RESUMO
Phytocytokines are endogenous danger peptides that are actively released after a pest or pathogen attack, triggering an amplification of plant immune responses. Here, we found that Systemin, a peptide from tomato, has a substantial impact at the molecular level in Arabidopsis plants that leads to induced resistance against Plectosphaerella cucumerina. Using transcriptional and metabolomics approaches, and loss-of-function mutants to analyse the molecular mechanisms underlying induced resistance against the necrotroph, we decipher the enhanced molecular responses in Systemin-treated plants following infection. Some protein complexes involved in the response to other damage signals, including the BAK1-BIK1 protein complex and heterotrimeric G proteins, as well as MPK activation, were among the early signalling events triggered by Systemin in Arabidopsis upon infection. Non-targeted analysis of the late responses underlying Systemin-Induced Resistance1 (Sys-IR) showed that phenolic and indolic compounds were the most representative groups in the Systemin metabolic fingerprint. Lack of flavonoids resulted in the impairment of Sys-IR. On the other hand, some indolic compounds showed a priming profile and were also essential for functional Sys-IR. Evidence presented here shows that plants can sense heterologous peptides from other species as danger signals driving the participation of common protein cascades activated in the PTI and promoting enhanced resistance against necrotrophic fungus.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , Solanum lycopersicum , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Ascomicetos , Peptídeos/metabolismo , Doenças das Plantas/microbiologia , Proteínas Serina-Treonina Quinases , Triptofano/metabolismoRESUMO
Indole-3-carboxylic acid (I3CA) is an indolic compound that induces resistance in Arabidopsis adult plants against the necrotrophic fungus Plectosphaerella cucumerina through primed callose accumulation. In this study, we confirm the relevance of ATL31 and SYP121 genes involved in vesicular trafficking in I3CA priming of defenses and we discard camalexin as a mediator of I3CA-induced resistance (IR) in adult plants. In addition, we observed that an intact I3CA biosynthetic pathway is necessary for I3CA-IR functionality.
Assuntos
Ascomicetos/fisiologia , Glucanos/metabolismo , Indóis/farmacologia , Arabidopsis/efeitos dos fármacos , Arabidopsis/imunologia , Arabidopsis/microbiologia , Ascomicetos/efeitos dos fármacos , Vias Biossintéticas/efeitos dos fármacos , Resistência à Doença/efeitos dos fármacos , Resistência à Doença/imunologia , Indóis/metabolismo , Doenças das Plantas/microbiologia , Tiazóis/metabolismoRESUMO
In low nutritive environments, the uptake of N by arbuscular mycorrhizal (AM) fungi may confer competitive advantages for the host. The present study aims to understand how mycorrhizal tomato plants perceive and then prepare for an N depletion in the root environment. Plants colonized by Rhizophagus irregularis displayed improved responses to a lack of N than nonmycorrhizal (NM) plants. These responses were accomplished by a complex metabolic and transcriptional rearrangement that mostly affected the gibberellic acid and jasmonic acid pathways involving DELLA and JAZ1 genes, which were responsive to changes in the C/N imbalance of the plant. N starved mycorrhizal plants showed lower C/N equilibrium in the shoots than starved NM plants and concomitantly a downregulation of the JAZ1 repressor and the increased expression of the DELLA gene, which translated into a more active oxylipin pathway in mycorrhizal plants. In addition, the results support a priorization in AM plants of stress responses over growth. Therefore, these plants were better prepared for an expected stress. Furthermore, most metabolites that were severely reduced in NM plants following the N depletion remained unaltered in starved AM plants compared with those normally fertilized, suggesting that the symbiosis buffered the stress, improving plant development in a stressed environment.
Assuntos
Micorrizas/metabolismo , Nitrogênio/metabolismo , Raízes de Plantas/microbiologia , Solanum lycopersicum/microbiologia , Clorofila/metabolismo , Ciclopentanos/metabolismo , Perfilação da Expressão Gênica , Giberelinas/metabolismo , Solanum lycopersicum/crescimento & desenvolvimento , Solanum lycopersicum/metabolismo , Solanum lycopersicum/fisiologia , Redes e Vias Metabólicas , Nitrogênio/análise , Oxilipinas/metabolismo , Fotossíntese , Raízes de Plantas/metabolismo , Raízes de Plantas/fisiologia , Reação em Cadeia da Polimerase em Tempo Real , Solo/químicaRESUMO
Being sessile organisms, plants must respond to various challenges in the environment. The priming process consists of three clear stages. The first stage includes all the cellular changes in the absence of the challenge so-called pre-challenge priming stage. These changes are expected to be rather subtle, affecting the preparation of the plant to properly manage subsequent responses to pathogens with no major fitness costs. Most of the research that has been conducted at this stage has been dedicated to the study of changes in gene expression and protein phosphorylation. However, the metabolic changes that occur during the pre-challenge priming stage are poorly understood. The second stage affects the early to late stages of the defence response, which occurs after the interaction with a pathogen has been established. Most studies involving priming are dedicated to the molecular events that take place during this stage. Most studies have shown that defence priming is strongly hormonally regulated; however, there is also evidence of the involvement of phenolic derivative compounds and many other secondary metabolites, leading to stronger and faster plant responses. The third priming phase ranges from long lasting defence priming to trans-generational acquired resistance. Long-term metabolic transitions, that occur in the offspring of primed plants, remain to be elucidated. Here we review existing information in the literature that relates to the metabolic changes that occur during all three defence priming stages and highlight the metabolic transitions that are associated with the stimulation of priming and the characteristics of the pathogens whenever possible.
Assuntos
Meio Ambiente , Fenômenos Fisiológicos Vegetais , Plantas/genética , Aminoácidos/metabolismo , Metabolômica , Plantas/metabolismoRESUMO
Arbuscular mycorrhizal (AM) symbioses are mutualistic associations between soil fungi and most vascular plants. Modulation of the hormonal and transcriptional profiles, including changes related to defense signalling, has been reported in many host plants during AM symbioses. These changes have been often related to the improved stress tolerance common in mycorrhizal plants. However, results on the alterations in phytohormones content and their role on the symbiosis are controversial. Here, an integrative analysis of the response of phylogenetically diverse plants (i.e., tomato, soybean, and maize) to two mycorrhizal fungi -Funneliformis mosseae and Rhizophagus irregularis- was performed. The analysis of the defense-related hormones salicylic acid, abscisic acid, and jasmonates, and the expression of marker genes of the pathways they regulate, revealed significant changes in the roots of mycorrhizal plants. These changes depended on both the plant and the AM fungus (AMF) involved. However, general trends can be identified: roots associated with the most effective colonizer R. irregularis showed fewer changes in these defense-related traits, while the colonization by F. mosseae led to significant modifications in all plants tested. The up-regulation of the jasmonate pathway by F. mosseae was found to be highly conserved among the different plant species, suggesting an important role of jasmonates during this AM interaction. Our study evidences a strong influence of the AMF genotype on the modulation of host defense signalling, and offers hints on the role of these changes in the symbiosis.
Assuntos
Glomeromycota/fisiologia , Glycine max/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Solanum lycopersicum/metabolismo , Zea mays/metabolismo , Ácido Abscísico/análise , Cromatografia Líquida de Alta Pressão , Ciclopentanos/análise , Genótipo , Glomeromycota/genética , Micorrizas/metabolismo , Oxilipinas/análise , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/química , Raízes de Plantas/metabolismo , Ácido Salicílico/análise , Simbiose , Espectrometria de Massas em Tandem , Regulação para CimaRESUMO
Plant resistance against the necrotrophic pathogen Plectosphaerella cucumerina is mediated by a combination of several hormonal-controlled signalling pathways. The priming agent ß-aminobutyric acid (BABA) is able to induce effective resistance against this pathogen by stimulating callose-rich cell wall depositions. In the present research it is demonstrated that BABA-Induced Resistance (BABA-IR) against P. cucumerina in Arabidopsis has additional components such as the induction of defences mediated by indolic derivatives. Chromatographic approach for the detection and characterization of metabolites enhanced by BABA compared with water-treated plants only when the challenge is present has been developed. The metabolites matching this criteria are considered to be primed by BABA. The analytic procedure is based on the combination of liquid chromatography (LC) with a triple quadrupole (TQD) detector in a precursor ion scanning mode. Using this analytical system a signal in negative electro-spray mode of 160 m/z is primed by BABA in infected plants. A subsequent exact mass analysis in a quadrupole time-of-flight mass spectrometer demonstrated that this ion was the indole-derivative metabolite indole-3-carboxylic acid (I3CA). The identity of indole-3-carboxilic acid was definitively confirmed by comparing its retention time and fragmentation spectra with a commercial standard. Quantification of I3CA in primed plants showed that this indolic metabolite is specifically primed by BABA upon P. cucumerina infection, while other indolic compounds such as IAA and camalexin are not. Taking together these observations with the known role of callose in priming against this pathogen, suggests that priming is not a single mechanism but rather a multicomponent defence.
Assuntos
Aminobutiratos/farmacologia , Arabidopsis/metabolismo , Resistência à Doença , Fungos , Indóis/metabolismo , Doenças das Plantas/microbiologia , Imunidade Vegetal , Arabidopsis/efeitos dos fármacos , Arabidopsis/microbiologia , Cromatografia Líquida/métodos , Glucanos/metabolismo , Espectrometria de Massas/métodos , Transdução de SinaisRESUMO
The effects of exogenous application of a chemical mixture consisting of adipic acid monoethyl ester, furfurylamine, and 1,2,3,4-tetra-O-acetyl-beta-D-glucopyranose (FGA) on various metabolic pathways and the plant-fungus interaction have been studied in Solanaceae plants. Tomato and pepper plants were sprayed with the FGA mixture, and different biochemical parameters such as gas exchange, chlorophyll concentration, protein, cell wall sugar and phenolics contents, and peroxidase and phenylalanine ammonia lyase (PAL) activities were measured. FGA-treated plants showed, in general, an increase in cell wall sugar content and decreases in the chlorophyll degrading rate and the peroxidase activity. These results suggest that FGA (a possible synthetic regulator) could act as a retardant--antisenescence agent in Solanaceae plants. The FGA mixture increased the PAL activity and promoted an overall rise in the concentration of flavonoids and phenolic compounds. Therefore, FGA induced the synthesis of compounds that could give protection to plants against pathogens or insects. To further verify this putative protection, several fungi were inoculated in intact plants. Exogenous FGA applications on intact plants delayed fungus-provoked lesion development. In addition, data also showed that applications of 1,2,3,4-tetra-O-acetyl-beta-D-glucopyranose inhibited fungal growth in vitro. These results confirm that FGA can activate protective mechanisms in plants upon contact with invaders such as fungi.
Assuntos
Fungicidas Industriais/farmacologia , Reguladores de Crescimento de Plantas/farmacologia , Solanaceae/efeitos dos fármacos , Solanaceae/fisiologia , Adipatos , Ésteres/farmacologia , Furanos/farmacologia , Glucosídeos/farmacologia , Fatores de TempoRESUMO
Three-month old citrange Troyer (hybrid of Citrus sinensis x Poncirus trifoliata) seedlings were grown hydroponically and, after a period of NO(3)(-) starvation, plants were transferred to solutions enriched with K(15)NO(3) (96% atoms 15N excess) to measure 15NO(3)(-) uptake rates as a function of external 15NO(3)(-) concentrations. Two different NO(3)(-) uptake systems were found. Between 1 and 50 mM 15NO(3)(-) in the uptake solution medium, the uptake rate increased linearly due to the low affinity transport system (LATS). Nitrate reductase activity showed the same response to external [NO(3)(-)], and also appears to be regulated by the rate of nitrate uptake. Nitrate pre-treatments had a represive effect on NO(3)(-) uptake rate measured at 5 or 30 mM external [15NO(3)(-)]. The extent of the inhibition depended on the [NO(3)(-)] during the pre-treatment and in the uptake solution. These results suggest that the LATS of Citrus seedlings is under feedback control by the N status of the plant. Accordingly, addition of amino acids (Glu, Asp, Asn, Gln) to the uptake solution resulted in a decrease in 15NO(3)(-) uptake rate. However, the inactivation of nitrate reductase activity after treatment of the seedlings with either 100 or 500 µM WO(4)(2-) did not affect the activity of the LATS. Metabolic uncouplers, 2,4-DNP and KCN, reduced the uptake rate by 43.3% and 41.4% respectively at 5mM external [15NO(3)(-)]. However, these compounds had little effect when 15NO(3)(-) uptake was assayed at 30 mM external concentration. The ATPase inhibitors DCCD and DES reduced 15NO(3)(-) uptake by 68.8%-35.6%, at both external [15NO(3)(-)]. Nitrate uptake by the LATS declined with the increase of the solution pH beyond pH 4. The data presented are discussed in the context of the kinetics, energy dependence and regulation of NO(3)(-) uptake.