Exploration of the potential of a boosted sterile insect technique to control fruit flies in mango orchards.

BACKGROUND: An innovative version of the sterile insect technique (SIT) for pest control, called boosted SIT, relies on the use of sterile males coated with a biocide to control a target wild pest population of the same species. The objective of the present study was to assess the relevance of such technology to control the fruit fly Bactrocera dorsalis and fruit losses in mango orchards using. An agent-based simulation model named BOOSTIT was used to explore the reduction of fruit losses thank to sterile male fruit flies control and economic benefits according to different strategies of sterile male release. The simulation considered a landscape of 30.25 ha made up of four mango orchards. RESULTS: The SIT and the boosted SIT reduced fruit losses when releases were made before the mango fruiting period. According to model simulations, releases should be performed at least seven times at 2-week intervals and with a sterile/wild male ratio of at least 10:1. Considering the benefit/cost ratio (BCR), few releases should be done with a late start date. The BCR showed economic gains from the two control methods, the number of saved fruits and BCR being higher for SIT. CONCLUSION: Our simulations showed that SIT would have better results than the boosted SIT to contribute to an effective control of Bactrocera dorsalis at the scale of a small landscape. We highlight the need for laboratory studies of other types of pathogen to find a suitable one with higher incubation time and lower cost. © 2024 Society of Chemical Industry.

The evolutionary process of invasion in the fall armyworm (Spodoptera frugiperda).

The fall armyworm (FAW; Spodoptera frugiperda) is one of the major agricultural pest insects. FAW is native to the Americas, and its invasion was first reported in West Africa in 2016. Then it quickly spread through Africa, Asia, and Oceania, becoming one of the main threats to corn production. We analyzed whole genome sequences of 177 FAW individuals from 12 locations on four continents to infer evolutionary processes of invasion. Principal component analysis from the TPI gene and whole genome sequences shows that invasive FAW populations originated from the corn strain. Ancestry coefficient and phylogenetic analyses from the nuclear genome indicate that invasive populations are derived from a single ancestry, distinct from native populations, while the mitochondrial phylogenetic tree supports the hypothesis of multiple introductions. Adaptive evolution specific to invasive populations was observed in detoxification, chemosensory, and digestion genes. We concluded that extant invasive FAW populations originated from the corn strain with potential contributions of adaptive evolution.

Genetic studies of fall armyworm indicate a new introduction into Africa and identify limits to its migratory behavior.

The fall armyworm, Spodoptera frugiperda (J.E. Smith) is native to the Americas and a major pest of corn and several other crops of economic importance. The species has characteristics that make it of particular concern as an invasive pest, including broad host range, long-distance migration behavior, and a propensity for field-evolved pesticide resistance. The discovery of fall armyworm in western Africa in 2016 was followed by what was apparently a remarkably rapid spread throughout sub-Saharan Africa by 2018, causing economic damage estimated in the tens of billions USD and threatening the food security of the continent. Understanding the history of the fall armyworm invasion of Africa and the genetic composition of the African populations is critical to assessing the risk posed to different crop types, the development of effective mitigation strategies, and to make Africa less vulnerable to future invasions of migratory moth pests. This paper tested and expanded on previous studies by combining data from 22 sub-Saharan nations during the period from 2016 to 2019. The results support initial descriptions of the fall armyworm invasion, including the near absence of the strain that prefers rice, millet, and pasture grasses, while providing additional evidence that the magnitude and extent of FAW natural migration on the continent is more limited than expected. The results also show that a second entry of fall armyworm likely occurred in western Africa from a source different than that of the original introduction. These findings indicate that western Africa continues to be at high risk of future introductions of FAW, which could complicate mitigation efforts.

Geographic Monitoring of Insecticide Resistance Mutations in Native and Invasive Populations of the Fall Armyworm.

Field evolved resistance to insecticides is one of the main challenges in pest control. The fall armyworm (FAW) is a lepidopteran pest species causing severe crop losses, especially corn. While native to the Americas, the presence of FAW was confirmed in West Africa in 2016. Since then, the FAW has been detected in over 70 countries covering sub-Saharan Africa, the Middle East, North Africa, South Asia, Southeast Asia, and Oceania. In this study, we tested whether this invasion was accompanied by the spread of resistance mutations from native to invasive areas. We observed that mutations causing Bt resistance at ABCC2 genes were observed only in native populations where the mutations were initially reported. Invasive populations were found to have higher gene numbers of cytochrome P450 genes than native populations and a higher proportion of multiple resistance mutations at acetylcholinesterase genes, supporting strong selective pressure for resistance against synthetic insecticides. This result explains the susceptibility to Bt insecticides and resistance to various synthetic insecticides in Chinese populations. These results highlight the necessity of regular and standardized monitoring of insecticide resistance in invasive populations using both genomic approaches and bioassay experiments.

DNA Metabarcoding as a Tool for Disentangling Food Webs in Agroecosystems.

Better knowledge of food webs and related ecological processes is fundamental to understanding the functional role of biodiversity in ecosystems. This is particularly true for pest regulation by natural enemies in agroecosystems. However, it is generally difficult to decipher the impact of predators, as they often leave no direct evidence of their activity. Metabarcoding via high-throughput sequencing (HTS) offers new opportunities for unraveling trophic linkages between generalist predators and their prey, and ultimately identifying key ecological drivers of natural pest regulation. Here, this approach proved effective in deciphering the diet composition of key predatory arthropods (nine species.; 27 prey taxa), insectivorous birds (one species, 13 prey taxa) and bats (one species; 103 prey taxa) sampled in a millet-based agroecosystem in Senegal. Such information makes it possible to identify the diet breadth and preferences of predators (e.g., mainly moths for bats), to design a qualitative trophic network, and to identify patterns of intraguild predation across arthropod predators, insectivorous vertebrates and parasitoids. Appropriateness and limitations of the proposed molecular-based approach for assessing the diet of crop pest predators and trophic linkages are discussed.

Assessing the multi-pathway threat from an invasive agricultural pest: Tuta absoluta in Asia.

Modern food systems facilitate rapid dispersal of pests and pathogens through multiple pathways. The complexity of spread dynamics and data inadequacy make it challenging to model the phenomenon and also to prepare for emerging invasions. We present a generic framework to study the spatio-temporal spread of invasive species as a multi-scale propagation process over a time-varying network accounting for climate, biology, seasonal production, trade and demographic information. Machine learning techniques are used in a novel manner to capture model variability and analyse parameter sensitivity. We applied the framework to understand the spread of a devastating pest of tomato, Tuta absoluta, in South and Southeast Asia, a region at the frontier of its current range. Analysis with respect to historical invasion records suggests that even with modest self-mediated spread capabilities, the pest can quickly expand its range through domestic city-to-city vegetable trade. Our models forecast that within 5-7 years, Tuta absoluta will invade all major vegetable growing areas of mainland Southeast Asia assuming unmitigated spread. Monitoring high-consumption areas can help in early detection, and targeted interventions at major production areas can effectively reduce the rate of spread.

Deciphering host-parasitoid interactions and parasitism rates of crop pests using DNA metabarcoding.

An accurate estimation of parasitism rates and diversity of parasitoids of crop insect pests is a prerequisite for exploring processes leading to efficient natural biocontrol. Traditional methods such as rearing have been often limited by taxonomic identification, insect mortality and intensive work, but the advent of high-throughput sequencing (HTS) techniques, such as DNA metabarcoding, is increasingly seen as a reliable and powerful alternative approach. Little has been done to explore the benefits of such an approach for estimating parasitism rates and parasitoid diversity in an agricultural context. In this study, we compared the composition of parasitoid species and parasitism rates between rearing and DNA metabarcoding of host eggs and larvae of the millet head miner, Heliocheilus albipunctella De Joannis (Lepidoptera, Noctuidae), collected from millet fields in Senegal. We first assessed the detection threshold for the main ten endoparasitoids, by sequencing PCR products obtained from artificial dilution gradients of the parasitoid DNAs in the host moth. We then assessed the potential of DNA metabarcoding for diagnosing parasitism rates in samples collected from the field. Under controlled conditions, our results showed that relatively small quantities of parasitoid DNA (0.07 ng) were successfully detected within an eight-fold larger quantity of host DNA. Parasitoid diversity and parasitism rate estimates were always higher for DNA metabarcoding than for host rearing. Furthermore, metabarcoding detected multi-parasitism, cryptic parasitoid species and differences in parasitism rates between two different sampling sites. Metabarcoding shows promise for gaining a clearer understanding of the importance and complexity of host-parasitoid interactions in agro-ecosystems, with a view to improving pest biocontrol strategies.

Plant training for induced defense against insect pests: a promising tool for integrated pest management in cotton.

Enhancing cotton pest management using plant natural defenses has been described as a promising way to improve the management of crop pests. We here reviewed various studies on cotton growing systems to illustrate how an ancient technique called plant training, which includes plant topping and pruning, may contribute to this goal. Using examples from cotton crops, we show how trained plants can be brought to a state of enhanced defense that causes faster and more robust activation of their defense responses. We revisit the agricultural benefits associated with this technique in cotton crops, with a focus on its potential as a supplementary tool for integrated pest management (IPM). In particular, we examine its role in mediating plant interactions with conspecific neighboring plants, pests and associated natural enemies. We propose a new IPM tool, plant training for induced defense, which involves inducing plant defense through artificial injury. Experimental evidence from various studies shows that cotton training is a promising technique, particularly for smallholders, which can be used as part of an IPM program to significantly reduce insecticide use and to improve productivity in cotton farming. © 2018 Society of Chemical Industry.

Life-History Traits of Macrolophus pygmaeus with Different Prey Foods.

Macrolophus pygmaeus Rambur (Hemiptera: Miridae) is a generalist predatory mirid widely used in augmentative biological control of various insect pests in greenhouse tomato production in Europe, including the invasive tomato leafminer, Tuta absoluta (Meyrick) (Lepidoptera, Gelechiidae). However, its biocontrol efficacy often relies on the presence of alternative prey. The present study aimed at evaluating the effect of various prey foods (Ephestia kuehniella eggs, Bemisia tabaci nymphs, Tuta absoluta eggs and Macrosiphum euphorbiae nymphs) on some life history traits of M. pygmaeus. Both nymphal development and adult fertility of M. pygmaeus were significantly affected by prey food type, but not survival. Duration of nymphal stage was higher when M. pygmaeus fed on T. absoluta eggs compared to the other prey. Mean fertility of M. pygmaeus females was greatest when fed with B. tabaci nymphs, and was greater when offered M. euphorbiae aphids and E. kuehniella eggs than when offered T. absoluta eggs. Given the low quality of T. absoluta eggs, the efficacy of M. pygmaeus to control T. absoluta may be limited in the absence of other food sources. Experiments for assessing effectiveness of generalist predators should involve the possible impact of prey preference as well as a possible prey switching.

A seed mixture increases dominance of resistance to Bt cotton in Helicoverpa zea.

Widely grown transgenic crops producing insecticidal proteins from Bacillus thuringiensis (Bt) can benefit agriculture, but adaptation by pests threatens their continued success. Refuges of host plants that do not make Bt toxins can promote survival of susceptible insects and delay evolution of resistance, particularly if resistance is inherited as a recessive trait. However, data have been lacking to compare the dominance of resistance when Bt and non-Bt seeds are planted in random mixtures versus separate blocks. Here we report results from greenhouse experiments with transgenic cotton producing Bt toxin Cry1Ac and the bollworm, Helicoverpa zea, showing that the dominance of resistance was significantly higher in a seed mixture relative to a block of Bt cotton. The proportion of larvae on non-Bt cotton plants in the seed mixture was also significantly higher than expected under the null hypothesis of random distribution. In simulations based on observed survival, resistance evolved 2- to 4.5-fold faster in the seed mixture relative to separate blocks of Bt and non-Bt cotton. These findings support previous modelling results indicating that block refuges may be more effective than seed mixtures for delaying resistance in pests with mobile larvae and inherently low susceptibility to the toxins in Bt crops.

Insect resistance to Bt crops: lessons from the first billion acres.

Evolution of resistance in pests can reduce the effectiveness of insecticidal proteins from Bacillus thuringiensis (Bt) produced by transgenic crops. We analyzed results of 77 studies from five continents reporting field monitoring data for resistance to Bt crops, empirical evaluation of factors affecting resistance or both. Although most pest populations remained susceptible, reduced efficacy of Bt crops caused by field-evolved resistance has been reported now for some populations of 5 of 13 major pest species examined, compared with resistant populations of only one pest species in 2005. Field outcomes support theoretical predictions that factors delaying resistance include recessive inheritance of resistance, low initial frequency of resistance alleles, abundant refuges of non-Bt host plants and two-toxin Bt crops deployed separately from one-toxin Bt crops. The results imply that proactive evaluation of the inheritance and initial frequency of resistance are useful for predicting the risk of resistance and improving strategies to sustain the effectiveness of Bt crops.

Potential shortfall of pyramided transgenic cotton for insect resistance management.

To delay evolution of pest resistance to transgenic crops producing insecticidal proteins from Bacillus thuringiensis (Bt), the "pyramid" strategy uses plants that produce two or more toxins that kill the same pest. In the United States, this strategy has been adopted widely, with two-toxin Bt cotton replacing one-toxin Bt cotton. Although two-toxin plants are likely to be more durable than one-toxin plants, the extent of this advantage depends on several conditions. One key assumption favoring success of two-toxin plants is that they kill insects selected for resistance to one toxin, which is called "redundant killing." Here we tested this assumption for a major pest, Helicoverpa zea, on transgenic cotton producing Bt toxins Cry1Ac and Cry2Ab. Selection with Cry1Ac increased survival on two-toxin cotton, which contradicts the assumption. The concentration of Cry1Ac and Cry2Ab declined during the growing season, which would tend to exacerbate this problem. Furthermore, analysis of results from 21 selection experiments with eight species of lepidopteran pests indicates that some cross-resistance typically occurs between Cry1A and Cry2A toxins. Incorporation of empirical data into simulation models shows that the observed deviations from ideal conditions could greatly reduce the benefits of the pyramid strategy for pests like H. zea, which have inherently low susceptibility to Bt toxins and have been exposed extensively to one of the toxins in the pyramid before two-toxin plants are adopted. For such pests, the pyramid strategy could be improved by incorporating empirical data on deviations from ideal assumptions about redundant killing and cross-resistance.

Assessing the role of non-cotton refuges in delaying Helicoverpa armigera resistance to Bt cotton in West Africa.

Non-cotton host plants without Bacillus thuringiensis (Bt) toxins can provide refuges that delay resistance to Bt cotton in polyphagous insect pests. It has proven difficult, however, to determine the effective contribution of such refuges and their role in delaying resistance evolution. Here, we used biogeochemical markers to quantify movement of Helicoverpa armigera moths from non-cotton hosts to cotton fields in three agricultural landscapes of the West African cotton belt (Cameroon) where Bt cotton was absent. We show that the contribution of non-cotton hosts as a source of moths was spatially and temporally variable, but at least equivalent to a 7.5% sprayed refuge of non-Bt cotton. Simulation models incorporating H. armigera biological parameters, however, indicate that planting non-Bt cotton refuges may be needed to significantly delay resistance to cotton producing the toxins Cry1Ac and Cry2Ab. Specifically, when the concentration of one toxin (here Cry1Ac) declined seasonally, resistance to Bt cotton often occurred rapidly in simulations where refuges of non-Bt cotton were rare and resistance to Cry2Ab was non-recessive, because resistance was essentially driven by one toxin (here Cry2Ab). The use of biogeochemical markers to quantify insect movement can provide a valuable tool to evaluate the role of non-cotton refuges in delaying the evolution of H. armigera resistance to Bt cotton.

Insecticide resistance in field populations of Bemisia tabaci (Hemiptera: Aleyrodidae) in West Africa.

BACKGROUND: The tobacco whitefly, Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae), has developed a high degree of resistance to several chemical classes of insecticides throughout the world. To evaluate the resistance status in West Africa, eight insecticides from different chemical families were tested using the leaf-dip method on four field populations collected from cotton in Benin, Togo and Burkina Faso. RESULTS: Some field populations showed a significant loss of susceptibility to pyrethroids such as deltamethrin [resistance ratio (RR) 3-5] and bifenthrin (RR 4-36), to organophosphates (OPs) such as dimethoate (RR 8-15) and chlorpyrifos (RR 5-7) and to neonicotinoids such as acetamiprid (RR 7-8) and thiamethoxam (RR 3-7). Bemisia tabaci was also resistant to pymetrozine (RR 3-18) and to endosulfan (RR 14-30). CONCLUSION: The resistance of B. tabaci to pyrethroids and OPs is certainly due to their systematic use in cotton treatments for more than 30 years. Acetamiprid has been recently introduced for the control of whiteflies. Unfortunately, B. tabaci populations from Burkina Faso seem to be already resistant. Because cross-resistance between these compounds has never been observed elsewhere, resistance to neonicotinoids could be due to the presence of an invasive B. tabaci biotype recently detected in the region.

Interaction between visual and olfactory cues during host finding in the tomato fruit fly Neoceratitis cyanescens.

Herbivorous insects searching for a host plant need to integrate a sequence of multimodal sensory inputs. We conducted a series of no-choice experiments in a laboratory wind tunnel to examine the behavioral response of the specialist fruit fly, Neoceratitis cyanescens (Diptera: Tephritidae), to host visual and olfactory stimuli presented singly or in combination (e.g., colored fruit model with or without host fruit odor). We also studied the influence of wind flow, age, and sex on the response of flies. In two-choice experiments, we evaluated the ability of mature females to discriminate between two fruit models emitting host vs. non-host fruit odor or clean air. Neoceratitis cyanescens mature females can use independently or interactively olfactory and visual stimuli to locate their host, whereas immature females and males respond primarily to host fruit odor. In the absence of wind, mature females mainly use visual information to locate the host fruit. In wind, host fruit odor significantly increases the probability and speed of locating the host fruit. In a two-choice situation between two bright orange spheres, flies accurately detected the sphere emitting host fruit odor vs. non-host fruit odor or odorless air. Nevertheless, they preferred to land on the bright orange sphere when the sphere emitting host fruit odor was blue. Furthermore, when odor source and fruit model were spatially decoupled (90 or 180 degrees ), >50% flies that landed on the fruit model initially performed an oriented flight toward the odor source, then turned back to the fruit model while in flight or after one landing, thus suggesting visual information to be the ultimate indicator of host fruit.

Insecticide resistance traits differ among and within host races in Aphis gossypii.

BACKGROUND: The polyphagous cotton-melon aphid Aphis gossypii Glover is structured into geographically widespread host races comprising a few clones specialised on Cucurbitaceae, cotton, eggplant or pepper. To assess insecticide resistance among and within host races, leaf disc bioassays were conducted on aphid clones collected from Cucurbitaceae (genotypes C4 and C9), cotton (genotypes Burk and Ivo), eggplant (genotype Auber) and pepper (genotype PsP4). Molecular diagnostic (PCR-RFLP) and enzyme assays were also performed to detect the basic mechanisms underlying insecticide resistance. RESULTS: All six clones were susceptible to acetamiprid (neonicotinoid) or carbosulfan (carbamate). Conversely, all clones were resistant to dimethoate (organophosphate) (RF = 4.1-38.1) and carried mutation S431F in the acetylcholinesterase gene. Auber, PsP4 and Burk also carried mutation A302S in this gene, which possibly conferred moderate resistance (RF = 3.7-6.8) to profenofos and monocrotophos (organophosphates). Auber and Burk were highly resistant (RF = 41.2 and 473 respectively) to cypermethrin (pyrethroid). This resistance was likely associated with point mutation super-kdr (M918L) in the voltage-gated sodium channel gene (para gene) or metabolic detoxification mediated by esterase and oxidase enzymes. CONCLUSION: Multiple resistance to a broad range of insecticides and multiple mechanisms of resistance in some clones could explain to some extent the low genetic diversity observed within A. gossypii host races.

Inheritance and stability of pyrethroid resistance in the cotton bollworm Helicoverpa armigera (Lepidoptera: Noctuidae) in Central Africa.

BACKGROUND: The cotton bollworm Helicoverpa armigera (Hübner) has developed esterase-mediated resistance to pyrethroids in Central Africa. To gain a better understanding of how quickly this resistance can evolve, its genetic basis and stability were examined in a field-derived strain of H. armigera (474-fold resistant to cypermethrin). RESULTS: Genetic crosses between susceptible, resistant and F(1) hybrids indicated that resistance was inherited as a dominant trait (D(LD) = 0.86) and conferred by a single autosomal gene. The dominance of resistance decreased as the cypermethrin dose increased, so that resistance was incompletely dominant (D(ML) = 0.73) at the highest dose tested. Resistance (DL(50)) of the F(1) hybrid progeny significantly decreased over five generations in the absence of pyrethroid exposure. CONCLUSION: Rapid selection of resistance alleles due to dominance supports the ability of H. armigera to develop resistance to pyrethroids in Central Africa. However, associated fitness costs provide useful information for managing the evolution of resistance.

Esterase-mediated resistance to pyrethroids in field populations of Helicoverpa armigera (Lepidoptera: Noctuidae) from Central Africa.

BACKGROUND: Evolution of pyrethroid resistance in the cotton bollworm Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) threatens continued cotton production in Central Africa. Dose-response bioassays were conducted on area-wide collection of bollworm populations from major host plants, while biochemical techniques were used to evaluate basic mechanisms underlying resistance. RESULTS: Pyrethroid resistance is primarily associated with detoxification by enhanced esterase activity. High resistance to cypermethrin (RF = 67-1771), cross-resistance to deltamethrin (RF = 60-2972) and lack of cross-resistance to the non-ester pyrethroid etofenprox (RF = 2-10) were observed among H. armigera field populations and laboratory-selected strains. Enzymatic assays showed that esterase activity, but not oxidase content or glutathione-S-transferase activity, was positively correlated with resistance to cypermethrin. Pretreatment with piperonyl butoxide (PBO) resulted in significant synergism with cypermethrin in 6/10 field populations, but not in the laboratory-selected strain, indicating that additional mechanisms such as mixed-function oxidase (MFO) may be involved in field resistance. The absence of cross-resistance to DDT ruled out a possible target-site modification. CONCLUSION: Knowledge of the mechanisms involved in pyrethroid resistance and the lack of cross-resistance to spinosad and indoxacarb is a key to devising new resistance management strategies aimed at restoring the efficacy of pyrethroid-based programmes.

Genetic structure of the polyphagous pest Helicoverpa armigera (Lepidoptera: Noctuidae) across the Sub-Saharan cotton belt.

The cotton bollworm Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) is a key pest of various cropping systems in West and Central Africa, and developed insecticide resistance recently. To understand how such insecticide resistance expands across the region, the genetic structure of bollworm populations was studied using microsatellite markers. At first, the study was performed within several populations from Northern Cameroon: during one year, 19 populations (504 larvae) were sampled in different locations, dates and host plants (6 villages, 6 dates, 5 host plants). Their genetic relationship was analysed using 10 polymorphic microsatellite markers. Despite the high polymorphism (5 to 50 alleles per locus), results reveal low level of genetic distances among locations, collection dates and host plants. The estimated values of F(ST) were very low across all populations and reveal a high level of gene flow. Moreover, all the loci presented heterozygote deficiency. This may arise either from inbreeding (sampling methodology) or from the presence of null alleles. Subsequently, larval sampling was performed at a higher scale, in five locations from Africa (Senegal, Mali, Burkina-Faso, Togo and Cameroon), to detect population differentiations according to geographic distance/isolation. Two other samples, from Madagascar and from Australia, were added to this experiment. F(ST) values and heterozygote frequency data were identical to the first one, indicating a high level of gene flow between these locations and the high migration capacity of the pest. Samples from Thailand, China, Pakistan and France were added to this study but it has been impossible to infer the presence of distinct populations. The opportunity to use neutral markers as microsatellites to understand population dynamics of H. armigera is discussed.

Status of self-pollen in bee pollination efficiency of white clover (Trifolium repens L.).

Flowers of white clover (Trifolium repens L.) are hermaphrodite and self-incompatible; their cross-pollination depends entirely on insect visitors, mainly bees (Apoidea). Because self-pollination of white clover occurs before flower anthesis, we determined whether selfing affected the pollination efficiency of a honeybee visit. We compared pollen deposition in emasculated and intact flowers following (1) a single honeybee visit, (2) open-pollination for a day and (3) enclosure in a cloth bag to prevent insect visits. In emasculated flowers, open-pollination resulted in more pollen deposited than after one visit (+30%) which is consistent with flowers being visited more than once by pollinators during the course of a day. On intact flowers, saturation of the stigma was achieved after the first visit of a honeybee (near 280 grains) because of self-pollination. Additional visits did not increase pollen deposits, but they improved pollen efficiency in terms of numbers of pollen tubes reaching the ovules. In such a context of easily saturated stigmas, self-pollen does not inhibit cross-pollen activity, but represents a constraint for pollination which demands multiple bee visits to each flower to achieve maximum fertilization.