ABSTRACT
In wheat, the transition of the inflorescence meristem to a terminal spikelet (IMâTS) determines the spikelet number per spike (SNS), an important yield component. In this study, we demonstrate that the plant-specific transcription factor LEAFY (LFY) physically and genetically interacts with WHEAT ORTHOLOG OF APO1 (WAPO1) to regulate SNS and floret development. Loss-of-function mutations in either or both genes result in significant and similar reductions in SNS, as a result of a reduction in the rate of spikelet meristem formation per day. SNS is also modulated by significant genetic interactions between LFY and the SQUAMOSA MADS-box genes VRN1 and FUL2, which promote the IMâTS transition. Single-molecule fluorescence in situ hybridization revealed a downregulation of LFY and upregulation of the SQUAMOSA MADS-box genes in the distal part of the developing spike during the IMâTS transition, supporting their opposite roles in the regulation of SNS in wheat. Concurrently, the overlap of LFY and WAPO1 transcription domains in the developing spikelets contributes to normal floret development. Understanding the genetic network regulating SNS is a necessary first step to engineer this important agronomic trait.
Subject(s)
Gene Expression Regulation, Plant , Meristem , Plant Proteins , Transcription Factors , Triticum , Triticum/genetics , Triticum/metabolism , Triticum/growth & development , Plant Proteins/genetics , Plant Proteins/metabolism , Transcription Factors/metabolism , Transcription Factors/genetics , Meristem/metabolism , Meristem/genetics , Meristem/growth & development , MADS Domain Proteins/genetics , MADS Domain Proteins/metabolism , Flowers/genetics , Flowers/growth & development , Flowers/metabolism , Mutation/genetics , Inflorescence/genetics , Inflorescence/growth & development , Inflorescence/metabolismABSTRACT
Plants deploy cell-surface and intracellular leucine rich-repeat domain (LRR) immune receptors to detect pathogens1. LRR receptor kinases and LRR receptor proteins at the plasma membrane recognize microorganism-derived molecules to elicit pattern-triggered immunity (PTI), whereas nucleotide-binding LRR proteins detect microbial effectors inside cells to confer effector-triggered immunity (ETI). Although PTI and ETI are initiated in different host cell compartments, they rely on the transcriptional activation of similar sets of genes2, suggesting pathway convergence upstream of nuclear events. Here we report that PTI triggered by the Arabidopsis LRR receptor protein RLP23 requires signalling-competent dimers of the lipase-like proteins EDS1 and PAD4, and of ADR1 family helper nucleotide-binding LRRs, which are all components of ETI. The cell-surface LRR receptor kinase SOBIR1 links RLP23 with EDS1, PAD4 and ADR1 proteins, suggesting the formation of supramolecular complexes containing PTI receptors and transducers at the inner side of the plasma membrane. We detected similar evolutionary patterns in LRR receptor protein and nucleotide-binding LRR genes across Arabidopsis accessions; overall higher levels of variation in LRR receptor proteins than in LRR receptor kinases are consistent with distinct roles of these two receptor families in plant immunity. We propose that the EDS1-PAD4-ADR1 node is a convergence point for defence signalling cascades, activated by both surface-resident and intracellular LRR receptors, in conferring pathogen immunity.
Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/immunology , Carboxylic Ester Hydrolases/metabolism , DNA-Binding Proteins/metabolism , Plant Immunity , Protein Serine-Threonine Kinases/metabolism , Arabidopsis Proteins/chemistry , Carboxylic Ester Hydrolases/chemistry , DNA-Binding Proteins/chemistry , Protein Domains , Protein Kinases/chemistry , Protein Kinases/metabolism , Protein Multimerization , Protein Serine-Threonine Kinases/chemistry , Receptors, Cell Surface/chemistry , Receptors, Cell Surface/metabolismABSTRACT
The photoperiodic response is critical for plants to adjust their reproductive phase to the most favorable season. Wheat heads earlier under long days (LD) than under short days (SD) and this difference is mainly regulated by the PHOTOPERIOD1 (PPD1) gene. Tetraploid wheat plants carrying the Ppd-A1a allele with a large deletion in the promoter head earlier under SD than plants carrying the wildtype Ppd-A1b allele with an intact promoter. Phytochromes PHYB and PHYC are necessary for the light activation of PPD1, and mutations in either of these genes result in the downregulation of PPD1 and very late heading time. We show here that both effects are reverted when the phyB mutant is combined with loss-of-function mutations in EARLY FLOWERING 3 (ELF3), a component of the Evening Complex (EC) in the circadian clock. We also show that the wheat ELF3 protein interacts with PHYB and PHYC, is rapidly modified by light, and binds to the PPD1 promoter in planta (likely as part of the EC). Deletion of the ELF3 binding region in the Ppd-A1a promoter results in PPD1 upregulation at dawn, similar to PPD1 alleles with intact promoters in the elf3 mutant background. The upregulation of PPD1 is correlated with the upregulation of the florigen gene FLOWERING LOCUS T1 (FT1) and early heading time. Loss-of-function mutations in PPD1 result in the downregulation of FT1 and delayed heading, even when combined with the elf3 mutation. Taken together, these results indicate that ELF3 operates downstream of PHYB as a direct transcriptional repressor of PPD1, and that this repression is relaxed both by light and by the deletion of the ELF3 binding region in the Ppd-A1a promoter. In summary, the regulation of the light mediated activation of PPD1 by ELF3 is critical for the photoperiodic regulation of wheat heading time.
Subject(s)
Phytochrome B , Triticum , Phytochrome B/genetics , Phytochrome B/metabolism , Triticum/genetics , Flowers/genetics , Flowers/metabolism , Circadian Rhythm/genetics , Photoperiod , Gene Expression Regulation, Plant , Plant Proteins/genetics , Plant Proteins/metabolismABSTRACT
In this article, we describe the development of the plant immunity field, starting with efforts to understand the genetic basis for disease resistance, which â¼30 y ago led to the discovery of diverse classes of immune receptors that recognize and respond to infectious microbes. We focus on knowledge gained from studies of the rice XA21 immune receptor that recognizes RaxX (required for activation of XA21 mediated immunity X), a sulfated microbial peptide secreted by the gram-negative bacterium Xanthomonas oryzae pv. oryzae. XA21 is representative of a large class of plant and animal immune receptors that recognize and respond to conserved microbial molecules. We highlight the complexity of this large class of receptors in plants, discuss a possible role for RaxX in Xanthomonas biology, and draw attention to the important role of sulfotyrosine in mediating receptor-ligand interactions.
Subject(s)
Disease Resistance/immunology , Oryza/immunology , Plant Proteins/immunology , Protein Serine-Threonine Kinases/immunology , Agriculture/history , Allergy and Immunology/history , Allergy and Immunology/trends , Bacterial Infections/genetics , Bacterial Proteins/genetics , Disease Resistance/genetics , History, 19th Century , History, 20th Century , History, 21st Century , Peptides/chemistry , Plant Diseases/microbiology , Plant Immunity/immunology , Plant Proteins/genetics , Plant Proteins/metabolism , Protein Serine-Threonine Kinases/genetics , Protein Serine-Threonine Kinases/metabolismABSTRACT
A hallmark of multicellular organisms is their ability to maintain physiological homeostasis by communicating among cells, tissues, and organs. In plants, intercellular communication is largely dependent on plasmodesmata (PD), which are membrane-lined channels connecting adjacent plant cells. Upon immune stimulation, plants close PD as part of their immune responses. Here, we show that the bacterial pathogen Pseudomonas syringae deploys an effector protein, HopO1-1, that modulates PD function. HopO1-1 is required for P. syringae to spread locally to neighboring tissues during infection. Expression of HopO1-1 in Arabidopsis (Arabidopsis thaliana) increases the distance of PD-dependent molecular flux between neighboring plant cells. Being a putative ribosyltransferase, the catalytic activity of HopO1-1 is required for regulation of PD. HopO1-1 physically interacts with and destabilizes the plant PD-located protein PDLP7 and possibly PDLP5. Both PDLPs are involved in bacterial immunity. Our findings reveal that a pathogenic bacterium utilizes an effector to manipulate PD-mediated host intercellular communication for maximizing the spread of bacterial infection.
Subject(s)
Arabidopsis/microbiology , Plasmodesmata/microbiology , Pseudomonas syringae/growth & development , Pseudomonas syringae/pathogenicity , Adenosine Diphosphate Ribose/metabolism , Arabidopsis/immunology , Arabidopsis/metabolism , Arabidopsis Proteins/chemistry , Arabidopsis Proteins/metabolism , Bacterial Proteins/metabolism , Cell Membrane/metabolism , Plant Immunity , Protein Binding , Protein Domains , Protein Stability , Protein Transport , Pseudomonas syringae/immunology , VirulenceABSTRACT
The rice immune receptor XA21 is activated by the sulfated microbial peptide required for activation of XA21-mediated immunity X (RaxX) produced by Xanthomonas oryzae pv. oryzae (Xoo). Mutational studies and targeted proteomics revealed that the RaxX precursor peptide (proRaxX) is processed and secreted by the protease/transporter RaxB, the function of which can be partially fulfilled by a noncognate peptidase-containing transporter component B (PctB). proRaxX is cleaved at a Gly-Gly motif, yielding a mature peptide that retains the necessary elements for RaxX function as an immunogen and host peptide hormone mimic. These results indicate that RaxX is a prokaryotic member of a previously unclassified and understudied group of eukaryotic tyrosine sulfated ribosomally synthesized, posttranslationally modified peptides (RiPPs). We further demonstrate that sulfated RaxX directly binds XA21 with high affinity. This work reveals a complete, previously uncharacterized biological process: bacterial RiPP biosynthesis, secretion, binding to a eukaryotic receptor, and triggering of a robust host immune response.
Subject(s)
Bacterial Proteins/metabolism , Peptide Hydrolases/metabolism , Peptides/metabolism , Plant Proteins/metabolism , Protein Serine-Threonine Kinases/metabolism , ATP-Binding Cassette Transporters/chemistry , ATP-Binding Cassette Transporters/genetics , ATP-Binding Cassette Transporters/metabolism , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Genes, Bacterial/genetics , Host-Pathogen Interactions/genetics , Host-Pathogen Interactions/immunology , Metabolic Networks and Pathways/genetics , Oryza/immunology , Oryza/metabolism , Oryza/microbiology , Peptide Hydrolases/chemistry , Peptide Hydrolases/genetics , Peptides/chemistry , Peptides/genetics , Plant Proteins/chemistry , Plant Proteins/immunology , Protein Serine-Threonine Kinases/chemistry , Protein Serine-Threonine Kinases/immunology , Xanthomonas/genetics , Xanthomonas/metabolism , Xanthomonas/pathogenicityABSTRACT
Upon encountering a susceptible plant host, a bacterial pathogen expresses specific virulence factors. For example, in planta, the Xanthomonas HrpX protein activates transcription of roughly 150 genes encoding components of the type III secretion system or its translocated effectors, as well as other secreted proteins implicated in pathogenesis. Here, we show that X. oryzae pv. oryzae growth in planta or in HrpX-inducing XOM2 media resulted in HrpX-dependent transcription of the raxX and raxST genes that control production of the RaxX sulfopeptide, exported through a type I secretion system. The RaxX protein is required for activation of XA21-mediated immunity in Xa21+ rice lines. We identified potential plant-inducible promoter elements upstream of the likely 5' ends of the raxX and raxST transcripts. Deletions and nucleotide substitutions confirmed that these elements are required for HrpX-dependent expression of raxX and raxST. We conclude that raxX-raxST gene expression is induced by HrpX during growth in planta and, therefore, is coordinately expressed with other genes required for pathogenesis.[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.
Subject(s)
Oryza , Plant Diseases , Plant Immunity , Xanthomonas , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Gene Expression Regulation, Bacterial , Oryza/metabolism , Oryza/microbiology , Plant Diseases/immunology , Plant Diseases/microbiology , Type III Secretion Systems/genetics , Type III Secretion Systems/metabolism , Xanthomonas/pathogenicityABSTRACT
The occurrence of potentially pathogenic free-living amoebae (FLA) in natural hot springs is considered a public health concern. FLAs are known to cause serious health outcomes to a wide spectrum of mammalian hosts. The present study aimed to provide the distribution of isolated cases of FLAs in hot springs through a systematic review process of available published articles online. Relevant studies are published between January 2010 and January 2020 involving the isolation of Naegleria spp., Acanthamoeba spp., Balamuthia spp., Sappinia spp., and Vermamoeba spp. in natural hot springs in the United States, South America, North America, Europe, Asia, and Africa. Articles were identified through a search of PubMed and Google Scholar databases. Out of 94 articles screened, a total of 20 articles are included in the study with consideration of established inclusion and exclusion criteria. The most common FLAs isolated in hot springs are Acanthamoeba spp. (134; 48.5%) and Naegleria spp. (127; 46.0%). Other FLAs isolated in hot springs include Balamuthia spp. (2; 0.7%) and Vermamoeba spp. (13; 4.7%). FLA in hot springs used for recreational and medical purposes is a potential source of infection. It is recommended that strict surveillance and maintenance of hot springs be implemented to prevent potential future infection.
Subject(s)
Acanthamoeba , Amoeba , Hot Springs , Naegleria , Animals , Humans , PrevalenceABSTRACT
Background: Pathogens often secrete molecules that mimic those present in the plant host. Recent studies indicate that some of these molecules mimic plant hormones required for development and immunity. Scope and Conclusion: This Viewpoint reviews the literature on microbial molecules produced by plant pathogens that functionally mimic molecules present in the plant host. This article includes examples from nematodes, bacteria and fungi with emphasis on RaxX, a microbial protein produced by the bacterial pathogen Xanthomonas oryzae pv. oryzae. RaxX mimics a plant peptide hormone, PSY (plant peptide containing sulphated tyrosine). The rice immune receptor XA21 detects sulphated RaxX but not the endogenous peptide PSY. Studies of the RaxX/XA21 system have provided insight into both host and pathogen biology and offered a framework for future work directed at understanding how XA21 and the PSY receptor(s) can be differentially activated by RaxX and endogenous PSY peptides.
Subject(s)
Host-Pathogen Interactions , Molecular Mimicry/physiology , Plant Immunity/physiology , Plant Diseases/immunology , Plant Diseases/microbiologyABSTRACT
Pathogens target important components of host immunity to cause disease. The Pseudomonas syringae type III-secreted effector HopU1 is a mono-ADP-ribosyltransferase required for full virulence on Arabidopsis thaliana. HopU1 targets several RNA-binding proteins including GRP7, whose role in immunity is still unclear. Here, we show that GRP7 associates with translational components, as well as with the pattern recognition receptors FLS2 and EFR. Moreover, GRP7 binds specifically FLS2 and EFR transcripts in vivo through its RNA recognition motif. HopU1 does not affect the protein-protein associations between GRP7, FLS2 and translational components. Instead, HopU1 blocks the interaction between GRP7 and FLS2 and EFR transcripts in vivo. This inhibition correlates with reduced FLS2 protein levels upon Pseudomonas infection in a HopU1-dependent manner. Our results reveal a novel virulence strategy used by a microbial effector to interfere with host immunity.
Subject(s)
ADP Ribose Transferases/metabolism , Arabidopsis Proteins/metabolism , Arabidopsis/microbiology , Plant Diseases/immunology , Protein Kinases/metabolism , Pseudomonas syringae/immunology , RNA, Messenger/genetics , RNA-Binding Proteins/metabolism , Receptors, Pattern Recognition/metabolism , Virulence/immunology , ADP Ribose Transferases/genetics , Arabidopsis/immunology , Arabidopsis/metabolism , Arabidopsis Proteins/genetics , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Blotting, Western , Cell Membrane , Electrophoretic Mobility Shift Assay , Immunity, Innate , Immunoprecipitation , Plant Diseases/genetics , Plant Diseases/microbiology , Protein Biosynthesis , Protein Kinases/genetics , Pseudomonas syringae/genetics , Pseudomonas syringae/pathogenicity , RNA, Plant/genetics , RNA-Binding Proteins/genetics , Real-Time Polymerase Chain Reaction , Receptors, Pattern Recognition/genetics , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
The biotrophic pathogen Xanthomonas oryzae pv. oryzae (Xoo) produces a sulfated peptide named RaxX, which shares similarity to peptides in the PSY (plant peptide containing sulfated tyrosine) family. We hypothesize that RaxX mimics the growth-stimulating activity of PSY peptides. Root length was measured in Arabidopsis and rice treated with synthetic RaxX peptides. We also used comparative genomic analyses and reactive oxygen species burst assays to evaluate the activity of RaxX and PSY peptides. Here we found that a synthetic sulfated RaxX derivative comprising 13 residues (RaxX13-sY), highly conserved between RaxX and PSY, induces root growth in Arabidopsis and rice in a manner similar to that triggered by PSY. We identified residues that are required for activation of immunity mediated by the rice XA21 receptor but that are not essential for root growth induced by PSY. Finally, we showed that a Xanthomonas strain lacking raxX is impaired in virulence. These findings suggest that RaxX serves as a molecular mimic of PSY peptides to facilitate Xoo infection and that XA21 has evolved the ability to recognize and respond specifically to the microbial form of the peptide.
Subject(s)
Bacterial Proteins/pharmacology , Peptides/pharmacology , Plant Proteins/chemistry , Xanthomonas/pathogenicity , Arabidopsis/drug effects , Arabidopsis/growth & development , Arabidopsis/microbiology , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Bacterial Proteins/chemistry , Host-Pathogen Interactions , Molecular Mimicry , Oryza/drug effects , Oryza/growth & development , Oryza/microbiology , Peptides/chemistry , Plant Diseases/microbiology , Plant Growth Regulators/chemistry , Plant Growth Regulators/pharmacology , Plant Proteins/pharmacology , Plant Roots/drug effects , Plant Roots/growth & development , Plants, Genetically Modified , Signal Transduction , Tyrosine/chemistry , Xanthomonas/geneticsABSTRACT
The Pseudomonas syringae type III effector HopU1 is a mono-ADP-ribosyltransferase that is injected into plant cells by the type III protein secretion system. Inside the plant cell it suppresses immunity by modifying RNA-binding proteins including the glycine-rich RNA-binding protein GRP7. The crystal structure of HopU1 at 2.7-Å resolution reveals two unique protruding loops, L1 and L4, not found in other mono-ADP-ribosyltransferases. Site-directed mutagenesis demonstrates that these loops are essential for substrate recognition and enzymatic activity. HopU1 ADP-ribosylates the conserved arginine 49 of GRP7, and this reduces the ability of GRP7 to bind RNA in vitro. In vivo, expression of GRP7 with Arg-49 replaced with lysine does not complement the reduced immune responses of the Arabidopsis thaliana grp7-1 mutant demonstrating the importance of this residue for GRP7 function. These data provide mechanistic details how HopU1 recognizes this novel type of substrate and highlights the role of GRP7 in plant immunity.
Subject(s)
ADP Ribose Transferases/metabolism , Arabidopsis/metabolism , Plant Immunity/immunology , Plant Proteins/metabolism , RNA, Plant/metabolism , ADP Ribose Transferases/genetics , Arabidopsis/genetics , Arabidopsis/immunology , Arabidopsis/microbiology , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Plant Immunity/genetics , Plant Proteins/genetics , Plants, Genetically Modified/genetics , Plants, Genetically Modified/immunology , Plants, Genetically Modified/metabolism , Plants, Genetically Modified/microbiology , Protein Binding , RNA-Binding Proteins/genetics , RNA-Binding Proteins/metabolismABSTRACT
Activation of plant pattern-triggered immunity (PTI) relies on the recognition of microbe-derived structures, termed patterns, through plant-encoded surface-resident pattern recognition receptors (PRRs). We show that proteobacterial translation initiation factor 1 (IF1) triggers PTI in Arabidopsis thaliana and related Brassicaceae species. Unlike for most other immunogenic patterns, IF1 elicitor activity cannot be assigned to a small peptide epitope, suggesting that tertiary fold features are required for IF1 receptor activation. We have deployed natural variation in IF1 sensitivity to identify Arabidopsis leucine-rich repeat (LRR) receptor-like protein 32 (RLP32) as IF1 receptor using a restriction site-associated DNA sequencing approach. RLP32 confers IF1 sensitivity to rlp32 mutants, IF1-insensitive Arabidopsis accessions and IF1-insensitive Nicotiana benthamiana, binds IF1 specifically and forms complexes with LRR receptor kinases SOBIR1 and BAK1 to mediate signaling. Similar to other PRRs, RLP32 confers resistance to Pseudomonas syringae, highlighting an unexpectedly complex array of bacterial pattern sensors within a single plant species.
Subject(s)
Arabidopsis Proteins , Arabidopsis , Prokaryotic Initiation Factors , Receptors, Pattern Recognition , Arabidopsis/metabolism , Arabidopsis Proteins/metabolism , Genotype , Plant Diseases/microbiology , Plant Immunity/genetics , Proteobacteria/metabolism , Pseudomonas syringae/metabolism , Receptors, Pattern Recognition/genetics , Receptors, Pattern Recognition/metabolismABSTRACT
Pseudomonas fluorescens Q8r1-96 represents a group of rhizosphere strains responsible for the suppressiveness of agricultural soils to take-all disease of wheat. It produces the antibiotic 2,4-diacetylphloroglucinol and aggressively colonizes the roots of cereal crops. In this study, we analyzed the genome of Q8r1-96 and identified a type III protein secretion system (T3SS) gene cluster that has overall organization similar to that of the T3SS gene cluster of the plant pathogen Pseudomonas syringae. We also screened a collection of 30 closely related P. fluorescens strains and detected the T3SS genes in all but one of them. The Q8r1-96 genome contained ropAA and ropM type III effector genes, which are orthologs of the P. syringae effector genes hopAA1-1 and hopM1, as well as a novel type III effector gene designated ropB. These type III effector genes encoded proteins that were secreted in culture and injected into plant cells by both P. syringae and Q8r1-96 T3SSs. The Q8r1-96 T3SS was expressed in the rhizosphere, but mutants lacking a functional T3SS were not altered in their rhizosphere competence. The Q8r1-96 type III effectors RopAA, RopB, and RopM were capable of suppressing the hypersensitive response and production of reactive oxygen species, two plant immune responses.
Subject(s)
Bacterial Proteins/metabolism , Gene Expression Regulation, Bacterial/physiology , Pseudomonas fluorescens/classification , Pseudomonas fluorescens/metabolism , Bacterial Proteins/genetics , Chromosomes, Bacterial , Mutation , Phosphotransferases (Phosphate Group Acceptor) , Phylogeny , Plant Diseases/microbiology , Plant Diseases/prevention & control , Plant Roots/microbiology , Pseudomonas fluorescens/genetics , Signal Transduction , Soil Microbiology , Triticum/microbiologyABSTRACT
Plants perceive microorganisms by recognizing microbial molecules known as pathogen-associated molecular patterns (PAMPs) inducing PAMP-triggered immunity (PTI) or by recognizing pathogen effectors inducing effector-triggered immunity (ETI). The hypersensitive response (HR), a programmed cell death response associated with ETI, is known to be inhibited by PTI. Here, we show that PTI-induced HR inhibition is due to direct or indirect restriction of the type III protein secretion system's (T3SS) ability to inject type III effectors (T3Es). We found that the Pseudomonas syringae T3SS was restricted in its ability to inject a T3E-adenylate cyclase (CyaA) injection reporter into PTI-induced tobacco (Nicotiana tabacum) cells. We confirmed this restriction with a direct injection assay that monitored the in planta processing of the AvrRpt2 T3E. Virulent P. syringae strains were able to overcome a PAMP pretreatment in tobacco or Arabidopsis (Arabidopsis thaliana) and continue to inject a T3E-CyaA reporter into host cells. In contrast, ETI-inducing P. syringae strains were unable to overcome PTI-induced injection restriction. A P. syringae pv tomato DC3000 mutant lacking about one-third of its T3E inventory was less capable of injecting into PTI-induced Arabidopsis plant cells, grew poorly in planta, and did not cause disease symptoms. PTI-induced transgenic Arabidopsis expressing the T3E HopAO1 or HopF2 allowed higher amounts of the T3E-CyaA reporter to be injected into plant cells compared to wild-type plants. Our results show that PTI-induced HR inhibition is due to direct or indirect restriction of T3E injection and that T3Es can relieve this restriction by suppressing PTI.
Subject(s)
Arabidopsis/immunology , Bacterial Proteins/metabolism , Nicotiana/immunology , Plant Immunity/immunology , Pseudomonas syringae/metabolism , Arabidopsis/cytology , Arabidopsis/genetics , Arabidopsis/microbiology , Flagellin/pharmacology , Mutation/genetics , Plant Diseases/immunology , Plant Diseases/microbiology , Plant Immunity/drug effects , Plants, Genetically Modified , Pseudomonas fluorescens/drug effects , Pseudomonas fluorescens/metabolism , Pseudomonas syringae/drug effects , Pseudomonas syringae/growth & development , Receptors, Pattern Recognition/metabolism , Time Factors , Nicotiana/cytology , Nicotiana/metabolism , Nicotiana/microbiologyABSTRACT
RNA-binding proteins (RBP) can control gene expression at both transcriptional and post-transcriptional levels. Plants respond to pathogen infection with rapid reprogramming of gene expression. However, little is known about how plant RBP function in plant immunity. Here, we describe the involvement of an RBP, Arabidopsis thaliana RNA-binding protein-defense related 1 (AtRBP-DR1; At4g03110), in resistance to the pathogen Pseudomonas syringae pv. tomato DC3000. AtRBP-DR1 loss-of-function mutants showed enhanced susceptibility to P. syringae pv. tomato DC3000. Overexpression of AtRBP-DR1 led to enhanced resistance to P. syringae pv. tomato DC3000 strains and dwarfism. The hypersensitive response triggered by P. syringae pv. tomato DC3000 avrRpt2 was compromised in the Atrbp-dr1 mutant and enhanced in the AtRBP-DR1 overexpression line at early time points. AtRBP-DR1 overexpression lines showed higher mRNA levels of SID2 and PR1, which are salicylic acid (SA) inducible, as well as spontaneous cell death in mature leaves. Consistent with these observations, the SA level was low in the Atrbp-dr1 mutant but high in the overexpression line. The SA-related phenotype in the overexpression line was fully dependent on SID2. Thus, AtRBP-DR1 is a positive regulator of SA-mediated immunity, possibly acting on SA signaling-related genes at a post-transcriptional level.
Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/immunology , Arabidopsis/microbiology , Plant Diseases/immunology , RNA-Binding Proteins/metabolism , Salicylic Acid/metabolism , Arabidopsis Proteins/genetics , Gene Expression Regulation, Plant/immunology , Plant Leaves , Protein Transport , Pseudomonas syringae , RNA-Binding Proteins/geneticsABSTRACT
Bacterial species in the genus Xanthomonas infect virtually all crop plants. Although many genes involved in Xanthomonas virulence have been identified through molecular and cellular studies, the elucidation of virulence-associated regulatory circuits is still far from complete. Functional gene networks have proven useful in generating hypotheses for genetic factors of biological processes in various species. Here, we present a genome-scale co-functional network of Xanthomonas oryze pv. oryzae (Xoo) genes, XooNet (www.inetbio.org/xoonet/), constructed by integrating heterogeneous types of genomics data derived from Xoo and other bacterial species. XooNet contains 106,000 functional links, which cover approximately 83% of the coding genome. XooNet is highly predictive for diverse biological processes in Xoo and can accurately reconstruct cellular pathways regulated by two-component signaling transduction systems (TCS). XooNet will be a useful in silico research platform for genetic dissection of virulence pathways in Xoo.
Subject(s)
Gene Regulatory Networks , Genes, Bacterial , Signal Transduction , Xanthomonas/genetics , Gene Expression Regulation, Bacterial , Internet , Signal Transduction/genetics , Xanthomonas/immunologyABSTRACT
The rice XA21-mediated immune response is activated on recognition of the RaxX peptide produced by the bacterium Xanthomonas oryzae pv. oryzae (Xoo). The 60-residue RaxX precursor is post-translationally modified to form a sulfated tyrosine peptide that shares sequence and functional similarity with the plant sulfated tyrosine (PSY) peptide hormones. The 5-kb raxX-raxSTAB gene cluster of Xoo encodes RaxX, the RaxST tyrosylprotein sulfotransferase, and the RaxA and RaxB components of a predicted type I secretion system. To assess raxX-raxSTAB gene cluster evolution and to determine its phylogenetic distribution, we first identified rax gene homologues in other genomes. We detected the complete raxX-raxSTAB gene cluster only in Xanthomonas spp., in five distinct lineages in addition to X. oryzae. The phylogenetic distribution of the raxX-raxSTAB gene cluster is consistent with the occurrence of multiple lateral (horizontal) gene transfer events during Xanthomonas speciation. RaxX natural variants contain a restricted set of missense substitutions, as expected if selection acts to maintain peptide hormone-like function. Indeed, eight RaxX variants tested all failed to activate the XA21-mediated immune response, yet retained peptide hormone activity. Together, these observations support the hypothesis that the XA21 receptor evolved specifically to recognize Xoo RaxX.
Subject(s)
Inheritance Patterns/genetics , Multigene Family , Oryza/immunology , Oryza/microbiology , Plant Immunity/genetics , Plant Proteins/metabolism , Protein Serine-Threonine Kinases/metabolism , Xanthomonas/genetics , Amino Acid Sequence , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Conserved Sequence , Gene Transfer, Horizontal/genetics , Genome, Bacterial , Mutation, Missense/genetics , Phylogeny , Plant Roots/growth & development , Plant Roots/microbiology , Recombination, Genetic/geneticsABSTRACT
Xanthomonas axonopodis pv. manihotis (Xam) causes cassava bacterial blight, the most important bacterial disease of cassava. Xam, like other Xanthomonas species, requires type III effectors (T3Es) for maximal virulence. Xam strain CIO151 possesses 17 predicted T3Es belonging to the Xanthomonas outer protein (Xop) class. This work aimed to characterize nine Xop effectors present in Xam CIO151 for their role in virulence and modulation of plant immunity. Our findings demonstrate the importance of XopZ, XopX, XopAO1 and AvrBs2 for full virulence, as well as a redundant function in virulence between XopN and XopQ in susceptible cassava plants. We tested their role in pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) and effector-triggered immunity (ETI) using heterologous systems. AvrBs2, XopR and XopAO1 are capable of suppressing PTI. ETI suppression activity was only detected for XopE4 and XopAO1. These results demonstrate the overall importance and diversity in functions of major virulence effectors AvrBs2 and XopAO1 in Xam during cassava infection.
Subject(s)
Xanthomonas axonopodis/pathogenicity , Xanthomonas/pathogenicity , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Host-Pathogen Interactions/genetics , Host-Pathogen Interactions/physiology , Plant Immunity/genetics , Plant Immunity/physiology , Virulence/genetics , Virulence/physiologyABSTRACT
Posttranslational modification (PTM) of proteins and peptides is important for diverse biological processes in plants and animals. The paucity of heterologous expression systems for PTMs and the technical challenges associated with chemical synthesis of these modified proteins has limited detailed molecular characterization and therapeutic applications. Here we describe an optimized system for expression of tyrosine-sulfated proteins in Escherichia coli and its application in a bio-based crop protection strategy in rice.