New Monoterpenoid as the Sex Pheromone of Spanish Populations of the Longtailed Mealybug Pseudococcus Longispinus (Hemiptera: Pseudococcidae).

Pseudococcus longispinus (Targioni-Tozzetti) (Hemiptera: Coccoidea: Pseudococcidae), a polyphagous and cosmopolitan pest native to Australia, is a highly damaging pest for numerous crops of economic importance. The sex pheromone of this species (2-(1,5,5-trimethylcyclopent-2-en-1-yl)ethyl acetate), currently used for pest monitoring purposes, was not attractive to males in field experiments conducted in Spanish persimmon orchards infested with this mealybug. The virgin and mated female volatile profiles of these P. longispinus populations were studied by the volatile collection of effluvia in Porapak-Q. The resulting extracts were analyzed by gas chromatography coupled to mass spectrometry (GC-MS), revealing a new compound specific to virgin females and different from the previously described sex pheromone. Based on GC-MS data and nuclear magnetic resonance experiments, we envisaged monoterpene 2-(1,5-dimethyl-4-methylenecyclopent-2-en-1-yl)ethyl acetate as the new sex pheromone candidate, which was synthesized and shown to be attractive in the field to P. longispinus males of the Spanish population.

Control of Leiodes cinnamomeus (Coleoptera: Leiodidae) in Cultivated Black Truffle Orchards by Kairomone-Based Mass Trapping.

The monoculture situation of truffle cultivation is favoring the appearance of pests that would not be economically important in naturally balanced forest ecosystems. The most prominent of them is the European truffle beetle Leiodes cinnamomeus (Panzer) (Coleoptera: Leiodidae), for which there are no effective control methods capable of reducing its populations. The potential of the mass trapping technique against this beetle, based on adapted pitfall traps and the semiochemical methyl disulfide as an attractant, is explored in the present work. Two trap densities (40 and 80 traps/ha) were tested in 2-yr field trials carried out in the region of Teruel (Spain) with black truffle cultivation tradition. Kairomone dispensers were placed in the field immediately before adult outbreak and remained active there throughout the season. The efficacy of each treatment was measured according to the reduction in beetle populations and the damaged truffles in the center of the treated areas. The results showed that both trapping densities reduced adult populations (mean 57% catch reduction), but 80 traps/ha were needed to significantly lower damage parameters (>40% reduction), percentage of attacked truffles and number of galleries/g truffle. The cost effectiveness of these treatments and possible improvements are discussed.

Production of Volatile Moth Sex Pheromones in Transgenic Nicotiana benthamiana Plants.

Plant-based bioproduction of insect sex pheromones has been proposed as an innovative strategy to increase the sustainability of pest control in agriculture. Here, we describe the engineering of transgenic plants producing (Z)-11-hexadecenol (Z11-16OH) and (Z)-11-hexadecenyl acetate (Z11-16OAc), two main volatile components in many Lepidoptera sex pheromone blends. We assembled multigene DNA constructs encoding the pheromone biosynthetic pathway and stably transformed them into Nicotiana benthamiana plants. The constructs contained the Amyelois transitella AtrΔ11 desaturase gene, the Helicoverpa armigera fatty acyl reductase HarFAR gene, and the Euonymus alatus diacylglycerol acetyltransferase EaDAct gene in different configurations. All the pheromone-producing plants showed dwarf phenotypes, the severity of which correlated with pheromone levels. All but one of the recovered lines produced high levels of Z11-16OH, but very low levels of Z11-16OAc, probably as a result of recurrent truncations at the level of the EaDAct gene. Only one plant line (SxPv1.2) was recovered that harboured an intact pheromone pathway and which produced moderate levels of Z11-16OAc (11.8 µg g-1 FW) and high levels of Z11-16OH (111.4 µg g-1). Z11-16OAc production was accompanied in SxPv1.2 by a partial recovery of the dwarf phenotype. SxPv1.2 was used to estimate the rates of volatile pheromone release, which resulted in 8.48 ng g-1 FW per day for Z11-16OH and 9.44 ng g-1 FW per day for Z11-16OAc. Our results suggest that pheromone release acts as a limiting factor in pheromone biodispenser strategies and establish a roadmap for biotechnological improvements.

Airborne Pheromone Quantification in Treated Vineyards with Different Mating Disruption Dispensers against Lobesia botrana.

Mating disruption (MD) is widely used against the European grapevine moth (EGVM), Lobesia botrana (Denis and Schiffermüller; Lepidoptera: Tortricidae), by installing passive dispensers or aerosol devices. The present work reports a new sampling and quantification methodology to obtain absolute data about field airborne pheromone concentration based on air samplings and sensitive chromatographic-spectroscopic methods. Samplings were performed in fields treated with passive dispensers or aerosol devices at different moments throughout the crop cycle to study how they act and how the disruption is triggered. Moreover, pheromone adsorption and releasing capacity of vine leaves were studied to elucidate their role in the disruption. Although both types of dispensers were effective in limiting the damage inflicted by EGVM, they performed differently and provided different airborne pheromone concentration profiles. Results also proved that leaves were able to adsorb and release part of the airborne pheromone acting as subsequent and additional pheromone sources. This fact could explain the different concentration profiles. Moreover, our results suggest that lower pheromone emission than that of the current passive dispensers still could provide an adequate performance in the field. Competitive mechanisms involved in MD using both dispensers, the dynamics of the airborne pheromone throughout the time and the importance of the canopy are discussed.

Survey on Drosophila suzukii Natural Short-Term Dispersal Capacities Using the Mark-Release-Recapture Technique.

Spotted wing drosophila, Drosophila suzukii Matsumura (Diptera: Drosophilidae), has become a key pest for soft fruits and cherries in Europe in less than a decade since the first outbreak in 2007. Although this pest's passive dispersal ability has been observed over more than 1400 km in 1 year, active spread has not yet been extensively studied. A mark-release-recapture (MRR) method based on protein-marked flies was employed to determine the flight capacity of D. suzukii. Sterile marked flies were released and recaptured in a trap grid at increasing distances from 10 to 250 m from the releasing point to study flight distance during periods ranging from 3 h to 1 week. MRR experiments were replicated in the presence and absence of host fruits to study how they could affect dispersal behavior. The dispersal capacity of the Mediterranean fruit fly, Ceratitis capitata Wiedemann (Diptera: Tephritidae) was also studied under the same conditions. The results showed a low dispersal ability for D. suzukii, with a daily flight distance below 100 m with no predominant wind. The implications on natural dispersion and control methods based on attractants are discussed.

Sex Pheromone of the Invasive Mealybug Citrus Pest, Delottococcus aberiae (Hemiptera: Pseudococcidae). A New Monoterpenoid with a Necrodane Skeleton.

Native to sub-Saharan Africa, Delottococcus aberiae De Lotto (Hemiptera: Pseudoccidae) is an invasive mealybug that has been recently reported in Europe, seriously damaging citrus production in eastern Spain. In this study, we isolated and determined the structure of the sex pheromone of D. aberiae, to provide a highly specific and effective lure for detecting, monitoring, and potentially controlling this pest. The volatile profile of D. aberiae virgin and mated females was studied by aeration and collection of effluvia in Porapak-Q. The resulting extracts were analyzed by gas chromatography coupled to mass spectrometry (GC-MS), revealing a candidate compound specific of virgin females. GC-MS and nuclear magnetic resonance spectroscopy data evidenced a new compound, (4,5,5-trimethyl-3-methylenecyclopent-1-en-1-yl)methyl acetate, with an unusual ß-necrodol skeleton. This compound was synthesized and shown to be attractive to male D. aberiae in both laboratory and field experiments. A GC analysis using an enantioselective stationary phase and polarimetry analyses of the synthetic enantiomers showed the natural compound emitted by virgin females to be the - enantiomer.

Synthesis of the Sex Pheromone of the Oleander Scale ( Aspidiotus nerii).

A total synthesis of the oleander scale [ Aspidiotus nerii (Bouche)] sex pheromone, the unique sesquiterpenoid containing a cyclobutane moiety of this class of compounds, has been developed. In order to implement this sex pheromone as a new environmentally friendly tool to manage this pest, a more cost-effective, multigram synthesis was required. This new synthetic route, having a Blaise reaction, iron-catalyzed carbon-carbon coupling, and [2 + 2] photocycloaddition reactions as key steps, provides a general access to 4-alkyl lactones as well as a robust access to the target sex pheromone. Starting from readily available compounds as 3-hydroxypropanenitrile, ethyl bromoacetate, and 2-acetyl butyrolactone, the synthetic sequence afforded the A. nerii sex pheromone with minimum intermediate purification and good overall yield in nine linear steps.

Improvements in Rhynchophorus ferrugineus (Coleoptera: Dryophthoridae) Trapping Systems.

Improved trap efficacy is crucial for implementing control methods for red palm weevil, Rhynchophorus ferrugineus (Olivier; Coleoptera: Dryophthoridae), based on trapping systems, such as mass trapping, attract and infect or attract and sterilize techniques. Although new trap designs have been proposed and aggregation pheromone dispensers have been optimized, aspects such as the use of co-attractants (molasses) and trap placement are still not well defined and standardized. The efficacy of three concentrations of molasses and different formulations to reduce water evaporation in traps was studied in different field trials to improve trapping systems and to prolong trap servicing periods. In addition, the performance of installing groups of traps or single traps was also evaluated with the aim of improving the attracted/captured weevils ratio. Our results showed that captures increased when molasses were added at 15% to the water contained in the trap and that a thin layer of oil, created by adding 2-3% of paraffinic oil to water, was able to effectively reduce evaporation and prolong trap servicing periods. Moreover, 3.5-fold more weevils were captured when placing five traps instead of one at the same trapping point. Results obtained allow improved efficacy and may have an impact in the economic viability of trapping systems and, therefore, in integrated pest management programs.

Identification, introgression, and validation of fruit volatile QTLs from a red-fruited wild tomato species.

Volatile organic compounds (VOCs) are major determinants of fruit flavor, a primary objective in tomato breeding. A recombinant inbred line (RIL) population consisting of 169 lines derived from a cross between Solanum lycopersicum and a red-fruited wild tomato species Solanum pimpinellifolium accession (SP) was characterized for VOCs in three different seasons. Correlation and hierarchical cluster analyses were performed on the 52 VOCs identified, providing a tool for the putative assignation of individual compounds to metabolic pathways. Quantitative trait locus (QTL) analysis, based on a genetic linkage map comprising 297 single nucleotide polymorphisms (SNPs), revealed 102 QTLs (75% not described previously) corresponding to 39 different VOCs. The SP alleles exerted a positive effect on most of the underlying apocarotenoid volatile QTLs-regarded as desirable for liking tomato-indicating that alleles inherited from SP are a valuable resource for flavor breeding. An introgression line (IL) population developed from the same parental genotypes provided 12 ILs carrying a single SP introgression and covering 85 VOC QTLs, which were characterized at three locations. The results showed that almost half of the QTLs previously identified in the RILs maintained their effect in an IL form, reinforcing the value of these QTLs for flavor/aroma breeding in cultivated tomato.

Identification of the Male-Produced Aggregation Pheromone of the Four-Spotted Coconut Weevil, Diocalandra frumenti.

The four-spotted coconut weevil, Diocalandra frumenti Fabricius (Coleoptera: Dryophthoridae), is a small weevil found attacking economically important palm species, such as coconut, date, oil, and Canary palms. Given the scarcity of detection and management tools for this pest, the availability of a pheromone to be included in trapping protocols would be a crucial advantage. Previous laboratory experiments showed evidence for aggregation behavior; thus, our main goal was to identify the aggregation pheromone in this species. The volatile profile of D. frumenti individuals was studied by aeration and collection of effluvia in Porapak-Q and also by solid phase microextraction (SPME) techniques. Moreover, solvent extraction of previously frozen crushed individuals was also performed. All resulting extracts and SPME fibers were analyzed by gas chromatography coupled to mass spectrometry (GC-MS). The comparison of male and female samples provided the candidate compound, 5-ethyl-2,4-dimethyl-6,8-dioxabicyclo[3.2.1]octane (multistriatin), whose biological activity was evaluated in olfactometer and field assays.

Lures for red palm weevil trapping systems: aggregation pheromone and synthetic kairomone.

BACKGROUND: The optimisation of the lure is essential for the implementation of trapping systems to control insect pests. In this work, the response of the red palm weevil (RPW), Rhynchophorus ferrugineus Olivier, to increasing emission rates of its aggregation pheromone (ferrugineol) and the efficacy of a convenient synthetic kairomone based on fermentation odours (ethyl acetate and ethanol) have been evaluated in different years and locations along the Mediterranean basin. RESULTS: In general, although capture data and emission had noticeable variability among locations, significantly fewer RPW were captured in pyramidal Picusan® traps with the lowest ferrugineol emission rates tested (0.6-3.8 mg day-1 ). Captures increased rapidly with ferrugineol emission up to 4-5 mg day-1 ; then, higher emission rates did not improve or reduce captures, up to the highest emission rate tested of 50.9 mg day-1 . Thus, there is no evidence of an optimum release rate corresponding to a maximum of RPW catches. Traps baited with the synthetic kairomone (1:3 ethyl acetate/ethanol) captured 1.4-2.2 times more total weevils than traps baited only with ferrugineol. Moreover, in most of the locations, the synthetic blend was at least as effective as the local coattractants used (plant material + molasses). CONCLUSIONS: Ferrugineol emission rate can vary in a wide range without significantly affecting RPW response. Coattractants based on fermenting compounds, ethyl acetate and ethanol, are able to improve the attractant level of ferrugineol and could be employed to replace non-standardised natural kairomones in RPW trapping systems after further optimisation of their proportions and doses. © 2016 Society of Chemical Industry.

Single-Layered Hybrid Materials Based on 1D Associated Metalorganic Nanoribbons for Controlled Release of Pheromones.

A new family of stable layered organic-inorganic materials has been prepared, in one-step solvothermal process. They are based on an ordered nickel cluster-type nanoribbons separated from each other by specific alkyl (heptyl- or dodecyl-) arylic mono-carboxylate moieties acting as molecular spacers, perpendicular to the 1D inorganic chains. These organic spacers contain hydrocarbon tails with different length which control the separation level between inorganic 1D sub-units, inhibiting the 3D growth of conventional DUT-8-type metal-organic frameworks (MOFs). The lamellar nature of the materials formed was studied and confirmed by different characterization techniques, showing the structural location of individual organic and inorganic building units. They have been successfully used as a long-lasting biodegradable and water-proof materials for controlled release of chemicals, such as pheromones for sustainable treatment of insect plagues.

Bait station devices can improve mass trapping performance for the control of the Mediterranean fruit fly.

BACKGROUND: The use of traps and other attract-and-kill devices in pest management strategies to reduce Mediterranean fruit fly populations has proved to be efficient. Nevertheless, many farmers are concerned about the effect of these devices on the trees where they are hung. Direct field observations have revealed that fruit damage is higher in trees with traps than in trees without them. This work evaluates the efficacy of different types of attract-and-kill device to protect fruit of the single tree on which the device is placed. RESULTS: Results suggested that trees with traps had at least the same fruit damage than trees without them. When traps were baited with protein hydrolysate, fruit damage was even higher than in trees without traps. However, fruit damage was significantly diminished when efficient bait station devices were used. CONCLUSION: Although mass trapping is able to control fruit fly populations as a control method, trees with some types of trap and bait are more susceptible to fly puncture. However, bait station devices reduce fruit damage in the single trees where they are hung. Bait stations are more efficient in fruit protection because fruit flies are affected as soon as they contact the device. Some recommendations for the use of the different attract-and-kill devices are discussed.

Role of α-copaene in the susceptibility of olive fruits to Bactrocera oleae (Rossi).

The influence of α-copaene as a fruit volatile in the susceptibility of Olea europaea L. to the olive fruit fly Bactrocera oleae (Rossi) has been investigated. By studies on the relative area of volatile components from different cultivars, a positive correlation was found between the abundance of α-copaene in the samples and the corresponding degree of fruit infestation. SPME-GC-MS analysis of volatiles from uninfested fruits of O. europaea L. cv. Serrana were performed over two years to determine the variation of α-copaene throughout the different phenological stages. The results suggested that this sesquiterpene has a significant effect on cultivar susceptibility and may act as an oviposition promoter. Further analysis by chiral GC showed that olive fruits release both α-copaene enantiomers. Bioassays on each enantiomer revealed that fruits with increased amounts of (+)-α-copaene favor oviposition of B. oleae females, whereas the increase of (-)-α-copaene affords no statistically significant differences in host preference.

Identification of pheromone synergists for Rhynchophorus ferrugineus trapping systems from Phoenix canariensis palm volatiles.

Trapping systems for the red palm weevil, Rhynchophorus ferrugineus Olivier, rely on the use of natural plant odor sources to boost the attractiveness of the aggregation pheromone. The identification of the key odorants involved in attraction is essential in the development of a synthetic pheromone synergist to replace the nonstandardized use of plant material in traps. Canary Islands date palms (Phoenix canariensis) have become preferred hosts for R. ferrugineus in Europe; thus, the volatile profile of different P. canariensis plant materials, including healthy and infested tissues, is investigated in the present work by means of solid phase microextraction (SPME-GC-MS), aimed to identify pheromone synergists. The electroantennography (EAG) response of the compounds identified was recorded, as well as the preliminary field response of several EAG-active compounds. The so-called "palm esters" (ethyl acetate, ethyl propionate, ethyl butyrate, and propyl butyrate) elicit the strongest EAG responses but performed poorly in the field. Mixtures of esters and alcohols give evidence of better performance, but release rates need further optimization.

Response of Tuta absoluta (Lepidoptera: Gelechiidae) to different pheromone emission levels in greenhouse tomato crops.

The response of Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) to different emission rates of its pheromone, (3E, 8Z, 11Z)-tetradecatrienyl acetate, was measured in two greenhouse trials with traps baited with mesoporous dispensers. For this purpose, weekly moth trap catches were correlated with increasing pheromone emission levels by multiple regression analysis. Pheromone release profiles of the dispensers were obtained by residual pheromone extraction and gas chromatography quantification. In the first trial carried out in summer 2010, effect of pheromone emission was significant as catches increased linearly with pheromone release rates up to the highest studied level of 46.8 µg/d. A new trial was carried out in spring 2011 to measure the effect of the emission factor when pheromone release rates were higher. Results demonstrated that trap catches and pheromone emission fitted to a quadratic model, with maximum catches obtained with a release level of 150.3 µg/d of (3E, 8Z, 11Z)-tetradecatrienyl acetate. This emission value should provide enhanced attraction of T. absoluta and improve mass trapping, attract-and-kill, or monitoring techniques under greenhouse conditions in the Mediterranean area.

Efficacy of attract-and-kill devices for the control of Ceratitis capitata.

BACKGROUND: The control of Ceratitis capitata Wiedemann traditionally has relied on chemical control with organophosphate insecticides. The use of many of these substances has been banned by new European directives; therefore, the development of new control methods is essential to manage this pest. Bait sprays with spinosad, mass trapping and lure-and-kill techniques have been the base for new integrated pest management programmes. In this study, a 2 year field trial was conducted in two citrus areas to test the efficacy of attract-and-kill devices against mass trapping and spinosad-plus-bait treatments. RESULTS: The Magnet(®) MED attract-and-kill device, Spintor(®) treatments and mass trapping achieved good control of C. capitata populations, as confirmed by low percentages of damaged fruit in the assessments performed during the harvest period. On the other hand, fly population levels on plots treated with other attract-and-kill prototype devices increased more than threefold by comparison with the populations recorded in the rest of the treated plots. The same effect was observed for fruit damage, with 6-8 times less damage with Magnet(®) MED and spinosad treatments, respectively, than with the attract-and-kill prototype devices. CONCLUSION: By using an effective attractant, conventional trapping systems can be replaced with cheaper and easier-to-handle attract-and-kill devices. The efficacy of these devices and their advantages over conventional mass trapping systems are discussed.

Identification of the sex pheromone of the mealybug Dysmicoccus grassii Leonardi.

Studies about the sex pheromone of the mealybug Dysmicoccus grassii , a main pest of Canary Islands banana cultivars, showed strong evidence that (-)-(R)-lavandulyl propionate and acetate in a 6:1 ratio are principal components of it. Volatile collection and GC-MS analysis from aeration of virgin females allowed the structural elucidation of the compounds. The activity and stereochemistry of both substances were established by means of relative attraction of mealybug males to synthetic standards in competitive Petri dish bioassays. (R)-Lavandulyl propionate induced a stronger attractive effect when compared to (R)-lavandulyl acetate. The attractiveness of the mixture of the two compounds at the original source ratio showed no statistically significant difference from that of the sum of each of the single compounds alone, suggesting that both components are not synergistic but additive.

Synthesis of new antimicrobial pyrrolo[2,1-a]isoquinolin-3-ones.

The attractive structure of the pyrroloisoquinoline moiety, together with its potential antimicrobial activity, encouraged us to prepare six 8-substituted and seven 8,9-disubstituted-1,2,3,5,6,10b-hexahydropyrrolo[2,1-a]isoquinolin-3-ones in a few steps with good yields. We applied a convenient methodology via double intramolecular cyclization conducted by a Bischler-Napieralski cyclodehydration-imine reduction sequence, which is widely employed in the synthesis of isoquinoline alkaloids. Therefore, we synthesized three series of these pyrrolo[2,1-a]isoquinolin-3-ones characterized by the substituent at the 8-position or 8,9-positions of the aromatic ring: (a) different side chains are attached to an 8-OH group (series 1); (b) a chlorine atom is attached to the 8-position (series 2); and (c) 8- and 9-carbons are bearing an identical group (series 3). The compounds bearing a benzylic moiety at the 8-position, for example, 8-benzyloxy-pyrrolo[2,1-a]isoquinolin-3-one (1a) and 8-(4-fluorobenzyloxy)-pyrrolo[2,1-a]isoquinolin-3-one (1e), as well as, a 8-chloro-9-methoxy moiety including the 8-chloro-9-methoxy-pyrrolo[2,1-a]isoquinolin-3-one (2a), provided the most fungicide and bactericide agents, respectively.

Life history parameters and scale-cover surface area of Aonidiella aurantii are altered in a mating disruption environment: implications for biological control.

BACKGROUND: In recent years, environmentally safe measures to control the California red scale (CRS), Aonidiella aurantii (Maskell), have been successfully implemented. These measures include mating disruption (MD) and biological control. The goal of this study was to examine the effect of high concentrations of the CRS sex pheromone on its life history parameters and scale-cover surface area under controlled laboratory conditions. RESULTS: The developmental time of both males and females of CRS increased with exposure to airborne pheromone. MD had an effect on both the total number of progeny and on the crawler production period for females. Accordingly, demographic parameters such as net fecundity (R(0) ) and intrinsic rate of increase (r(m) ) were significantly lower in the pheromone-treated populations. The largest scale-cover surface areas were observed in the CRS reared in the pheromone environment. CONCLUSION: A clear influence of airborne pheromone on the biology of CRS has been demonstrated. In addition to the classical mating disruption benefits of this technique, additional benefits, such as increase in the duration of exposure to natural enemies and increase in size, which benefits some species of parasitoids, have been confirmed.