Visual inspections of nursery stock fail to protect new plantings from Blueberry scorch virus infection

Blueberry scorch virus (BlScV) is one of the most pervasive pathogens of highbush blueberry. The virus is aphid-vectored and exhibits a latent period between infection and symptom expression in the host plant of up to 5 years. In many cases, we have observed BlScV symptom expression in new fields that appears inconsistent with aphid-vectored introduction and spread. It was, therefore, speculated that the virus may be introduced through infected nursery stock. To examine this possibility, we first surveyed selected nurseries to determine if mother plants, used for propagation by cuttings, were BlScV-infected. Two nurseries were found to harbor symptomless, infected, mother plants (cv. Duke). Cuttings from one nursery were collected from infected and non-infected plants and rooted in propagation beds. The survival and infection of cohorts from each mother plant were determined one year after planting. A significantly greater proportion of cuttings survived from non-infected mother plants (0.7) than from infected mother plants (0.5). Of the cohort from infected mother plants that survived, 40% tested positive for BlScV. We also surveyed the distribution of infected, symptomatic plants in recent ‘Duke’ plantings that originated from nurseries with BlScV-infected mother plants and compared distribution with older plantings with more advanced BlScV outbreaks. In all cases, the distribution of BlScV symptom development in young fields was random, which is consistent with introduction from planting stock. Older plantings showed a strong clustered distribution, which is consistent with aphid transmission. This study identifies infected nursery stock as an important source of BlScV dissemination and underscores the importance of having symptomless mother plants virus tested.     

Field evaluation of yield effects on the U.S.A. heirloom sweetpotato cultivars infected by Sweet potato leaf curl virus

The incidence of Sweet potato leaf curl virus (SPLCV), a Begomovirus, on sweetpotato, Ipomoea batatas (L.) Lam. (Convolvulaceae), in South Carolina, U.S.A. has increased rapidly in recent years. This is likely due to the use of infected propagating materials and the increasing population of its vector, the sweetpotato whitefly, Bemisia tabaci (Gennadius). In this study, we demonstrated in field experiments that SPLCV infection reduced the yields of most heirloom sweetpotato lines relative to the yields of non-infected plants. Real-time polymerase chain reaction (PCR) technology specific to several common sweetpotato viruses was used to determine the virus infection status in 69 selected accessions of heirloom sweetpotato lines. Meristem tip culture technology was used to regenerate virus-free plants from these materials. To ensure the virus-free status, each mericlone was evaluated using real-time PCR and graft bio-indexing on the indicator species, Ipomoea setosa Ker Gawl. Mericlones of 27 cultivars were found to be free of the viruses. The 27 cultivars were included in a field test to determine the effect of SPLCV infection on yield. Yields of virus-free plants of the cultivars ranged from 10 to 80% greater than the yields of SPLCV-infected plants. However, the yield differences between virus-free and infected plants were diminished in the second year of the field experiment due to a rapid re-infection by SPLCV. These results demonstrate the importance of using certified, virus-tested seed roots or cuttings. The rapid re-infection of the virus-tested sweetpotato plants with SPLCV observed in these studies suggests that management of the whitefly population should be a critical element in control of this important virus.     

Insecticidal activities of crude extracts and phospholipids from Chenopodium ficifolium against melon and cotton aphid, Aphis gossypii

Several organic solvent extracts of Chenopodium ficifolium were tested for their insecticidal activity against melon and cotton aphid, Aphis gossypii, on cucumber plants. Both methanol and ethanol extracts, at 5000 μg ml⁻¹, were highly active giving over 80% control. The other crude extracts displayed moderate or weak insecticidal activity giving control in the range of 16–69%. Two phospholipids were isolated as insecticidal active substances from C. ficifolium. Their chemical structures were identified as 1-palmitoyl-2-(3-trans)-hexadecenoyl-sn-glycero-3-phosphoglycerol and 1-palmitoyl-2-linoleoyl-3-glycerophosphocholine by GC–MS, EDS, mass and NMR spectral analyses. Both compounds displayed a dose-dependent mortality of A. gossypii. Furthermore, the liquid formulation that was obtained by partitioning with n-hexane from the methanol extract of C. ficifolium controlled melon and cotton aphid on cucumber plants effectively. These results indicate that extracts of C. ficifolium have potential for development as botanical insecticides for controlling A. gossypii infesting cucumber plants.     

Comparative susceptibility of Ostrinia furnacalis, Ostrinia nubilalis and Diatraea saccharalis (Lepidoptera: Crambidae) to Bacillus thuringiensis Cry1 toxins

Transgenic corn hybrids that express toxins from Bacillus thuringiensis (Bt) are highly effective against the European corn borer, Ostrinia nubilalis (Hübner), and the closely related Asian corn borer, Ostrinia furnacalis (Guenée). Since the registration of Bt corn hybrids in the U.S. in 1996, there has been a great deal of information generated on O. nubilalis. However, relatively little information exists for O. furnacalis. To help determine whether the information generated for O. nubilalis can be leveraged for decisions regarding the use of transgenic Bt corn against O. furnacalis, experiments were designed to determine whether the pattern of sensitivity to various Bt Cry1 toxins is similar between the two species. Test insects included laboratory-reared O. furnacalis originating from Malaysia, a Bt-susceptible laboratory colony of O. nubilalis maintained at the University of Nebraska-Lincoln (UNL) and an out-group consisting of the sugarcane borer, Diatraea saccharalis (F.), from Louisiana which represents a different genus from the same family. O. furnacalis and O. nubilalis exhibited a similar pattern of susceptibility to all the Cry1 toxins and were highly susceptible to the range of Bt toxins tested including Cry1Aa, Cry1Ab, Cry1Ac and Cry1F. Both of the Ostrinia species were more tolerant to Cry1Ba compared with D. saccharalis, although sensitivity of O. furnacalis was intermediate and did not differ significantly from that of O. nubilalis and D. saccharalis. D. saccharalis was also susceptible to the range of toxins tested but unlike the two Ostrinia species, was more tolerant to Cry1F and more susceptible to Cry1Ba. These results indicate that both of the Ostrinia corn borer species are similar in sensitivity to the Cry1Aa, Cry1Ab, Cry1Ac, Cry1Ba and Cry1F toxins, thus suggesting shared toxin receptors and mechanisms of toxicity for the two species.     

Control of virus diseases in intensively cultivated vanilla plots of French Polynesia

The results of long term virus surveys in intensively cultivated vanilla plots in the Society Islands (French Polynesia), between 1999 and 2007 are reported here. The data confirmed a potential for high incidence of aphid borne viruses in particular Cucumber mosaic virus (CMV) and Watermelon mosaic virus (WMV) as well as the non vectored Cymbidium mosaic virus (CymMV). CMV had a particularly high prevalence (over 30% of the plots) and could severely damage up to 50% of the vines before blossom. Severe outbreaks of CMV were correlated to the presence of the weed Commelina diffusa as a reservoir of virus and aphid vectors. The application in 2003 of simple prophylactic measures resulted in a sharp reduction of virus incidence in new plantations, compared to levels of virus diseases recorded in the previous decade. Indeed, at the beginning of fruiting, the incidence of aphid borne viruses did not exceeded 1.6% of vines for the 29 shade houses which adopted the virus prophylaxis. The three remaining shade houses were severely (20–50%) infected by WMV or CymMV because of the planting of virus-infected cuttings. These results demonstrate the benefit of implementing prophylaxis at the scale of an archipelago, and widen the possibilities of developing intensive cultivation of vanilla in other areas confronted with similar virus constraints.     

Endophytic Lecanicillium lecanii and Beauveria bassiana reduce the survival and fecundity of Aphis gossypii following contact with conidia and secondary metabolites

Lecanicillium lecanii and Beauveria bassiana are important entomopathogens of Aphis gossypii. Their capacity to colonize crop plants is also becoming widely recognized. Their presence in crop plants indicates the possibility of a much greater potential for contact between insect and fungus than previously recognized. The present experiment aimed to study the effects of endophytic strains of these fungi on the survival, and reproduction of A. gossypii. Contact with conidia of both fungi significantly reduced the rate and period of reproduction of A. gossypii. The culture filtrates of L. lecanii and B. bassiana significantly increased mortality and feeding-choice experiments indicate that insects may be able to detect metabolites of the fungi. The culture filtrate of L. lecanii also significantly reduced the reproduction of the aphid. The ethyl acetate and methanolic fractions of the culture filtrate and of mycelia of L. lecanii also caused significant mortality and reduced fecundity of A. gossypii. The methanolic fractions of mycelia of B. bassiana caused significant mortality of A. gossypii. The present investigations indicated that A. gossypii is affected by contact with both conidia and fungal metabolites. This broad influence indicates that these fungi may have a role in regulating insect pest populations.     

High level of resistance to Sclerotinia sclerotiorum in introgression lines derived from hybridization between wild crucifers and the crop Brassica species B. napus and B. juncea

Sclerotinia rot caused by the fungus Sclerotinia sclerotiorum is one of the most serious and damaging diseases of oilseed rape and there is keen worldwide interest to identify Brassica genotypes with resistance to this pathogen. Complete resistance against this pathogen has not been reported in the field, with only partial resistance being observed in some Brassica genotypes. Introgression lines were developed following hybridization of three wild crucifers (viz. Erucastrum cardaminoides, Diplotaxis tenuisiliqua and E. abyssinicum) with B. napus or B. juncea. Their resistance responses were characterized by using a stem inoculation test. Seed of 54 lines of B. napus and B. juncea obtained from Australia, India and China through an Australian Centre for International Agricultural Research (ACIAR) collaboration programme were used as susceptible check comparisons. Introgression lines derived from E. cardaminoides, D. tenuisiliqua and E. abyssinicum had much higher levels (P < 0.001) of resistance compared with the ACIAR germplasm. Median values of stem lesion length of introgression lines derived from the wild species were 1.2, 1.7 and 2.0 cm, respectively, as compared with the ACIAR germplasm where the median value for stem lesion length was 8.7 cm. This is the first report of high levels of resistance against S. sclerotiorum in introgression lines derived from E. cardaminoides, D. tenuisiliqua and E. abyssinicum. The novel sources of resistance identified in this study are a highly valuable resource that can be used in oilseed Brassica breeding programmes to enhance resistance in future B. napus and B. juncea cultivars against Sclerotinia stem rot.     

Growth and accumulation of caffeic acid derivatives in Echinacea angustifolia DC. var. angustifolia grown in hydroponic culture

In separate experiments conducted in 2007 and 2008, growth and accumulation of selected caffeic acid derivatives (CADs; i.e., caftaric acid, chlorogenic acid, echinacoside, caffeic acid, cynarin, p-coumaric acid, ferulic acid and cichoric acid) were determined in Echinacea angustifolia DC. var. angustifolia seedlings grown in hydroponic culture (floating raft system) at a density of 122 plant m⁻² (at planting). Plants were harvested 11 (2007) or 16 (2008) weeks after transplanting (i.e., 15 or 20 weeks after sowing). In both years, plants grew vigorously and at harvest approximately half of the plants under observation had developed one to three inflorescences. In 2008, the root yield (2940 kg ha⁻¹) harvested in nearly eight months from two consecutive hydroponic cultures was within the yield reported in the literature for field cultivations lasting two to four years. None of the selected CADs was found in the leaves, while the inflorescences (stem and capitulum) contained only caftaric acid and echinacoside at concentrations higher than the detection limits (0.05 mg g⁻¹ dry weight). Echinacoside, cynarin and chlorogenic acid were found in root tissues at concentrations ranging from 0.36 to 5.25 mg g⁻¹ dry weight. In all plant samples, echinacoside, which is the marker compound for E. angustifolia material, did not reach the minimum quality standard (10 mg g⁻¹ dry weight) for the production of standardized extract. We concluded that short-cycle, high-density greenhouse hydroponic culture stimulates plant growth and root production in E. angustifolia, but it does not ensure sufficient CADs accumulation in dried roots.     

Mycoparasitic Trichoderma viride as a biocontrol agent against Fusarium oxysporum f. sp. adzuki and Pythium arrhenomanes and as a growth promoter of soybean

Trichoderma viride was proved as an effective biocontrol agent against two fungal pathogens, Fusarium oxysporum f. sp. adzuki and Pythium arrhenomanes, infecting soybean. During an in vitro biocontrol test, Trichoderma showed mycoparasitism and destructive control against the tested fungal pathogens. Both the pathogens significantly influence the germination and P. arrhenomanes had a severe effect (only 5% germination). The root system of the soybean plant was poorly developed due to the infection and it exerted a negative influence on the nodulation and further growth phases of the plant. During pot assay along with biocontrol activity, Trichoderma showed growth promoting action on the soybean plant. Trichoderma enhanced growth of shoot and root systems and fruit yield after 12 weeks of growth. Pythium and Fusarium infected plants treated with Trichoderma had ∼194% and 141% more height than pathogens alone. The fruit yield treated with Trichoderma was ∼66 per plant whereas the yield was only 41 for a control plant. The plants infected with Pythium and Fusarium and treated with Trichoderma had fruit yields of 43 and 53 respectively and those were 5 and 1.6 times higher than plants infected with pathogens.     

Characterization of Solanum chomatophilum resistance to 2 aphid potato pests, Macrosiphum euphorbiae (Thomas) and Myzus persicae (Sulzer)

The aphids Macrosiphum euphorbiae (Thomas) and Myzus persicae (Sulzer) (Hemiptera: Aphididae) are responsible for yield reduction in potato (Solanum tuberosum) production by direct phloem feeding and by spreading viruses. Breeding resistant traits from Solanum chomatophilum into the potato germplasm provides alternative means to control aphid infestations. Integrated pest management strategy, using plant resistance, benefits from the characterization of the resistance and of its impact on aphid biology. Our objective was to characterize the resistance of S. chomatophilum by assessing the effects of accessions, plant parts on aphid performance, and by assessing the impact of the resistance factors on different aphid developmental stages and on alate morph production. Detailed aphid performance was obtained by measuring fecundity, survival, percentage of nymphs that reached adult moult, and population growth using whole plant and clip cage experimental designs. Accession and plant physiological age, but not aphid developmental stage, influenced all life-history parameters, except for alate morph production which was not induced on the resistant accessions. Plant part influence was independent of plant species and accession. Both experimental designs resulted in congruent resistance levels at the accession level for each of the two aphid species, supporting the use of any of them in S. chomatophilum resistance screening. PI243340 was resistant to both aphid species, while PI365324 and PI310990 were also resistant to M. euphorbiae and M. persicae, respectively.     

Disinfection using ozone microbubbles to inactivate Fusarium oxysporum f. sp. melonis and Pectobacterium carotovorum subsp. carotovorum

To establish a method for disinfecting hydroponic culture solutions using ozone microbubbles (OMB), we examined the disinfectant activity of OMB against Fusarium oxysporum f. sp. melonis and Pectobacterium carotovorum subsp. carotovorum in infected plant roots. OMB had a higher solubility and remained in the water for a longer period than ozone millibubbles, resulting in extremely high disinfecting activity against both phytopathogens. Furthermore, disinfectant activity and durability of OMB-treated water against both phytopathogens increased with an increase in the initial concentration of dissolved ozone. Therefore, these results suggest that OMB may be suitable for use as a new disinfectant against phytopathogens in hydroponic culture solutions.     

Differential response of pea (Pisum sativum) to Orobanche crenata, Orobanche foetida and Phelipanche aegyptiaca

Several broomrape species including Orobanche crenata, Orobanche foetida and Phelipanche aegyptiaca are reported to infect various grain and forage legumes in the Mediterranean and West Asia. Pea (Pisum sativum) is severely damaged by O. crenata, but there are no reports on O. foetida or P. aegyptiaca infection. We report here that pea can induce high germination of seeds of O. crenata, O. foetida and P. aegyptiaca but only O. crenata success infecting pea roots and developing further. Some differences in levels of infection by O. crenata were observed among pea accessions what can be exploited in pea resistance breeding programmes. On the contrary, all pea accessions studied were highly resistant to infection by both O. foetida and P. aegyptiaca, preventing any tubercle attachment and development. This makes pea a promising candidate as trap crop for O. foetida and P. aegyptiaca seed bank demise in infested soils.     

Effects of crude seed extracts of Annona atemoya and Annona squamosa L. against the cabbage looper, Trichoplusia ni in the laboratory and greenhouse

Antifeedant, growth inhibitory and toxic effects of crude seed extracts of Annona squamosa and Annona atemoya from Fazenda Viveiro Bona, Parasisópolis – Minas Gerais, Brazil, were evaluated against the cabbage looper, Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) using different bioassays. Crude methanolic seed extracts deterred feeding of third instar T. ni larvae in a leaf disc choice bioassay. A. squamosa was ∼10 times more active as a feeding deterrent than A. atemoya (DC₅₀ = 2.3 mg/ml vs. 20.1 mg/ml). A. squamosa was ∼three times more active as a growth inhibitor than A. atemoya (EC₅₀ = 38.0 ppm vs. 117.0 ppm). Methanolic seed extracts of A. squamosa and A. atemoya were toxic to third instar T. ni larvae both through topical and oral application. A. squamosa was more toxic through feeding (LC₅₀ = 167.5 ppm vs. 382.4 ppm) whereas, A. atemoya exerted greater toxicity via topical application (LC₅₀ = 301.3 μg/larva vs. 197.7 μg/larva). Both A. squamosa and A. atemoya extracts reduced leaf area consumption and larval growth in a greenhouse experiment. Our results indicate that both A. squamosa and A. atemoya have potential for development as botanical insecticides, especially for local use in Brazil.     

Biocontrol of Rhizoctonia solani and Sclerotium rolfsii on tomato by delivering antagonistic bacteria through a drip irrigation system

In a 2-year assessment carried out on tomato crops of Central and Southern Italy, a high incidence of Rhizoctonia solani and Sclerotium rolfsii was found. These fungal pathogens attack horticultural crops and are responsible for severe crown and stem rot. Because of technical, economical and environmental issues their chemical control is an arduous task. To find alternative and eco-compatible control methods, the effectiveness of two new antagonistic bacterial isolates (Burkholderia cepacia, T1A-2B, and Pseudomonas sp., T4B-2A), previously selected from suppressive organic amendments, were tested on tomato plants grown under both growth chamber and field conditions. The potential antagonists were compared with two commercial biofungicides, based on Bacillus subtilis (BSF4) and Trichoderma asperellum (TV1), and four synthetic fungicides (tolclofos-methyl, azoxystrobin, fosetyl-Al and fosetyl-Al + propamocarb). In 2-year field experiments carried out on tomato plants, the biocontrol bacteria as well as the other treatments were applied to the soil, proximal to the plant crowns and main roots, by means of an effective and specific system of drip irrigation. In all the experiments the novel selected biocontrol bacteria significantly reduced both incidence and severity of the diseases caused by S. rolfsii or R. solani, with results demonstrating effectiveness equal to TV1, better than BSF4 and comparable with the synthetic fungicides, except for tolclofos-methyl which was the most effective treatment. In field experiments, carried out for two consecutive years, isolate T1A-2B reduced up to 58.33% and up to 63.8% the severity of the diseases caused by S. rolfsii and R. solani respectively; whereas isolate T4B-2A gave reduction of S. rolfsii and R. solani diseases severity up to 73.2% and up to 62.7%, respectively.     

Insecticidal effects of Ungernia severtzovii bulb extracts against the grain aphid Schizaphis graminum (Rondani)

Insecticidal effect of Ungernia severtzovii bulbs extract has been investigated against the grain aphid Schizaphis graminum. LC₅₀ value of the ethanolic extract of U. severtzovii against the female aphids was 0.235% (2.35 g/l). There was 100% mortality of the subsequent nymphs of the treated females at 0.25-1.0%. The extract was equally active against the nymphs at the highest concentrations. Hexane and ethanolic extracts were the most active against the aphids of all the extracts. Fractionation of the hexane extract by TLC yielded three fractions. Fraction II was the most active (>90% mortality) against the aphids. Xanthoxylin was identified as the major constituent in fraction II of the hexane extract and may be responsible for the insecticidal effect of U. severtzovii.     

Population dynamics of Xanthomonas campestris pv. vitians on different plant species and management of bacterial leaf spot of lettuce under greenhouse conditions

The population dynamics of Xanthomonas campestris pv. vitians (Xcv) was studied both externally and internally in lettuce, tomato and pepper plants. In addition, the application of bactericides during transplant production period was carried out for the management of bacterial leaf spot of lettuce under greenhouse conditions. Epiphytic populations of Xcv were recovered on leaves of lettuce plants (10⁵ CFU/g) 5 weeks after sprayed than the other plant species when inoculated with 10⁸ CFU/ml of Xcv. When plants of each crop species infiltrated with the bacterium at 10⁵ CFU/ml, the highest populations were developed in lettuce (10⁸ CFU/cm²) followed by pepper with 10⁶ CFU/cm² and tomato with 10⁵ CFU/cm² 10-days after infiltration. Application of a mixture of Maneb and Kocide or Kocide alone as a foliar spray on lettuce significantly reduced the incidence and disease severity of bacterial leaf spot by 29 and 27% respectively. Spread of the bacterium and development of the disease may partly be managed by avoiding intercropping of these plants commonly grown in close proximity to lettuce. For the management of leaf-associated populations of Xcv in lettuce, use of a mixture of Maneb and Kocide is advocated to minimize the effect of attacks.     

Biological control of Macrophomina phaseolina by chemotactic fluorescent Pseudomonas aeruginosa PN1 and its plant growth promotory activity in chir-pine

Ten strains of Pseudomonas aeruginosa (PN1 ˜ PN10) isolated from rhizosphere of chir-pine were tested for their plant growth promontory properties and antagonistic activities against Macrophomina phaseolina in vitro and in vivo. P. aeruginosa PN1 produced siderophore, IAA, cyanogen and solubilized phosphorus, besides producing chitinase and β-1,3-glucanase. In dual culture, P. aeruginosa PN1 caused 69% colony growth inhibition. However, cell free culture filtrate also posed inhibitory effect but to a lesser extent. After 90 days, P. aeruginosa PN1 increased plant growth and biomass in pots trial containing M. phaseolina-infested soil. PN1 showed the strong chemotaxis toward root exudates resulting in effective root colonization. Moreover, increased population in rhizosphere of these bacteria was also recorded after 90 days of treatment. Thus, chemotactic fluorescent P. aeruginosa PN1 exhibited strong antagonistic property against M. phaseolina, suppressed the disease and improved plant growth of the seedlings of chir-pine proving potential biocontrol agent.     

Isolation and characterization of EMS-induced Dy10 and Ax1 high molecular weight glutenin subunit deficient mutant lines of elite hexaploid wheat (Triticum aestivum L.) cv. Summit

The mixing properties of the dough are critical in the production of bread and other food products derived from wheat. The high molecular weight glutenin subunits (HMW-GS) are major determinants of wheat dough processing qualities. The different alleles of the HMW-GS genes in hexaploid wheat vary in their effect on dough quality. To determine the contribution of the individual HMW-GS alleles, lines deficient in HMW-GS proteins were generated by chemical mutagenesis in the elite bread wheat Triticum aestivum cv. Summit. In this report we describe the identification and characterization of Dy10 and Ax1 deficient lines. Examination of the effect of Dy10 and Ax1 deficiency on dough rheological properties by mixography showed shorter mixing time to reach peak resistance, and weaker and less extensible doughs relative to the wild type control. This is the first time that the role of Dy10 in vivo has been examined apart from the Dx5 + Dy10 allelic pair combination.     

Identification of an insecticidal polyacetylene derivative from Chrysanthemum macrotum leaves

Compounds responsible for insecticidal properties of Chrysanthemum macrotum (D.R.) Ball. leaves against Spodoptera littoralis Boiduval caterpillars have been investigated. The screening of the insecticidal activity was performed by incorporating methanol, buthanol or ethyl acetate extracts, or some chromatographic fractions to the caterpillars’ artificial diet. It was noted that extracts and fractions ameliorated or disturbed nutritional indexes, being not always toxic for caterpillars. Among the tested fractions, one pure compound with a high insecticidal activity (percentage of mortality 66.7%) was purified. The nuclear magnetic resonance study allowed its identification as a polyacetylene derivative, in particular a spiroketal enol ether one.     

Response of two tephritid species, Bactrocera oleae and Ceratitis capitata, to different emission levels of pheromone and parapheromone

Attractants and pheromones are commonly used in integrated pest management programs in crop systems. However, pheromone dispensers employed in monitoring traps and lure and kill devices are not usually well studied and attractants are released at uncontrolled rates leading to low treatment efficacies and misleading monitoring estimations. Fruit flies are pests of economic importance and monitoring is essential in order to program insecticidal treatments. Moreover, lure and kill techniques are being increasingly used, but the cost of these techniques depends on the number of required traps and, therefore, on the efficacy of the attractants. Ceratitis capitata and Bactrocera oleae are the two main fruit flies in Mediterranean countries, and the effect of different doses of trimedlure and spiroacetal on fly attraction has been studied. Results showed that a release rate over 1.28 mg/day of spiroacetal reduces B. oleae attraction and emission values over 2.4 mg of trimedlure per day did not increase C. capitata catches. Under the environmental conditions of our study, an optimum release rate for pheromone attraction in B. oleae was determined. Emission values over this optimum level reduced B. oleae attraction. However, when a parapheromone was used with C. capitata, a fruit fly of the same family, the optimum emission value was not found and higher quantities of parapheromone attracted the same number of flies. The saturation effect of high concentrations of pheromone and parapheromone is discussed.