that when the natural population is uniformly sensitive to the meiotic drive gene, the antipathogen gene may be driven close to fixation if the fitness costs of the drive gene, the insensitive response allele, and the antipathogen gene are low. However, when the natural population has a small proportion of an X-linked insensitive response allele or an autosomal gene that strongly reduces the effect of the drive gene, the antipathogen gene does not spread if it has an associated fitness cost. Our modeling results provide a theoretical foundation for further experimental tests.

Introducing transgenes into insect populations using combined gene-drive strategies: modeling and analysis.

Engineered underdominance (EU), meiotic drive (MD) and Wolbachia have been proposed as mechanisms for driving anti-pathogen transgenes into natural populations of insect vectors of human diseases. EU can drive transgenes to high and stable frequencies but requires the release of sizeable numbers of engineered insects. MD and Wolbachia either cannot maintain high frequencies of transgenes or lack appropriate expression in critical tissues, but both can drive the transgenes to spread from very low initial frequencies. Here we use mathematical models to assess the utility of combining EU with MD or with Wolbachia. Under some conditions, the combination of EU and MD results in a more efficient transgene-drive strategy than either mechanism alone. This combined strategy could drive the transgenes to stable fixation and would require fewer released insects than EU alone, especially when only males are released. However, a combination of EU and Wolbachia does not work better than EU alone because it requires the release of even more engineered insects.

Age-structured effects and disease interference in childhood infections.

Recent studies have demonstrated that ecological interference among some childhood diseases may have important dynamic consequences. An interesting question is, when would we expect the interference effect to be pronounced? To address the issue, here we develop a seasonally forced two-disease age-structured model, using empirically derived age-specific force of infection (ASFOI) for numerous infections of childhood. Our comparative numerical analysis shows that when the ASFOIs for the two diseases largely overlap, the dynamics predicted by the two-disease model are generally different from those predicted by the analogous single-disease model, suggesting strong fingerprints of disease interference. When the ASFOIs overlap less, on the other hand, both diseases behave as predicted by the single-disease model, suggesting weak interference. We conclude that age structure is an important factor that should be taken into account in order to explore the underlying mechanisms of disease interference.

How do cross-migration models arise?

In this paper we present a general method to derive spatio-temporal population models mechanistically. We consider a system of multiple species living in a patchy habitat in which the local population of each species consists of some behavioural groups. We then formulate a continuous-time model where a small positive parameter is present, measuring the time scale of behavioural transitions relative to that of giving birth, death and migration among patches. By the singular perturbation method the model is reduced to a lower dimensional one in which the migration terms are, in general, nonlinear and related to the reaction terms describing the local dynamics. Two examples demonstrating the emergence of cross-migration models, i.e., the models in which the per-capita migration rate of one species depends on the density of some other species, are given.

Diminishing returns from increased percent Bt cotton: the case of pink bollworm.

Regional suppression of pests by transgenic crops producing insecticidal proteins from Bacillus thuringiensis (Bt) has been reported in several cropping systems, but little is known about the functional relationship between the ultimate pest population density and the pervasiveness of Bt crops. Here we address this issue by analyzing 16 years of field data on pink bollworm (Pectinophora gossypiella) population density and percentage of Bt cotton in the Yangtze River Valley of China. In this region, the percentage of cotton hectares planted with Bt cotton increased from 9% in 2000 to 94% in 2009 and 2010. We find that as the percent Bt cotton increased over the years, the cross-year growth rate of pink bollworm from the last generation of one year to the first generation of the next year decreased. However, as the percent Bt cotton increased, the within-year growth rate of pink bollworm from the first to last generation of the same year increased, with a slope approximately opposite to that of the cross-year rates. As a result, we did not find a statistically significant decline in the annual growth rate of pink bollworm as the percent Bt cotton increased over time. Consistent with the data, our modeling analyses predict that the regional average density of pink bollworm declines as the percent Bt cotton increases, but the higher the percent Bt cotton, the slower the decline in pest density. Specifically, we find that 95% Bt cotton is predicted to cause only 3% more reduction in larval density than 80% Bt cotton. The results here suggest that density dependence can act against the decline in pest density and diminish the net effects of Bt cotton on suppression of pink bollworm in the study region. The findings call for more studies of the interactions between pest density-dependence and Bt crops.

The halo effect: suppression of pink bollworm on non-Bt cotton by Bt cotton in China.

In some previously reported cases, transgenic crops producing insecticidal proteins from Bacillus thuringiensis (Bt) have suppressed insect pests not only in fields planted with such crops, but also regionally on host plants that do not produce Bt toxins. Here we used 16 years of field data to determine if Bt cotton caused this "halo effect" against pink bollworm (Pectinophora gossypiella) in six provinces of the Yangtze River Valley of China. In this region, the percentage of cotton hectares planted with Bt cotton increased from 9% in 2000 to 94% in 2009 and 2010. We found that Bt cotton significantly decreased the population density of pink bollworm on non-Bt cotton, with net decreases of 91% for eggs and 95% for larvae on non-Bt cotton after 11 years of Bt cotton use. Insecticide sprays targeting pink bollworm and cotton bollworm (Helicoverpa armigera) decreased by 69%. Previously reported evidence of the early stages of evolution of pink bollworm resistance to Bt cotton in China has raised concerns that if unchecked, such resistance could eventually diminish or eliminate the benefits of Bt cotton. The results reported here suggest that it might be possible to find a percentage of Bt cotton lower than the current level that causes sufficient regional pest suppression and reduces the risk of resistance.

Increased frequency of pink bollworm resistance to Bt toxin Cry1Ac in China.

Transgenic crops producing insecticidal proteins from Bacillus thuringiensis (Bt) kill some key insect pests, but evolution of resistance by pests can reduce their efficacy. The main approach for delaying pest adaptation to Bt crops uses non-Bt host plants as "refuges" to increase survival of susceptible pests. To delay evolution of pest resistance to transgenic cotton producing Bt toxin Cry1Ac, the United States and some other countries have required refuges of non-Bt cotton, while farmers in China have relied on "natural" refuges of non-Bt host plants other than cotton. The "natural" refuge strategy focuses on cotton bollworm (Helicoverpa armigera), the primary target of Bt cotton in China that attacks many crops, but it does not apply to another major pest, pink bollworm (Pectinophora gossypiella), which feeds almost entirely on cotton in China. Here we report data showing field-evolved resistance to Cry1Ac by pink bollworm in the Yangtze River Valley of China. Laboratory bioassay data from 51 field-derived strains show that the susceptibility to Cry1Ac was significantly lower during 2008 to 2010 than 2005 to 2007. The percentage of field populations yielding one or more survivors at a diagnostic concentration of Cry1Ac increased from 0% in 2005-2007 to 56% in 2008-2010. However, the median survival at the diagnostic concentration was only 1.6% from 2008 to 2010 and failure of Bt cotton to control pink bollworm has not been reported in China. The early detection of resistance reported here may promote proactive countermeasures, such as a switch to transgenic cotton producing toxins distinct from Cry1A toxins, increased planting of non-Bt cotton, and integration of other management tactics together with Bt cotton.

Gene-drive into insect populations with age and spatial structure: a theoretical assessment.

The potential benefits and risks of genetically engineered gene-drive systems for replacing wild pest strains with more benign strains must be assessed prior to any field releases. We develop a computer simulation model to assess the feasibility of using engineered underdominance constructs to drive transgenes into age- and spatially structured mosquito populations. Our practical criterion for success is the achievement of a transgene frequency of at least 0.80 within 3 years of release. The impacts of a number of parameters that may affect the success of gene-drive, such as the release area, release age, density-dependent larval survival, fitness cost of the engineered genes, and migration probability of adults, are examined. Results show that patchy release generally requires the release of fewer engineered insects to achieve success than central release. When the fitness cost is very low, central release covering 25% of the total area can be more effective than a completely uniform release over the whole area. This study demonstrates that to determine the best method of spatial release, and the total number of engineered insects that must be released, it is important to take into account the age and sex of the released insects and spatial structure of the population.

Medea selfish genetic elements as tools for altering traits of wild populations: a theoretical analysis.

One strategy for controlling transmission of insect-borne disease involves replacing the native insect population with transgenic animals unable to transmit disease. Population replacement requires a drive mechanism to ensure the rapid spread of linked transgenes, the presence of which may result in a fitness cost to carriers. Medea selfish genetic elements have the feature that when present in a female, only offspring that inherit the element survive, a behavior that can lead to spread. Here, we derive equations that describe the conditions under which Medea elements with a fitness cost will spread, and the equilibrium allele frequencies are achieved. Of particular importance, we show that whenever Medea spreads, the non-Medea genotype is driven out of the population, and we estimate the number of generations required to achieve this goal for Medea elements with different fitness costs and male-only introduction frequencies. Finally, we characterize two contexts in which Medea elements with fitness costs drive the non-Medea allele from the population: an autosomal element in which not all Medea-bearing progeny of a Medea-bearing mother survive, and an X-linked element in species in which X/Y individuals are male. Our results suggest that Medea elements can drive population replacement under a wide range of conditions.

Gene-drive in age-structured insect populations.

To date, models of gene-drive mechanisms proposed for replacing wild-type mosquitoes with transgenic strains that cannot transmit diseases have assumed no age or mating structure. We developed a more detailed model to analyze the effects of age and mating-related factors on the number of engineered insects that must be introduced into a wild population to achieve successful gene-drive based on the Medea and engineered underdominance mechanisms. We found that models without age-structure and mating details can substantially overestimate or underestimate the numbers of engineered insects that must be introduced. In general, introduction thresholds are lowest when young adults are introduced. When both males and females are introduced, assortative mating by age has little impact on the introduction threshold unless the introduced females have diminished reproductive ability because of their age. However, when only males are introduced, assortative mating by age is generally predicted to increase introduction thresholds. In most cases, introduction thresholds are much higher for male-only introductions than for both-sex introductions, but when mating is nearly random and the introduced insects are adults with Medea constructs, male-only introductions can have somewhat lower thresholds than both-sex introductions. Results from this model suggest specific parameters that should be measured in field experiments.

The dynamical implications of disease interference: correlations and coexistence.

Ecological interference between unrelated diseases, caused by the temporary or permanent removal of individuals susceptible to one disease following infection with another, might be an important mechanism underlying epidemics. In this paper, we explore the potential dynamic consequences of interference by analyzing a two-disease model. By studying the stability domain of the model's equilibria, we find that the stable region of the two-disease endemic state becomes increasingly smaller as the strength of interference (largely determined by the disease-induced mortality) increases. When seasonal changes are included in the transmission rates, the bifurcation structure of the model's periodic cycles reveals that when the two diseases have similar mean transmission rates, multiple attractors in which the two diseases are strongly correlated can coexist, and that when the two diseases have very different mean transmission rates, the one with higher mean transmission rate may determine the dynamics of the system, with the other infection mimicking the behavior. We conclude that ecological interference can have important effects on the dynamical pattern of interacting diseases, the extent of which is determined by the epidemiological features of the diseases, their mean transmission rates in particular.