Claudine Balagué

Lesion mimic mutants: keys for deciphering cell death and defense pathways in plants?

The identification of several lesion mimic mutants (LMM) that misregulate cell death constitutes a powerful tool to unravel programmed cell death (PCD) pathways in plants, particularly the hypersensitive response (HR), a form of PCD associated with resistance to pathogens. Recently, the characterization of novel LMM has enabled genes that might regulate cell death programmes to be identified as well as the dissection of defense signaling pathways and of crosstalk between multiple pathways in ways that might not be possible by studying the responses of wild-type plants to pathogens.

Cinnamyl alcohol dehydrogenases-C and D, key enzymes in lignin biosynthesis, play an essential role in disease resistance in Arabidopsis

The deposition of lignin during plant-pathogen interactions is thought to play a role in plant defence. However, the function of lignification genes in plant disease resistance is poorly understood. In this article, we provide genetic evidence that the primary genes involved in lignin biosynthesis in Arabidopsis, CAD-C and CAD-D, act as essential components of defence to virulent and avirulent strains of the bacterial pathogen Pseudomonas syringae pv. tomato, possibly through the salicylic acid defence pathway. Thus, in contrast with cellulose synthesis, whose alteration leads to an increase in disease resistance, alteration of the cell wall lignin content leads directly or indirectly to defects in some defence components.

Secretome analysis reveals effector candidates associated with broad host range necrotrophy in the fungal plant pathogen Sclerotinia sclerotiorum.

BackgroundThe white mold fungus Sclerotinia sclerotiorum is a devastating necrotrophic plant pathogen with a remarkably broad host range. The interaction of necrotrophs with their hosts is more complex than initially thought, and still poorly understood.ResultsWe combined bioinformatics approaches to determine the repertoire of S. sclerotiorum effector candidates and conducted detailed sequence and expression analyses on selected candidates. We identified 486 S. sclerotiorum secreted protein genes expressed in planta, many of which have no predicted enzymatic activity and may be involved in the interaction between the fungus and its hosts. We focused on those showing (i) protein domains and motifs found in known fungal effectors, (ii) signatures of positive selection, (iii) recent gene duplication, or (iv) being S. sclerotiorum-specific. We identified 78 effector candidates based on these properties. We analyzed the expression pattern of 16 representative effector candidate genes on four host plants and revealed diverse expression patterns.ConclusionsThese results reveal diverse predicted functions and expression patterns in the repertoire of S. sclerotiorum effector candidates. They will facilitate the functional analysis of fungal pathogenicity determinants and should prove useful in the search for plant quantitative disease resistance components active against the white mold.

The hypersensitive response. A programmed cell death associated with plant resistance

In plants, the hypersensitive response (HR) is defined as a rapid cell death occurring in response to pathogen attack, and is closely related to active resistance. Initiation of the HR process begins with the recognition of the pathogen by the plant, which is mediated mainly by the pathogen avirulence genes and the plant resistance genes. Then, complex signal transduction pathways intervene, involving changes in protein phosphorylation, production of reactive oxygen species and modification of ion fluxes. Components required for the regulation of the HR cell death are now being identified genetically by the isolation of mutants, in contrast to those involved in the execution of the cell death programme, which are still largely unknown. Further genetic and biochemical analyses will undoubtedly answer the question as to whether this form of programmed cell death (PCD) can be compared with other forms of PCD in plants and with apoptosis in animals.

Delayed symptom development in ein2-1, an Arabidopsis ethylene-insensitive mutant, in response to bacterial wilt caused by Ralstonia solanacearum

ABSTRACT Wilt disease caused by the phytopathogenic bacterium Ralstonia solanacearum is poorly understood at the molecular level. The possible roles of salicylic acid, jasmonic acid, and ethylene, compounds commonly associated with the plant response to pathogens, in wilt symptom development were investigated using various Arabidopsis thaliana mutants in a Col-0 background, an ecotype that develops wilt symptoms in response to the virulent GMI1000 strain. Following root inoculation, wilt symptoms were delayed in ein2-1, an ethylene-insensitive mutant, in response to several virulent strains of the pathogen. In ein2-1, bacteria invade the plant and multiply, reaching concentrations slightly lower than those detected in susceptible plants but 1 to 2 logs higher than in Nd-1, an A. thaliana ecotype resistant to strain GMI1000. This delay in disease symptom development of ein2-1 plants suggests that ethylene signaling plays a critical role in wilt disease development. Furthermore, a strong accumulation of transcripts corresponding to PR-3 and PR-4, two ethylene-responsive genes, was observed in susceptible Col-0 plants, but not in ein2-1 and Nd-1 plants, providing additional evidence for a role of ethylene in wilt symptom production. However, this hormone is probably not involved in the establishment of resistance to R. solanacearum, because homozygous ein2-1 plants in a resistant background remain fully resistant to strain GMI1000.

Resistance to phytopathogens e tutti quanti: placing plant quantitative disease resistance on the map

Plant disease resistance can be seen as a process of a dual nature: both qualitative and quantitative.The nature of non-self molecules perceived by plants has led to the depiction of plant immunity as a two-layer defence system. The first layer is mediated by cell surface and intracellular pattern recognition receptors (PRRs) which perceive conserved microbial elicitors, termed pathogenassociated molecular patterns (PAMPs). The perception of these conserved elicitors initiates cascades of signalling and transcription events, known as PAMP-triggered immunity (PTI). Adapted pathogens secrete effector molecules able to suppress PTI, but which may also be recognized by plant intracellular resistance (R) proteins. This initiates effector-triggered immunity (ETI), the second layer of plant defence. ETI typically yields complete disease resistance phenotypes against pathogens containing the recognized effector, a process designated as qualitative resistance. By contrast, perception of a single PAMP typically has a weaker contribution to overall plant resistance. More generally, in the absence of qualitative resistance, an incomplete resistance phenomenon is often observed, leading to a reduction rather than absence of disease. This is usually referred to as quantitative disease resistance (QDR). The definition of QDR varies in the literature according to the interests of the authors, so that the QDR concept may be puzzling. We settled to define QDR based on the simultaneous occurrence of two observations: (i) the disease resistance phenotype follows a continuous distribution from susceptibility to resistance in host populations, as opposed to the ‘susceptibility or resistance’ binary phenotype observed in qualitative resistance (Fig. 1A); and (ii) the disease resistance phenotype is determined by multiple quantitative trait loci (QTLs) as revealed by crossing host parents with contrasted phenotypes. As a consequence, full disease resistance can be achieved by the introduction of a single R gene, but not by the introduction of a single QDR gene, in a susceptible genetic background. Based on this definition, the molecular architecture of QDR can be hypothesized as an intricate network integrating multiple response pathways to several pathogen molecular determinants and environmental cues (Fig. 1B). THE QUALITATIVE–QUANTITATIVE DUALITY OF PLANT DISEASE RESISTANCE

The hnRNP-Q Protein LIF2 Participates in the Plant Immune Response

Eukaryotes have evolved complex defense pathways to combat invading pathogens. Here, we investigated the role of the Arabidopsis thaliana heterogeneous nuclear ribonucleoprotein (hnRNP-Q) LIF2 in the plant innate immune response. We show that LIF2 loss-of-function in A. thaliana leads to changes in the basal expression of the salicylic acid (SA)- and jasmonic acid (JA)- dependent defense marker genes PR1 and PDF1.2, respectively. Whereas the expression of genes involved in SA and JA biosynthesis and signaling was also affected in the lif2-1 mutant, no change in SA and JA hormonal contents was detected. In addition, the composition of glucosinolates, a class of defense-related secondary metabolites, was altered in the lif2-1 mutant in the absence of pathogen challenge. The lif2-1 mutant exhibited reduced susceptibility to the hemi-biotrophic pathogen Pseudomonas syringae and the necrotrophic ascomycete Botrytis cinerea. Furthermore, the lif2-1 sid2-2 double mutant was less susceptible than the wild type to P. syringae infection, suggesting that the lif2 response to pathogens was independent of SA accumulation. Together, our data suggest that lif2-1 exhibits a basal primed defense state, resulting from complex deregulation of gene expression, which leads to increased resistance to pathogens with various infection strategies. Therefore, LIF2 may function as a suppressor of cell-autonomous immunity. Similar to its human homolog, NSAP1/SYNCRIP, a trans-acting factor involved in both cellular processes and the viral life cycle, LIF2 may regulate the conflicting aspects of development and defense programs, suggesting that a conserved evolutionary trade-off between growth and defense pathways exists in eukaryotes.

Identification and phylogenetic analyses of VASt, an uncharacterized protein domain associated with lipid-binding domains in Eukaryotes

BackgroundSeveral regulators of programmed cell death (PCD) in plants encode proteins with putative lipid-binding domains. Among them, VAD1 is a regulator of PCD propagation harboring a GRAM putative lipid-binding domain. However the function of VAD1 at the subcellular level is unknown and the domain architecture of VAD1 has not been analyzed in details.ResultsWe analyzed sequence conservation across the plant kingdom in the VAD1 protein and identified an uncharacterized VASt (VAD1 Analog of StAR-related lipid transfer) domain. Using profile hidden Markov models (profile HMMs) and phylogenetic analysis we found that this domain is conserved among eukaryotes and generally associates with various lipid-binding domains. Proteins containing both a GRAM and a VASt domain include notably the yeast Ysp2 cell death regulator and numerous uncharacterized proteins. Using structure-based phylogeny, we found that the VASt domain is structurally related to Bet v1-like domains.ConclusionWe identified a novel protein domain ubiquitous in Eukaryotic genomes and belonging to the Bet v1-like superfamily. Our findings open perspectives for the functional analysis of VASt-containing proteins and the characterization of novel mechanisms regulating PCD.

The Arabidopsis thaliana lectin receptor kinase LecRK-I.9 is required for full resistance to Pseudomonas syringae and affects jasmonate signalling

On microbial attack, plants can detect invaders and activate plant innate immunity. For the detection of pathogen molecules or cell wall damage, plants employ receptors that trigger the activation of defence responses. Cell surface proteins that belong to large families of lectin receptor kinases are candidates to function as immune receptors. Here, the function of LecRK-I.9 (At5g60300), a legume-type lectin receptor kinase involved in cell wall-plasma membrane contacts and in extracellular ATP (eATP) perception, was studied through biochemical, gene expression and reverse genetics approaches. In Arabidopsis thaliana, LecRK-I.9 expression is rapidly, highly and locally induced on inoculation with avirulent strains of Pseudomonas syringae pv. tomato (Pst). Two allelic lecrk-I.9 knock-out mutants showed decreased resistance to Pst. Conversely, over-expression of LecRK-I.9 led to increased resistance to Pst. The analysis of defence gene expression suggests an alteration of both the salicylic acid (SA) and jasmonic acid (JA) signalling pathways. In particular, LecRK-I.9 expression during plant-pathogen interaction was dependent on COI1 (CORONATINE INSENSITIVE 1) and JAR1 (JASMONATE RESISTANT 1) components, and JA-responsive transcription factors (TFs) showed altered levels of expression in plants over-expressing LecRK-I.9. A similar misregulation of these TFs was obtained by JA treatment. This study identified LecRK-I.9 as necessary for full resistance to Pst and demonstrated its involvement in the control of defence against pathogens through a regulation of JA signalling components. The role of LecRK-I.9 is discussed with regard to the potential molecular mechanisms linking JA signalling to cell wall damage and/or eATP perception.

Parallel evolution of the POQR prolyl oligo peptidase gene conferring plant quantitative disease resistance

Plant pathogens with a broad host range are able to infect plant lineages that diverged over 100 million years ago. They exert similar and recurring constraints on the evolution of unrelated plant populations. Plants generally respond with quantitative disease resistance (QDR), a form of immunity relying on complex genetic determinants. In most cases, the molecular determinants of QDR and how they evolve is unknown. Here we identify in Arabidopsis thaliana a gene mediating QDR against Sclerotinia sclerotiorum, agent of the white mold disease, and provide evidence of its convergent evolution in multiple plant species. Using genome wide association mapping in A. thaliana, we associated the gene encoding the POQR prolyl-oligopeptidase with QDR against S. sclerotiorum. Loss of this gene compromised QDR against S. sclerotiorum but not against a bacterial pathogen. Natural diversity analysis associated POQR sequence with QDR. Remarkably, the same amino acid changes occurred after independent duplications of POQR in ancestors of multiple plant species, including A. thaliana and tomato. Genome-scale expression analyses revealed that parallel divergence in gene expression upon S. sclerotiorum infection is a frequent pattern in genes, such as POQR, that duplicated both in A. thaliana and tomato. Our study identifies a previously uncharacterized gene mediating QDR against S. sclerotiorum. It shows that some QDR determinants are conserved in distantly related plants and have emerged through the repeated use of similar genetic polymorphisms at different evolutionary time scales.