Isoforms of the heteropteran Nezara viridula ecdysone receptor: protein characterisation, RH5992 insecticide binding and homology modelling

BACKGROUND: Certain bisacylhydrazine compounds such as tebufenozide (RH5992) have been shown to act as order‐specific insecticides. Their compatibility with predatory Heteroptera, which are used as biological control agents, has also been demonstrated. However, the molecular mode of action of these ecdysone agonists has not been explored in a heteropteran, much less one that is a significant agricultural pest, such as Nezara viridula. RESULTS: Alternatively spliced ligand‐binding regions of the N. viridula ecdysone receptor were expressed, purified and characterised by 2D gel analysis, mass spectrometry, homology modelling and competitive binding of a bisacylhydrazine insecticidal compound (RH5992) and various ecdysteroids. Ligand binding by the two splice isoforms was indistinguishable, and relative affinities were found to occur in the order muristerone A > ponasterone A > 20‐hydroxyecdysone > inokosterone > RH5992 > α‐ecdysone. CONCLUSION: The predicted difference in amino acid sequence between the ligand‐binding domains of the N. viridula ecdysone receptor splice variants was verified by mass spectrometry. Both splice variant isoforms exhibit a greater affinity for the bisacylhydrazine insecticide RH5992 than do the other hemipteran ecdysone receptors characterised to date. Their affinities for a range of ecdysteroids also distinguish them from the ecdysone receptors of other Hemiptera characterised thus far. Homology models of both N. viridula receptor isoforms provide further insight into the bisacylhydrazine‐ and ecdysteroid‐binding properties of these receptors, including their similar affinity for 20‐hydroxyecdysone and the postulated pentatomomorphan moulting hormone makisterone A. Copyright © 2011 Society of Chemical Industry     

Plant growth inhibitors in turmeric (Curcuma longa) and their effects on Bidens pilosa

Turmeric (Curcuma spp.) has numerous biological activities, including anticancer, antibacterial, antifungal and insecticidal properties. Here, we evaluated the plant growth‐inhibitory activities of two cultivars of Curcuma longa (C. longa ; Ryudai gold and Okinawa ukon) against radish, cress, lettuce and Bidens pilosa (B. pilosa ). The methanol extracts of both turmeric varieties inhibited the seed germination and seedling growth of all the tested plants. Ryudai gold had a significantly higher inhibitory effect on the seed germination and root and shoot growth of the plants than Okinawa ukon. Therefore, Ryudai gold was chosen for the isolation of plant growth‐inhibitory compounds using a silica gel column and high‐performance liquid chromatography. The structural identification of the compounds was carried out using ¹H NMR, ¹³C NMR and liquid chromatography–tandem mass spectrometry. The growth inhibitors were identified as four curcuminoids; dihydrobisdemethoxycurcumin ( 1 ), bisdemethoxycurcumin ( 2 ), demethoxycurcumin ( 3 ) and curcumin ( 4 ). The IC₅₀ of the curcuminoids against the root and shoot growth of B. pilosa ranged from 8.7 ± 1.7 to 12.9 ± 1.8 and from 15.5 ± 1.8 to 38.9 ± 2.8 μmol L^?1, respectively. Compound 1 showed the lowest IC₅₀ against the root and shoot growth of B. pilosa . These results suggested that Ryudai gold has a potential growth‐inhibitory effect against B. pilosa .     

RNAi‐mediated plant protection against aphids

Aphids (Aphididae) are major agricultural pests that cause significant yield losses of crop plants each year by inflicting damage both through the direct effects of feeding and by vectoring harmful plant viruses. Expression of double‐stranded RNA (dsRNA) directed against suitable insect target genes in transgenic plants has been shown to give protection against pests through plant‐mediated RNA interference (RNAi). Thus, as a potential alternative and effective strategy for insect pest management in agricultural practice, plant‐mediated RNAi for aphid control has received close attention in recent years. In this review, the mechanism of RNAi in insects and the so far explored effective RNAi target genes in aphids, their potential applications in the development of transgenic plants for aphid control and the major challenges in this regard are reviewed, and the future prospects of using plant‐mediated RNAi for aphid control are discussed. This review is intended to be a helpful insight into the generation of aphid‐resistant plants through plant‐mediated RNAi strategy. © 2016 Society of Chemical Industry     

Ecdysteroid receptor docking suggests that dibenzoylhydrazine-based insecticides are devoid of any deleterious effect on the parasitic wasp Psyttalia concolor (Hym. Braconidae)

BACKGROUND: The moulting accelerating compounds (MACs) or ecdysteroid agonists represent a selective group of insecticides acting upon binding to the ecdysteroid receptor (EcR) and leading to lethal premature moulting in larval stages and aborted reproduction in adults. Psyttalia concolor Szèpl. is a useful parasitic wasp attacking important tephritid pests such as the medfly and olive fruit fly. RESULTS: Contact and oral exposure in the laboratory of female parasitic wasps to the dibenzoylhydrazine‐based methoxyfenozide, tebufenozide and RH‐5849 did not provoke negative effects. No mortality and no reduction in beneficial capacity were observed. The ligand‐binding domain (LBD) of the EcR of P. concolor was sequenced, and a homology protein model was constructed which confirmed a cavity structure with 12 α‐helices, harbouring the natural insect moulting hormone 20‐hydroxyecdysone. However, a steric clash occurred for the MAC insecticides owing to a restricted extent of the ligand‐binding cavity of the PcLBD‐EcR, while they did dock well in that of susceptible insects. CONCLUSIONS: The insect toxicity assays demonstrated that MACs are selective for P. concolor. The modelling/docking experiments are indications that these insecticides do not bind with the LBD‐EcR of P. concolor and support the theory that they show no biological effects in the parasitic wasp. These data may help in explaining the compatible use of MACs together with parasitic wasps in IPM programmes. Copyright © 2012 Society of Chemical Industry     

Temporal dynamics of the arthropod community in pear orchards intercropped with aromatic plants

BACKGROUND: Increasing attention has been paid to enhancing biological control through habitat management in agricultural systems for enhanced pest management. Pest management benefits can be realised by intercropping, which can increase natural enemy abundance and, in turn, reduce pest abundance. In this study, the composition and temporal dynamics of arthropod communities in pear orchards when intercropped with aromatic plants were investigated, and the effectiveness and applicability of aromatic plants as intercrops for enhancing insect control were assessed. RESULTS: When compared with natural grasses or clean tillage, intercropping significantly reduced pest abundance and increased the ratio of natural enemies to pests. Intercropping also shortened the occurrence duration and depressed the incidence peak in annual dynamics curves of the pest subcommunity and the arthropod community, mainly because of the repellent effects of aromatic plants. Equally important, intercropping significantly reduced the numbers of major pests, such as Psylla chinensis, Aphis citricola and Pseudococcus comstocki, while their incidence period was delayed to varying degrees, and the numbers of their dominant natural enemies (Coccinella septempunctata, Phytoseiulus persimilis and Chrysoperla sinica) increased. CONCLUSION: Intercropping with aromatic plants led to a considerable improvement in arthropod pest management by enhancing the activity of the beneficial arthropod community within the pear orchard ecosystem. Copyright © 2011 Society of Chemical Industry     

Per os efficacy of Ajuga extracts against sucking insects

We studied the efficacy of water-soluble extracts from four Ajuga spp on the post-embryonic development of two exopterygota (sucking insect) species. To allow comparison between different Ajuga species, results are expressed in terms of quantity of plant extracted per litre of test solution. Crude methanolic extracts of all Ajuga plants tested, with the exception of A genevensis, showed considerable per os efficacy against larvae of both Dysdercus cingulatus F and Acyrthosiphon pisum (Harris) even at 1 g litre(-1). In the aphid tests the order of efficacy was A bracteosa Wallich ex Benth > A chamaepitys Schreber > A reptans L > A genevensis L. On D cingulatus the order of efficacy was: A reptans > A bracteosa > A chamaepitys > A genevensis. Extracts were fractionated on SepPak using a range of methanol/water mixtures. Results are expressed in terms of the initial weight of plant extracted. The 100% methanolic fraction of A chamaepitys was highly effective on A pisum (100% mortality at 1 g litre(-1)) and less effective on D cingulatus (about 60% mortality at 5 g litre(-1)). The entire 60 methanol + 40 water fraction was effective against test insects but showed different efficacies according to test species and concentration applied. 20-Hydroxyecdysone (20E), cyasterone (Cy) and ajugalactone (Ajl) were identified in the fractions from all Ajuga species, but the remaining phytoecdysteroid profile was quite different between Ajuga species. Capitasterone (Cap) and 28-epi-sengosterone (5Cy28') were found only in A reptans, makisterone A (MaA) and 29-norcyasterone (29NCy) were only in A chamaepitys, while 22-acetylcyasterone (Cy22A), 3-epi-cyasterone (Cy') and 3-epi-22-acetylcyasterone (Cy'22A) were only in A bracteosa. The total amount of phytoecdysteroids was 2053 mgkg(-1) for A bracteosa, 1892 mgkg(-1) for A reptans and 95 mg kg(-1) for A chamaepitys.     

Assessment of the high-dose concept and level of control provided by MON 87701 × MON 89788 soybean against Anticarsia gemmatalis and Pseudoplusia includens (Lepidoptera: Noctuidae) in Brazil

BACKGROUND: Genetically modified MON 87701 × MON 89788 soybean (Glycine max), which expresses the Cry1Ac and EPSP‐synthase proteins, has been registered for commercial use in Brazil. To develop an Insect Resistance Management (IRM) program for this event, laboratory and field studies were conducted to assess the high‐dose concept and level of control it provides against Anticarsia gemmatalis and Pseudoplusia includens. RESULTS: The purified Cry1Ac protein was more active against A. gemmatalis [LC₅₀ (FL 95%) = 0.23 (0.15–0.34) µg Cry1Ac mL^?1 diet] than P. includens [LC₅₀ (FL 95%) = 3.72 (2.65–4.86) µg Cry1Ac mL^?1 diet]. In bioassays with freeze‐dried MON 87701 × MON 89788 soybean tissue diluted 25 times in an artificial diet, there was 100% mortality of A. gemmatalis and up to 95.79% mortality for P. includens. In leaf‐disc bioassays and under conditions of high artificial infestation in the greenhouse and natural infestation in the field, MON 87701 × MON 89788 soybean showed a high level of efficacy against both target pests. CONCLUSIONS: The MON 87701 × MON 89788 soybean provides a high level of control against A. gemmatalis and P. includes, but a high‐dose event only to A. gemmatalis. Copyright © 2012 Society of Chemical Industry     

Increased toxicity of Bacillus thuringiensis Cry3Aa against Crioceris quatuordecimpunctata,Phaedon brassicae and Colaphellus bowringi by a Tenebrio molitor cadherin fragment

BACKGROUND: Biopesticides containing Cry insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) are effective against many lepidopteran pests, but there is a lack of Bt‐based pesticides for efficient control of important coleopteran pests. Based on the reported increase in Bt toxin oligomerization by a polypeptide from the Cry3Aa receptor cadherin in Tenebrio molitor (Coleoptera: Tenebrionidae), it was hypothesized that this cadherin peptide, rTmCad1p, would enhance Cry3Aa toxicity towards coleopteran larvae. To test this hypothesis, the relative toxicity of Cry3Aa, with or without rTmCad1p, against damaging chrysomelid vegetable pests of China was evaluated. RESULTS: Cry3Aa toxicity was evaluated in the spotted asparagus beetle (Crioceris quatuordecimpunctata), cabbage leaf beetle (Colaphellus bowringi) and daikon leaf beetle (Phaedon brassicae). To assess the effect of rTmCad1p on Cry3Aa toxicity, neonate larvae were fed Cry3Aa toxin alone or in combination with increasing amounts of rTmCad1p. The data demonstrated that Cry3Aa toxicity was significantly increased in all three vegetable pests, resulting in as much as a 15.3‐fold increase in larval mortality. CONCLUSION: The application of rTmCad1p to enhance Cry3Aa insecticidal activity has potential for use in increasing range and activity levels against coleopteran pests displaying low susceptibility to Bt‐based biopesticides. Copyright © 2011 Society of Chemical Industry     

Extraction of allyl isothiocyanate from horseradish (Armoracia rusticana) and its fumigant insecticidal activity on four stored‐product pests of paddy

BACKGROUND: Isothiocyanates (ITCs) extracted from Armoracia rusticana Gaertn., May & Scherb. have been shown previously to have insecticidal activity. Allyl isothiocyanate (AITC), a major component of ITCs with high volatility, was therefore extracted using different methods and tested as a fumigant against four major pest species of stored products, maize weevil Sitophilus zeamais (Motsch.), lesser grain borer Rhizopertha dominica (F.), Tribolium ferrugineum (F.) and book louse Liposcelis entomophila (Enderlein). RESULTS: Whereas there was no significant difference between hydrodistillation and supercritical carbon dioxide fluid extraction in extraction rate for AITC from A. rusticana, both methods resulted in higher extraction efficiency than water extraction. AITC fumigation showed strong toxicity to the four species of stored‐product pests. Adult mortality of 100% of all four pest species, recorded after 72 h exposure to AITC fumes at an atmospheric concentration of 3 µg mL^?1, showed no significant difference from that of insects exposed to phosphine at 5 µg mL^?1, the recommended dose for phosphine. CONCLUSIONS: The results suggest good insecticidal efficacy of AITC against the four stored‐product pests, with non‐gaseous residuals on stored products. AITC obtained from A. rusticana may be an alternative to phosphine and methyl bromide against the four pest species. Copyright © 2009 Society of Chemical Industry     

Immobilization of the pesticide 2,4‐dichlorophenoxyacetic acid on a silica gel surface

A route for the immobilization of 2,4‐dichlorophenoxyacetic acid (2,4‐D) on silica gel has been developed. In the first step the precursor was prepared by reacting the silylating agent 3‐(trimethoxysilyl)propylamine with silica gel. Nitrogen analysis of this anchored compound showed the presence of 1.16 mmol of amine groups per gram of support. The herbicide was covalently bonded to the amine groups previously anchored onto silica gel. Infrared, [¹³C] and [²⁹Si] NMR spectra supported the reaction between the nitrogen of the amine group of the anchored silica with the carbon on the para‐position of the aromatic ring of 2,4‐D. The reaction yield, 90.3%, was confirmed through elemental analysis. © 2000 Society of Chemical Industry     

Ovicidal and oviposition deterrent activities of the flavonoid pectolinaringenin from Clerodendrum phlomidis against Earias vittella

Fractions derived from Clerodendrum phlomidis Linn. F. and isolated compounds from chloroform extract were evaluated against Earias vittella Fab. for their ovicidal and oviposition deterrent activities. There were 12 fractions obtained with silica gel column chromatographic techniques using different solvent systems. Among the fractions, fraction 5 recorded maximum ovicidal activity of 68.49% and oviposition deterrent activity of 100% at 1000 ppm concentration followed by 500 ppm concentration. The active fraction was further purified using HPLC. The purified fraction was subjected to UV, ¹HNMR, ¹³CNMR, IR and EI-Mass spectroscopy and their spectral data confirmed the compound pectolinaringenin which belongs to the flavonoids. The compound pectolinaringenin was screened for its ovicidal and oviposition deterrent activities. It showed maximum ovicidal activity of 64.29% and oviposition deterrent activity of 100% at 100 ppm concentration against E. vittella. To the best of our knowledge, this is the first report of fractions and isolated compounds from C. phlomidis evaluated against E. vittella for their ovicidal and oviposition deterrent activities. The results from the present investigation clearly suggest that the active fraction or the isolated compound could be used to develop a novel pesticidal formulation to control economically important agricultural pests.     

Effects of cycloxaprid,a novel cis‐nitromethylene neonicotinoid insecticide,on the feeding behaviour of Sitobion avenae

BACKGROUND: Cycloxaprid with cis‐configuration is a novel neonicotinoid insecticide, developed in China, that has good industrialisation prospects for its high activity against imidacloprid‐resistant pests. Studies were carried out to investigate the biological activity of cycloxaprid and its effect on the feeding behaviour of Sitobion avenae. RESULTS: The results demonstrated that cycloxaprid had good contact and root‐systemic activity. The electrical penetration graph (EPG) revealed that cycloxaprid significantly increased the total time of non‐probing periods and greatly inhibited phloem ingestion of treated aphids because it significantly reduced the total time and the frequency of phloem ingestion. Furthermore, cycloxaprid can increase the phloem salivation of treated aphids by comparison with aphids treated with imdacloprid and distilled water. Consequently, a sublethal dose of cycloxaprid impaired aphid ingestion of phloem sap and thereby reduced the weight of aphids. On the other hand, EPG data showed that a sublethal dose of cycloxaprid had no significant effect on xylem sap ingestion, in contrast to imidacloprid. CONCLUSION: All these tests indicated that cycloxaprid had both contact and root‐systemic activity, with sublethal effects resulting in reduction in Sitobion avenae phloem‐feeding behaviour and growth rate. Copyright © 2012 Society of Chemical Industry     

Selectivity of diacylhydrazine insecticides to the predatory bug Orius laevigatus: in vivo and modelling/docking experiments

BACKGROUND: Knowledge of pesticide selectivity to natural enemies is necessary for a successful implementation of biological and chemical control methods in integrated pest management (IPM) programmes. Diacylhydrazine (DAH)‐based ecdysone agonists, also known as moulting‐accelerating compounds (MACs), are considered to be a selective group of insecticides, and their compatibility with predatory Heteroptera, which are used as biological control agents, is known. However, their molecular mode of action has not been explored in beneficial insects such as Orius laevigatus (Fieber) (Hemiptera: Anthocoridae). RESULTS: In this project, in vivo toxicity assays demonstrated that the DAH‐based RH‐5849, tebufenozide and methoxyfenozide have no toxic effect against O. laevigatus. The ligand‐binding domain (LBD) of the ecdysone receptor (EcR) of O. laevigatus was sequenced, and a homology protein model was constructed that confirmed a cavity structure with 12 α‐helices, harbouring the natural insect moulting hormone 20‐hydroxyecdysone. However, docking studies showed that a steric clash occurred for the DAH‐based insecticides owing to a restricted extent of the ligand‐binding cavity of the EcR of O. laevigatus. CONCLUSIONS: The insect toxicity assays demonstrated that MACs are selective for O. laevigatus. The modelling/docking experiments are indications that these pesticides do not bind with the LBD‐EcR of O. laevigatus and support the supposition that they show no biological effects in the predatory bug. These data help in explaining the compatible use of MACs together with predatory bugs in IPM programmes. Copyright © 2012 Society of Chemical Industry     

Field trial measuring the compatibility of methoxyfenozide and flonicamid with Orius laevigatus Fieber (Hemiptera: Anthocoridae) and Amblyseius swirskii (Athias-Henriot) (Acari: Phytoseiidae) in a commercial pepper greenhouse

BACKGROUND: Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) and Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) are among the most serious pests of sweet peppers in greenhouses. Chemical control is difficult because of their high reproductive rates and insecticide resistance, and seasonal inoculative releases of Orius laevigatus (Fieber) and Amblyseius swirskii (Athias‐Henriot) are commonly used to reduce their populations. As chemical treatments are often needed in the crop against other pests, the side effects of methoxyfenozide (an insect growth regulator against lepidopteran pests) and flonicamid (a selective feeding inhibitor against sucking insects) were studied in both beneficial organisms in a commercial greenhouse. RESULTS: Orius laevigatus and A. swirskii were released at commercial rates (4–5 and 100 m^?2), and a strong establishment and a very homogeneous distribution were reached. One pesticide treatment with the maximum field recommended concentration of methoxyfenozide and flonicamid (96 and 100 mg AI L^?1) was done when they were well established, and their population levels were not affected either immediately or up to 30 days after treatment. CONCLUSION: The results are indicative of no impact of methoxyfenozide and flonicamid on the two natural enemies in the field, and both can be considered as potential alternatives to be included in IPM programmes in sweet pepper. Copyright © 2011 Society of Chemical Industry     

Inheritance of sulfonylurea resistance in Monochoria vaginalis

The inheritance of sulfonylurea (SU) resistance in Monochoria vaginalis was investigated based on the bensulfuron‐methyl response phenotypes of F₁ plants between SU‐resistant (R) and ‐susceptible (S) and segregation analysis in F₂ progenies. Differences of SU resistance between SU‐R biotypes and F₁ plants at the recommended field dose were also investigated by comparing shoot dry weight. All F₁ plants survived the treatment with 25 g a.i. ha^?1 bensulfuron‐methyl, one‐third of the recommended field dose, and showed similar responses to SU‐R plants. Conversely, all F₁ plants died or showed extreme necrosis at 225 g a.i. ha^?1, three times the recommended field dose, as SU‐S plants. F₂ plants were classified as either R or S phenotype. Segregation for resistance to bensulfuron‐methyl in F₂ families did not differ from the expected 3:1 (R:S) ratio at 25 g a.i. ha^?1. At 225 g a.i. ha^?1, the F₂ families segregated in a 1:3 (R:S) ratio. These results suggest that SU resistance in M. vaginalis is controlled by a single nuclear allele with resistance being dominant at low dose and susceptibility dominant at high dose. Moreover, F₁ plants died or were extremely injured after application of bensulfuron‐methyl at the recommended field dose, although SU‐R biotypes grew normally.     

Antifeedant effect of commercial chemicals and plant extracts against Schistocerca americana (Orthoptera: Acrididae) and Diaprepes abbreviatus (Coleoptera: Curculionidae)

BACKGROUND: The deterrent effect of seven commercial compounds against Schistocerca americana (Drury) nymphs and Diaprepes abbreviatus L. adults, two important citrus pests, was investigated. Extracts of three Florida local plants were also tested as deterrents. Antifeedant activity was assayed using a leaf‐disk bioassay in choice and no‐choice tests. The residual activity of the commercial compounds that significantly deterred the insects was assayed by applying them to foliage of citrus plants exposed to three time intervals of sunlight. RESULTS: Sabadilla, azadirachtin and ryanodine effectively deterred S. americana, whereas rotenone, sabadilla and ryanodine reduced the feeding activity of D. abbreviatus. Methanol and methylene chloride extracts of Ceratiola ericoides Michaux deterred D. abbreviatus, but only methylene chloride extract dissuaded S. americana. Methanol extract of Ardisia crenata Sims functioned as a feeding deterrent against both S. americana and D. abbreviatus, but only the latter species was deterred by methylene chloride extract of A. crenata. Extracts of Illicium parviflorum Michaux ex Ventetat only dissuaded the insects in choice bioassays. Rapid loss of effectiveness was observed under field conditions. Sabadilla was the only compound that maintained its antifeedant properties in the field, but only against S. americana. CONCLUSION: Based on their deterrency, some of the commercial products and plant extracts have potential for use as substitute crop protectants against the pest species tested. Copyright © 2011 Society of Chemical Industry     

Mode of action of methoprene in affecting female reproduction in the African malaria mosquito,Anopheles gambiae

BACKGROUND: One of the most studied actions of juvenile hormone (JH) is its ability to modulate ecdysteroid signaling during insect development and metamorphosis. Previous studies in mosquitoes showed that 20‐hydroxyecdysone (20E) regulates vitellogenin synthesis. However, the action of JH and its mimics, e.g. methoprene, on female reproduction of mosquitoes remains unknown. RESULTS: Here, a major malaria vector, Anopheles gambiae Giles, was used as a model insect to study the action of methoprene on female reproduction. Ecdysteroid titers and expression profiles of ecdysone‐regulated genes were determined before and after a blood meal. An ecdysteroid peak was detected at 12 h post blood meal (PBM). The maximum expression of ecdysone‐regulated genes, such as ecdysone receptor (EcR), hormone receptor 3 (HR3) and vitellogenin (Vg) gene, coincided with the ecdysteroid peak. Interestingly, topical application of methoprene at 6 h PBM delayed ovarian development and egg maturation by suppressing the expression of ecdysone‐regulated genes in female mosquitoes. CONCLUSION: The data suggest that ecdysteroid titers are correlated with Vg synthesis, and methoprene affects vitellogenesis by modulating ecdysteroid action in A. gambiae. Copyright © 2010 Society of Chemical Industry     

Long‐lasting β‐aminobutyric acid‐induced resistance protects tomato fruit against Botrytis cinerea

Minimizing losses to pests and diseases is essential for producing sufficient food to feed the world's rapidly growing population. The necrotrophic fungus Botrytis cinerea triggers devastating pre‐ and post‐harvest yield losses in tomato (Solanum lycopersicum). Current control methods are based on the pre‐harvest use of fungicides, which are limited by strict legislation. This investigation tested whether induction of resistance by β‐aminobutyric acid (BABA) at different developmental stages provides an alternative strategy to protect post‐harvest tomato fruit against B. cinerea. Soil‐drenching plants with BABA once fruit had already formed had no impact on tomato susceptibility to B. cinerea. However, BABA application to seedlings significantly reduced post‐harvest infection of fruit. This resistance response was not associated with a yield reduction; however, there was a delay in fruit ripening. Untargeted metabolomics revealed differences between fruit from water‐ and BABA‐treated plants, demonstrating that BABA triggered a defence‐associated metabolomics profile that was long lasting. Targeted analysis of defence hormones suggested a role of abscisic acid (ABA) in the resistance phenotype. Post‐harvest application of ABA to the fruit of water‐treated plants induced susceptibility to B. cinerea. This phenotype was absent from the ABA‐exposed fruit of BABA‐treated plants, suggesting a complex role of ABA in BABA‐induced resistance. A final targeted metabolomic analysis detected trace residues of BABA accumulated in the red fruit. Overall, it was demonstrated that BABA induces post‐harvest resistance in tomato fruit against B. cinerea with no penalties in yield.     

A protocol for analysing the costs and benefits of phytosanitary measures*

This paper describes a protocol developed within the PRATIQUE project for applying cost/benefit analysis to select appropriate phytosanitary measures for use against quarantine pests. The protocol consists of nine steps, each underpinned by clear guidelines for collating and structuring the relevant data, to assist risk managers with the challenging task of assessing the benefits and costs of phytosanitary measures in a consistent, transparent and reproducible manner. Together with the decision‐support scheme that generates contingency plants and prioritizes action during pest outbreaks, it provides a framework for decision making on phytosanitary measures and helps to provide economic justification for selecting appropriate measures. The practical application of the protocol is demonstrated using two examples: cost/benefit analysis of eradication measures against Anoplophora glabripennis in Northern Italy; and eradication and containment measures against Diabrotica virgifera virgifera in Germany.