acibenzolar-S-methyl Research Articles

Mitogen-Activated Protein Kinase Kinase OsMEK2 Positively Regulates Ca2+ Influx and Ferroptotic Cell Death during Rice Immune Responses.

Mitogen-activated protein (MAP) kinase (MAPK) signaling pathway is important in plant immune responses, involved in iron- and reactive oxygen species (ROS)-dependent ferroptotic cell death mediated by Ca2+. High Ca2+ influx triggered iron-dependent ROS accumulation, lipid peroxidation, and subsequent hypersensitive response (HR) cell death in rice (Oryza sativa). Apoplastic Ca2+ chelation by EGTA during avirulent Magnaporthe oryzae infection altered Ca2+, ROS, and Fe2+ accumulation, increasing rice susceptibility to infection. By contrast, acibenzolar-S-methyl (ASM), a plant defense activator, significantly enhanced Ca2+ influx, and H2O2 accumulation, triggering rice ferroptotic cell death during virulent Magnaporthe oryzae infection. Here, we report a novel role of the MAPK signaling pathway in regulating cytoplasmic Ca2+ increase during ferroptotic cell death in rice immunity, using the ΔOsmek2 knockout mutant rice. The knockout of rice OsMEK2 impaired the ROS accumulation, lipid peroxidation, and iron accumulation during avirulent M. oryzae infection. This study has shown that OsMEK2 could positively regulate iron- and ROS-dependent ferroptotic cell death in rice by modulating the expression of OsNADP-ME, OsRBOHB, OsPLC, and OsCNGC. This modulation indicates a possible mechanism for how OsMEK2 participates in Ca2+ regulation in rice ferroptotic cell death, suggesting its broader role in plant immune responses in response to M. oryzae infection.

Fungicides, bio-controllers and resistance inducers affect bioactive compounds and oxidative metabolism in bell pepper plants inoculated with Phytophthora capsici

ABSTRACT The rational use of chemical fungicides in chilli pepper (Capsicum annuum L.) for controlling Phytophthora capsici Leo. should include molecules that promote induced systemic resistance in the plant. The study objective was to assess how the application of fungicides, bio-controllers and resistance inducers affects bioactive compounds production and oxidative metabolism in pepper plants when inoculated with P. capsici zoospores. Statistical analyses show significant differences in total flavonoids, total phenols and salicylic acid concentrations in plants treated with Trichoderma harzianum, Acibenzolar-S-Methyl (ASM) and phosphonic acid. In addition, the application of ASM and Bacillus subtilis showed the highest values of antioxidant capacity. On the other hand, ASM application favoured increases in superoxide dismutase and phenyl ammonium lyase activity with values of 4.42 ± 0.016 (units · min−1 · g−1) and 3.90 ± 0.22 U · g−1, respectively. However, hydrogen peroxide production was similar to that of phosphonic acid treatments. Similarly, plants treated with B. subtilis and Metalaxyl-M showed increases in peroxidase enzyme activity with values ranging from 2.86 ± 0.05 to 4.52 ± 0.02 (μmol · [mg protein]−1 · min−1). Likewise, phosphonic acid-treated plants exhibited high catalase enzyme activity with values ranging from 2.38 ± 0.032 μmol H2O2 · min−1 · g−1 to 4.30 ± 0.024 μmol H2O2 · min−1 · g−1. The complementary use of products with biological and resistance-inducing functions could help to mitigate the environmental impacts that occur due to the use of chemical fungicides for controlling P. capsici on chilli pepper.

Acibenzolar-S-methyl promotes wound healing of harvested sweet potatoes (Ipomoea batatas) by regulation of reactive oxygen species metabolism and phenylpropanoid pathway.

Rapid wound healing is crucial in protecting sweet potatoes (Ipomoea batatas ) against infection, water loss and quality deterioration during storage. The current study investigated how acibenzolar-S-methyl (ASM) treatment influenced wound healing in harvested sweet potatoes by investigating the underlying mechanism. It was found that ASM treatment of wounded sweet potatoes induced a significant accumulation of lignin at the wound sites, which effectively suppressed weight loss. After 4days of healing, the lignin content of ASM-treated sweet potatoes was 41.8% higher than that of untreated ones, and the weight loss rate was 20.4% lower. Moreover, ASM treatment increased the ability of sweet potatoes to defend against wounding stress through enhancing processes such as increased production of reactive oxygen species (ROS), activation of enzymes involved in the ROS metabolism (peroxidase, superoxide dismutase and catalase) and phenylpropanoid pathway (phenylalanine ammonia lyase, cinnamate-4-hydroxylase, 4-coumarate-CoA ligase and cinnamyl alcohol dehydrogenase), and intensive synthesis of phenolics and flavonoids. These results suggest that treating harvested sweet potatoes with ASM promotes wound healing through the activation of the ROS metabolism and phenylpropanoid pathway.

Cytoplasmic Ca2+ influx mediates iron- and reactive oxygen species-dependent ferroptotic cell death in rice immunity.

Iron- and reactive oxygen species (ROS)-dependent ferroptosis occurs in plant cells. Ca2+ acts as a conserved key mediator to control plant immune responses. Here, we report a novel role of cytoplasmic Ca2+ influx regulating ferroptotic cell death in rice immunity using pharmacological approaches. High Ca2+ influx triggered iron-dependent ROS accumulation, lipid peroxidation, and subsequent hypersensitive response (HR) cell death in rice (Oryza sativa). During Magnaporthe oryzae infection, 14 different Ca2+ influx regulators altered Ca2+, ROS and Fe2+ accumulation, glutathione reductase (GR) expression, glutathione (GSH) depletion and lipid peroxidation, leading to ferroptotic cell death in rice. High Ca2+ levels inhibited the reduction of glutathione isulphide (GSSG) to GSH in vitro. Ca2+ chelation by ethylene glycol-bis (2-aminoethylether)-N, N, N', N'-tetra-acetic acid (EGTA) suppressed apoplastic Ca2+ influx in rice leaf sheaths during infection. Blocking apoplastic Ca2+ influx into the cytoplasm by Ca2+ chelation effectively suppressed Ca2+-mediated iron-dependent ROS accumulation and ferroptotic cell death. By contrast, acibenzolar-S-methyl (ASM), a plant defense activator, significantly enhanced Ca2+ influx, as well as ROS and iron accumulation to trigger ferroptotic cell death in rice. The cytoplasmic Ca2+ influx through calcium-permeable cation channels, including the putative resistosomes, could mediate iron- and ROS-dependent ferroptotic cell death under reduced GR expression levels in rice immune responses.

Ecologically Based Management of Pineapple Mealybug Wilt: Controlling Dysmicoccus brevipes Mealybug Populations with Salicylic Acid Analogs and Plant Extracts

Mealybug wilt of pineapple (MWP) is a destructive disease worldwide caused by a parasitic complex that includes Pineapple Mealybug Wilt-associated Viruses (PMWaVs) and mealybugs (Dysmicoccus brevipes), which concurrently act as vectors for these viruses. Reducing the mealybug population is key to managing MWP, which is achieved in intensive production systems through the use of insecticides. SA (salicylic acid), ASM (acibenzolar-S-methyl), BABA (β-aminobutyric acid), and MeSA (methyl salicylate) are key components of systemic acquired resistance (SAR), the defense mechanism of plants against biotrophic agents such as mealybugs. In this study, these compounds were applied either as pure chemicals and/or as a major constituent of plant extracts. Both the Hawaiian hybrid MD-2 and Queen Victoria tissue culture plants, as well as suckers used for vegetative propagation, were treated with these compounds by direct application on the soil of pineapple pots. Subsequently, five mealybugs were released on each plant or each daughter plant in case of a transgenerational experiment; then, after 45 days, the number of mealybugs was counted. Exogenous SA, ASM, and MeSA reduced the population of mealybugs by a minimum of 50% and up to 80%. These SAR-inducing treatments could be an interesting alternative for controlling mealybugs and are already used in other pathosystems. The SAR mechanisms behind this effect are yet to be confirmed by molecular and enzymatic markers. ASM and MeSA are promising treatments for pineapples using tissue culture plants or traditional shoots.

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.

Kiwifruit Resistance to Sclerotinia sclerotiorum and Pseudomonas syringae pv. actinidiae and Defence Induction by Acibenzolar-S-methyl and Methyl Jasmonate Are Cultivar Dependent.

Pathogen susceptibility and defence gene inducibility were compared between the Actinidia arguta cultivar 'Hortgem Tahi' and the two cultivars of A. chinensis 'Hayward' and 'Zesy002'. Plants were treated with acibenzolar-s-methyl (ASM) or methyl jasmonate (MeJA) one week before inoculation with Pseudomonas syringae pv. actinidiae (Psa biovar3) or Sclerotinia sclerotiorum, or secondary induction with chitosan+glucan (Ch-Glu) as a potential pathogen proxy. Defence expression was evaluated by measuring the expression of 18 putative defence genes. 'Hortgem Tahi' was highly susceptible to sclerotinia and very resistant to Psa, whereas 'Zesy002' was highly resistant to both, and 'Hayward' was moderately susceptible to both. Gene expression in 'Hayward' and 'Zesy002' was alike but differed significantly from 'Hortgem Tahi' which had higher basal levels of PR1-i, PR5-i, JIH1, NPR3 and WRKY70 but lower expression of RD22 and PR2-i. Treatment with ASM caused upregulation of NIMIN2, PR1-i, WRKY70, DMR6 and PR5-i in all cultivars and induced resistance to Psa in 'Zesy002' and 'Hayward' but decreased resistance to sclerotinia in 'Zesy002'. MeJA application caused upregulation of LOX2 and downregulation of NIMIN2, DMR6 and PR2-i but did not affect disease susceptibility. The Ch-Glu inducer induced PR-gene families in each cultivar, highlighting its possible effectiveness as an alternative to actual pathogen inoculation. The significance of variations in fundamental and inducible gene expression among the cultivars is explored.

Evaluation of systemic acquired resistance inducers for the management of copper and streptomycin resistant strains of Xanthomonas perforans on tomato

Bacterial spot (Xanthomonas spp.) is one of the most detrimental diseases of tomato (Solanum lycopersicum L.) worldwide. Bacterial spot Xanthomonads are either endemic or spread by contaminated seed or transplant material into tomato production areas. Due to limited disease management options, transplant producers rely on copper- and streptomycin-based bactericides that are often ineffective due to the predominance of resistant Xanthomonas strains. In this study, transplant production facilities located across eight counties in western North Carolina were surveyed for bacterial spot symptoms from 2018 to 2019. In total, 29 strains (n = 6 in 2018 and n = 23 in 2019) were isolated and identified as Xanthomonas perforans. All strains were resistant to copper, while only 24% of the strains were resistant to streptomycin. Therefore, the ability of three systemic acquired resistance (SAR) inducers, methyl salicylate, Bacillus mycoides isolate J, and acibenzolar-S-methyl (ASM), alone or in combination with copper or streptomycin were evaluated as potential alternatives to copper-mancozeb (grower standard) to manage bacterial spot under greenhouse and field conditions. Under greenhouse conditions, applications of ASM at 0.4 g/L alone, or in combination with copper or streptomycin, significantly reduced bacterial spot disease severity when compared to the grower standard and nontreated controls (P < 0.05). However, none of the SAR inducers significantly reduced disease severity or improved yield when compared to copper-mancozeb-ASM under field conditions (P < 0.05). This study highlights the potential of using ASM for the management of bacterial spot of tomato transplant production to reduce bacterial spot introduction into the field.

Effect of Acibenzolar-S-Methyl on severity of Fusarium wilt and fruit yield of two tomato cultivars

Tomato wilt caused by Fusarium oxysporum f.sp. lycopersici (Fol) is one of the limiting factors to tomato production in the whole world because of its negative effect on the fruit yield of tomato. The study investigated the application of Acibenzolar-S-Methyl (ASM), a plant activator that stimulates plants’ defence mechanisms, at different concentrations (0, 25, 35 and 45 mg/L) on two tomato genotypes (Delila and Kerewa) using drenching method. Factorial experiments were laid out in a randomized complete block design and completely randomized design in both field and the screenhouse, respectively. Results showed that the application of ASM significantly reduced (p≤0.05) disease severity. At 9 weeks after transplanting Delila recorded severity scores of 3.00 and 3.33(screenhouse) and 0.33 (field) while Kerewa had severity scores of 1.33 and 1.50 (screenhouse) and 0.33 (field) when ASM was apllied at 35 and 45 mg/L concentrations, respectively. These values were significantly lower (p≤0.05) than those recorded in untreated plots. Delila yielded 151.23 and 151.73 kg/ha (screenhouse), and 754.70 and 795.8 kg/ha (field) while Kerewa yielded 109.60 and 124.03 kg/ha (screenhouse) and 601.10 and 279.8 kg/ha (field) when ASM was apllied at 35 and 45 mg/l concentrations, respectively. Yields in both screenhouse and the field were significantly higher (p≤0.05) than yields in the untreated control plots. The study concluded that application of ASM at 35 and 45 mg/L to tomato could enhance its resistance which would translate to fruit yield increase.

Acibenzolar-S-methyl induces resistance against ambrosia beetle attacks in dogwoods exposed to simulated flood stress.

Xylosandrus spp. ambrosia beetles (Coleoptera: Curculionidae: Scolytinae) are important wood-boring pests of nursery trees weakened by abiotic and biotic stressors. Acibenzolar-S-methyl (ASM), a plant defense elicitor, was tested for inhibiting Xylosandrus spp. tunneling (i.e., attacks) into flood-stressed flowering dogwoods (Cornus florida L. (Cornales: Cornaceae)). Container-grown dogwoods were treated with ASM substrate drench + flooding, ASM foliar spray + flooding, ASM drench + no flooding, ASM foliar + no flooding, no ASM + flooding, or no ASM + no flooding at 3 days before flood stress in a completely randomized design under field conditions. Trees were flooded for 14 days and then drained and watered as needed. Attacks were counted every 2 days for 28 days. Plant tissue samples were collected at 7 and 14 days after flooding to determine ethanol content using solid-phase microextraction-gas chromatography-mass spectrometry. Trees were dissected to determine gallery formation and depth, fungal colonization, and the presence of eggs, larvae, and adults. The highest number of Xylosandrus beetle species attacks were recorded from plants exposed to no ASM + flooding, but attacks were reduced in ASM treated trees (drench or foliar) + flooding. Trees treated with drenches had fewer attacks than foliar sprays. Plants assigned to no flood had the fewest beetle attacks. Moreover, ASM reduced Xylosandrus spp. gallery formation and depth, fungal colonization, and presence of eggs, larvae, and adults. All flooded trees produced ethanol. In conclusion, ASM induced a plant defense response to Xylosandrus spp. tunneling in dogwoods under flood stress conditions.

Economic versus technical efficiency in using ASM combined with fungicides to elicit wheat varieties with different disease susceptibilities

Despite the positive results of using elicitors to induce resistance against plant diseases, some factors have inhibited the popularization of their use in agriculture. There is an energetic cost related to the elicitors' induced response which can cause undesired effects on growth under low-pressure disease conditions. Elicitors can create phytotoxicity and show high variation in their efficiency between different genotypes within the same crop; in addition, the positive results related to the induced resistance may not repeat in field treatments, adding to the possibility that they are not economically viable. Thus, we carried out two experiments to investigate the technical and economic efficiency of acibenzolar-S-methyl (ASM) and its association with fungicides in the control of leaf diseases of susceptible and resistant wheat varieties, and as how it reflects on the photosynthetic and production performance of wheat. This study showed the limitations of incorporating ASM into foliar fungal disease control in economic terms. However, it was evident that ASM effectively induced plant resistance against Leaf Rust and Powdery Mildew in the field and can be considered a sustainable option for wheat cultivation. Even though its association with chemical control was not the best economic strategy the use of ASM is a tool that can be incorporated into wheat cultivation to minimize the emergence of fungicide-resistant pathogens due to the diversification of modes of action employed and reduce the toxic residue deposition to the environment and human health.

Azelaic and hexanoic acids‐inducing resistance in soybean against Phakopsora pachyrhizi infection

AbstractRust in soybean, caused by Phakopsora pachyrhizi, has been controlled using different fungicide molecules. This study pinpoints the use of inducers of resistance as a promising alternative by investigating the hypothesis that reduction in rust symptoms on soybean sprayed with azelaic acid (AzA) and hexanoic acid (HxA) is linked to a fungistatic effect against urediniospore germination of P. pachyrhizi and/or through the potentiation of host defence reactions. A 4 × 2 factorial experiment was designed consisting of plants sprayed with water (control), acibenzolar‐S‐methyl (ASM), AzA or HxA and either non‐inoculated or inoculated with P. pachyrhizi. Both AzA and HxA significantly inhibited urediniospore germination in vitro. The area under disease progress curve significantly decreased for ASM, HxA (5 and 20 mM) and AzA (0.1 and 1 mM), and fewer hyphae of P. pachyrhizi colonized tissues of plants sprayed with ASM, HxA (20 mM) and AzA (1 mM), compared to control. Host defence genes were strongly up‐regulated for infected plants sprayed with AzA, ASM and HxA compared to water. Most of these genes were expressed earlier for infected and HxA‐sprayed plants than for infected and AzA‐sprayed plants and at greater expression levels than infected and ASM‐sprayed plants. The physiological, biochemical and molecular responses obtained with AzA‐ and HxA‐sprayed plants were comparable to ASM, a well‐known inducer of resistance. This study highlights the potential of using AzA and HxA for rust management considering their fungistatic effect against urediniospores and the capacity to provide soybean plants with a more efficient defence against P. pachyrhizi infection.

Chemical composition and antioxidant properties of beetroot parasitized by Meloidogyne javanica and treated with elicitors

Beetroot is rich in nutrients and bioactive compounds, such as, betalains and phenolics. The objective of this work was to analyze the chemical composition and antioxidant properties of beetroot parasitized by Meloidogyne javanica and treated with elicitors. Beet seedlings were inoculated with 1000 eggs and second-stage juveniles of M. javanica and treated with elicitors based on mannan oligosaccharides, citrus biomass, or acibenzolar-S-methyl (ASM). At 60 days after inoculation, plants were evaluated for nematode reproduction factor (RF), proximate composition, betalains, phenolic compounds, and antioxidant activity. Beetroot was found to be susceptible to M. javanica, with RF values ranging from 15.26 to 27.94. ASM and citrus biomass treatments increased ash content by 15–25% in nematode-inoculated plants. There was no treatment effect on moisture or protein content, but uninoculated plants had higher total phenolic content (~35%) than inoculated plants. Nematode infection was found to compromise betalain production but did not impact antioxidant activity. Nematode-inoculated plants showed reduced sucrose content and increased glucose and fructose levels in all treatments. Regardless of the elicitor used, beetroots inoculated M. javanica exhibit deformities that render them unmarketable in fresh form. However, the results indicate that nematode-infected beetroot may be suitable for industrial processing and compound extraction.

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.

Optimizing the application of acibenzolar-S-methyl and copper to control bacterial spot on fresh-market tomato

This study aimed to adjust the number of applications of acibenzolar-S-methyl (ASM) and copper in an integrated program for the control of bacterial spot on fresh-market tomato. Two field trials were carried out during 2018 and 2019. Four sets of number of weekly applications of ASM were evaluated (7, 9, 11, and 13 applications). In addition to these, four other treatments were included: two as a tank mixture compound [13 weekly applications of ASM + biostimulant and nine applications of ASM + copper oxychloride]. Copper oxychloride was applied after ASM treatments in order to complete 13 applications per growing season. Copper oxychloride and untreated plots were used as a control. The variables evaluated were bacterial spot severity, total yield, and yield per classes of fruit according to the Brazilian market standards. Benefit-cost ratio (B/C) was calculated considering the income increase of a given treatment in relation to the untreated control. The disease ratings and yield data were subjected to ANOVA and the means tested for significance by Fisher, Dunnett, and orthogonal contrasts comparison tests. Regression analysis were carried out with the disease ratings to evaluate the ASM number of applications. As the number of ASM applications increased, disease severity decreased, although, the best B/C was obtained with seven applications of ASM, followed by copper applications until the end of the cycle. The tank mixture compound treatments did not result in economic gain increase. Moreover, disease severity was significantly high when ASM was applied in tank mixture with copper.

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

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. 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. 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.

Bioactive copper and Bacillus subtilis for the control and resistance induction against citrus canker in sweet orange [Citrus sinensis (L.) Osbeck] orchard establishment

• The biological product based on Bacillus subtilis lineage QST 731 applied both via drench and foliar sprays reduces the incidence of citrus canker in citrus trees. • The application of Bacillus subtilis lineage QST 731 via drench in combination with foliar sprays of copper hydroxide or bioactive copper increases the efficiency in the control of citrus canker. • Foliar applications of Bacillus subtilis lineage QST 731 reduces the incidence of citrus leafminer injure. • The copper-based product with amino acids, the bioactive copper, increases the expression of the pr2 gene, which encodes the protein β−1,3 glucanase. Resistance is the plant ability to prevent or delay the infection and the development of a pathogen. The objective of this study was to determine the efficiency of treatments with chemical and biological products for the control of citrus canker during the establishment of citrus orchards through resistance induction. The treatments consisted of a combination of foliar and drench applications of products based on Bacillus subtilis (Bs), bioactive copper (BioCu), copper hydroxide (HCu) and acibenzolar-S-methyl (ASM), at the transplanting of the nursery trees to the field and throughout the first year of establishment of the citrus orchard. Evaluations were carried out on mature branches determining the incidence of citrus canker, defoliation, and citrus leafminer (CLM) injures. The vegetative growth of the citrus trees, i.e. tree height and trunk and canopy diameters, were also determined. Under controlled conditions, studies were carried out to determine the activation of the systemic acquired resistance (SAR) by treatments with ASM and Bs applied via drench, and by BioCu and copper sulfate (SCu) applied by spraying on the leaves of the citrus trees. The expression of the PR-2 gene was evaluated by RT-qPCR at 24, 42, and 72 h, and seven days after the application of the treatments. In the field, the ‘IAPAR 73′ sweet orange trees treated via drench with Bs and foliar applications of Bs + HCu, Bs + BioCu and HCu with and without ASM showed reductions in the incidence of citrus canker. HCu treatments reduced defoliation, while foliar treatments with Bs decreased the incidence of CLM injures. The ‘IAPAR 73′ sweet orange trees treated with ASM showed a reduction in the vegetative growth. The BioCu and Bs products were effective for the control of citrus canker in young citrus trees during the orchard establishment. In addition, the BioCu product increased the expression of the PR-2 gene, which encodes the β-1,3 glucanase, a protein associated with the systemic acquired resistance (SAR) in plants.

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.

Potentiation of soybean resistance against Phakopsora pachyrhizi infection using phosphite combined with free amino acids

AbstractConsidering the importance of Asian soybean rust (ASR), caused by Phakopsora pachyrhizi, in the decrease in soybean yield, this study investigated the potential of using phosphite combined with l‐α‐free amino acids (referred to as induced resistance [IR] stimulus hereafter) to boost defence responses of soybean plants against P. pachyrhizi infection. Plants were sprayed with water (control), acibenzolar‐S‐methyl (ASM) or IR stimulus and noninoculated or inoculated with P. pachyrhizi. Urediniospore germination was not affected by the IR stimulus in vitro. Reduced ASR severity, lower malondialdehyde concentration and less colonization of leaf tissues by P. pachyrhizi (lower TEF‐1α expression from 1 to 15 days after inoculation [dai]) occurred for IR stimulus‐sprayed plants. The pattern of gene expression for IR stimulus‐sprayed and infected plants was strikingly similar but sometimes more remarkable than that in ASM‐sprayed and infected plants. Higher production of phenolics and lignin along with stronger up‐regulation of PAL1.3 (5 and 10 dai), PAL2.2 (3 dai), PAL3.1 (1, 3 and 5 dai), ICS1 (5 dai), CHIA1 (1, 5 and 10 dai), CHI1B1 (5 dai), PR‐1A (5 and 10 dai), NR1‐2 (5 and 10 dai) and INR‐2 (5 and 10 dai) for IR stimulus‐sprayed plants increased their resistance against ASR. In addition, IR stimulus‐sprayed and infected plants showed less impairment of the photosynthetic apparatus and maintained high concentrations of chlorophyll a + b and carotenoids. These findings highlight the potential of using this IR stimulus for developing a well‐tuned and effective defensive strategy in soybean plants against P. pachyrhizi infection.