Turnip crinkle virus (TCV) has been implicated in the suppression of the hypersensitive response (HR), a type of programmed cell death induced during active resistance in Arabidopsis thaliana. In order to investigate the involvement of individual viral components in mediating suppression, TCV genes were cloned for use in an Agrobacterium tumefaciens mediated transient expression in Nicotiana benthamiana. Agroinfiltration of the HR-inducing avrPto/Pto system in conjunction with individual TCV genes has identified the p38 gene, which encodes the viral coat protein, as the gene responsible for the cell death suppression phenotype. The extent of cell death suppression by coat protein was quantified and found to be equal to the level of suppression by the whole virus and AvrPtoB, another cell death inhibitor from bacteria. Thus, the coat protein alone is sufficient to inhibit the HR in plants. Further, the effect of TCV on HR initiation by an avirulence factor from an unrelated bacterial pathogen was investigated. The presence of TCV does not affect the production, secretion or cellular processing of the bacterial avirulence factor.

La RH est une réaction de défense. L’interaction entre G.hirsutum et Xcm repose sur le concept gène-à-gène. L’infection du cultivar Réba B50 possédant les gènes R B2B3 par Xcm18 conduit à une RH associée à une activité LOX, responsable de la peroxydation des lipides, et à la transcription du GhLOX1. Premièrement, 6 génotypes de G.hirsutum contenant divers gènes R ont été retenus pour analyser la variabilité de la réponse LOX suite à l’infection par Xcm1, 18 ou 20. Notre étude a porté sur plusieurs critères, le phénotype, la perte en eau, l’activité LOX et la transcription du GhLOX1. Les résultats montrent une variabilité du phénotype RH en fonction des sources de résistances. Pour chaque type d’interaction incompatible, l’activité LOX et la transcription du GhLOX1 révèlent une augmentation significative corrélée à l’apparition des symptômes RH et à la diminution de la teneur en eau. La réponse LOX est conservée lors de la RH, quelle que soit la race de Xcm ou le génotype. Le GhLOX1 considère comme un marqueur moléculaire de la résistance spécifique du cotonnier à Xcm. Deuxièmement, le rôle du GhLOX1dans la mise en place de la RH en analysant sa fonction potentielle par surexpression. Des cotylédons ont été transformés avec la séquence codante GhLOX1 fusionnée au CaMV35S. Ces cotylédons transformés ont révélé (i) une activité LOX significativement supérieure à celle des cotylédons témoins montrant que le GhLOX1 code pour une protéine active et (ii) un phénotype sans modifications apparentes par rapport à celui des cotylédons non transformées, sauf dans certains contextes d’interactions cotonnier/Xcm où la surexpression de ce gène induit l’apparition de symptômes de type RH. L’effet de l’agro-infiltration sur l’expression de certains gènes pendant la transformation a révélé l’induction précoce et non spécifique de l’expression de gènes de défense. Ces travaux constituent une première étape dans l’analyse fonctionnelle du GhLOX1 dans la résistance spécifique du cotonnier à Xcm / The HR is a defense strategy. The interaction between G.hirsutum and Xcm is governed by the gene-for-gene concept. The infection of the cultivar Reba B50 that contains B2B3 R genes by race Xcm18 leads to a HR associated with a LOX activity response involved in peroxidation of lipids and with transcription of GhLOX1. First, 6 genotypes of G. hirsutum containing various R genes were tested to analyze the variability of the LOX response following the infection by Xcm1, 18 or 20. Several criteria were investigated including the phenotype, the water loss, the LOX activity and GhLOX1 transcription. The results showed variation in HR phenotype according to the tested R genes. For each type of the incompatible interaction, LOX activity and transcription of GhLOX1 were always significantly increased paralleled the apparition of the HR symptoms and the decrease in the water content. LOX response (enzymatic activity and GhLOX1 transcription) is associated with HR whatever the genotype of both Xcm races and cotton plant. Thus, the GhLOX1 consider as a molecular marker of the cotton specific resistance to Xcm. Second, the role of the GhLOX1 gene in the execution of the cotton HR to Xcm by analyzing its possible function by over-expression, the cotyledons were transformed with the GhLOX1 coding sequence fused to the CaMV35S. These transformed cotyledons revealed (i) a LOX activity significantly higher than that detected in the control, showing that the GhLOX1 encodes for an active protein and (ii) that the phenotype of these cotyledons was indistinguishable as compared to the non transformed cotyledons, except when the HR symptoms were induced in some GhLOX1-over-expressed cotyledons. The effect of agro-infiltration on expression of some plant genes during the transformation revealed early and nonspecific induction of the expression of defense genes. This work constitutes a preliminary investigation for the functional analysis of the GhLOX1 in order to assess its role in the cotton specific resistance to Xcm

<p>  </p> <p>Forward genetics discovery of the molecular basis of induced mutants has fundamentally contributed to our understanding of basic biological processes such as metabolism, cell dynamics, growth, and development. Advances in Next-Generation Sequencing (NGS) technologies enabled rapid genome sequencing but also come with limitations such as sequencing errors, dependence on reference genome accuracy, and alignment errors. By incorporating pedigree information to help correct for some errors I optimized variant calling and filtering strategies to respond to experimental design. This led to the identification of multiple causative alleles, the detection of pedigree errors, and an ability to explore the mutational spectrum of multiple mutagens in Arabidopsis. Similar to the problems in forward genetic discovery of mutant alleles, variation in genomes complicates the analysis of gene expression affected by natural variation. The plant hypersensitive response (HR) is a highly localized and rapid form of programmed cell death that plants use to contain biotrophic pathogens. Substantial natural variation exists in the mechanisms that trigger and control HR, yet a complete understanding of the molecular mechanisms modulating HR is lacking. I explored the gene expression consequences of the plant HR in maize using a semi-dominant mutant encoding a constitutively active HR-inducing Nucleotide Binding Site Leucine Rich Repeat protein, <em>Rp1-D21,</em> derived from the receptor responsible for perceiving certain strains of the common rust <em>Puccinia sorghi</em>. Differentially expressed genes (DEG) in response to <em>Rp1-D21</em> were identified in different genetic backgrounds and hybrids that exhibit divergent enhancing (NC350) or suppressing (H95, B73) effects on the visual manifestations of HR. To enable this analysis, I created anonymized reference genomes for each comparison, so that the reference genome induced less bias in the mapping steps. Comprehensive identification of DEG corroborated the visual phenotypes and provided the identities of genes influential in plant hypersensitive response for further studies. The locations of expression quantitative trait loci (eQTL) that determined the differential response of NC350 and B73 were identified using 198 F1 families generated by crossing B73 x NC350 RIL population and <em>Rp1-D21</em>/+ in H95. This identified 3514 eQTL controlling the variability in differential expression between mutant versus wild-type. <em>Trans-</em>eQTL were dramatically arranged in the genome and identified 17 hotspots with more than 200 genes influenced by each locus. A single locus significantly affected expression variation in 5700 genes, 5396 (94.7%) of which were DGE. An allele specific expression analysis of NC350 x H95 and B73 x H95 F1 hybrids with and without <em>Rp1-D21</em> identified <em>cis-</em>eQTL and ASE at a subset of these genes. Bias in the confirmation of eQTL by ASE was still present despite the anonymized reference genomes indicating that additional efforts to improve signal processing in these experiments is needed.</p>

Variation at regulatory elements, identified through hypersensitivity to digestion by Deoxyribonuclease I (DNase I), is believed to contribute to variation in complex traits, but the extent and consequences of this variation are poorly characterized. To investigate the relationship between sequence variation, and the functional consequences of variation in chromatin accessibility, genome-wide DNase I hypersensitive sites (DHS) of terminally differentiated erythroblasts were studied in eight inbred strains of mice studied (A/J, AKR/J, BALBc/J, C3H/HeJ, C57BL/6J, CBA/J, DBA/2J, and LP/J). These strains were selected because of the availability of their genome sequence and quantitative trait loci (QTL) data. After confirming that next generation sequencing could identify DNase I hypersensitive sites with high sensitivity and specificity, and that differential peaks could be found, an automated peak calling pipeline was developed and optimized. 36,693 DHS peaks were identified covering 9.1 Mb (0.29%) of mouse genome. There was no indication of within strain variation. Between strains reproducible variation was observed at approximately 5% of DNase hypersensitive sites (1,397 DHSs). Variable DHSs were more likely to be enhancers than promoters and less likely to occur at conserved regions of the genome. Only 36% of such variable DHSs contain a sequence variant predictive of site variation and 12% contain at least one variant that disrupts transcription factor binding sites. The majority (86%) of variable DHSs differ in size/shape and the remaining 14% demonstrate discrete variation in single peak or cluster of peaks. Sequence variants within variable DHS are more likely to be associated with complex traits than those in non-variant DHS, and variants associated with complex traits preferentially occur in enhancer-like elements. Changes at a small proportion (7%) of discretely variable DHS are associated with changes in nearby transcriptional activity. Our results show that whilst DNA sequence variation is not the major determinant of variation in open chromatin, where such variants exist they are likely to be causal for complex traits.

A interacao planta-patogeno desencadeia uma serie de sinais que ainda nao estao completamente elucidados. Uma das respostas e a reacao de hipersensibilidade (RH), onde ocorre a morte celular programada no sitio da infeccao, impedindo a proliferacao do patogeno. Acredita que a morte celular e provocada pelo aumento do ERO, principalmente peroxido de hidrogenio (H2O2) e com o acumulo de acido salicilico (AS) que inibe a catalase, enzima responsavel pela transformacao de H2O2 em H2O e O2. Alem disso, ocorre o aumento da sintese e liberacao dos compostos fenolicos e alteracao da parede celular dos vegetais, com o aumento das atividades de diversas enzimas, capazes de degradar a parede celular da planta e do microrganismo invasor, liberando fragmentos que podem atuar como moleculas sinalizadoras, tornando as plantas mais resistentes. Nesse trabalho as celulas de Rubus fruticosus (amora-preta) foram tratadas, separadamente, com tres diferentes moleculas elicitoras, ou seja, moleculas capazes de ativar o mecanismo de defesa das plantas, o acido salicilico (AS), o metil jasmonato (MeJA) e ramnoglucuronogalactana (F-I), na concentracao de 1 Êmol/L durante 1h, para o estudo dos componentes extracelulares liberados e das modificacoes dos monossacarideos da parede celular durante resposta de hipersensibilidade. A concentracao de proteinas totais extracelulares foi aumentada com os indutores F-I e MeJA. A atividade enzimatica de -D-xilosidase nao se alterou na presenca de F-I, AS e MeJA. Entretanto, o MeJA tem a capacidade de aumentar as atividades das enzimas -D-galactosidase, -Dglucosidase, quitinase e laminarinase e inibir as atividades das enzimas galacturonase e -Lfucosidase na concentracao e tempo usado. O AS e F-I provocaram um aumento nas atividades de galacturonase e quitinase e inibiram a laminarinase. A aplicacao exogena de F-I e AS induziram a liberacao de compostos fenolicos para meio extracelular, que provavelmente, foi decorrente da tentativa das celulas de se protegerem de microrganismos invasores, com um decrescimo desses compostos no meio intracelular. O MeJA nao foi capaz de alterar a sintese de compostos fenolicos totais intracelulares e extracelulares e de acucares extracelulares, em tais condicoes. Tambem F-I e AS nao alteraram o teor de acucar redutor extracelular. O MeJA foi mais efetivo na producao de ERO durante 30 minutos de incubacao na concentracao de1 Êmol/L . F-I foi tambem ativador na liberacao de ERO, no entanto, o AS provocou inibicao. Os principais monossacarideos neutros que constitui a parede celular de suspensao de celulas de Rubus fruticosus sao as glucose (55-61%), arabinose (22-29%) e manose (13,8-15%). Ocorrendo em menor concentracoes os monossacarideos de fucose (0,65-1,2%), galactose (0,5-0,8%), xilose (0,5-0,8%) e ramnose (aproximadamente 0,5%).Os monossacarideos ramnose, fucose, xilose e galactose de parede celular tiveram um decrescimo na presenca do AS e um aumento na presenca de MeJA. Entretanto, o AS e o MeJA nao alteraram o percentual de arabinose, manose e glucose. O F-I foi capaz de aumentar o percentual dos monossacarideos ramnose e fucose e diminuir de glucose. Os resultados obtidos demonstram que a via de ativacao dos mecanismos de defesa da celula vegetal, induzida pelo MeJA, difere das vias ativadas pelo AS e F-I, pois o F-I e o AS induziram a liberacao de compostos fenolicos e o MeJA provocou aumento nas atividades enzimaticas, principalmente que atuam na parede celular da propria planta. O AS e o F-I foram mais efetivos no aumento das atividades enzimaticsa relacionadas a defesa da planta, as quais agem nas paredes de diversos fitopatogenos, sendo que as enzimas que podem atuar na parede celular da propria planta foram inibidas ou nao sofreram alteracao. / The plant-pathogen interactions trigger a series of signals that are not yet completely understood. One of the mechanisms is the hypersensitive response (HR), which is characterized by cell death in the infection site in order to prevent pathogen proliferation. Our previous studies with different elicitors demonstrated the correlation between the formation of reactive oxygen species (ROS) and cell wall degradation. Here, the cells were elicited with 1 mol/L salicylic acid (SA), methyl jasmonate (MeJA) or acid polysaccharide (rhamnoglucuronogalactan, F-I) (1mol/L) from characterization the extracellular components released and the modifications of the monosaccharide composition in cell wall during a hypersensitive response in Rubus fruticosus (blackberry-black).The extracellular proteins released to the extracellular were increased with the inducers molecules F-I and MeJA. The -D-xylosidase enzymatic activities didnt change in the presence of F-I, SA and MeJA. The time-course curves for -D-galactosidase, -D-glucosidase activities in fraction E were most effective for MeJA, while F-I and AS inhibited -Dgalactosidase. Also, the MeJA has ability to activate laminarinase and chitinase enzymatic activities and inhibit galacturonase and -L-fucosidase enzymatic activities. After 1h, the SA and F-I caused an increase galacturonase and chitinase activities and inhibited laminarinase enzymatic activity. Also, the time-course curves chitinase in the fraction increased with SA.The F-I and SA increased extracellular phenolic compounds, although they decreased them in the fraction I. MeJA was unable to change the synthesis of either intracellular or extracellular phenolic compounds. The data suggest that F-I and AS modulate the defense responses of plants through a via different that of MeJA. The extracellular reducing sugar didnt change with F-I, SA and MeJA.The MeJA was more effective in the release ROS incubation of 30 minutes at concentration of 1 mol/L. However, the presence of SA caused inhibition and F-I activated of ROS by cells.The main constituents of neutral sugars in the cell wall of Rubus fruticosus were glucose (55-61%), arabinose (22-29%) and mannose (13.8-15%). Minor constituents were fucose (0.65-1.2%), galactose (0.5- 0.8%), xylose (0.5-0.8%) and rhamnose (~0.5%). SA decreased the rhamnose and fucose concentrations; F-I both decreased the percentage of mannose and glucose and increased rhamnose and fucose. MeJA, in turn, increased the percentage of rhamnose, xylose and galactose. The data suggest that F-I and SA modulate the defense responses of plants through a mechanism unrelated to the MeJA via. Since the F-I and the SA induced the release phenolic compounds and the MeJA increased in enzymatic activities, mainly age in the own plant cell wall. The SA and F-I were more effective in the increasing defense enzyme-related activity of the plant that acts on the walls of several phytopathogens, and the enzymes that can act in the cell wall of the plant were inhibited or did not change.

Plant programmed cell death (PCD) is an essential process in plant-pathogen interactions. Importantly, PCD can have contrasting effects on the outcome depending on context. For example, plant PCD in plant-biotroph interactions is clearly beneficial to plants, whereas it could be detrimental to plants in plant-necrotroph interactions. Sclerotinia sclerotiorum is an agriculturally and economically important necrotrophic pathogen. Previous studies have shown that S. sclerotiorum secretes oxalic acid (OA) to enhance Sclerotinia virulence by various mechanisms including induction of PCD in plants. A recent study has also shown that reactive oxygen species (ROS) generation correlates with induction of PCD during disease development. These studies focus on links between ROS, oxalate, and PCD, and how they impact S. sclerotiorum disease development. I examined the involvement of ROS in pathogenic development of S. sclerotiorum. I identified and functionally characterized two predicted S. sclerotiorum NADPH oxidases (Nox1 and Nox2) by RNAi. Both nox genes appear to have roles in sclerotial development, while only Nox1-silenced mutants showed reduced virulence. Interestingly, the reduced virulence of the Nox1-silenced mutant correlated with decreased production of OA in the mutant. This observation suggests that regulation of ROS by S. sclerotiorum Nox1 may be linked to OA. The next study details the phenotype of plants inoculated with an S. sclerotiorum oxalate deficient mutant (A2), which showed restricted growth at the infected site. This response resembles the hypersensitive response (HR), and is associated with plant resistance responses including cell wall strengthening, plant oxidative burst, and induction of defensin genes. Conversely, leaves infected with wild type showed unrestricted spreading of cell death and were not associated with these resistant responses. Furthermore, previous work had shown that a Caenorhabditis elegans anti-apoptotic gene (ced-9) conferred resistance to wild type S. sclerotiorum, while this gene had negligible effects on the phenotype of plant leaves inoculated with A2 mutants. These findings suggest that HR-like cell death by A2 and PCD by wild type S. sclerotiorum may be regulated by different pathways. As a whole, these results reveal the importance of ROS, oxalate, and PCD in Sclerotinia disease development as well as the significance of interplay between them. These studies contribute to the understanding of the underlying mechanisms of Sclerotinia disease.

The objective of this study was to transform sweet pepper ferredoxin-like protein (PFLP) gene, which has antimicrobial properties, to tobacco and investigate the disease resistance abilities of transgenic tobacco. This protein interacts with another protein, harpin that is produced by the bacteria which is invading the plant tissues, and stimulates hypersensitivity response in plants, thus the spreading of disease is limited. Gene transfer was achieved to tobacco by Agrobacterium- mediated method and with indirect organogenesis / the explants were grown on selective media and then transferred to jars and pots respectively. Molecular and genetic analyses such as PCR, RT-PCR, Sequence Analysis and Northern Blot, were performed with plants which their seeds survived and grew on selective medium and also gave positive reactions for GUS histochemical assay. Finally, with putative transgenic plants, some hypersensitive response assays were carried out with Pseudomonas syringae and it was observed that the recovered plants showed hypersensitive response (HR) in the preliminary tests. These results indicated that putative transgenic tobacco plants which carry pflp transgene, can be used in disease resistance studies.

No description available.

A interacao planta-patogeno desencadeia uma serie de sinais que ainda nao estao completamente elucidados. Uma das respostas e a reacao de hipersensibilidade (RH), onde ocorre a morte celular programada no sitio da infeccao, impedindo a proliferacao do patogeno. Acredita que a morte celular e provocada pelo aumento do ERO, principalmente peroxido de hidrogenio (H2O2) e com o acumulo de acido salicilico (AS) que inibe a catalase, enzima responsavel pela transformacao de H2O2 em H2O e O2. Alem disso, ocorre o aumento da sintese e liberacao dos compostos fenolicos e alteracao da parede celular dos vegetais, com o aumento das atividades de diversas enzimas, capazes de degradar a parede celular da planta e do microrganismo invasor, liberando fragmentos que podem atuar como moleculas sinalizadoras, tornando as plantas mais resistentes. Nesse trabalho as celulas de Rubus fruticosus (amora-preta) foram tratadas, separadamente, com tres diferentes moleculas elicitoras, ou seja, moleculas capazes de ativar o mecanismo de defesa das plantas, o acido salicilico (AS), o metil jasmonato (MeJA) e ramnoglucuronogalactana (F-I), na concentracao de 1 Êmol/L durante 1h, para o estudo dos componentes extracelulares liberados e das modificacoes dos monossacarideos da parede celular durante resposta de hipersensibilidade. A concentracao de proteinas totais extracelulares foi aumentada com os indutores F-I e MeJA. A atividade enzimatica de -D-xilosidase nao se alterou na presenca de F-I, AS e MeJA. Entretanto, o MeJA tem a capacidade de aumentar as atividades das enzimas -D-galactosidase, -Dglucosidase, quitinase e laminarinase e inibir as atividades das enzimas galacturonase e -Lfucosidase na concentracao e tempo usado. O AS e F-I provocaram um aumento nas atividades de galacturonase e quitinase e inibiram a laminarinase. A aplicacao exogena de F-I e AS induziram a liberacao de compostos fenolicos para meio extracelular, que provavelmente, foi decorrente da tentativa das celulas de se protegerem de microrganismos invasores, com um decrescimo desses compostos no meio intracelular. O MeJA nao foi capaz de alterar a sintese de compostos fenolicos totais intracelulares e extracelulares e de acucares extracelulares, em tais condicoes. Tambem F-I e AS nao alteraram o teor de acucar redutor extracelular. O MeJA foi mais efetivo na producao de ERO durante 30 minutos de incubacao na concentracao de1 Êmol/L . F-I foi tambem ativador na liberacao de ERO, no entanto, o AS provocou inibicao. Os principais monossacarideos neutros que constitui a parede celular de suspensao de celulas de Rubus fruticosus sao as glucose (55-61%), arabinose (22-29%) e manose (13,8-15%). Ocorrendo em menor concentracoes os monossacarideos de fucose (0,65-1,2%), galactose (0,5-0,8%), xilose (0,5-0,8%) e ramnose (aproximadamente 0,5%).Os monossacarideos ramnose, fucose, xilose e galactose de parede celular tiveram um decrescimo na presenca do AS e um aumento na presenca de MeJA. Entretanto, o AS e o MeJA nao alteraram o percentual de arabinose, manose e glucose. O F-I foi capaz de aumentar o percentual dos monossacarideos ramnose e fucose e diminuir de glucose. Os resultados obtidos demonstram que a via de ativacao dos mecanismos de defesa da celula vegetal, induzida pelo MeJA, difere das vias ativadas pelo AS e F-I, pois o F-I e o AS induziram a liberacao de compostos fenolicos e o MeJA provocou aumento nas atividades enzimaticas, principalmente que atuam na parede celular da propria planta. O AS e o F-I foram mais efetivos no aumento das atividades enzimaticsa relacionadas a defesa da planta, as quais agem nas paredes de diversos fitopatogenos, sendo que as enzimas que podem atuar na parede celular da propria planta foram inibidas ou nao sofreram alteracao. / The plant-pathogen interactions trigger a series of signals that are not yet completely understood. One of the mechanisms is the hypersensitive response (HR), which is characterized by cell death in the infection site in order to prevent pathogen proliferation. Our previous studies with different elicitors demonstrated the correlation between the formation of reactive oxygen species (ROS) and cell wall degradation. Here, the cells were elicited with 1 mol/L salicylic acid (SA), methyl jasmonate (MeJA) or acid polysaccharide (rhamnoglucuronogalactan, F-I) (1mol/L) from characterization the extracellular components released and the modifications of the monosaccharide composition in cell wall during a hypersensitive response in Rubus fruticosus (blackberry-black).The extracellular proteins released to the extracellular were increased with the inducers molecules F-I and MeJA. The -D-xylosidase enzymatic activities didnt change in the presence of F-I, SA and MeJA. The time-course curves for -D-galactosidase, -D-glucosidase activities in fraction E were most effective for MeJA, while F-I and AS inhibited -Dgalactosidase. Also, the MeJA has ability to activate laminarinase and chitinase enzymatic activities and inhibit galacturonase and -L-fucosidase enzymatic activities. After 1h, the SA and F-I caused an increase galacturonase and chitinase activities and inhibited laminarinase enzymatic activity. Also, the time-course curves chitinase in the fraction increased with SA.The F-I and SA increased extracellular phenolic compounds, although they decreased them in the fraction I. MeJA was unable to change the synthesis of either intracellular or extracellular phenolic compounds. The data suggest that F-I and AS modulate the defense responses of plants through a via different that of MeJA. The extracellular reducing sugar didnt change with F-I, SA and MeJA.The MeJA was more effective in the release ROS incubation of 30 minutes at concentration of 1 mol/L. However, the presence of SA caused inhibition and F-I activated of ROS by cells.The main constituents of neutral sugars in the cell wall of Rubus fruticosus were glucose (55-61%), arabinose (22-29%) and mannose (13.8-15%). Minor constituents were fucose (0.65-1.2%), galactose (0.5- 0.8%), xylose (0.5-0.8%) and rhamnose (~0.5%). SA decreased the rhamnose and fucose concentrations; F-I both decreased the percentage of mannose and glucose and increased rhamnose and fucose. MeJA, in turn, increased the percentage of rhamnose, xylose and galactose. The data suggest that F-I and SA modulate the defense responses of plants through a mechanism unrelated to the MeJA via. Since the F-I and the SA induced the release phenolic compounds and the MeJA increased in enzymatic activities, mainly age in the own plant cell wall. The SA and F-I were more effective in the increasing defense enzyme-related activity of the plant that acts on the walls of several phytopathogens, and the enzymes that can act in the cell wall of the plant were inhibited or did not change.

Les réactions de défense végétales sont souvent associées au développement de la réponse hypersensible (HR), une forme de mort cellulaire programmée qui confine l'agent pathogène au niveau du site d'infection. La frontière nette de la HR suggère l'existence de mécanismes efficaces qui contrôlent la frontière entre mort cellulaire et survie. Le facteur de transcription d'Arabidopsis AtMYB30 régule positivement la HR et les réponses de défense de la plante en augmentant la synthèse des acides gras à très longue chaîne (VLCFA) après infection bactérienne. L'activité d’AtMYB30 est étroitement contrôlée à l'intérieur des cellules végétales par des interactions protéine-protéine et des modifications post-traductionnelles. Au cours de mes travaux de thèse, nous avons identifié une protéase de la famille des subtilases (AtSBT5.2) en tant que partenaire protéique d’AtMYB30. Chose intéressante, nous avons montré que le transcrit d’AtSBT5.2 est épissée de façon alternative, conduisant à la production de deux produits de gènes distincts codant soit pour une isoforme sécrétée [AtSBT5.2 (a)] soit une isoforme intracellulaire [AtSBT5.2 (b)]. L'interaction spécifique d’AtMYB30 avec AtSBT5.2(b), mais pas avec AtSBT5.2(a), conduit à une rétention d’AtMYB30 à l'extérieur du noyau au sein de petites vésicules intracellulaires. Des plantes d’Arabidopsis mutantes atsbt5.2, ne montrant ni expression d’AtSBT5.2(a) ni d’AtSBT5.2(b), présentent des réactions de défense et de HR accrues. Ce phénotype étant abolie dans un fond génétique mutant atmyb30, AtSBT5.2 est donc un régulateur négatif de la résistance aux maladies induites par AtMYB30. Fait important, la surexpression de l’isoforme AtSBT5.2(b), mais pas celle de l’isoforme AtSBT5.2(a), dans le fond mutant atsbt5.2 rétablit les phénotypes présentés par les plantes mutantes atsbt5.2, ce qui suggère qu’AtSBT5.2(b) réprime spécifiquement la réponse de défense induite par AtMYB30. / Plants defence responses are often associated with the development of the so-called hypersensitive response (HR), a form of PCD that confines the pathogen to the infection site. The sharp boundary of the HR suggests the existence of efficient mechanisms that control cell death and survival. The Arabidopsis transcription factor AtMYB30 positively regulates plant defence and HR responses by enhancing the synthesis of sphingolipid-containing Very Long Chain Fatty Acids (VLCFA) after bacterial infection. The activity of AtMYB30 is tightly controlled inside plant cells through protein-protein interactions and post-translational modifications. During my PhD, we identified a protease of the subtilase family (AtSBT5.2) as a AtMYB30-interacting partner. Interestingly, we have shown that the AtSBT5.2 transcript is alternatively spliced, leading to the production of two distinct gene products that encode either a secreted [AtSBT5.2(a)] or an intracellular [AtSBT5.2(b)] protein. The specific interaction between AtMYB30 and AtSBT5.2(b), but not AtSBT5.2(a), leads to AtMYB30 specific retention outside of the nucleus in small intracellular vesicles. atsbt5.2 Arabidopsis mutant plants, in which both AtSBT5.2(a) and AtSBT5.2(b) expression was abolished, displayed enhanced HR and defence responses. The fact that this phenotype is abolished in an atmyb30 mutant background suggests that AtSBT5.2 is a negative regulator of AtMYB30-mediated disease resistance. Importantly, overexpression of the AtSBT5.2(b), but not the AtSBT5.2(a), isoform in the atsbt5.2 mutant background reverts the phenotypes displayed by atsbt5.2 mutant plants, suggesting that AtSBT5.2(b) specifically represses AtMYB30-mediated defence.