Climatic factors on entomopathogenic hyphomycetes infection of Trialeurodes vaporariorum (Homoptera: Aleyrodidae) in Mediterranean glasshouse tomato

Collaborative research was conducted at the INRA Research Centers to assess the microbial control potential of Beauveria bassiana- and Lecanicillium lecanii-based formulations against whiteflies in protected crops under Mediterranean conditions. Four series of small-scale glasshouse trials were performed in 1999 and 2000 in southern France. Two applications at 4–5 day intervals of Naturalis-L and Mycotal were conducted on young larvae of the greenhouse whitefly, Trialeurodes vaporariorum, at rates recommended by the manufacturers. Because of the expectation that environmental conditions prevailing in Mediterranean greenhouse crops may lead to greater climatic constraints for mycoinsecticide efficacy than in more temperate areas, manipulation of the greenhouse climate has been used to aim at optimizing mycoinsecticide efficacy. The climatic management strategy was mainly based on closing the ridge vents 2 h more at night-time in so-called “humid” glasshouse compartment than in a “dry” one. Thus, the daily period at high humidity (>90% RH) was two or three times longer in the “humid” compartment than in the “dry” one. In spite of this differential, mycoinsecticide treatments reduced numbers of surviving whitefly larvae by >85% in the “humid” compartment as expected as favorable, as well as in the “dry” compartment, expected as unfavorable. The results indicated clearly that both B. bassiana- and L. lecanii-based mycoinsecticides have a strong potential for microbial control of whitefly larvae infesting tomato crops at moderate ambient humidity in Mediterranean glasshouses. Our investigations provided strong arguments for explaining these unexpected results. The RH conditions prevailing in the targeted insect habitat should be greatly disconnected from that of the ambient glasshouse air. We suggest that strategies of mycoinsecticide optimization against phyllophagous insects in protected crops have to take into account factors acting on the leaf transpiration activity.     

The effects of mullein plants (Verbascum thapsus) on the population dynamics of Dicyphus hesperus (Heteroptera: Miridae) in tomato greenhouses

The response of Dicyphus hesperus Knight (Heteroptera: Miridae) to whitefly populations in tomato greenhouses was measured in the presence and absence of mullein (Verbascum thapsus L.) as an alternative host plant. The dynamics of the D. hesperus population on tomato (Lycopersicon esculentum Mill.) and on mullein plants were followed through an entire growing season. In houses with mullein plants, more predators occurred on mullein when whitefly density was low on tomato. A mark-release-recapture experiment where rabbit IgG was used as an external marker showed that D. hesperus adults moved from mullein plants to tomato plants. D. hesperus was always more abundant in houses with mullein than in the houses with tomato plants alone. Movements between tomato and mullein plants are discussed as a strategy to optimize predator foraging. The use of mullein as an alternative host plant may contribute to the establishment of D. hesperus and help to preserve the predator population when prey on tomato crops is scarce.     

Effect of microclimate heterogeneity and ventilation system on entomopathogenic hyphomycete infection of Trialeurodes vaporariorum (Homoptera: Aleyrodidae) in Mediterranean greenhouse tomato

Collaborative research was conducted in the south of France to assess constraints related to both climate heterogeneity and ventilation systems on the control potential of a Lecanicillium muscarium-based formulation against whiteflies in Mediterranean greenhouses. Four series of small-scale greenhouse trials were performed in 2001 and 2002. Two applications at 4–5 day intervals of Mycotal were conducted on young larvae of the greenhouse whitefly, Trialeurodes vaporariorum, at the rate recommended by the manufacturer (ca. 10¹⁰ viable spores per liter of water suspension). The climatic heterogeneity was taken into account by comparing the fungus-induced mortality of nymphs located on lateral row plants to that of nymphs on center row plants. In spite of significant differences in air flows (0.7–1.2 and 0.3 ms⁻¹, respectively) there was no effect on fungus efficacy (53–76% mortality). When comparing the influence of greenhouse equipment (sophisticated glasshouse vs. polyethylene-covered greenhouse), the fungus was not affected (89–96% mortality) in spite of significant differences in ventilation rates. The results confirmed that entomopathogenic Hyphomycetes have a strong potential for microbial control of whitefly larvae infesting tomato crops at moderate ambient humidity in Mediterranean greenhouses in spite of windy periods. These investigations confirmed that microclimatic conditions prevailing in the targeted insect habitat (under-leaf surface boundary layer) are greatly disconnected from that of both outside and inside the greenhouse. In northern Mediterranean greenhouses, non-stressed tomato crops provide unexpected favorable conditions for mycoinsecticide use against a phyllophagous insect.     

Predation by <Emphasis Type="Italic">Nesidiocoris tenuis</Emphasis> on <Emphasis Type="Italic">Bemisia tabaci</Emphasis> and injury to tomato

Tomato is the most important vegetable crop in Spain. The mirid bug Nesidiocoris tenuis (Reuter) commonly appears in large numbers in protected and open-air tomato crops where little or no broad-spectrum insecticides are used. Nesidiocoris tenuis is known to be a predator of whiteflies, thrips and several other pest species. However, it is also considered a pest because it can feed on tomato plants, causing necrotic rings on stems and flowers and punctures in fruits. Our objectives were to evaluate predation by N. tenuis on sweetpotato whitefly Bemisia tabaci Gennadius under greenhouse conditions and establish its relationship to N. tenuis feeding on tomato. Two different release rates of N. tenuis were compared with an untreated control (0, 1 and 4 N. tenuis plant⁻¹) in cages of 8 m². Significant reductions of greater than 90% of the whitefly population and correspondingly high numbers of N. tenuis were observed with both release rates. Regression analysis showed that necrotic rings on foliage caused by N. tenuis were best explained by the ratio of B. tabaci nymphs:N. tenuis as predicted by the equation y = 15.086x − 0.6359.

Establishment of papaya banker plant system for parasitoid, Encarsia sophia (Hymenoptera: Aphilidae) against Bemisia tabaci (Hemiptera: Aleyrodidae) in greenhouse tomato production

The silverleaf whitefly, Bemisia tabaci biotype B (Gennadius) (Hemiptera: Aleyrodidae), is a key pest of tomato (Solanum lycopersicum L.) and other vegetable crops worldwide. To combat this pest, a non-crop banker plant system was evaluated that employs a parasitoid, Encarsia sophia (Girault & Dodd) (Hymenoptera: Aphelinidae) with whitefly, Trialeurodes variabilis (Quaintance) (Hemiptera: Aleyrodidae), as an alternative host for rearing and dispersal of the parasitoid to the target pest. (a) Multi-choice and no-choice greenhouse experiments were conducted to determine host specificity of T. variabilis to papaya (Carica papaya L.) and three vegetable crops including tomato, green bean (Phaseolus vulgaris L.), and cabbage (Brassica oleracea L.). The result showed that papaya was an excellent non-crop banker plant for supporting the non-pest alternative host, T. variabilis, whose adults had a strong specificity to papaya plants for feeding and oviposition in both multi-choice and no-choice tests. (b) The dispersal ability of E. sophia was investigated from papaya banker plants to tomato and green bean plants infested with B. tabaci, as well as to papaya control plants infested with T. variabilis; and (c) the percent parasitism by E. sophia on T. variabilis reared on papaya plants and on B. tabaci infested on tomato plants was also evaluated. These data proved that E. sophia was able to disperse at least 14.5 m away from papaya plants to target tomato, bean or papaya control plants within 48–96 h. Furthermore, E. sophia was a strong parasitoid of both T. variabilis and B. tabaci. There was no significant difference in percent parasitism by E. sophia on T. variabilis (36.2–47.4%) infested on papaya plants or B. tabaci (29–45.9%) on tomato plants. Thus, a novel banker plant system for the potential management of B. tabaci was established using papaya as a non-crop banker plant to support a non-pest alternative host, T. variabilis for maintaining the parasitoid to control B. tabaci. The established banker plant system should provide growers with a new option for long-term control of B. tabaci in greenhouse vegetable production. Ongoing studies on the papaya banker plant system are being performed in commercial greenhouses.     

Zoophytophagy in the plantbug Nesidiocoris tenuis

1 The zoophytophagy of Nesidiocoris tenuis (Reuter) (Heteroptera: Miridae) was characterized in relation to prey availability and environmental factors by: (i) monitoring its population dynamics in tomato greenhouses; (ii) analysis of the influence of N. tenuis and whitefly density, temperature and humidity on the intensity of N. tenuis plant feeding; and (iii) laboratory assays under controlled conditions to determine the intensity of plant feeding in relation to prey availability, temperature and humidity. 2 A negative relationship was found between plant feeding and predated whiteflies in tomato greenhouses. Plant feeding was directly related to N. tenuis density and temperature and inversely related to whitefly density. The significance of prey availability and temperature was corroborated in laboratory assays. The intensification of plant feeding at low prey density indicates switching from zoophagy to phytophagy as prey become scarce. 3 Nesidiocoris tenuis showed a typical predator dynamic in relation to variance in prey density. Populations increased after whitefly outbreaks and decreased after whitefly had been depleted. The rapid decrease of N. tenuis populations after whitefly decreased, however, suggests that plants are a poorer nutrient source than whitefly for this species.     

Parasitism rates and sex ratios of a parasitoid wasp: effects of herbivore and plant quality

Summary We studied interactions among collards, Brassica oleracea var. acephala, the diamondback moth (DBM), Plutella xylostella (Lepidoptera: Yponomeutidae) and its parasitoid Diadegma insulare (Hymenoptera: Ichneumonidae) by manipulating plant nitrogen (N) concentrations in field and laboratory experiments. Parasitoid abundance strongly reflected DBM abundance and was related to total leaf N. Parasitism rates were high (70.7%) and density-independent. Wasp sex ratios varied markedly (3–93% female) in response to the herbivores, the plants, or both. Higher proportions of female wasps emerged from DBM larvae on plants with high leaf N than on unfertilized plants. More female wasps also emerged from larvae parasitized as larger instars. We suggest that wasps have the potential to control DBM populations through long-term numerical responses mediated by variable sex ratios.     

Impact of natural epizootics of the fungal pathogen Neozygites floridana (Zygomycetes: Entomophthorales) on population dynamics of Tetranychus evansi (Acari: Tetranychidae) in tomato and nightshade

The tomato red spider mite, Tetranychus evansi (Acari: Tetranychidae) was recently introduced in Africa and Europe, where there is an increasing interest in using natural enemies to control this pest on solanaceous crops. Two promising candidates for the control of T. evansi were identified in South America, the fungal pathogen, Neozygites floridana and the predatory mite Phytoseiulus longipes. In this study, population dynamics of T. evansi and its natural enemies together with the influence of environmental conditions on these organisms were evaluated during four crop cycles in the field and in a protected environment on nightshade and tomato plants with and without application of chemical pesticides. N. floridana was the only natural enemy found associated with T. evansi in the four crop cycles under protected environment but only in the last crop cycle in the field. In the treatments where the fungus appeared, reduction of mite populations was drastic. N. floridana appeared in tomato plants even when the population density of T. evansi was relatively low (less than 10 mites/3.14 cm² of leaf area) and even at this low population density, the fungus maintained infection rates greater than 50%. The application of pesticides directly affected the fungus by delaying epizootic initiation and contributing to lower infection rates than unsprayed treatments. Rainfalls did not have an apparent impact on mite populations. These results indicate that the pathogenic fungus, N. floridana can play a significant role in the population dynamics of T. evansi, especially under protected environment, and has the potential to control this pest in classical biological control programs.     

Effect of Host Plant on the Potential ofPaecilomyces fumosoroseus(Deuteromycotina: Hyphomycetes) for Controlling the Silverleaf Whitefly,Bemisia argentifolii(Homoptera: Aleyrodidae) in Greenhouses

We determined host plant effect on susceptibility of the silverleaf whitefly,Bemisia argentifolii, to the entomopathogenic fungusPaecilomyces fumosoroseus. Whiteflies were reared on three vegetable species (cucumber, cabbage, and tomato) and three cultivars of tomato (Heatwave, Better Boy, and Rutgers). Second instars were sprayed with 5 × 10⁴conidia/cm²ofPfr97, aP. fumosoroseusstrain, used as a microbial control agent of whiteflies. Trials were conducted in an experimental greenhouse, where temperature and relative humidity were adjusted to favor infection (22–33°C, and 68–100% RH). Larval susceptibility to fungal infection was high and not significantly affected by the host plant. Mortality was > 70% 1 week after treatment and increased further during the second week. Percentages of cadavers with subsequent production of conidia observed in the greenhouse did not vary significantly either with the host vegetable species (85–93% 7 days after treatment and 99–100% 14 days after treatment), or with the cultivar of tomato (96–97% 7 days after treatment and 99–100% 14 days after treatment). After incubation under optimal laboratory conditions, the percentages based on the total number of sporulating cadavers (includingin situsporulating individuals and cadavers sporulating afterin vitroincubation) were not significantly influenced either by host vegetable or cultivar of tomato. According to the conditions prevailing in the series of experiments with the three vegetable species or in the series of experiments with the three cultivars of tomatoes, the production of newly formed conidia varied from approximately 10,000 to 18,000 conidia/cadaver. However, in both series, there was no significant influence of the host vegetable species or cultivar. The survival of the newly formed conidia harvested 7 days following treatment reached more than 50% but was not affected by host plant. These results indicate thatP. fumosoroseusshows potential as a microbial control agent for controllingB. argentifoliion greenhouse crops.     

Intraguild predation and cannibalism among Dicyphini: Dicyphus cerastii vs. two commercialized species

Dicyphine mirids are one of the most important groups of predators on tomato. In the Mediterranean region, several species in the genera Dicyphus, Macrolophus, and Nesidiocoris (Hemiptera: Miridae, Bryocorinae, Dicyphini) colonize protected horticultural crops. In Portugal, Nesidiocoris tenuis (Reuter) is increasingly abundant in the mirid species complex of tomato crops and appears to be displacing the native Dicyphus cerastii Wagner. In order to know whether intraguild predation (IGP) can explain the decreasing abundance of D. cerastii, we evaluated predatory interactions between adult females and first instars of D. cerastii vs. N. tenuis but also D. cerastii vs. Macrolophus pygmaeus (Rambur), as this species is also naturally present in horticultural crops in Portugal. Cannibalistic interactions were also tested for the same three species. All experiments were performed under laboratory conditions, in Petri dish arenas, in the presence or absence of Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) eggs as alternative prey. Predation on both heterospecific and conspecific nymphs occurred only in the absence of alternative food. Intraguild predation was mutual and symmetrical between D. cerastii and M. pygmaeus. However, IGP was asymmetrical between D. cerastii and N. tenuis, favouring the first. Cannibalism was not significantly different among these mirid species. Our results show that D. cerastii has a greater capacity to feed on intraguild prey than N. tenuis. Therefore, IGP on small nymphs does not explain the abundance shift between D. cerastii and N. tenuis.     

Life table and biological characteristics of Macrolophus pygmaeus when feeding on Myzus persicae and Trialeurodes vaporariorum

The life table and biological characteristics of the predatory bug Macrolophus pygmaeus Rambur (Hemiptera: Miridae) were studied when the bugs were fed with Myzus persicae (Sulzer) (Homoptera: Aphididae) feeding on eggplant and with Trialeurodes vaporariorum Westwood (Homoptera: Aleyrodidae) feeding on tomato plants. The tests were done at five temperatures between 15 and 30 °C, using a L16:D8 photoperiod and 65 ± 5% r.h. Most eggs (range 85 to 90%) hatched at 15 and 20 °C. Incubation period was shortest at 27.5 °C (8.45 and 8.38 days on eggplant and tomato, respectively). Preoviposition was also shortest at 27.5 °C (5.10 and 4.75 days on eggplant and tomato, respectively) whereas fecundity was highest at 20 °C (213.90 and 228.25 eggs on eggplant and tomato, respectively). Maximum longevity of females was at 15 °C (122.40 and 129.35 days on eggplant and tomato, respectively). Mean generation time was longest at 15 °C on both host plants (122.75 and 124.64 days, respectively). The intrinsic rate of increase of M. pygmaeus was highest at 27.5 °C with similar values on eggplant (0.0981 day^–1) and tomato (0.1040 day^–1). Doubling time was shortest at 27.5 °C (7.06 and 6.67 days on eggplant and tomato, respectively) and, also, finite rate of increase was highest at 27.5 °C (1.1031 and 1.1096 on eggplant and tomato, respectively). The results show that the predator M. pygmaeus develops well on the aphid M. persicae or on the whitefly T. vaporariorum, both of which are important pests of vegetable crops. This predator is also well adapted to the temperatures that occur both in greenhouses and in the open field in the Mediterranean region. Compared to other natural enemies of whiteflies, such as Encarsia formosa Gahan (Hymenoptera: Aphelinidae), Macrolophus pygmaeus can increase at relatively low temperatures.     

Dicyphus tamaninii as a beneficial insect and pest in tomato crops in Catalonia,Spain

The mirid bugDicyphus tamaninii Wagner, is common in Catalonia in vegetable crops which have not been sprayed with insecticide. To study the feeding habits of this bug in tomato crops, several treated and untreated tomato plots were set up at Cabrils Experimental Station (North of Barcelona) during 1983, 1984 and 1985. The results indicate thatTrialeurodes vaporariorum Westwood populations are greatly affected by the mirid bug when no, or only a few, insecticide sprays are applied. However, the bug heavily damaged tomato fruits in unsprayed plots, where bug numbers were high and whitefly numbers low. It seems that tomato fruit is only preferred when whitefly numbers are extremely low. Similar results were obtained in 1984 in a preliminary experiment using cages to exclude the bug.      

Previous and Present Diets of Mite Predators Affect Antipredator Behaviour of Whitefly Prey

Predator diet is known to influence antipredator behaviour in prey. Yet, it is not clear how antipredator behaviour is affected by diet changes of the predator. We studied the effect of previous and present diet of a predatory mite Typhlodromips swirskii on the antipredator response of its prey, the whitefly Bemisia tabaci. An earlier study showed that adult female whiteflies that had experienced predators, had learned to avoid ovipositing on plants with predators whose previous and present diet consisted of whitefly eggs and immatures. Here, we investigate whether adult whiteflies also avoid plants with predators whose present and/or previous diet consisted of a non-whitefly food source. Adult whiteflies were found not to avoid plants with predators whose present diet consisted of pollen and whose previous diet had consisted of either pollen or whitefly eggs and larvae. They did avoid plants with predators whose present diet consisted of whiteflies and whose previous diet had consisted of pollen, but to a lesser extent than when previous and present diet consisted of whiteflies. In a choice experiment, whiteflies discriminated between plants with predators whose present diet consisted of whiteflies, but that differed in previous diet. Our results show that both previous and present diets of predators are important in eliciting antipredator behaviour.     

Changes in leaf δC and δN for three Mediterranean tree species in relation to soil water availability

A rain exclusion experiment simulating drought conditions expected in Mediterranean areas for the following decades (15% decrease in soil moisture) was conducted in a Mediterranean holm oak forest to study the response of leaf δ¹³C, δ¹⁵N, and N concentrations to the predicted climatic changes for the coming decades. Plant material was sampled in 2000, 2003, 2004, and 2005 in eight plots: four of them were control plots and the other four plots received the rain exclusion treatment. Although there was a negative relationship between δ¹³C and soil moisture, for each species and year, the rain exclusion treatment did not have any significant effect on δ¹³C, and therefore on the intrinsic water use efficiency (iWUE) of the three dominant species: Phillyrea latifolia, Arbutus unedo, and Quercus ilex. On the other hand, rain exclusion clearly increased the δ¹⁵N values in the three species studied, probably indicating higher N losses at the soil level leading to a ¹⁵N enrichment of the available N. It suggested that rain exclusion exerted a greater effect on the nitrogen biogeochemical cycle than on the carbon assimilation process. δ¹⁵N values were inversely correlated with summer soil moisture in Q. ilex and A. unedo, but no relationship was observed in P. latifolia. This latter species showed the lowest iWUE values, but it was the only species with no decrease in annual basal increment in response to the rain exclusion treatment, and it also had the highest resistance to the hot and dry conditions projected for the Mediterranean basin in the coming decades. The different strategies to resist rain exclusion conditions of these species could induce changes in their competitive ability and future distribution. The losses of N from the ecosystem may further limit plant growth and ecosystem functioning.     

Colonization of tomato greenhouses by the predatory mirid bugs Macrolophus caliginosus and Dicyphus tamaninii

Colonization of tomato greenhouses by native predatory mirid bugs at the end of the spring cycle is common in the western Mediterranean area when no broad-spectrum insecticides are applied. Due to their polyphagy, these predators interact with pest populations and also with other natural enemies present in the crop. In this work we evaluate the abundance and timing of greenhouse colonization by these predators and their interaction with the greenhouse whitefly Trialeurodes vaporariorum, a key crop pest, and its introduced parasitoid Encarsia formosa. Although quite unpredictable, natural colonization of greenhouses by Macrolophus caliginosus and Dicyphus tamaninii, the two predominant species in our location, usually leads to the establishment of predator populations in the crop that subsequently prey on greenhouse whitefly. No preference for parasitized pupae was observed in greenhouse samples, while laboratory experiments revealed a marked tendency to avoid parasitoid pupae. In our area, IPM programs for greenhouse tomatoes and other vegetables should take advantage of the presence of this predator complex by allowing the immigration and establishment of its populations without disturbing them with highly toxic and non-selective insecticides.     

The production of fertile,triploid somatic hybrid plants (Nicotiana glutinosa (n) + N. tabacum (2n)) via gametic: somatic protoplast fusion

Summary The fusion of gametic protoplasts with somatic protoplasts giving rise to gametosomatic hybrid plants was investigated. Gametosomatic hybrid plants were regenerated following the fusion of nitrate reductase deficient (Nr^–) Nicotiana tabacum Nia-130 leaf mesophyll protoplasts with N. glutinosa tetrad protoplasts. The resulting plants were confirmed as hybrids, based on leaf and floral morphology, chromosome number, leaf esterase and leaf callus peroxidase zymograms and Fraction-1-protein analysis. The five gametosomatic hybrid plants had the expected pentaploid, but functionally triploid chromosome number of 3n=5x=60. The relevance of triploid gametosomatic hybrids in facilitating limited gene transfer, is discussed. The utilisation of tetrads as a generally available source of haploid protoplasts for fusion studies is proposed.     

Evaluation of formulations of Bacillus licheniformis for the biological control of tomato gray mold caused by Botrytis cinerea

Bacillus licheniformis N1, which has previously exhibited potential as a biological control agent, was investigated to develop a biofungicide to control the gray mold of tomato caused by Botrytis cinerea. Various formulations of B. licheniformis N1 were developed using fermentation cultures of the bacteria in Biji medium, and their ability to control gray mold on tomato plants was evaluated. The results of pot experiments led to the selection of the wettable powder formulation N1E, based on corn starch and olive oil, for evaluation of the disease control activity of this bacterium after both artificial infection of the pathogen and natural disease occurrence under production conditions. In plastic-house artificial infection experiments, a 100-fold diluted N1E treatment was found to be the optimum biofungicide spray formulation. This treatment resulted in the significant reduction of symptom development when N1E was applied before Bo. cinerea infection, but not after the infection. Both artificial infection experiments in a plastic house and natural infection experiments under production conditions revealed that the N1E significantly reduced disease severity on tomato plants and flowers. The disease control value of N1E on tomato plants was 90.5% under production conditions, as compared to the 77% conferred by a chemical fungicide, the mixture of carbendazim and diethofencarb (1:1). The prevention of flower infection by N1E resulted in increased numbers of tomato fruits on each plant. N1E treatment also had growth promotion activity, which showed the increased number of tomato fruits compared to fungicide treatment and non-treated control and the increased fruit size compared the non-treated control under production conditions. This study suggests that the corn starch-based formulation of B. licheniformis developed using liquid fermentation will be an effective tool in the biological control of tomato gray mold.     

Visualizing eggs of Nesidiocoris tenuis (Heteroptera: Miridae) embedded in tomato plant tissues

Nesidiocoris tenuis (Heteroptera: Miridae) is a predator of some major pests of Solanaceae crops, yet it is scarcely used in biological control because it also feeds on plants and may damage crops. The study of N. tenuis biology may promote the ability to use it as a biological control agent. Because N. tenuis, like some other insect taxa, oviposits into plant tissues, its eggs are hard to detect. This limits our ability to study N. tenuis – plant interactions. We therefore looked for a staining method for plant‐embedded eggs, which will allow their detection within tomato plants, a common host of N. tenuis. We first used lactophenol solution with acid fuchsin to stain eggs inside tomato foliage. Because of the high toxicity of lactophenol, we later substituted lactophenol with a lactoglycerol solution, which was found to be similarly efficient. Five minutes immersion in the staining solution at 80°C followed by a two‐minute soak in hot water made the eggs stain deep red, while the foliage became transparent and was stained weak red. Eggs within leaves were easily visible under 10–30× magnification with sub‐stage lighting; top‐lighting was needed for the detection of eggs embedded in less‐transparent tissues such as stems. This rapid staining method improves the ability to study some important biological aspects of N. tenuis, such as its fecundity. Also, the elimination of phenol made the method cheaper and safer to use. Finally, this method may be adapted for other arthropod–plant systems.     

Interactions and effects of multiple biological control insects on diffuse and spotted knapweed in the Front Range of Colorado

Abundances and interactions among biological control insects and their effects on target invasive plants were monitored within the flower heads and roots of diffuse knapweed, Centaurea diffusa, and in spotted knapweed, Centaurea stoebe, along the Colorado Front Range. Flower weevils, (Larinus species) and root-feeders (Cyphocleonus achates and Sphenoptera jugoslavica) were released on knapweed that already supported biological control gall flies (Urophora species). At a single monitoring site, seed production by C. diffusa declined from 4400 seeds m⁻² in 1997 to zero seeds m⁻² on the monitoring sites in 2006, while the flowering stem density of C. diffusa declined from a peak of almost 30 stems m⁻² in 2000 to zero stems m⁻² in 2006. The average abundance of Urophora and Larinus in flower heads fluctuated independently during the 2001–2006 interval, while the relative abundance of C. achates and S. jugoslavica in roots exhibited a weak inverse relationship that appeared driven by climate effects. The relative abundance of insects on a population of C. stoebe was monitored for five years as Larinus species and C. achates became established on spotted knapweed that already supported Urophora species. Spotted knapweed seed production on our monitoring site declined from 4600 seeds m⁻² in 2003 to zero seeds m⁻² in 2006. Unlike C. diffusa, substantial numbers of rosettes of C. stoebe remained present. Larinus consumed almost all Urophora encountered in C. diffusa, and consumed about 40% of the Urophora in co-infested flower heads of C. stoebe (ca. 10–15% of the total Urophora population). No negative correlations between the relative densities of flower head and root-feeding insects were observed. The effects of these insects on target plants have produced results consistent with the ‘cumulative stress hypothesis’ for biological control of Centaurea species.     

Influence of Hairiness ofGerbera jamesoniiLeaves on the Searching Efficiency of the ParasitoidEncarsia formosa

To characterize the relationship between the leaf surface ofGerbera jamesoniiHooker cultivars and the searching behavior of the parasitoidEncarsia formosaGahan on a leaf, the trichome density and shape were described, and the walking behavior was quantified. Leaf hair density varied from 80 to more than 1000 trichomes/cm²and the hair shape varied from single erect trichomes to tapestries of entangled trichomes above the leaf surface. The walking activity, speed, and pattern of the parasitoid were tested on leaves with different hair density and shape. In spite of the large differences in leaf surface structure, the walking activity was similar and around 75% on most cultivars. The walking speed was between 0.2 and 0.3 mm/s on all cultivars and was not significantly different from the speed on tomato. On hairless sweet pepper leaves the walking speed is much higher (0.73 mm/s), so the hairs onGerberaleaves do hamper parasitoid females and result in a strong reduction of the walking speed, but the variation in hair density and shape onGerberadoes not lead to differences in walking activity, pattern, and speed. It appears that the hairs ofGerberaare less of an obstacle forE. formosathan the stiff, large hairs occurring in a high density on cucumber, where the walking speed of the parasitoid is less than 0.2 mm/s. A rise of temperature of 5°C gave a significantly higher walking speed (0.39 mm/s) ofE. formosafemales on all cultivars tested. The relative straightness of the walking track was high and the same on all cultivars. Consequences of the results of the searching behavior ofE. formosaonG. jamesoniicultivars are discussed with respect to other host plants. As parasitoid walking speeds are the same onGerberaand tomato, and whitefly reproduction is also similar on these two host plants, we conclude that biological control of whiteflies onGerberais a realistic option. This conclusion is confirmed by the strong increase of commercial biological control onGerberawhich occurred during the past 5 years.