Local Application of Acibenzolar-S-Methyl Treatment Induces Antiviral Responses in Distal Leaves of Arabidopsis thaliana.

Systemic acquired resistance (SAR) is a plant defense mechanism that provides protection against a broad spectrum of pathogens in distal tissues. Recent studies have revealed a concerted function of salicylic acid (SA) and N-hydroxypipecolic acid (NHP) in the establishment of SAR against bacterial pathogens, but it remains unknown whether NHP is also involved in SAR against viruses. We found that the local application of acibenzolar-S-methyl (ASM), a synthetic analog of SA, suppressed plantago asiatica mosaic virus (PlAMV) infection in the distal leaves of Arabidopsis thaliana. This suppression of infection in untreated distal leaves was observed at 1 day, but not at 3 days, after application. ASM application significantly increased the expression of SAR-related genes, including PR1, SID2, and ALD1 after 1 day of application. Viral suppression in distal leaves after local ASM application was not observed in the sid2-2 mutant, which is defective in isochorismate synthase 1 (ICS1), which is involved in salicylic acid synthesis; or in the fmo1 mutant, which is defective in the synthesis of NHP; or in the SA receptor npr1-1 mutant. Finally, we found that the local application of NHP suppressed PlAMV infection in the distal leaves. These results indicate that the local application of ASM induces antiviral SAR against PlAMV through a mechanism involving NHP.

The Impact of a Commercial Biostimulant on the Grape Mycobiota of Vitis vinifera cv. Barbera.

Reducing the use of fungicides, insecticides, and herbicides in order to limit environmental pollution and health risks for agricultural operators and consumers is one of the goals of European regulations. In fact, the European Commission developed a package of measures (the European Green Deal) to promote the sustainable use of natural resources and strengthen the resilience of European agri-food systems. As a consequence, new plant protection products, such as biostimulants, have been proposed as alternatives to agrochemicals. Their application in agroecosystems could potentially open new scenarios regarding the microbiota. In particular, the vineyard microbiota and the microbiota on the grape surface can be affected by biostimulants and lead to different wine features. The aim of this work was to assess the occurrence of a possible variation in the mycobiota due to the biostimulant application. Therefore, our attention has been focused on the yeast community of grape bunches from vines subjected to the phytostimulant BION®50WG treatment. This work was carried out in the CREA-VE experimental vineyard of Vitis vinifera cv. Barbera in Asti (Piedmont, Italy). The composition of fungal communities on grapes from three experimental conditions such as IPM (integrated pest management), IPM+BION®50WG, and IPM+water foliar nebulization was compared by a metabarcoding approach. Our results revealed the magnitude of alpha and beta diversity, and the microbial biodiversity index and specific fungal signatures were highlighted by comparing the abundance of yeast and filamentous fungi in IPM and BION®50WG treatments. No significant differences in the mycobiota of grapevines subjected to the three treatments were detected.

Modification of the existing maximum residue levels for acibenzolar-S-methyl in grapes.

In accordance with Article 6 of Regulation (EC) No 396/2005, the applicant Syngenta Crop Protection AG submitted a request to the competent national authority in Italy to modify the existing maximum residue levels (MRLs) for the active substance acibenzolar-S-methyl in grapes. The data submitted in support of the request were found to be sufficient to derive MRL proposals for table and wine grapes. Adequate analytical methods for enforcement are available to control the residues of acibenzolar-S-methyl residues and of the metabolite acibenzolar acid (free and conjugated) in plant matrices at the validated limit of quantification (LOQ) of 0.01 mg/kg. Based on the risk assessment results, EFSA concluded that the short-term and long-term intake of residues resulting from the use of acibenzolar-S-methyl according to the reported agricultural practices is unlikely to present a risk to consumer health.

Evaluation of Plant Defense Inducers and Plant Growth Regulators for Fire Blight Management Using Transcriptome Studies and Field Assessments.

Fire blight, caused by Erwinia amylovora, is a destructive disease of pome fruit trees. In the United States, apple and pear growers rely on applications of copper and antibiotics during bloom to control fire blight, but such methods have already led to regional instances of resistance. In this study, we used transcriptome analyses and field trials to evaluate the effectiveness of three commercially available plant defense elicitors and one plant growth regulator for fire blight management. Our data indicated that foliar applications of acibenzolar-S-methyl (ASM; Actigard 50WG) triggered a strong defense-related response in apple leaves, whereas applications of Bacillus mycoides isolate J (LifeGard WG) or Reynoutria sachalinensis extract (Regalia) did not. Genes upregulated by ASM were enriched in the biological processes associated with plant immunity, such as defense response and protein phosphorylation. The expression of several pathogenesis-related (PR) genes was induced by ASM as well. Surprisingly, many differentially expressed genes in ASM-treated apple leaves overlapped with those induced by treatment with prohexadione-calcium (ProCa; Apogee), a plant growth regulator that suppresses shoot elongation. Further analysis suggested that ProCa likely acts similarly to ASM to stimulate plant immunity because genes involved in plant defense were shared and significantly upregulated (more than twofold) by both treatments. Our field trials agreed with the transcriptome study, demonstrating that ASM and ProCa exhibit the best control performance relative to the other biopesticides. Taken together, these data are pivotal for the understanding of plant response and shed light on future improvements of strategies for fire blight management.

Efficacy of Biological Control Agents and Resistance Inducer for Control of Mal Secco Disease.

Mal secco, caused by Plenodomus tracheiphilus, is an economically important fungal vascular disease in citrus-growing countries of the Mediterranean basin. Preventing fungal infections usually requires a high number of copper treatments but European legislation imposes the minimization of their accumulation in soil. In our study, biological control agents (BCAs) and a plant resistance inducer (PRI), tested in four different experiments on citrus seedlings under controlled conditions, have resulted in promising strategies to control mal secco disease. Foliar (Experiment I) and soil (Experiment II) applications of two formulations of Bacillus amyloliquefaciens strain D747 (Amylo-X® LC and Amylo-X® WG) provided similar performances in reducing the disease amount (incidence and symptoms severity) over time compared to the untreated control, whereas copper hydroxide (Kocide Opti®) used as standard was the most effective treatment over time. In the third experiment, Pythium oligandrum strain M1 (Polyversum®) and Trichoderma asperellum strain ICC012 + Trichoderma gamsii strain ICC080 (Remedier®) were able to reduce disease incidence and symptoms severity compared to the untreated control. Remedier® provided the best performances in reducing the disease amount, whereas the Polyversum® application was the least effective treatment over time. The effectiveness of the Trichoderma spp. formulation in reducing P. tracheiphilus infections did not significantly differ from the standard copper compound (Kocide Opti®). Comprehensively, in the last experiment (IV), acibenzolar-S-methyl (ASM) alone and in mixture with metalaxyl-M proved as effective as B. amyloliquefaciens strain FZB24, with no dose-response relationships observed. These findings provide important insight for the integrated management of mal secco disease.

Statement concerning the testing strategy and timelines proposed by the applicant for the assessment of the endocrine disruption properties of acibenzolar-S-methyl in the context of the review of the approval of the active substance.

On 8 July 2021, EFSA published a Conclusion on the peer review of the pesticide risk assessment for the active substance acibenzolar-S-methyl in light of confirmatory data submitted. EFSA concluded that based on the confirmatory information submitted by the applicant, the assessment of endocrine-disrupting properties could not be finalised for humans and non-target organisms and identified further data deemed necessary to finalise the assessment. Consequently, during the decision-making stage it could not be concluded by risk managers that acibenzolar-S-methyl still meets the approval criteria laid down in Article 4 of Regulation (EC) No 1107/2009 and therefore the European Commission decided to launch a review of the existing approval in accordance with Article 21 of that Regulation and on 6 July 2022 invited the applicant to submit comments on the findings in the EFSA Conclusion including any relevant information. On 14 December 2022, the European Commission requested EFSA to consider the proposal as submitted by the applicant in light of the EFSA Conclusion and to confirm whether the proposed studies are considered sufficient to complete the assessment of the endocrine disrupting properties of the substance in line with Commission Regulation (EU) 2018/605. The current statement contains EFSA's considerations as regards the testing strategy and associated timelines for additional data generation proposed by the applicant to complete the assessment of the endocrine disrupting properties of acibenzolar-S-methyl in line with Commission Regulation (EU) 2018/605.

Acibenzolar-S-methyl activates phenylpropanoid pathway to enhance resistance against Alternaria alternata in pear fruit.

BACKGROUND: Alternaria alternata is a causal agent of black spot rot of pear fruit after harvest. Acibenzolar-S-methyl (ASM) has been shown to be a potential elicitor of tolerance in several horticultural products. This work was performed to research the influence of ASM on black spot rot of Docteur Jules Guyot pears and vital enzyme activity and gene expression in the phenylpropanoid pathway. RESULTS: ASM remarkably decreased the lesion diameter of A. alternata-inoculated pears. ASM also increased phenylalanine ammonialyase, cinnamate 4-hydroxylase, cinnamyl alcohol dehydrogenase, peroxidase, polyphenol oxidase activities and gene expression, and enhanced 4-coumarate/coenzyme A ligase activity in pears. Moreover, ASM improved the content of phenylalanine, total phenolic compounds, caffeic acid, flavonoids, anthocyanin and lignin in pears. CONCLUSION: ASM could modulate vital enzyme activity and gene expression in the phenylpropanoid pathway to accelerate metabolite synthesis, thereby enhancing resistance against A. alternata in pears. © 2022 Society of Chemical Industry.

Acibenzolar-S-methyl activates calcium signalling to mediate lignin synthesis in the exocarp of Docteur Jules Guyot pears.

'Docteur Jules Guyot' pears were immersed in acibenzolar-S-methyl (ASM) and 0.01 mol L-1 ethyl glycol tetra acetic acid (EGTA) to investigate the changes of Ca2+ receptor proteins and phenylpropanoid pathway. Results showed that ASM treatment increased the activities of phenylalanine ammonia-lyase (PAL), cinnamate-4-hydroxylase (C4H), 4-coumarate coenzyme A ligase (4CL), polyphenol oxidase (PPO), and cinnamyl alcohol dehydrogenase (CAD) in the exocarp of pears, whereas EGTA pre-treatment inhibited the activities of these enzymes. ASM treatment also enhanced the transcription of PcPAL, PcC4H, Pc4CL, PcC3H, PcCOMT, PcCCoAOMT, PcCCR, PcPOD, PcCDPK1, PcCDPK2, PcCDPK5, PcCDPK11, PcCDPK13, PcCBL1, PcCBL9, PcCIPK14, and PcCML27 in pears. EGTA + ASM treatments inhibited the transcription of PcPAL, PcC4H, Pc4CL, PcC3H, PcCCR, PcF5H, PcCAD, PcCDPK11, PcCDPK26, PcCDPK32, PcCBL1, PcCIPK14, PcCIPK23, and PcCaM in the fruit. All these results indicated that ASM induced the gene expressions of Ca2+ receptor proteins, the key enzyme activities and gene expressions in phenylpropanoid pathway; Ca2+ mediated phenylpropane metabolism in pears after ASM treatment.

Distinct chemical resistance-inducing stimuli result in common transcriptional, metabolic, and nematode community signatures in rice root and rhizosphere.

Induced resistance (IR), a phenotypic state induced by an exogenous stimulus and characterized by enhanced resistance to future (a)biotic challenge, is an important component of plant immunity. Numerous IR-inducing stimuli have been described in various plant species, but relatively little is known about 'core' systemic responses shared by these distinct IR stimuli and the effects of IR on plant-associated microbiota. In this study, rice (Oryza sativa) leaves were treated with four distinct IR stimuli (ß-aminobutyric acid, acibenzolar-S-methyl, dehydroascorbic acid, and piperonylic acid) capable of inducing systemic IR against the root-knot nematode Meloidogyne graminicola and evaluated their effect on the root transcriptome and exudome, and root-associated nematode communities. Our results reveal shared transcriptional responses-notably induction of jasmonic acid and phenylpropanoid metabolism-and shared alterations to the exudome that include increased amino acid, benzoate, and fatty acid exudation. In rice plants grown in soil from a rice field, IR stimuli significantly affected the composition of rhizosphere nematode communities 3 d after treatment, but by 14 d after treatment these changes had largely reverted. Notably, IR stimuli did not reduce nematode diversity, which suggests that IR might offer a sustainable option for managing plant-parasitic nematodes.

DNP and ATP regulate the pulp breakdown development in Phomopsis longanae Chi-infected longan fruit through modulating the ROS metabolism.

Compared with the P. longanae-infected longan, the DNP-treated P. longanae-infected fruit represented a higher pulp breakdown index, a higher O2 -. production rate, and a higher MDA content, but the lower activities of APX, SOD and CAT, the lower transcript levels of DlAPX6, DlSOD1, DlSOD2, DlSOD3 and DlCAT1, the lower values of AsA, GSH, flavonoid and total phenolics, a lower scavenging ability of DPPH radical, and a lower value of reducing power. Whereas, the ATP-treated P. longanae-infected samples showed the contrary results. The above findings indicated that the DNP-promoted the pulp breakdown in P. longanae-infected longan was because DNP weakened the capacity of scavenging ROS, raised the O2 -. level, and accelerated the membrane lipids peroxidation. However, the ATP-suppressed the pulp breakdown in P. longanae-infected longan was because ATP improved the capacity of scavenging ROS, reduced the O2 -. level, and reduced the membrane lipids peroxidation.

Effects of Acibenzolar-S-methyl on the Probing Behaviour and Mortality of Cacopsylla pyri on Pear Plants.

European pear psylla, Cacopsylla pyri, is one of the worst pests of pear in Europe. We investigated whether acibenzolar-S-methyl (ASM) application on pear plants might affect the behaviour in C. pyri. The elicitor was applied on pear potted plants, and after 48 h, we confirmed the ASM-mediated induction of several Pathogenesis-Related protein (PR) coding genes. At the same time, an in-depth analysis was performed on the probing behaviour of adults and nymphs of C. pyri on ASM-treated pear plants by the EPG-DC system, as well as the assessment of young nymphs' survival 7 days after the ASM application. The elicitor application weakly interfered with C. pyri nymphs probing behaviour and survival, while it did not affect adult stages. These data confirm previous observations obtained on C. pyricola and suggest that the elicitor does not represent a viable tool in the control of pear psylla species, especially if used alone, but it might be used in integrated management strategies focused on other plant pathogens such as Erwinia amylovora.

Ellman's method-based determination of acibenzolar-S-methyl in tobacco by transesterification with methanol and SABP2-catalyzed hydrolysis.

Acibenzolar-S-methyl (ASM) is the most commercially successful biological antibacterial agent used for crop through systemic acquired resistance (SAR). In this study, a reproducible and accurate procedure, based on the spectrophotometric/microplate reader analysis, has been developed to detect ASM in tobacco leaves. This method involves oxidation of methyl mercaptan by the Ellman's reagent 5,5'-dithio-bis (2-nitrobenzoic acid) (DTNB) to form the yellow derivative 5'-thio-2-nitrobenzoic acid (TNB2-), measurable at 412 nm. Methyl mercaptan can be produced by either the ASM transesterification with methanol or the SA-binding protein 2 (SABP2)-catalyzed ASM hydrolysis. The proportions of methanol, reaction time, temperature, the concentrations of EDTA and DTNB were optimized in a 96-well plate. The calibration curve of ASM was linear over the range of 25.2-315 µg g-1. The results of the intra- and inter-day accuracy and precision data were within the FDA acceptance criteria. With ASM as substrate, the turnover number of SABP2 was determined, with the kcat value of 31.1 min-1 using the Michaelis-Menten equation. In tobacco plants treated with 100 µM ASM, it was decreased as time elapsed in treated tobacco, reaching negligible values 72 h after treatment. The optimized method was applied for the determination of ASM transesterification with methanol and the kinetic data determination of SABP2-catalyzed ASM hydrolysis.

Acibenzolar-S-methyl activates mitogen-activated protein kinase cascade to mediate chlorophyll and carotenoid metabolisms in the exocarp of Docteur Jules Guyot pears.

BACKGROUND: Acibenzolar-S-methyl (ASM), a well-known plant activator, has been used to protect fruit and vegetable from fungal invasion and maintain quality. However, little is known about the molecular mechanism of ASM in regulating chlorophyll and carotenoid metabolisms. Therefore, Docteur Jules Guyot pears were used as the materials to study the changes of hydrogen peroxide (H2 O2 ) production, mitogen-activated protein kinase (MAPK) cascade, transcription factors, chlorophyll, and carotenoid metabolisms after ASM and PD98059 (a MAPK cascade blocker) treatments. RESULTS: ASM increased NADPH oxidase (NOX) and superoxide dismutase (SOD) activities, and H2 O2 content, promoted PcMAPKKK1, PcMAPKK3, and PcMAPK6 expressions, and down-regulated PcMYC2, PcPIF1, PcPIF3, and PcPIF4 expressions in exocarp of pears. ASM also delayed the decrease of chlorophyll a and b contents, and inhibited the accumulation of ß-carotene, lycopene and lutein, PcNYC1, PcHCAR, PcPPH, PcSGR1/2, PcPAO, PcPSY, PcLCYB, PcCRTZ2, PcCCS1 expressions, and promoted PcLCYE expression. PD98059 + ASM treatments depressed SOD and NOX activities and H2 O2 content, inhibited PcMAPKKK1, PcMAPKK3, PcMAPK6, PcPIF1, and PcPIF3 expressions, and promoted PcMYC2 and PcPIF4 expressions in exocarp of pears. Additionally, PD98059 + ASM accelerated PcNYC1, PcHCAR, PcPPH, PcSGR1/2, PcPAO, PcPSY, PcCYB, PcCRTZ2, and PcCCS1 expressions, thereby reducing chlorophyll a and b contents, and promoting ß-carotene, lycopene and lutein contents. CONCLUSIONS: Postharvest ASM treatment promoted the production of H2 O2 to activate the MAPK cascade, then phosphorylated/dephosphorylated transcription factors expression, and delayed chlorophyll decomposition and carotenoid synthesis in pears. © 2022 Society of Chemical Industry.

Acibenzolar-S-methyl-mediated restriction of loading of plantago asiatica mosaic virus into vascular tissues of Nicotiana benthamiana.

Long-distance movement via vascular tissues is an essential step for systemic infection by plant viruses. We previously reported that pre-treatment of Nicotiana benthamiana with acibenzolar-S-methyl (ASM) both suppressed the accumulation of plantago asiatica mosaic virus (PlAMV) in inoculated leaves and delayed the long-distance movement to uninoculated upper leaves. These two effects occurred independently of each other. However, it remained unclear where and when the viral long-distance movement is inhibited upon ASM treatment. In this study, we found that ASM treatment restricted the loading of GFP-expressing PlAMV (PlAMV-GFP) into vascular tissues in the inoculated leaves. This led to delays in viral translocation to the petiole and the main stem, and to untreated upper leaves. We used cryohistological fluorescence imaging to show that ASM treatment affected the viral localization and reduced its accumulation in the phloem, xylem, and mesophyll tissues. A stem girdling experiment, which blocked viral movement downward through phloem tissues, demonstrated that ASM treatment could inhibit viral systemic infection to upper leaves, which occurred even with viral downward movement restricted. Taken together, our results showed that ASM treatment affects the loading of PlAMV-GFP into the vascular system in the inoculated leaf, and that this plays a key role in the ASM-mediated delay of viral long-distance movement.

Peer review of the pesticide risk assessment for the active substance acibenzolar-S-methyl in light of confirmatory data submitted.

The conclusions of the EFSA following the peer review of the initial risk assessment carried out by the competent authority of the rapporteur Member State, France, for the pesticide active substance acibenzolar-S-methyl are reported. The context of the peer review was that requested by the European Commission following the submission and evaluation of confirmatory information with regard to the endocrine disruption potential of the substance. The conclusions were reached on the basis of the evaluation of the representative uses of acibenzolar-S-methyl as a plant activator on pome fruit, tomato and tobacco. The reliable endpoints concluded as being appropriate for use in regulatory risk assessment, derived from the available studies and literature in the dossier peer reviewed, are presented. Assessments not finalised together with the missing information identified as being required by the regulatory framework are listed.

Proof of Concept for Shoot Blight and Fire Blight Canker Management with Postinfection Spray Applications of Prohexadione-Calcium and Acibenzolar-S-Methyl in Apple.

To reduce the severity of shoot blight and prevent the resulting development of cankers on perennial apple wood, we evaluated eight fire blight postinfection spray programs of prohexadione-calcium (PCA) alone or with acibenzolar-S-methyl (ASM) over 2 years. On mature trees of cultivar Royal Court, a single application of the high PCA rate (247 mg/liter) at 2 to 3 days after inoculation resulted in 89.5 and 69.5% reduction of shoot blight severity after inoculation. Two applications of PCA 247 mg/liter 12 or 14 days apart, with the first one applied 2 to 3 days after inoculation, resulted in 78.8 and 74.5% reduction of shoot blight severity in both years. A 100% control of canker incidence on perennial wood from infected shoots in both years was achieved with a single application of PCA (247 mg/liter) applied at 2 or 3 days after the inoculation, and three applications of PCA (125 mg/liter) + ASM (25 mg/liter) 12 to 16 days apart reduced canker incidence by 83.5 and 69% in the 2 years. The other programs with lower PCA rates and frequencies of application reduced shoot blight severity 50.8 and 51.8% (PCA) and 62.6 to 72% and 59.3% (PCA + ASM) over 2 years, respectively. Reduction of canker incidence on wood by the other programs was 66.5% and 69 to 90.4% in the two years, respectively. As fire blight cankers lead to death of dwarf apple trees and serve as primary sources of inoculum, our effective PCA and PCA + ASM programs could serve as viable postinfection management options. These treatments can reduce or prevent canker development and thus significantly abate tree losses in high-density apple orchards after fire blight epidemics occur.

In vitro inhibition of shikimate hydroxycinnamoyltransferase by acibenzolar acid, the first metabolite of the plant defence inducer acibenzolar-S-methyl.

Acibenzolar acid, the first metabolite formed in planta from the defence inducer acibenzolar-S-methyl (ASM), has been shown to be an inhibitor of the enzyme shikimate hydroxycinnamoyltransferase (HST), extracted from grapevine or tobacco cell suspension cultures. Using a purified recombinant Arabidopsis thaliana HST, the inhibition was found to be competitive, acibenzolar acid binding reversibly to the shikimate binding site of the HST:p-coumaroyl-CoA complex, with a Ki value of 250 µM. The other hydroxycinnamoyltransferases tested in the course of this study, using either hydroxypalmitic acid, putrescine, tyramine, or quinic acid as acyl acceptors were not, or only slightly, inhibited by acibenzolar acid. To understand the specificity of the interaction of acibenzolar acid with HST, we analyzed the structure-activity relationship of a series of benzoic or acibenzolar acid analogues, tested either as AtHST substrates or as inhibitors. This analysis confirmed previously published data on the substrate flexibility of HST and demonstrated that both the carboxyl group and the thiadiazole moiety of acibenzolar acid are playing an important role in the interaction with the shikimate binding site. Acibenzolar acid, which cannot form an ester bond with p-coumaric acid, was however a less potent inhibitor than protocatechuic or 3-hydroxybenzoic acids, which are used as acyl acceptors by HST. Our results show that the interaction of acibenzolar acid with HST, which is probably directly linked to the substrate promiscuity of HST, is unlikely to play a direct role in the defence-inducing properties of ASM in plants.

Modification of the existing maximum residue levels for acibenzolar-S-methyl in beans with pods and peas with pods.

In accordance with Article 6 of Regulation (EC) No 396/2005, the applicant Syngenta France SAS submitted a request to the competent national authority in France to amend the residue definition for risk assessment and to modify the existing maximum residue levels (MRLs) for the active substance acibenzolar-S-methyl in beans (with pods) and peas (with pods). The toxicological data submitted in support of this MRL application were found to be sufficient to conclude that the plant metabolite 4-OH acibenzolar acid is not genotoxic in vitro, however lacking investigation of the general toxicity, a definitive conclusion cannot be derived whether 4-OH acibenzolar acid (free and conjugated) should be included or excluded from the residue definition for risk assessment for all commodities belonging to the groups of leafy crops and pulses/oilseeds. The data submitted in support of the request were found to be sufficient to derive MRL proposals on the basis of the current residue definition (sum of acibenzolar-S-methyl and acibenzolar acid (free and conjugated), expressed as acibenzolar-S-methyl) for the intended uses on lettuces and salad plants, beans (with pods) and peas (with pods). The available residue trials are sufficient to derive MRL proposals of 1 mg/kg for beans with pods and peas with pods. The submitted information did not provide evidence that the existing MRL has to be changed for the intended uses on lettuces and other salad plants. For herbs and edible flowers, data gaps were identified which precluded the derivation of MRL proposals. The consumer risk assessment for the current residue definition did not identify an unacceptable risk for consumers. The indicative exposure calculations performed for the metabolite 4-OH acibenzolar indicate that exposure to this metabolite is lower than the exposure to acibenzolar-S-methyl and acibenzolar acid (free and conjugated). However, considering the data gaps identified, a definitive conclusion cannot be derived by EFSA on the risk for consumers related to the intake of residues of 4-OH acibenzolar acid. The reliable end points appropriate for use in regulatory risk assessment are presented.

Roles of calcium-dependent protein kinases mediated reactive oxygen species homeostasis in inducing resistance of apples by acibenzolar-S-methyl.

This study was carried out to investigate the effect of acibenzolar-S-methyl (ASM) and ethylenebis (oxyethylenenitrilo) tetraacetic acid (EGTA) treatments on calcium-dependent protein kinases (CDPKs) and reactive oxygen species (ROS) metabolism in apples. Postharvest ASM treatment increased H2O2 content, reduced glutathione and ascorbic acid contents, and NADPH oxidase, peroxidase, ascorbate peroxidase, superoxide dismutase and glutathione reductase activities and retarded catalase activity and MdCAT expression in apples. ASM treatment enhanced MdSOD, MdPOD, MdAPX, MdGR, MdCDPK1, MdCDPK4, MdCDPK5, MdCDPK7, and MdCDPK21 expressions in apples. However, EGTA + ASM treatments suppressed H2O2, glutathione and ascorbic acid contents, NADPH oxidase, peroxidase, superoxide dismutase, ascorbate peroxidase and glutathione reductase activities. EGTA + ASM treatments suppressed the selected genes expressions in ROS metabolism and CDPKs, but up-regulated MdCAT expression in apples. These findings suggest that CDPKs play a vital role in regulating ROS metabolism and involve in inducing resistance in apples by ASM.

Alternative products to control powdery mildew in soybeans culture in field

The occurrence of powdery mildew (Microsphaera diffusa) in soybean (Glycine max L.) has increased in the last harvests. In order to study the efficiency of powdery mildew control due to the application of alternative products and conventional fungicide, trials were conducted in Ponta Grossa, PR, Brazil, during the 2013/2014 and 2014/2015 growing seasons. The design used was randomized blocks with four replications. The treatments for the experiments were: 1 - control; 2 - acibenzolar-S-methyl (Bion 500 WG®); 3 - calcium (Max Fruit®); 4 - Micronutrients: copper, manganese and zinc (Wert Plus®); 5 - Micronutrients: manganese, zinc and molybdenum (V6®); 6 - NK fertilizer (Hight Roots®); 7 - Ascophyllum nodosum (Acadian®) and 8 - fungicide (azoxystrobin + cyproconazole) (Priori XTRA®) with the addition of the adjuvant. Four applications of alternative products (phenological stages V3, V6, R1 and R5.1) and two of fungicide (phenological stages R1 and R5.1) were carried out. The parameters evaluated were powdery mildew severity and productivity. The severity data made it possible to calculate the area under the disease progress curve (AUDPG). Alternative products didn't reduce powdery mildew in the two harvests. The conventional fungicide treatment was the only one that controlled powdery mildew and didn't reduce the productivity in both experiments.