RESUMO
In this study, we used virus-mediated gene silencing technology and found that the HSP17.4 gene-silenced cultivar Sweet Charlie plants were more susceptible to Colletotrichum gloeosporioides than the wild-type Sweet Charlie, and the level of infection was even higher than that of the susceptible cultivar Benihopp. The results of differential quantitative proteomics showed that after infection with the pathogen, the expression of the downstream response genes NPR1, TGA, and PR-1 of the salicylic acid (SA) signalling pathway was fully up-regulated in the wild-type Sweet Charlie, and the expression of the core transcription factor MYC2 of the jasmonic acid (JA) pathway was significantly down-regulated. The expression of the proteins encoded by these genes did not change significantly in the HSP17.4-silenced Sweet Charlie, indicating that the expression of HSP17.4 activated the up-regulation of downstream signals of SA and inhibited the JA signal pathway. The experiments that used SA, methyl jasmonate, and their inhibitors to treat plants provide additional evidence that the antagonism between SA and JA regulates the resistance of strawberry plants to C. gloeosporioides.
Assuntos
Colletotrichum/fisiologia , Resistência à Doença , Fragaria/genética , Proteínas de Choque Térmico/metabolismo , Doenças das Plantas/imunologia , Reguladores de Crescimento de Plantas/metabolismo , Transdução de Sinais , Acetatos/metabolismo , Ciclopentanos/metabolismo , Fragaria/imunologia , Fragaria/microbiologia , Proteínas de Choque Térmico/genética , Oxilipinas/metabolismo , Doenças das Plantas/microbiologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Ácido Salicílico/metabolismoRESUMO
Microbial-root associations are important to help plants cope with abiotic and biotic stressors. Managing these interactions offers an opportunity for improving the efficiency and sustainability of agricultural production. By characterizing the bacterial and archaeal community (via 16S rRNA sequencing) associated with bulk and rhizosphere soil of sixteen strawberry cultivars in two controlled field studies, we explored the relationships between the soil microbiome and plant resistance to two soil-borne fungal pathogens (Verticillium dahliae and Macrophomina phaseolina). Overall, the plants had a distinctive and genotype-dependent rhizosphere microbiome with higher abundances of known beneficial bacteria such as Pseudomonads and Rhizobium. The rhizosphere microbiome played a significant role in the resistance to the two soil-borne pathogens as shown by the differences in microbiome between high and low resistance cultivars. Resistant cultivars were characterized by higher abundances of known biocontrol microorganisms including actinobacteria (Arthrobacter, Nocardioides and Gaiella) and unclassified acidobacteria (Gp6, Gp16 and Gp4), in both pathogen trials. Additionally, cultivars that were resistant to V. dahliae had higher rhizosphere abundances of Burkholderia and cultivars resistant to M. phaseolina had higher abundances of Pseudomonas. The mechanisms involved in these beneficial plant-microbial interactions and their plasticity in different environments should be studied further for the design of low-input disease management strategies.
Assuntos
Ascomicetos/genética , Resistência à Doença , Fragaria/microbiologia , Doenças das Plantas/microbiologia , Acidobacteria/classificação , Acidobacteria/genética , Acidobacteria/crescimento & desenvolvimento , Actinobacteria/classificação , Actinobacteria/genética , Actinobacteria/crescimento & desenvolvimento , Ascomicetos/crescimento & desenvolvimento , Ascomicetos/patogenicidade , Fragaria/imunologia , Metagenoma , Doenças das Plantas/imunologia , Pseudomonas/genética , Pseudomonas/crescimento & desenvolvimento , RNA Ribossômico 16S/genética , Rhizobium/genética , Rhizobium/crescimento & desenvolvimento , Rizosfera , Microbiologia do SoloRESUMO
Strawberry cultivation is associated with high mineral fertilizer doses and extensive use of chemical plant protection products. Based on previous research, we expected that chitin application to peat substrate would increase the nutrient availability and activate the plant systemic defense response, resulting in higher strawberry yields and fewer disease symptoms. We set up two experiments in which the temporal variability and differences in initial nutrient concentrations of the growing media were taken into account. Chitin treatment resulted in the attraction of plant growth-promoting fungi toward the plant root, such as species from genera Mortierella and Umbelopsis. In addition, by the end of the experiments 87 mg of mineral nitrogen (N) per liter of substrate was mineralized, which can be related to the observed increase in plant shoot biomass. This, however, led to nutrient imbalances in plant shoots and fruit; N concentration in the leaves increased over 30%, exceeding the optimal range, while phosphorous (P) and potassium (K) deficiencies occurred, with concentrations lower than 50% of the optimal range. This may explain the decreased fruit yield and disease resistance of the fruit toward Botrytis cinerea. In contrast, chitin caused a clear defense priming effect in the strawberry leaves, with a strong induction of the jasmonic acid response, resulting in fewer foliar disease symptoms. Chitin causes positive effects on shoot growth and foliar disease resistance, but caution needs to be taken for nutrient imbalances leading to negative influences on root growth, fruit production, and disease susceptibility toward B. cinerea.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.
Assuntos
Resistência à Doença , Fragaria , Frutas , Fenômenos Fisiológicos da Nutrição , Botrytis/fisiologia , Quitina/farmacologia , Resistência à Doença/efeitos dos fármacos , Fragaria/efeitos dos fármacos , Fragaria/crescimento & desenvolvimento , Fragaria/imunologia , Fragaria/microbiologia , Frutas/crescimento & desenvolvimento , Frutas/microbiologia , Nitrogênio/metabolismo , Nutrientes/metabolismo , Fósforo/metabolismo , Folhas de Planta/químicaRESUMO
Acremonium strictum Elicitor Subtilisin (AsES) is a fungal elicitor that activates innate immunity, conferring disease resistance in strawberry (Fragaria × ananassa Duch.), Arabidopsis and other plant species. The aim of the present work was to evaluate the involvement of the ethylene (ET) signalling pathway in AsES-mediated immune response in strawberry. Ethylene production and expression of the genes responsible for ET synthesis, perception and response were measured after AsES treatment. ROS (H2 O2 ) accumulation and immunity induced by AsES were studied after ET perception was blocked by 1-methylcyclopropene (1-MCP). Biochemical and molecular results showed that AsES induced a marked increase in local and systemic biosynthesis of ET, both in a biphasic manner. Blocking of ET perception by 1-MCP prior to AsES induction reduced production of ROS (H2 O2 ) and prevented AsES from eliciting defence against fungal pathogens having different lifestyles, such as Botrytis cinerea (necrotrophic) and Colletotrichum acutatum (hemibiotrophic). These findings contribute to elucidate the mode of action of the novel elicitor subtilase, AsES, specifically regarding the role of ET signalling in the activation of plant innate immunity, in addition to the multitude of processes regulated by ET in plants.
Assuntos
Etilenos , Fragaria , Transdução de Sinais , Subtilisina , Etilenos/metabolismo , Fragaria/efeitos dos fármacos , Fragaria/imunologia , Fragaria/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Hypocreales/química , Transdução de Sinais/efeitos dos fármacos , Subtilisina/farmacologiaRESUMO
This study aimed to reveal the effects and possible mechanism of terpinen-4-ol, the main component of tea tree oil (TTO), on the disease resistance of strawberry fruit. When the effects of TTO and its components were compared on the decay development in fruit inoculated with Botrytis cinerea after treatment, strawberry treated with terpinen-4-ol showed the lowest disease incidence (44.4%) after 48 h and also the smallest lesion diameter during the whole storage. This indicates that terpinen-4-ol induces the highest disease resistance in strawberry compared with TTO and other components. Untargeted metabolomic analysis showed that terpinen-4-ol treatment strongly activated phenylpropanoid biosynthesis and flavonoid metabolism pathway by increasing the accumulation of cinnamaldehyde, coniferyl aldehyde, naringenin, taxifolin, quercetin, and quercitrin in fruit at 12 h after treatment. In addition, terpinen-4-ol treatment also caused the accumulation of total phenolics and lignin by enhancing activities and relative gene expression of key enzymes in the phenylpropanoid metabolism pathway. These results suggest that terpinen-4-ol, as the key component of TTO, is the most important contributor to the effectiveness of TTO in improving disease resistance of strawberry fruit through activating the phenylpropanoid metabolism pathway.
Assuntos
Antifúngicos/farmacologia , Fragaria/microbiologia , Fenilpropionatos/imunologia , Doenças das Plantas/imunologia , Óleo de Melaleuca/farmacologia , Terpenos/farmacologia , Botrytis/efeitos dos fármacos , Botrytis/crescimento & desenvolvimento , Resistência à Doença , Fragaria/imunologia , Frutas/imunologia , Frutas/microbiologia , Testes de Sensibilidade Microbiana , Doenças das Plantas/microbiologiaAssuntos
Alérgenos/imunologia , Hipersensibilidade Alimentar/diagnóstico , Fragaria/imunologia , Administração Oral , Criança , Reações Cruzadas , Diagnóstico Diferencial , Feminino , Hipersensibilidade Alimentar/epidemiologia , Humanos , Imunização , Imunoglobulina E/sangue , Masculino , Região do Mediterrâneo/epidemiologia , Profilinas/imunologia , AutorrelatoRESUMO
WRKY genes and jasmonic acid (JA) play a crucial role in plants' responses against biotic and abiotic stress. However, the regulating mechanism of WRKY genes on strawberry fruits' resistance against Botrytis cinerea is largely unknown, and few studies have been performed on their effect on the JA-mediated defense mechanism against B. cinerea. This study explored the effect of FaWRKY25 on the JA-mediated strawberry resistance against B. cinerea. Results showed that the JA content decreased significantly as the fruits matured, whereas the FaWRKY25 expression rose substantially, which led to heightened susceptibility to B. cinerea and in strawberries. External JA treatment significantly increased the JA content in strawberries and reduced the FaWRKY25 expression, thereby enhancing the fruits' resistance against B. cinerea. FaWRKY25 overexpression significantly lowered the fruits' resistance against B. cinerea, whereas FaWRKY25 silencing significantly increased resistance. Moreover, FaWRKY25 overexpression significantly lowered the JA content, whereas FaWRKY25 silencing significantly increased it. FaWRKY25 expression level substantially affects the expression levels of genes related to JA biosynthesis and metabolism, other members of the WRKY family, and defense genes. Accordingly, FaWRKY25 plays a crucial role in regulating strawberries' resistance against B. cinerea and may negatively regulate their JA-mediated resistance mechanism against B. cinerea.
Assuntos
Botrytis/patogenicidade , Ciclopentanos/farmacologia , Fragaria/genética , Interações Hospedeiro-Patógeno/genética , Oxilipinas/farmacologia , Reguladores de Crescimento de Plantas/farmacologia , Proteínas de Plantas/genética , Fatores de Transcrição/genética , Sequência de Aminoácidos , Botrytis/crescimento & desenvolvimento , Ciclopentanos/metabolismo , Resistência à Doença/genética , Fragaria/classificação , Fragaria/imunologia , Fragaria/microbiologia , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Interações Hospedeiro-Patógeno/imunologia , Oxilipinas/metabolismo , Filogenia , Doenças das Plantas/genética , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Reguladores de Crescimento de Plantas/metabolismo , Imunidade Vegetal/genética , Proteínas de Plantas/antagonistas & inibidores , Proteínas de Plantas/imunologia , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Fatores de Transcrição/antagonistas & inibidores , Fatores de Transcrição/imunologiaRESUMO
Although strawberries are highly appreciated fruits, their intake can induce allergic reactions in atopic patients. These reactions can be due to the patient's previous sensitization to the major birch pollen allergen Bet v 1, by which IgE generated in response to Bet v 1 cross-reacts with the structurally related strawberry Fra a 1 protein family. Fra a 1.02 is the most expressed paralog in ripe strawberries and is highly allergenic. To better understand the molecular mechanisms regulating this allergic response, we have determined the three-dimensional structure of Fra a 1.02 and four site-directed mutants that were designed based on their positions in potential epitopes. Fra a 1.02 and mutants conform to the START fold. We show that the cross-reactivity of all the mutant variants to IgE from patients allergic to Bet v 1 was significantly reduced without altering the conserved structural fold, so that they could potentially be used as hypoallergenic Fra a 1 variants for the generation of vaccines against strawberry allergy in atopic patients.
Assuntos
Antígenos de Plantas/química , Antígenos de Plantas/imunologia , Fragaria/imunologia , Proteínas de Plantas/química , Proteínas de Plantas/imunologia , Antígenos de Plantas/genética , Reações Cruzadas , Hipersensibilidade Alimentar/imunologia , Fragaria/química , Fragaria/genética , Frutas/química , Frutas/imunologia , Humanos , Imunoglobulina E/imunologia , Simulação de Acoplamento Molecular , Proteínas de Plantas/genéticaRESUMO
MAIN CONCLUSION: Colletotrichum acutatum M11 produces a diffusible compound that suppresses the biochemical, physiological, molecular and anatomical events associated with the defence response induced by the plant defence elicitor AsES. The fungal pathogen Colletotrichum acutatum, the causal agent of anthracnose disease, causes important economical losses in strawberry crop worldwide and synthetic agrochemicals are used to control it. In this context, the control of the disease using bioproducts is gaining reputation as an alternative of those toxic and pollutant agrochemicals. However, the success of the strategies using bioproducts can be seriously jeopardized in the presence of biological agents exerting a defence suppression effect. In this report, we show that the response defence induced in plant by the elicitor AsES from the fungus Acremonium strictum can be suppressed by a diffusible compound produced by isolate M11 of C. acutatum. Results revealed that strawberry plants treated with conidia of the isolated M11 or the culture supernatant of the isolate M11 suppress: ROS accumulation (e.g., H2O2, O2·- and NO), cell wall reinforcement (e.g., lignin and callose), and the up-regulation of defence-related genes (e.g., FaPR1, FaCHI23, FaPDF1.2, FaCAT, FaCDPK, FaCML39) induced by the elicitor AsES. Additionally, we show that the defence suppressing effect causes a systemic sensitization of plants. Results presented here highlights the necessity to make an integral study of the microbiome present in soils and plant biosphere before applying defence activation bioproducts to control crop diseases.
Assuntos
Colletotrichum/patogenicidade , Resistência à Doença , Controle Biológico de Vetores , Doenças das Plantas/microbiologia , Parede Celular/metabolismo , Colletotrichum/química , Fragaria/genética , Fragaria/imunologia , Fragaria/microbiologia , Glucanos/metabolismo , Peróxido de Hidrogênio/metabolismo , Lignina/metabolismo , Doenças das Plantas/imunologiaRESUMO
BACKGROUND: The soil-borne vascular pathogen Verticillium dahliae causes severe wilt symptoms in a wide range of plants including strawberry (Fragaria × ananassa). To enhance our understanding of the effects of V. dahliae on the growth and development of F. × ananassa, the expression patterns of 21 PR-10 genes were investigated by qPCR analysis and metabolite changes were determined by LC-MS in in vitro F. × ananassa plants upon pathogen infection. RESULTS: The expression patterns of the 21 isoforms showed a wide range of responses. Four PR-10 genes were highly induced in leaves upon pathogen infection while eight members were significantly up-regulated in roots. A simultaneously induced expression in leaves and roots was detected for five PR-10 genes. Interestingly, two isoforms were expressed upon infection in all three tissues (leaves, roots and stems) while no induction was detected for two other members. Accumulation of antifungal catechin and epicatechin was detected upon pathogen infection in roots and stems at late stages, while caffeic acid and citric acid were observed only in infected roots. Production of abscisic acid, salicylic acid, jasmonic acid (JA), gibberellic acid and indole acetic acid (IAA) was induced in infected leaves and stems at early stages. IAA and JA were the sole hormones to be ascertained in infected roots at late stages. CONCLUSIONS: The induction of several PR-10 genes upon infection of strawberry plants with V. dahliae suggest a role of PR-10 genes in the defense response against this pathogen. Production of phytohormones in the early stages of infection and antifungal metabolites in late stages suppose that they are implicated in this response. The results may possibly improve the control measures of the pathogen.
Assuntos
Fragaria/genética , Interações Hospedeiro-Patógeno , Doenças das Plantas/imunologia , Reguladores de Crescimento de Plantas/metabolismo , Proteínas de Plantas/metabolismo , Verticillium/fisiologia , Ácido Abscísico/metabolismo , Ciclopentanos/metabolismo , Fragaria/imunologia , Fragaria/metabolismo , Fragaria/microbiologia , Oxilipinas/metabolismo , Doenças das Plantas/microbiologia , Folhas de Planta/genética , Folhas de Planta/imunologia , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , Raízes de Plantas/microbiologia , Ácido Salicílico/metabolismoRESUMO
Hormonal modulation plays a central role in triggering various resistant responses to biotic and abiotic stresses in plants. In cultivated strawberry (Fragaria x ananassa), the salicylic acid (SA)-dependent defense pathway has been associated with resistance to Colletotrichum spp. and the other pathogens. To better understand the SA-mediated defense mechanisms in strawberry, we analyzed two strawberry cultivars treated with SA for their resistance to anthracnose and gene expression profiles at 6, 12, 24, and 48 hr post-treatment. Strawberry genes related to SA biosynthesis, perception, and signaling were identified from SA-responsive transcriptomes of the two cultivars, and the induction of 17 candidate genes upon SA treatment was confirmed by qRT-PCR. Given the pivotal role of the non-expressor of pathogenesis-related (NPR) family in controlling the SA-mediated defense signaling pathway, we then analyzed NPR orthologous genes in strawberry. From the expression profile, FaNPRL-1 [ortholog of FvNPRL-1 (gene20070 in F. vesca)] was identified as an NPR-like gene significantly induced after SA treatment in both cultivars. With a conserved BTB/POZ domain, ankyrin repeat domain, and nuclear localization signal, FvNPRL-1 was found phylogenetically closer to NPR3/NPR4 than NPR1 in Arabidopsis. Ectopic expression of FvNPRL-1 in the Arabidopsis thaliana wild type suppressed the SA-mediated PR1 expression and the resistance to Pseudomonas syringae pv. tomato DC3000. Transient expression of FvNPRL-1 fused with green fluorescent protein in Arabidopsis protoplasts showed that SA affected nuclear translocation of FvNPRL-1. FvNPRL-1 likely functions similar to Arabidopsis NPR3/NPR4 as a negative regulator of the SA-mediated defense.
Assuntos
Fragaria/genética , Genes de Plantas/imunologia , Reguladores de Crescimento de Plantas/metabolismo , Imunidade Vegetal/genética , Ácido Salicílico/metabolismo , Arabidopsis , Resistência à Doença/genética , Resistência à Doença/imunologia , Fragaria/imunologia , Fragaria/metabolismo , Filogenia , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , TranscriptomaRESUMO
Anthracnose is a destructive disease of strawberry caused by several Colletotrichum species including C. acutatum, C. fragariae, and C. gloeosporioides. Identification of anthracnose resistant strawberry germplasm has commonly relied on inoculation of whole plants with isolates of these pathogens. In this study, whole plants and detached leaves from 81 germplasm lines were inoculated with a conidial suspension of isolates of C. acutatum, C. fragariae, and C. gloeosporioides, incubated in the dark at 30°C, 100% relative humidity, for 48 h, and assessed for disease severity based on symptoms on inoculated petioles and leaves. The correlation between the disease severity ratings of the whole plants rated 30 days after inoculation and detached leaves rated 5 days after inoculation was determined. Based on leaf symptoms and petiole lesions, the association between the whole plant leaf disease severity rating (DSR) and detached leaf DSR was positive (rp = 0.70), and the association between the whole plant DSR and the detached leaf DSR was also positive (rp = 0.66). Whole plant and detached leaf DSRs were used to assign each germplasm line to a resistance category, and a posthoc Tukey's test showed that the whole plant DSR means and the detached leaf DSR means for each resistance category differed significantly at p < 0.05. This research was used to develop a strawberry detached leaf assay which can reliably and quickly determine the degree of resistance of strawberry germplasm to anthracnose.
Assuntos
Colletotrichum/fisiologia , Resistência à Doença/genética , Fragaria/genética , Doenças das Plantas/imunologia , Fragaria/imunologia , Fragaria/microbiologia , Filogenia , Doenças das Plantas/microbiologia , Folhas de Planta/genética , Folhas de Planta/imunologia , Folhas de Planta/microbiologiaRESUMO
Birch pollen allergic patients show cross-reactivity to vegetables and fruits, including strawberries (Fragaria × ananassa). The objective of this study was to quantify the level of the Fra a 1 protein, a Bet v 1-homologous protein in strawberry fruits by a newly developed ELISA, and determine the effect of genotype, cultivation and food processing on the allergen amount. An indirect competitive ELISA using a specific polyclonal anti-Fra a 1.02 antibody was established and revealed high variability in Fra a 1 levels within 20 different genotypes ranging from 0.67 to 3.97 µg/g fresh weight. Mature fruits of red-, white- and yellow-fruited strawberry cultivars showed similar Fra a 1 concentrations. Compared to fresh strawberries, oven and solar-dried fruits contained slightly lower levels due to thermal treatment during processing. SDS-PAGE and Western blot analysis demonstrated degradation of recombinant Fra a 1.02 after prolonged (>10 min) thermal treatment at 99 °C. In conclusion, the genotype strongly determined the Fra a 1 quantity in strawberries and the color of the mature fruits does not relate to the amount of the PR10-protein. Cultivation conditions (organic and conventional farming) do not affect the Fra a 1 level, and seasonal effects were minor.
Assuntos
Antígenos de Plantas/imunologia , Betula/imunologia , Hipersensibilidade Alimentar/imunologia , Fragaria/imunologia , Pólen/imunologia , Rinite Alérgica Sazonal/imunologia , Antígenos de Plantas/genética , Antígenos de Plantas/metabolismo , Temperatura Baixa , Produção Agrícola , Reações Cruzadas , Dessecação , Ensaio de Imunoadsorção Enzimática , Manipulação de Alimentos/métodos , Fragaria/genética , Fragaria/crescimento & desenvolvimento , Fragaria/metabolismo , Liofilização , Frutas/imunologia , Genótipo , Temperatura Alta , Humanos , Estabilidade Proteica , Estações do AnoRESUMO
Chitin has not been extensively used in agriculture owing to its handling difficulties despite its utilizable functions such as induction of disease resistance and growth promotion in plants. Chitin nanofiber (CNF), which has an elicitor activity to induce plant disease resistance, can be handled like a water-soluble material, because of its high dispersibility. To determine the potential use of CNF in agriculture, the nanofibrillation degree of chitin for elicitor activity and its effect on the disease resistance against pathogens were examined in cabbage and strawberry plants. The similarity in thickness and length of CNF to that of polymeric chitin was sufficient to induce elicitor activity in both plants. Cabbage and strawberry plants, which were grown in a mixture of soil and CNF with optimized specification, challenged with fungal pathogens showed a reduction in the number of spots caused by Alternaria brassicicola and lesion size by Colletotrichum fructicola, respectively. Gene expression analysis revealed that the defense-related genes in cabbage plant grown in CNF-containing soil were significantly upregulated before and after pathogen infection. These results indicate that CNF can systemically induce disease resistance in cabbage and strawberry plants and is a promising natural-based material to control diseases in cultivated plants.
Assuntos
Brassica/imunologia , Quitina/química , Resistência à Doença , Fragaria/imunologia , Nanofibras/química , Doenças das Plantas/imunologia , Animais , Brassica/genética , Brassica/microbiologia , Fragaria/crescimento & desenvolvimento , Fragaria/microbiologia , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Folhas de Planta/microbiologia , Espécies Reativas de Oxigênio/metabolismoRESUMO
The elicitor AsES (Acremonium strictum elicitor subtilisin) is a 34-kDa subtilisin-like protein secreted by the opportunistic fungus Acremonium strictum. AsES activates innate immunity and confers resistance against anthracnose and gray mold diseases in strawberry plants (Fragaria × ananassa Duch.) and the last disease also in Arabidopsis. In the present work, we show that, upon AsES recognition, a cascade of defense responses is activated, including: calcium influx, biphasic oxidative burst (O2â - and H2O2), hypersensitive cell-death response (HR), accumulation of autofluorescent compounds, cell-wall reinforcement with callose and lignin deposition, salicylic acid accumulation, and expression of defense-related genes, such as FaPR1, FaPG1, FaMYB30, FaRBOH-D, FaRBOH-F, FaCHI23, and FaFLS. All these responses occurred following a spatial and temporal program, first induced in infiltrated leaflets (local acquired resistance), spreading out to untreated lateral leaflets, and later, to distal leaves (systemic acquired resistance). After AsES treatment, macro-HR and macro-oxidative bursts were localized in infiltrated leaflets, while micro-HRs and microbursts occurred later in untreated leaves, being confined to a single cell or a cluster of a few epidermal cells that differentiated from the surrounding ones. The differentiated cells initiated a time-dependent series of physiological and anatomical changes, evolving to idioblasts accumulating H2O2 and autofluorescent compounds that blast, delivering its content into surrounding cells. This kind of systemic cell-death process in plants is described for the first time in response to a single elicitor. All data presented in this study suggest that AsES has the potential to activate a wide spectrum of biochemical and molecular defense responses in F. ananassa that may explain the induced protection toward pathogens of opposite lifestyle, like hemibiotrophic and necrotrophic fungi.
Assuntos
Acremonium/fisiologia , Resistência à Doença , Fragaria/imunologia , Fragaria/microbiologia , Proteínas Fúngicas/metabolismo , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Explosão Respiratória , Subtilisina/metabolismo , Morte Celular/genética , Parede Celular/metabolismo , Fluorescência , Fragaria/genética , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Lignina/metabolismo , Necrose , Moléculas com Motivos Associados a Patógenos/metabolismo , Doenças das Plantas/genética , Folhas de Planta/microbiologia , Ácido Salicílico/metabolismoRESUMO
BACKGROUND: Plant immune response is associated with a large-scale transcriptional reprogramming, which is regulated by numerous transcription regulators such as the Elongator complex. Elongator is a multitasking protein complex involved in diverse cellular processes, including histone modification, DNA methylation, and tRNA modification. In recent years, Elongator is emerging as a key regulator of plant immune responses. However, characterization of Elongator's function in plant immunity has been conducted only in the model plant Arabidopsis thaliana. It is thus unclear whether Elongator's role in plant immunity is conserved in higher plants. The objective of this study is to characterize transgenic woodland strawberry (Fragaria vesca L.) overexpressing the Arabidopsis Elongator (AtELP) genes, AtELP3 and AtELP4, and to determine whether F. vesca carries a functional Elongator complex. METHODS: Transgenic F. vesca and Arabidopsis plants were produced via Agrobacterium-mediated genetic transformation and characterized by morphology, PCR, real-time quantitative PCR, and disease resistance test. The Student's t test was used to analyze the data. RESULTS: Overexpression of AtELP3 and AtELP4 in F. vesca impacts plant growth and development and confers enhanced resistance to anthracnose crown rot, powdery mildew, and angular leaf spot, which are caused by the hemibiotrophic fungal pathogen Colletotrichum gloeosporioides, the obligate biotrophic fungal pathogen Podosphaera aphanis, and the hemibiotrophic bacterial pathogen Xanthomonas fragariae, respectively. Moreover, the F. vesca genome encodes all six Elongator subunits by single-copy genes with the exception of FvELP4, which is encoded by two homologous genes, FvELP4-1 and FvELP4-2. We show that FvELP4-1 complemented the Arabidopsis Atelp4/elo1-1 mutant, indicating that FvELP4 is biologically functional. CONCLUSIONS: This is the first report on overexpression of Elongator genes in plants. Our results indicate that the function of Elongator in plant immunity is most likely conserved in F. vesca and suggest that Elongator genes may hold potential for helping mitigate disease severity and reduce the use of fungicides in strawberry industry.
Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Resistência à Doença/genética , Fragaria/genética , Histona Acetiltransferases/genética , Doenças das Plantas/imunologia , Proteínas de Arabidopsis/fisiologia , Fragaria/imunologia , Fragaria/microbiologia , Genes de Plantas , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Plantas Geneticamente ModificadasRESUMO
The strawberry fruit Fra a 1-proteins are homologues of the major birch pollen allergen Bet v 1 and have essential biological functions in pigment formation during fruit ripening. Patients affected by allergy against birch pollen tolerated fruits of a naturally occurring white-fruited F.×ananassa genotype, which showed reduced levels of Fra a 1 proteins along with enzymes of the anthocyanin pigment pathway. We evaluated the cross-reactive allergenic potential of a number of naturally occurring white- and red-fruited strawberry varieties to detect genotypes with low allergenic reactivity, whose fruit might be tolerated by patients with mild allergy. Protein extracts of 51 different strawberry varieties (Fragaria×ananassa, F. vesca, and F. nilgerensis) were screened by Western blot analysis with a polyclonal Fra a 1.02 antibody. Besides, activation of basophils of eight atopic patients allergic to birch pollen were studied using Bet v 1a and different concentrations of 15 selected strawberry protein extracts out of the 51 strawberry genotypes. Median percentages of activated basophils stimulated by extracts from white- and red-fruited genotypes ranged from 36 to 84% and 44 to 76%, respectively indicating that white-fruited strawberry are not per se hypoallergenic. Protein extracts from white-fruited F. vesca cv. Yellow Wonder showed the lowest cross-reactivity but high biological variability. The knowledge about the allergenic potential of different strawberry genotypes may help to improve food safety and can serve as starting point for the development of red-fruited hypoallergenic strawberry cultivars.
Assuntos
Alérgenos , Antígenos de Plantas , Hipersensibilidade Alimentar , Fragaria , Alérgenos/genética , Alérgenos/imunologia , Antígenos de Plantas/genética , Antígenos de Plantas/imunologia , Basófilos/imunologia , Células Cultivadas , Hipersensibilidade Alimentar/imunologia , Hipersensibilidade Alimentar/prevenção & controle , Fragaria/classificação , Fragaria/genética , Fragaria/imunologia , Frutas , Genótipo , HumanosRESUMO
The Chilean strawberry (Fragaria chiloensis) fruit has interesting organoleptic properties, but its postharvest life is affected by gray mold decay caused by Botrytis cinerea. The effect of preharvest applications of methyl jasmonate (MeJA) or chitosan on the molecular defense-related responses and protection against gray mold decay were investigated in Chilean strawberry fruit during postharvest storage. Specifically, we inoculated harvested fruit with B. cinerea spores and studied the expression of genes encoding for the pathogenesis-related (PR) proteins ß-1,3-glucanases (FcBG2-1, FcBG2-2 and FcBG2-3) and chitinases (FcCHI2-2 and FcCHI3-1), and for polygalacturonase inhibiting proteins (FcPGIP1 and FcPGIP2) at 0, 2, 24, 48, and 72 h post inoculation (hpi). Remarkably, MeJA- and chitosan-treated fruit exhibited a lower incidence of B. cinerea infection than the control-treated at 48 and 72 hpi. At the molecular level, both are efficient elicitors for priming in F. chiloensis fruit since we observed an upregulation of the FcBG2-1, FcBG2-3, FcPGIP1, and FcPGIP2 at 0 hpi. Moreover, a chitosan-mediated upregulation of FcPGIPs at early times post inoculation (2-24 hpi) and MeJA upregulated FcBGs (24-72 hpi) and FcPGIP1 at later times could contribute to reduce B. cinerea incidence by differential upregulation of defense genes. We concluded that preharvest applications of MeJA or chitosan had a long-lasting effect on the reduction of B. cinerea incidence during postharvest as well as an enhancer effect on the induction of PR and PGIP gene expression.
Assuntos
Acetatos/farmacologia , Botrytis/fisiologia , Quitosana/farmacologia , Ciclopentanos/farmacologia , Fragaria/genética , Fragaria/microbiologia , Frutas/microbiologia , Oxilipinas/farmacologia , Doenças das Plantas/microbiologia , Regulação para Cima/efeitos dos fármacos , Botrytis/efeitos dos fármacos , Fragaria/efeitos dos fármacos , Fragaria/imunologia , Frutas/efeitos dos fármacos , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genes de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Fatores de TempoRESUMO
Tetranychus urticae Koch (Acari: Tetranychidae) is considered the main pest of strawberry. Several factors can favor its development, among them the genotype susceptibility and cropping system. The aims of this study were to evaluate the agronomic performance of strawberry cultivars under different managements and to identify strawberry cultivars that meet tolerance to T. urticae and high fruit yield. Thirteen cultivars of strawberry ('Albion', 'Aleluia', 'Aromas', 'Camarosa', 'Camino Real', 'Campinas', 'Diamante', 'Dover', 'Festival', 'Seascape', 'Toyonoka', 'Tudla', and 'Ventana') under three managements (open field, low tunnel, and high tunnel) were evaluated. The T. urticae attack to different cultivars was influenced by managements, being low tunnel the one that provided higher infestations in the most evaluated cultivars. 'Camarosa' was the cultivar with the lower incidence of pest and 'Dover' had the higher infestation. The genotype most suitable for growing under different managements is the 'Festival' genotype, since it meets tolerance to T. urticae, high fruit yield, and phenotypic stability.
Assuntos
Fragaria/genética , Melhoramento Vegetal , Imunidade Vegetal/genética , Seleção Genética , Tetranychidae/patogenicidade , Animais , Fragaria/classificação , Fragaria/imunologia , Fragaria/parasitologia , FenótipoRESUMO
The specific forms of disease resistance induced by ß-aminobutyric acid (BABA) and their impacts on sucrose metabolism of postharvest strawberries were determined in the present research. Treatment with 10-500 mmol L(-1) BABA inhibited the Botrytis cinerea infection, possibly directly by suppressing the fungus growth and indirectly by triggering disease resistance. Moreover, BABA-induced resistance against B. cinerea infection in strawberries was associated with either one of two mechanisms, depending upon the concentration used: BABA at concentrations higher than 100 mmol L(-1) directly induced the defense response, including a H2O2 burst, modulation of the expression of PR genes, including FaPR1, FaChi3, Faßglu, and FaPAL, and increased activities of chitinase, ß-1,3-glucanase, and PAL, whereas BABA at 10 mmol L(-1) activated a priming response because the BABA-treated fruits exhibited an increased capacity to express molecular defense only when the fruits were inoculated with B. cinerea. Activation of the priming defense appeared almost as effective against B. cinerea as inducing direct defense. However, the primed strawberries maintained higher activities of SS synthesis and SPS and SPP enzymes) and lower level of SS cleavage during the incubation; these activities contributed to higher sucrose, fructose, and glucose contents, sweetness index, and sensory scores compared to fruits exhibiting the direct defense. Thus, it is plausible that the priming defense, which can be activated by BABA at relatively low concentrations, represents an optimal strategy for combining the advantages of enhanced disease protection and soluble sugar accumulation.