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Sheng Luan - One of the best experts on this subject based on the ideXlab platform.
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Receptor kinase FERONIA regulates flowering time in Arabidopsis.
BMC plant biology, 2020Co-Authors: Long Wang, Tao Yang, Bingqian Wang, Qinlu Lin, Sheng LuanAbstract:The receptor-like kinase FEROINA (FER) plays a crucial role in controlling plant vegetative growth partially by sensing the Rapid Alkalinization Factor (RALF) peptide. However, the role of RALF1-FER in the vegetative-reproductive growth transition remains unknown. Here, we analyze the mechanism through which FER affects the flowering time in Arabidopsis. We found that the FER mRNA levels exhibit an oscillating pattern with a diurnal rhythm and that the clock oscillator CIRCADIAN CLOCK-ASSOCIATED1 (CCA1) up-regulates the expression of FER by associating with its chromatin. In addition, FER expression is regulated by clock genes, and FER also modulates the expression patterns of clock genes. Consistent with its gene expression pattern, FER positively regulates flowering by modulating the transcript accumulation and mRNA alternative splicing of certain flowering-related genes, including FLOWERING LOCUS C (FLC) and its homolog MADS AFFECTING FLOWERING (MAF). However, the RALF1 ligand negatively regulates flowering compared with FER. We found that FER, which is up-regulated by CCA1, controls the flowering time by regulating the transcript accumulation and mRNA alternative splicing (AS) of some important flowering genes, and these findings link FER to the floral transition.
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EBP1 nuclear accumulation negatively feeds back on FERONIA-mediated RALF1 signaling
PLoS biology, 2018Co-Authors: Xuanming Liu, Long Wang, Sirui Zhu, Hongdong Liao, Xiaonan Qiang, Yuan Wang, Sheng LuanAbstract:FERONIA (FER), a plasma membrane receptor-like kinase, is a central regulator of cell growth that integrates environmental and endogenous signals. A peptide ligand Rapid Alkalinization Factor 1 (RALF1) binds to FER and triggers a series of downstream events, including inhibition of Arabidopsis H+-ATPase 2 activity at the cell surface and regulation of gene expression in the nucleus. We report here that, upon RALF1 binding, FER first promotes ErbB3-binding protein 1 (EBP1) mRNA translation and then interacts with and phosphorylates the EBP1 protein, leading to EBP1 accumulation in the nucleus. There, EBP1 associates with the promoters of previously identified RALF1-regulated genes, such as CML38, and regulates gene transcription in response to RALF1 signaling. EBP1 appears to inhibit the RALF1 peptide response, thus forming a transcription–translation feedback loop (TTFL) similar to that found in circadian rhythm control. The plant RALF1-FER-EBP1 axis is reminiscent of animal epidermal growth Factor receptor (EGFR) signaling, in which EGF peptide induces EGFR to interact with and phosphorylate EBP1, promoting EBP1 nuclear accumulation to control cell growth. Thus, we suggest that in response to peptide signals, plant FER and animal EGFR use the conserved key regulator EBP1 to control cell growth in the nucleus.
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feronia receptor kinase at the crossroads of hormone signaling and stress responses
Plant and Cell Physiology, 2017Co-Authors: Hongdong Liao, Xin Zhang, Renjie Tang, Sheng LuanAbstract:Plant receptor-like kinases (RLKs) are involved in nearly all aspects of plant life including growth, development and stress response. Recent studies show that FERONIA (FER), a CrRLK1L subfamily member, is a versatile regulator of cell expansion and serves as a signaling node mediating cross-talk among multiple phytohormones. As a receptor for the RALF (Rapid Alkalinization Factor) peptide ligand, FER triggers a downstream signaling cascade that leads to a Rapid cytoplasmic calcium increase and inhibition of cell elongation in plants. Moreover, FER recruits and activates small G proteins through the guanine nucleotide exchange Factor-Rho-like GTPase (GEF-ROP) network to regulate both auxin and ABA responses that cross-talk with the RALF signaling pathway. One of the downstream processes is NADPH oxidase-dependent ROS (reactive oxygen species) production that modulates cell expansion and responses to both abiotic and biotic stress responses. Intriguingly, some pathogenic fungi produce RALF-like peptides to activate the host FER-mediated pathway and thus increase their virulence and cause plant disease. Studies so far indicate that FER may serve as a central node of the cell signaling network that integrates a number of regulatory pathways targeting cell expansion, energy metabolism and stress responses. This review focuses on recent findings and their implications in the context of FER action as a modulator that is crucial for hormone signaling and stress responses.
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receptor kinase complex transmits ralf peptide signal to inhibit root growth in arabidopsis
Proceedings of the National Academy of Sciences of the United States of America, 2016Co-Authors: Jia Chen, Weijun Chen, Long Wang, Xiaoying Zhao, Jianzhong Lin, Xuanming Liu, Sheng LuanAbstract:A number of hormones work together to control plant cell growth. Rapid Alkalinization Factor 1 (RALF1), a plant-derived small regulatory peptide, inhibits cell elongation through suppression of rhizosphere acidification in plants. Although a receptor-like kinase, FERONIA (FER), has been shown to act as a receptor for RALF1, the signaling mechanism remains unknown. In this study, we identified a receptor-like cytoplasmic kinase (RPM1-induced protein kinase, RIPK), a plasma membrane-associated member of the RLCK-VII subfamily, that is recruited to the receptor complex through interacting with FER in response to RALF1. RALF1 triggers the phosphorylation of both FER and RIPK in a mutually dependent manner. Genetic analysis of the fer-4 and ripk mutants reveals RIPK, as well as FER, to be required for RALF1 response in roots. The RALF1-FER-RIPK interactions may thus represent a mechanism for peptide signaling in plants.
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feronia interacts with abi2 type phosphatases to facilitate signaling cross talk between abscisic acid and ralf peptide in arabidopsis
Proceedings of the National Academy of Sciences of the United States of America, 2016Co-Authors: Jia Chen, Xuanming Liu, Sirui Zhu, Ying Liu, Xianchun Wang, Wenzhi Lan, Pedro L Rodriguez, Liangbi Chen, Sheng LuanAbstract:Receptor-like kinase FERONIA (FER) plays a crucial role in plant response to small molecule hormones [e.g., auxin and abscisic acid (ABA)] and peptide signals [e.g., Rapid Alkalinization Factor (RALF)]. It remains unknown how FER integrates these different signaling events in the control of cell growth and stress responses. Under stress conditions, increased levels of ABA will inhibit cell elongation in the roots. In our previous work, we have shown that FER, through activation of the guanine nucleotide exchange Factor 1 (GEF1)/4/10-Rho of Plant 11 (ROP11) pathway, enhances the activity of the phosphatase ABA Insensitive 2 (ABI2), a negative regulator of ABA signaling, thereby inhibiting ABA response. In this study, we found that both RALF and ABA activated FER by increasing the phosphorylation level of FER. The FER loss-of-function mutant displayed strong hypersensitivity to both ABA and abiotic stresses such as salt and cold conditions, indicating that FER plays a key role in ABA and stress responses. We further showed that ABI2 directly interacted with and dephosphorylated FER, leading to inhibition of FER activity. Several other ABI2-like phosphatases also function in this pathway, and ABA-dependent FER activation required PYRABACTIN RESISTANCE (PYR)/PYR1-LIKE (PYL)/REGULATORY COMPONENTS OF ABA RECEPTORS (RCAR)-A-type protein phosphatase type 2C (PP2CA) modules. Furthermore, suppression of RALF1 gene expression, similar to disruption of the FER gene, rendered plants hypersensitive to ABA. These results formulated a mechanism for ABA activation of FER and for cross-talk between ABA and peptide hormone RALF in the control of plant growth and responses to stress signals.
Daniel S. Moura - One of the best experts on this subject based on the ideXlab platform.
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BAK1 is involved in AtRALF1-induced inhibition of root cell expansion.
PLoS genetics, 2017Co-Authors: Keini Dressano, Paulo H O Ceciliato, Aparecida Leonir Da Silva, Tabata Bergonci, Marcio C. Silva-filho, Juan Carlos Guerrero-abad, Fausto Andres Ortiz-morea, Marco Bürger, Daniel S. MouraAbstract:The Rapid Alkalinization Factor (RALF) peptide negatively regulates cell expansion, and an antagonistic relationship has been demonstrated between AtRALF1, a root-specific RALF isoform in Arabidopsis, and brassinosteroids (BRs). An evaluation of the response of BR signaling mutants to AtRALF1 revealed that BRI1-associated receptor kinase1 (bak1) mutants are insensitive to AtRALF1 root growth inhibition activity. BAK1 was essential for the induction of AtRALF1-responsive genes but showed no effect on the mobilization of Ca2+ and Alkalinization responses. Homozygous plants accumulating AtRALF1 and lacking the BAK1 gene did not exhibit the characteristic semi-dwarf phenotype of AtRALF1-overexpressors. Biochemical evidence indicates that AtRALF1 and BAK1 physically interact with a Kd of 4.6 μM and acridinium-labeled AtRALF1 was used to demonstrate that part of the specific binding of AtRALF1 to intact seedlings and to a microsomal fraction derived from the roots of Arabidopsis plants is BAK1-dependent. Moreover, AtRALF1 induces an increase in BAK1 phosphorylation, suggesting that the binding of AtRALF1 to BAK1 is functional. These findings show that BAK1 contains an additional AtRALF1 binding site, indicating that this protein may be part of a AtRALF1-containing complex as a co-receptor, and it is required for the negative regulation of cell expansion.
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Antagonistic relationship between AtRALF1 and brassinosteroid regulates cell expansion-related genes.
Plant signaling & behavior, 2014Co-Authors: Tabata Bergonci, Marcio C. Silva-filho, Daniel S. MouraAbstract:Rapid Alkalinization Factor (RALF) is a peptide signal that plays a role in plant cell expansion. We have recently proposed that AtRALF1 negatively regulates root cell elongation and lateral root formation by opposing the effects of brassinosteroid (BR). We reported 6 AtRALF1-inducible cell wall-related genes and 2 P450 monooxygenase -encoding genes involved in the BR biosynthetic pathway. The AtRALF1-inducible genes implicated in cell wall remodeling were not downregulated by brassinolide (BL) treatment alone; their induction was only compromised following simultaneous treatment with AtRALF1 and BL. We further examined the cell wall-remodeling gene EXPANSIN A5 (AtEXPA5), which is upregulated by BL and has been shown to positively affect root cell elongation. Herein, we report that AtEXPA5 expression is downregulated by AtRALF1 in a dose-dependent manner in the roots and hypocotyls of Arabidopsis plants. AtEXPA5 is also downregulated in plants that overexpress AtRALF1, and it is upregulated in plants in w...
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Arabidopsis thaliana RALF1 opposes brassinosteroid effects on root cell elongation and lateral root formation
Journal of experimental botany, 2014Co-Authors: Tabata Bergonci, Paulo H O Ceciliato, Marcio C. Silva-filho, Juan Carlos Guerrero-abad, Bianca Ribeiro, Daniel S. MouraAbstract:Rapid Alkalinization Factor (RALF) is a peptide signal that plays a basic role in cell biology and most likely regulates cell expansion. In this study, transgenic Arabidopsis thaliana lines with high and low levels of AtRALF1 transcripts were used to investigate this peptide’s mechanism of action. Overexpression of the root-specific isoform AtRALF1 resulted in reduced cell size. Conversely, AtRALF1 silencing increased root length by increasing the size of root cells. AtRALF1-silenced plants also showed an increase in the number of lateral roots, whereas AtRALF1 overexpression produced the opposite effect. In addition, four AtRALF1-inducible genes were identified: two genes encoding prolinerich proteins (AtPRP1 and AtPRP3), one encoding a hydroxyproline-rich glycoprotein (AtHRPG2), and one encoding a xyloglucan endotransglucosylase (TCH4). These genes were expressed in roots and involved in cell-wall rearrangement, and their induction was concentration dependent. Furthermore, AtRALF1-overexpressing plants were less sensitive to exogenous brassinolide (BL); upon BL treatment, the plants showed no increase in root length and a compromised increase in hypocotyl elongation. In addition, the treatment had no effect on the number of emerged lateral roots. AtRALF1 also induces two brassinosteroid (BR)-downregulated genes involved in the BR biosynthetic pathway: the cytochrome P450 monooxygenases CONSTITUTIVE PHOTOMORPHISM AND DWARFISM (CPD) and DWARF4 (DWF4). Simultaneous treatment with both AtRALF1 and BL caused a reduction in AtRALF1-inducible gene expression levels, suggesting that these signals may compete for components shared by both pathways. Taken together, these results indicate an opposing effect of AtRALF1 and BL, and suggest that RALF’s mechanism of action could be to interfere with the BR signalling pathway.
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SacRALF1, a peptide signal from the grass sugarcane (Saccharum spp.), is potentially involved in the regulation of tissue expansion
Plant Molecular Biology, 2010Co-Authors: Fabiana B. Mingossi, Marcio C. Silva-filho, Juliana L. Matos, Ana Paula Rizzato, Ane H. Medeiros, Maria C. Falco, Daniel S. MouraAbstract:Rapid Alkalinization Factor (RALF) is part of a growing family of small peptides with hormone characteristics in plants. Initially isolated from leaves of tobacco plants, RALF peptides can be found throughout the plant kingdom and they are expressed ubiquitously in plants. We took advantage of the small gene family size of RALF genes in sugarcane and the ordered cellular growth of the grass sugarcane leaves to gain information about the function of RALF peptides in plants. Here we report the isolation of two RALF peptides from leaves of sugarcane plants using the Alkalinization assay. SacRALF1 was the most abundant and, when added to culture media, inhibited growth of microcalli derived from cell suspension cultures at concentrations as low as 0.1 μM. Microcalli exposed to exogenous SacRALF1 for 5 days showed a reduced number of elongated cells. Only four copies of SacRALF genes were found in sugarcane plants. All four SacRALF genes are highly expressed in young and expanding leaves and show a low or undetectable level of expression in expanded leaves. In half-emerged leaf blades, SacRALF transcripts were found at high levels at the basal portion of the leaf and at low levels at the apical portion. Gene expression analyzes localize SacRALF genes in elongation zones of roots and leaves. Mature leaves, which are devoid of expanding cells, do not show considerable expression of SacRALF genes. Our findings are consistent with SacRALF genes playing a role in plant development potentially regulating tissue expansion.
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CHAPTER 7 – RALF Peptides
Handbook of Biologically Active Peptides, 2006Co-Authors: Daniel S. Moura, Gregory Pearce, Clarence A RyanAbstract:RALF (Rapid Alkalinization Factor) peptides are composed of 49 to 52 amino acids and are ubiquitous in plants. When added to suspension-cultured plant cells at low nM concentrations, the peptides cause a Rapid activation of a MAP kinase and the blockage of a membrane-bound ATP-dependent proton pump that results in the Rapid Alkalinization of the culture medium. When added to the growth medium of germinating seeds, RALF causes an immediate arrest of root growth and elongation that can be reversed by transferring the seeds to a medium lacking the peptide. RALF peptides are cleaved from larger precursor proteins that are synthesized through the Golgi and secretory pathway and sequestered in the apoplast. RALF precursor genes are members of gene families that exhibit tissue-specific expression in roots, stems, leaves, and flowers. RALF interacts strongly and reversibly with a cell surface receptor complex composed of 120 kD and 25 kD RALF-binding proteins. In Arabidopsis roots, an endogenous RALF gene is expressed at high levels in slowly dividing cells and at low levels in Rapidly dividing cells, suggesting that RALF peptides may play important tissue-specific roles in regulating cell division and cell expansion during growth and development.
Elena Baraldi - One of the best experts on this subject based on the ideXlab platform.
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genomic structure and transcript analysis of the Rapid Alkalinization Factor ralf gene family during host pathogen crosstalk in fragaria vesca and fragaria x ananassa strawberry
PLOS ONE, 2020Co-Authors: Francesca Negrini, Kevin Ogrady, M Hyvonen, Kevin M Folta, Elena BaraldiAbstract:Rapid Alkalinization Factors (RALFs) are cysteine-rich peptides ubiquitous within plant kingdom. They play multiple roles as hormonal signals in diverse processes, including root elongation, cell growth, pollen tube development, and fertilization. Their involvement in host-pathogen crosstalk as negative regulators of immunity in Arabidopsis has also been recognized. In addition, peptides homologous to RALF are secreted by different fungal pathogens as effectors during early stages of infection. Previous studies have identified nine RALF genes in the diploid strawberry (Fragaria vesca) genome. This work describes the genomic organization of the RALF gene families in commercial octoploid strawberry (Fragaria x ananassa) and the re-annotated genome of F. vesca, and then compares findings with orthologs in Arabidopsis thaliana. We reveal the presence of 15 RALF genes in F. vesca genotype Hawaii 4 and 50 in Fragaria x ananassa cv. Camarosa, showing a non-homogenous localization of genes among the different Fragaria x ananassa subgenomes. Expression analysis of Fragaria x ananassa RALF genes upon infection with Colletotrichum acutatum or Botrytis cinerea showed that FanRALF3-1 was the only fruit RALF gene upregulated after fungal infection. In silico analysis was used to identify distinct pathogen inducible elements upstream of the FanRALF3-1 gene. Agroinfiltration of strawberry fruit with deletion constructs of the FanRALF3-1 promoter identified a 5' region required for FanRALF3-1 expression in fruit, but failed to identify a region responsible for fungal induced expression.
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Rapid Alkalinization Factor ralf gene family genomic structure and transcriptional regulation during host pathogen crosstalk in fragaria vesca and fragaria x ananassa strawberry
bioRxiv, 2019Co-Authors: Francesca Negrini, Kevin Ogrady, M Hyvonen, Kevin M Folta, Elena BaraldiAbstract:Rapid Alkalinization Factor (RALF) are cysteins-rich peptides ubiquitous in plant kingdom. They play multiple roles as hormone signals, starting from root elongation, cell growth, pollen tube development and fertilization. Their involvement in host-pathogen crosstalk as negative regulator of immunity in Arabidopsis has also been recognized. In addition, RALF peptides are secreted by different fungal pathogens as effectors during early stages of infections. Campbell and Turner previously identified nine RALF genes inF. vesca v1 genome. Here, based on the recent release of Fragaria x ananassa genome and F. vesca reannotation, we aimed to characterize the genomic organization of the RALF gene family in both type of strawberry species according to tissue specific expression and homology withArabidopsis. We reveal the presence of 13 RALF genes in F. vesca and 50 in Fragaria x ananassa, showing a non-homogenous localization of genes among the different Fragaria x ananassa subgenomes associated with their different TE element contents and genome remodeling during evolution. Fragaria x ananassa RALF genes expression inducibility upon infection with C. acutatum or B. cinerea was assessed andshowed that, among fruit expressed RALFgenes, FaRALF3-1was the only one upregulated after fungal infection. In silico analysis and motif frequency analysis of the putative regulatory elements upstream of the FaRALF3gene was carried out in order to identify distinct pathogen inducible elements. Agroinfiltration of strawberry fruit with 5’ deletion constructs of theFaRALF3-1promoter identified a region required for FaRALF3 expression in fruit, but did not identify a region responsible for fungal induced expression.
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induced expression of the fragaria ananassa Rapid Alkalinization Factor 33 like gene decreases anthracnose ontogenic resistance of unripe strawberry fruit stages
Molecular Plant Pathology, 2019Co-Authors: Maria Cecilia Merino, Michela Guidarelli, Francesca Negrini, Dario De Biase, Annalisa Pession, Elena BaraldiAbstract:Rapid Alkalinization Factor (RALF) genes encode for ubiquitous small peptides that stimulate apoplastic Alkalinization through interaction with malectin-like receptor kinase. RALF peptides may act as negative regulators of plant immune response, inhibiting the formation of the signal receptor complex for immune activation. Recently RALF homologues were identified in different fungal pathogen genomes contributing to host infection ability. Here, FaRALF-33-like gene expression was evaluated in strawberry fruits inoculated with Colletotrichum acutatum, Botrytis cinerea, or Penicillium expansum after 24 and 48 h post-infection. To investigate the role of FaRALF-33-like in strawberry susceptibility, transient transformation was used to overexpress it in white unripe fruits and silence it in red ripe fruits. Agroinfiltrated fruits were inoculated with C. acutatum and expression, and histological analysis of infection were performed. Silencing of FaRALF-33-like expression in C. acutatum-inoculated red fruits led to a delay in fruit colonization by the fungal pathogen, and infected tissues showed less penetrated infective hyphae than in wild-type fruits. In contrast, C. acutatum-inoculated white unripe fruits overexpressing the FaRALF-33-like gene decreased the ontogenic resistance of these fruits, leading to the appearance of disease symptoms and penetrated subcuticular hyphae, normally absent in white unripe fruits. The different response of transfected strawberry fruits to C. acutatum supports the hypothesis that the FaRALF-33-like gene plays an important role in the susceptibility of fruits to the fungal pathogen C. acutatum.
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Induced expression of the Fragaria × ananassa Rapid Alkalinization Factor‐33‐like gene decreases anthracnose ontogenic resistance of unripe strawberry fruit stages
Molecular plant pathology, 2019Co-Authors: Maria Cecilia Merino, Michela Guidarelli, Francesca Negrini, Dario De Biase, Annalisa Pession, Elena BaraldiAbstract:Rapid Alkalinization Factor (RALF) genes encode for ubiquitous small peptides that stimulate apoplastic Alkalinization through interaction with malectin-like receptor kinase. RALF peptides may act as negative regulators of plant immune response, inhibiting the formation of the signal receptor complex for immune activation. Recently RALF homologues were identified in different fungal pathogen genomes contributing to host infection ability. Here, FaRALF-33-like gene expression was evaluated in strawberry fruits inoculated with Colletotrichum acutatum, Botrytis cinerea, or Penicillium expansum after 24 and 48 h post-infection. To investigate the role of FaRALF-33-like in strawberry susceptibility, transient transformation was used to overexpress it in white unripe fruits and silence it in red ripe fruits. Agroinfiltrated fruits were inoculated with C. acutatum and expression, and histological analysis of infection were performed. Silencing of FaRALF-33-like expression in C. acutatum-inoculated red fruits led to a delay in fruit colonization by the fungal pathogen, and infected tissues showed less penetrated infective hyphae than in wild-type fruits. In contrast, C. acutatum-inoculated white unripe fruits overexpressing the FaRALF-33-like gene decreased the ontogenic resistance of these fruits, leading to the appearance of disease symptoms and penetrated subcuticular hyphae, normally absent in white unripe fruits. The different response of transfected strawberry fruits to C. acutatum supports the hypothesis that the FaRALF-33-like gene plays an important role in the susceptibility of fruits to the fungal pathogen C. acutatum.
Francesca Negrini - One of the best experts on this subject based on the ideXlab platform.
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genomic structure and transcript analysis of the Rapid Alkalinization Factor ralf gene family during host pathogen crosstalk in fragaria vesca and fragaria x ananassa strawberry
PLOS ONE, 2020Co-Authors: Francesca Negrini, Kevin Ogrady, M Hyvonen, Kevin M Folta, Elena BaraldiAbstract:Rapid Alkalinization Factors (RALFs) are cysteine-rich peptides ubiquitous within plant kingdom. They play multiple roles as hormonal signals in diverse processes, including root elongation, cell growth, pollen tube development, and fertilization. Their involvement in host-pathogen crosstalk as negative regulators of immunity in Arabidopsis has also been recognized. In addition, peptides homologous to RALF are secreted by different fungal pathogens as effectors during early stages of infection. Previous studies have identified nine RALF genes in the diploid strawberry (Fragaria vesca) genome. This work describes the genomic organization of the RALF gene families in commercial octoploid strawberry (Fragaria x ananassa) and the re-annotated genome of F. vesca, and then compares findings with orthologs in Arabidopsis thaliana. We reveal the presence of 15 RALF genes in F. vesca genotype Hawaii 4 and 50 in Fragaria x ananassa cv. Camarosa, showing a non-homogenous localization of genes among the different Fragaria x ananassa subgenomes. Expression analysis of Fragaria x ananassa RALF genes upon infection with Colletotrichum acutatum or Botrytis cinerea showed that FanRALF3-1 was the only fruit RALF gene upregulated after fungal infection. In silico analysis was used to identify distinct pathogen inducible elements upstream of the FanRALF3-1 gene. Agroinfiltration of strawberry fruit with deletion constructs of the FanRALF3-1 promoter identified a 5' region required for FanRALF3-1 expression in fruit, but failed to identify a region responsible for fungal induced expression.
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Rapid Alkalinization Factor ralf gene family genomic structure and transcriptional regulation during host pathogen crosstalk in fragaria vesca and fragaria x ananassa strawberry
bioRxiv, 2019Co-Authors: Francesca Negrini, Kevin Ogrady, M Hyvonen, Kevin M Folta, Elena BaraldiAbstract:Rapid Alkalinization Factor (RALF) are cysteins-rich peptides ubiquitous in plant kingdom. They play multiple roles as hormone signals, starting from root elongation, cell growth, pollen tube development and fertilization. Their involvement in host-pathogen crosstalk as negative regulator of immunity in Arabidopsis has also been recognized. In addition, RALF peptides are secreted by different fungal pathogens as effectors during early stages of infections. Campbell and Turner previously identified nine RALF genes inF. vesca v1 genome. Here, based on the recent release of Fragaria x ananassa genome and F. vesca reannotation, we aimed to characterize the genomic organization of the RALF gene family in both type of strawberry species according to tissue specific expression and homology withArabidopsis. We reveal the presence of 13 RALF genes in F. vesca and 50 in Fragaria x ananassa, showing a non-homogenous localization of genes among the different Fragaria x ananassa subgenomes associated with their different TE element contents and genome remodeling during evolution. Fragaria x ananassa RALF genes expression inducibility upon infection with C. acutatum or B. cinerea was assessed andshowed that, among fruit expressed RALFgenes, FaRALF3-1was the only one upregulated after fungal infection. In silico analysis and motif frequency analysis of the putative regulatory elements upstream of the FaRALF3gene was carried out in order to identify distinct pathogen inducible elements. Agroinfiltration of strawberry fruit with 5’ deletion constructs of theFaRALF3-1promoter identified a region required for FaRALF3 expression in fruit, but did not identify a region responsible for fungal induced expression.
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induced expression of the fragaria ananassa Rapid Alkalinization Factor 33 like gene decreases anthracnose ontogenic resistance of unripe strawberry fruit stages
Molecular Plant Pathology, 2019Co-Authors: Maria Cecilia Merino, Michela Guidarelli, Francesca Negrini, Dario De Biase, Annalisa Pession, Elena BaraldiAbstract:Rapid Alkalinization Factor (RALF) genes encode for ubiquitous small peptides that stimulate apoplastic Alkalinization through interaction with malectin-like receptor kinase. RALF peptides may act as negative regulators of plant immune response, inhibiting the formation of the signal receptor complex for immune activation. Recently RALF homologues were identified in different fungal pathogen genomes contributing to host infection ability. Here, FaRALF-33-like gene expression was evaluated in strawberry fruits inoculated with Colletotrichum acutatum, Botrytis cinerea, or Penicillium expansum after 24 and 48 h post-infection. To investigate the role of FaRALF-33-like in strawberry susceptibility, transient transformation was used to overexpress it in white unripe fruits and silence it in red ripe fruits. Agroinfiltrated fruits were inoculated with C. acutatum and expression, and histological analysis of infection were performed. Silencing of FaRALF-33-like expression in C. acutatum-inoculated red fruits led to a delay in fruit colonization by the fungal pathogen, and infected tissues showed less penetrated infective hyphae than in wild-type fruits. In contrast, C. acutatum-inoculated white unripe fruits overexpressing the FaRALF-33-like gene decreased the ontogenic resistance of these fruits, leading to the appearance of disease symptoms and penetrated subcuticular hyphae, normally absent in white unripe fruits. The different response of transfected strawberry fruits to C. acutatum supports the hypothesis that the FaRALF-33-like gene plays an important role in the susceptibility of fruits to the fungal pathogen C. acutatum.
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Induced expression of the Fragaria × ananassa Rapid Alkalinization Factor‐33‐like gene decreases anthracnose ontogenic resistance of unripe strawberry fruit stages
Molecular plant pathology, 2019Co-Authors: Maria Cecilia Merino, Michela Guidarelli, Francesca Negrini, Dario De Biase, Annalisa Pession, Elena BaraldiAbstract:Rapid Alkalinization Factor (RALF) genes encode for ubiquitous small peptides that stimulate apoplastic Alkalinization through interaction with malectin-like receptor kinase. RALF peptides may act as negative regulators of plant immune response, inhibiting the formation of the signal receptor complex for immune activation. Recently RALF homologues were identified in different fungal pathogen genomes contributing to host infection ability. Here, FaRALF-33-like gene expression was evaluated in strawberry fruits inoculated with Colletotrichum acutatum, Botrytis cinerea, or Penicillium expansum after 24 and 48 h post-infection. To investigate the role of FaRALF-33-like in strawberry susceptibility, transient transformation was used to overexpress it in white unripe fruits and silence it in red ripe fruits. Agroinfiltrated fruits were inoculated with C. acutatum and expression, and histological analysis of infection were performed. Silencing of FaRALF-33-like expression in C. acutatum-inoculated red fruits led to a delay in fruit colonization by the fungal pathogen, and infected tissues showed less penetrated infective hyphae than in wild-type fruits. In contrast, C. acutatum-inoculated white unripe fruits overexpressing the FaRALF-33-like gene decreased the ontogenic resistance of these fruits, leading to the appearance of disease symptoms and penetrated subcuticular hyphae, normally absent in white unripe fruits. The different response of transfected strawberry fruits to C. acutatum supports the hypothesis that the FaRALF-33-like gene plays an important role in the susceptibility of fruits to the fungal pathogen C. acutatum.
Tabata Bergonci - One of the best experts on this subject based on the ideXlab platform.
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arabidopsis thaliana Rapid Alkalinization Factor 1 mediated root growth inhibition is dependent on calmodulin like protein 38
Journal of Biological Chemistry, 2017Co-Authors: Wellington F Campos, Keini Dressano, Paulo H O Ceciliato, Juan Carlos Guerreroabad, Aparecida Leonir Da Silva, Celso S Fiori, Amanda Morato Do Canto, Tabata Bergonci, Lucas A N Claus, Marcio C SilvafilhoAbstract:: Arabidopsis thaliana Rapid Alkalinization Factor 1 (AtRALF1) is a small secreted peptide hormone that inhibits root growth by repressing cell expansion. Although it is known that AtRALF1 binds the plasma membrane receptor FERONIA and conveys its signals via phosphorylation, the AtRALF1 signaling pathway is largely unknown. Here, using a yeast two-hybrid system to search for AtRALF1-interacting proteins in Arabidopsis, we identified calmodulin-like protein 38 (CML38) as an AtRALF1-interacting partner. We also found that CML38 and AtRALF1 are both secreted proteins that physically interact in a Ca2+- and pH-dependent manner. CML38-knockout mutants generated via T-DNA insertion were insensitive to AtRALF1, and simultaneous treatment with both AtRALF1 and CML38 proteins restored sensitivity in these mutants. Hybrid plants lacking CML38 and having high accumulation of the AtRALF1 peptide did not exhibit the characteristic short-root phenotype caused by AtRALF1 overexpression. Although CML38 was essential for AtRALF1-mediated root inhibition, it appeared not to have an effect on the AtRALF1-induced Alkalinization response. Moreover, acridinium-labeling of AtRALF1 indicated that the binding of AtRALF1 to intact roots is CML38-dependent. In summary, we describe a new component of the AtRALF1 response pathway. The new component is a calmodulin-like protein that binds AtRALF1, is essential for root growth inhibition, and has no role in AtRALF1 Alkalinization.
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BAK1 is involved in AtRALF1-induced inhibition of root cell expansion.
PLoS genetics, 2017Co-Authors: Keini Dressano, Paulo H O Ceciliato, Aparecida Leonir Da Silva, Tabata Bergonci, Marcio C. Silva-filho, Juan Carlos Guerrero-abad, Fausto Andres Ortiz-morea, Marco Bürger, Daniel S. MouraAbstract:The Rapid Alkalinization Factor (RALF) peptide negatively regulates cell expansion, and an antagonistic relationship has been demonstrated between AtRALF1, a root-specific RALF isoform in Arabidopsis, and brassinosteroids (BRs). An evaluation of the response of BR signaling mutants to AtRALF1 revealed that BRI1-associated receptor kinase1 (bak1) mutants are insensitive to AtRALF1 root growth inhibition activity. BAK1 was essential for the induction of AtRALF1-responsive genes but showed no effect on the mobilization of Ca2+ and Alkalinization responses. Homozygous plants accumulating AtRALF1 and lacking the BAK1 gene did not exhibit the characteristic semi-dwarf phenotype of AtRALF1-overexpressors. Biochemical evidence indicates that AtRALF1 and BAK1 physically interact with a Kd of 4.6 μM and acridinium-labeled AtRALF1 was used to demonstrate that part of the specific binding of AtRALF1 to intact seedlings and to a microsomal fraction derived from the roots of Arabidopsis plants is BAK1-dependent. Moreover, AtRALF1 induces an increase in BAK1 phosphorylation, suggesting that the binding of AtRALF1 to BAK1 is functional. These findings show that BAK1 contains an additional AtRALF1 binding site, indicating that this protein may be part of a AtRALF1-containing complex as a co-receptor, and it is required for the negative regulation of cell expansion.
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Antagonistic relationship between AtRALF1 and brassinosteroid regulates cell expansion-related genes.
Plant signaling & behavior, 2014Co-Authors: Tabata Bergonci, Marcio C. Silva-filho, Daniel S. MouraAbstract:Rapid Alkalinization Factor (RALF) is a peptide signal that plays a role in plant cell expansion. We have recently proposed that AtRALF1 negatively regulates root cell elongation and lateral root formation by opposing the effects of brassinosteroid (BR). We reported 6 AtRALF1-inducible cell wall-related genes and 2 P450 monooxygenase -encoding genes involved in the BR biosynthetic pathway. The AtRALF1-inducible genes implicated in cell wall remodeling were not downregulated by brassinolide (BL) treatment alone; their induction was only compromised following simultaneous treatment with AtRALF1 and BL. We further examined the cell wall-remodeling gene EXPANSIN A5 (AtEXPA5), which is upregulated by BL and has been shown to positively affect root cell elongation. Herein, we report that AtEXPA5 expression is downregulated by AtRALF1 in a dose-dependent manner in the roots and hypocotyls of Arabidopsis plants. AtEXPA5 is also downregulated in plants that overexpress AtRALF1, and it is upregulated in plants in w...
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Arabidopsis thaliana RALF1 opposes brassinosteroid effects on root cell elongation and lateral root formation
Journal of experimental botany, 2014Co-Authors: Tabata Bergonci, Paulo H O Ceciliato, Marcio C. Silva-filho, Juan Carlos Guerrero-abad, Bianca Ribeiro, Daniel S. MouraAbstract:Rapid Alkalinization Factor (RALF) is a peptide signal that plays a basic role in cell biology and most likely regulates cell expansion. In this study, transgenic Arabidopsis thaliana lines with high and low levels of AtRALF1 transcripts were used to investigate this peptide’s mechanism of action. Overexpression of the root-specific isoform AtRALF1 resulted in reduced cell size. Conversely, AtRALF1 silencing increased root length by increasing the size of root cells. AtRALF1-silenced plants also showed an increase in the number of lateral roots, whereas AtRALF1 overexpression produced the opposite effect. In addition, four AtRALF1-inducible genes were identified: two genes encoding prolinerich proteins (AtPRP1 and AtPRP3), one encoding a hydroxyproline-rich glycoprotein (AtHRPG2), and one encoding a xyloglucan endotransglucosylase (TCH4). These genes were expressed in roots and involved in cell-wall rearrangement, and their induction was concentration dependent. Furthermore, AtRALF1-overexpressing plants were less sensitive to exogenous brassinolide (BL); upon BL treatment, the plants showed no increase in root length and a compromised increase in hypocotyl elongation. In addition, the treatment had no effect on the number of emerged lateral roots. AtRALF1 also induces two brassinosteroid (BR)-downregulated genes involved in the BR biosynthetic pathway: the cytochrome P450 monooxygenases CONSTITUTIVE PHOTOMORPHISM AND DWARFISM (CPD) and DWARF4 (DWF4). Simultaneous treatment with both AtRALF1 and BL caused a reduction in AtRALF1-inducible gene expression levels, suggesting that these signals may compete for components shared by both pathways. Taken together, these results indicate an opposing effect of AtRALF1 and BL, and suggest that RALF’s mechanism of action could be to interfere with the BR signalling pathway.
