Suberin plasticity to developmental and exogenous cues is regulated by a set of MYB transcription factors

Suberin is a hydrophobic biopolymer that can be deposited at the periphery of cells, forming protective barriers against biotic and abiotic stress. In roots, suberin forms lamellae at the periphery of endodermal cells where it plays crucial roles in the control of water and mineral transport. Suberin formation is highly regulated by developmental and environmental cues. However, the mechanisms controlling its spatiotemporal regulation are poorly understood. Here, we show that endodermal suberin is regulated independently by developmental and exogenous signals to fine tune suberin deposition in roots. We found a set of four MYB transcription factors (MYB41, MYB53, MYB92 and MYB93), that are regulated by these two signals, and are sufficient to promote endodermal suberin. Mutation of these four transcription factors simultaneously through genome editing, lead to a dramatic reduction of suberin formation in response to both developmental and environmental signals. Most suberin mutants analyzed at physiological levels are also affected in another endodermal barrier made of lignin (Casparian strips), through a compensatory mechanism. Through the functional analysis of these four MYBs we generated plants allowing unbiased investigations of endodermal suberin function without accounting for confounding effects due to Casparian strip defects, and could unravel specific roles of suberin in nutrient homeostasis

Le système immunitaire des plantes : induction, mémoire et désamorçage des réponses de défense par des éliciteurs endogènes, exogènes et synthétiques.

As sessile organism, plants have to react quickly and strongly with defense responses to repel any invading pathogen. The plant immune system can be triggered by exogenous or endogenous elicitor molecules. Another class of elicitors are defense promoting compounds which are also known as synthetic elicitors. Here I describe the discovery and characterization of a novel family of potentially secreted small endogenous peptides (PROSCOOP) which members harbor small peptides (SCOOPs). I show that the SCOOP family is involved in plant defense and root development. Various SCOOP peptides induce short- and long-term defense responses. Moreover, treatments with the SCOOP12 peptide induce the resistance against Pseudomonas syringae in Arabidopsis. In the second part of this thesis, I show that treatments with a synthetic elicitor can lead to long-term transcriptional memory and that subsequent challenging of such plants with an exogenous elicitor reverted this transcriptional memory. In conclusion, my thesis shows (1) how diverse the function of these elicitors can be and (2) the impact the plant defense system and its triggers have on plant development and memory.En tant qu’organismes sessiles, les plantes doivent réagir rapidement et intensément, via des réponses défensives, pour repousser les pathogènes invasifs. Le système immunitaire des plantes peut être déclenché par des molécules élicitrices exogènes ou endogènes. Une autre classe d’éliciteurs, les éliciteurs synthétiques, contient également des composés promouvant une réponse défensive.Dans ce manuscrit, je décris la découverte et caractérisation d’une nouvelle famille de petits peptides endogènes potentiellement sécrétés(PROSCOOP), dont les membres incluent de petits peptides (SCOOP). Je démontre que les SCOOP sont impliqués dans les mécanismes de défense de la plante et le développement racinaire. Une variété de peptides SCOOP induit des réponses défensives de courtes et longues durées.De plus, des traitements avec le peptideSCOOP12 induisent une résistance à Pseudomonas syringae chez Arabidopsis.Dans la seconde partie de cette thèse, je démontre que le traitement des plantes avec un éliciteur synthétique peut mener à une mémoire transcriptionnelle à long terme, et que le challenge subséquent des plantes traitées par application d’un éliciteur exogène désactive cette mémoire transcriptionnelle. En conclusion, ma thèse présente (1) la diversité des fonctions que peuvent avoir ces éliciteurs et (2) l’impact sur les systèmes de défense de la plante et ses conséquences sur la mémoire et le développement de la plante

Perception of Arabidopsis AtPep peptides, but not bacterial elicitors, accelerates starvation-induced senescence

Members of the AtPep group of Arabidopsis endogenous peptides have frequently been reported to induce pattern-triggered immunity (PTI) and to increase resistance to diverse pathogens by amplifying the innate immune response. Here, we made the surprising observation that dark-induced leaf senescence was accelerated by the presence of Peps. Adult leaves as well as leaf discs of Col-0 wild type plants showed a Pep-triggered early onset of chlorophyll breakdown and leaf yellowing whereas pepr1 pepr2 double mutant plants were insensitive. In addition, this response was dependent on ethylene signaling and inhibited by the addition of cytokinins. Notably, addition of the bacterial elicitors flg22 or elf18, both potent inducers of PTI, did not provoke an early onset of leaf senescence. Continuous darkness leads to energy deprivation and starvation and therewith promotes leaf senescence. We found that continuous darkness also strongly induced PROPEP3 transcription. Moreover, Pep-perception led to a rapid induction of PAO, APG7, and APG8a, genes indispensable for chlorophyll degradation as well as autophagy, respectively, and all three hallmarks of starvation and senescence. Notably, addition of sucrose as a source of energy inhibited the Pep-triggered early onset of senescence. In conclusion, we report that Pep-perception accelerates dark/starvation-induced senescence via an early induction of chlorophyll degradation and autophagy. This represents a novel and unique characteristic of PEPR signaling, unrelated to PTI

Biotic Stress-Induced Priming and De-Priming of Transcriptional Memory in Arabidopsis and Apple

Under natural growth conditions, plants experience various and repetitive biotic and abiotic stresses. Salicylic acid (SA) is a key phytohormone involved in the response to biotic challenges. Application of synthetic SA analogues can efficiently prime defense responses, and leads to improved pathogen resistance. Because SA analogues can result in long-term priming and memory, we identified genes for which expression was affected by the SA analogue and explored the role of DNA methylation in this memorization process. We show that treatments with an SA analogue can lead to long-term transcriptional memory of particular genes in Arabidopsis. We found that subsequent challenging of such plants with a bacterial elicitor reverted this transcriptional memory, bringing their expression back to the original pre-treatment level. We also made very similar observations in apple (Malus domestica), suggesting that this expression pattern is highly conserved in plants. Finally, we found a potential role for DNA methylation in the observed transcriptional memory behavior. We show that plants defective in DNA methylation pathways displayed a different memory behavior. Our work improves our understanding of the role of transcriptional memory in priming, and has important implication concerning the application of SA analogues in agricultural settings

Impaired Cuticle Functionality and Robust Resistance to <i>Botrytis cinerea </i>in <i>Arabidopsis thaliana</i> Plants With Altered Homogalacturonan Integrity Are Dependent on the Class III Peroxidase AtPRX71

Pectin is a major cell wall component that plays important roles in plant development and response to environmental stresses. Arabidopsis thaliana plants expressing a fungal polygalacturonase (PG plants) that degrades homogalacturonan (HG), a major pectin component, as well as loss-of-function mutants for QUASIMODO2 (QUA2), encoding a putative pectin methyltransferase important for HG biosynthesis, show accumulation of reactive oxygen species (ROS), reduced growth and almost complete resistance to the fungal pathogen Botrytis cinerea. Both PG and qua2 plants show increased expression of the class III peroxidase AtPRX71 that contributes to their elevated ROS levels and reduced growth. In this work, we show that leaves of PG and qua2 plants display greatly increased cuticle permeability. Both increased cuticle permeability and resistance to B. cinerea in qua2 are suppressed by loss of AtPRX71. Increased cuticle permeability in qua2, rather than on defects in cuticle ultrastructure or cutin composition, appears to be dependent on reduced epidermal cell adhesion, which is exacerbated by AtPRX71, and is suppressed by the esmeralda1 mutation, which also reverts the adhesion defect and the resistant phenotype. Increased cuticle permeability, accumulation of ROS, and resistance to B. cinerea are also observed in mutants lacking a functional FERONIA, a receptor-like kinase thought to monitor pectin integrity. In contrast, mutants with defects in other structural components of primary cell wall do not have a defective cuticle and are normally susceptible to the fungus. Our results suggest that disrupted cuticle integrity, mediated by peroxidase-dependent ROS accumulation, plays a major role in the robust resistance to B. cinerea of plants with altered HG integrity

Suberin plasticity to developmental and exogenous cues is regulated by a set of MYB transcription factors

Suberin is a hydrophobic biopolymer that can be deposited at the periphery of cells, forming protective barriers against biotic and abiotic stress. In roots, suberin forms lamellae at the periphery of endodermal cells where it plays crucial roles in the control of water and mineral transport. Suberin formation is highly regulated by developmental and environmental cues. However, the mechanisms controlling its spatiotemporal regulation are poorly understood. Here, we show that endodermal suberin is regulated independently by developmental and exogenous signals to fine-tune suberin deposition in roots. We found a set of four MYB transcription factors (MYB41, MYB53, MYB92, and MYB93), each of which is individually regulated by these two signals and is sufficient to promote endodermal suberin. Mutation of these four transcription factors simultaneously through genome editing leads to a dramatic reduction in suberin formation in response to both developmental and environmental signals. Most suberin mutants analyzed at physiological levels are also affected in another endodermal barrier made of lignin (Casparian strips) through a compensatory mechanism. Through the functional analysis of these four MYBs, we generated plants allowing unbiased investigation of endodermal suberin function, without accounting for confounding effects due to Casparian strip defects, and were able to unravel specific roles of suberin in nutrient homeostasis

GDSL-domain proteins have key roles in suberin polymerization and degradation

Plant roots acquire nutrients and water while managing interactions with the soil microbiota. The root endodermis provides an extracellular diffusion barrier through a network of lignified cell walls called Casparian strips, supported by subsequent formation of suberin lamellae. Whereas lignification is thought to be irreversible, suberin lamellae display plasticity, which is crucial for root adaptative responses. Although suberin is a major plant polymer, fundamental aspects of its biosynthesis and turnover have remained obscure. Plants shape their root system via lateral root formation, an auxin-induced process requiring local breaking and re-sealing of endodermal lignin and suberin barriers. Here, we show that differentiated endodermal cells have a specific, auxin-mediated transcriptional response dominated by cell wall remodelling genes. We identified two sets of auxin-regulated GDSL lipases. One is required for suberin synthesis, while the other can drive suberin degradation. These enzymes have key roles in suberization, driving root suberin plasticity