Visualizing polymeric components that define distinct root barriers across plant lineages

Sexauer, Moritz; Shen, Deyan; Schön, Maria; Andersen, Tonni Grube; Markmann, Katharina

doi:10.1242/dev.199820

Cited by 22 publications

(22 citation statements)

References 28 publications

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“…Root segments (first 25 mm from the root apex) of WT and HvMPK3 KO plants 24 h after inoculation with F. graminearum were used for suberin and lignin histochemical detection. Suberin and lignin staining was carried out according to a previously published protocol (Sexauer et al ., 2021; Ursache et al ., 2018). Freehand cross-sections of fixed and cleared root segments were made in the root region between 15 and 25 mm from the root apex and stained with 0,01% (v/v) Fluorol Yellow 088 in absolute ethanol (stock solution: 1% (w/v) Fluorol Yellow in DMSO) for 30 min.…”

Section: Methodsmentioning

confidence: 99%

Knockout of mitogen-activated protein kinase 3 causes barley root resistance against Fusarium graminearum

Jasim

Vadovič

Šamajová

et al. 2022

Preprint

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The roles of mitogen-activated protein kinases (MAPKs) in plant-fungal pathogenic interactions are less understood in crops. Here, microscopic, phenotyping, proteomic and biochemical analyses revealed that independent TALEN-based knockout lines of Hordeum vulgare MITOGEN-ACTIVATED PROTEIN KINASE 3 (HvMPK3 KO) were resistant against Fusarium graminearum infection. When co-cultured with roots of the HvMPK3 KO lines, F. graminearum hyphae were excluded to the extracellular space, the growth pattern of hyphae was considerably deregulated, mycelia development was less efficient and number of appressoria and their penetration potential were significantly reduced. Intracellular penetration of hyphae was preceded by the massive production of reactive oxygen species (ROS) in attacked cells of the wild type, but it was mitigated in the HvMPK3 KO lines. Suppression of ROS production in these lines coincided with the elevated abundances of catalase and ascorbate peroxidase. Moreover, differential proteomic analysis revealed downregulation of defense-related proteins in wild type, and the upregulation of peroxidases, lipid transfer proteins, and cysteine proteases in HvMPK3 KO lines after 24h of F. graminearum inoculation. Consistently with proteomic analysis, microscopic observations showed an enhanced suberin accumulation in roots of HvMPK3 KO lines, most likely contributing to the arrested infection by F. graminearum. These results suggest that TALEN-based knockout of HvMPK3 leads to the barley root resistance against Fusarium root rot.

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Section: Methodsmentioning

confidence: 99%

Knockout of mitogen-activated protein kinase 3 causes barley root resistance against Fusarium graminearum

Jasim

Vadovič

Šamajová

et al. 2022

Preprint

View full text Add to dashboard Cite

show abstract

“…Root segments (first 25 mm from the root apex) of WT and HvMPK3 KO plants 24 h after inoculation with F. graminearum were used for suberin and lignin histochemical detection. Suberin and lignin staining was carried out according to a previously published protocol ( Ursache et al, 2018 ; Sexauer et al, 2021 ). Freehand cross-sections of fixed and cleared root segments were made in the root region between 15 and 25 mm from the root apex and stained with 0.01% (v/v) Fluorol Yellow 088 in absolute ethanol (stock solution: 1% (w/v) Fluorol Yellow 088 in DMSO) for 30 min.…”

Section: Methodsmentioning

confidence: 99%

Knockout of MITOGEN-ACTIVATED PROTEIN KINASE 3 causes barley root resistance against Fusarium graminearum

et al. 2022

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The roles of mitogen-activated protein kinases (MAPKs) in plant-fungal pathogenic interactions are poorly understood in crops. Here, microscopic, phenotypic, proteomic and biochemical analyses revealed that roots of independent transcription activator-like effector nuclease (TALEN)-based knockout lines of barley (Hordeum vulgare L.) MITOGEN-ACTIVATED PROTEIN KINASE 3 (HvMPK3 KO) were resistant against Fusarium graminearum infection. When co-cultured with roots of the HvMPK3 KO lines, F. graminearum hyphae were excluded to the extracellular space, the growth pattern of extracellular hyphae was considerably deregulated, mycelia development was less efficient, and number of appressoria-like structures and their penetration potential were substantially reduced. Intracellular penetration of hyphae was preceded by the massive production of reactive oxygen species (ROS) in attacked cells of the wild type, but ROS production was mitigated in the HvMPK3 KO lines. Suppression of ROS production in these lines coincided with elevated abundance of catalase and ascorbate peroxidase. Moreover, differential proteomic analysis revealed downregulation of several defense-related proteins in wild type, and the upregulation of pathogenesis-related protein 1 (PR-1) and cysteine proteases in HvMPK3 KO lines. Proteins involved in suberin formation, such as peroxidases, lipid transfer proteins, and the GDSL esterase/lipase were differentially regulated in HvMPK3 KO lines after F. graminearum inoculation. Consistent with proteomic analysis, microscopic observations showed enhanced suberin accumulation in roots of HvMPK3 KO lines, most likely contributing to the arrested infection by F. graminearum. These results suggest that TALEN-based knockout of HvMPK3 leads to barley root resistance against Fusarium root rot.

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“…It was also robustly correlated with the expression of reporters based on suberin biosynthetic genes (Naseer et al, 2012;Barberon et al, 2016;Andersen et al, 2021). Very recently, FY staining has been applied to previously fixed and ClearSee-treated samples, allowing confocal imaging of whole-mount sections while preserving the fluorescence of genetically encoded fluorophores, such as GFP, and co-staining with other compatible dyes such as Basic Fuchsin for lignin or Calcofluor White for polysaccharides (Kurihara et al, 2015;Ursache et al, 2018;Sexauer et al, 2021). Another fluorescent suberin dye, Nile Red, is also compatible with ClearSee, but appears to only stain older, more mature suberin in the Arabidopsis endodermis, compared with FY.…”

Section: In Situ Visualisation Techniquesmentioning

confidence: 99%

The making of suberin

Serra

Geldner

2022

New Phytologist

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Summary Outer protective barriers of animals use a variety of bio‐polymers, based on either proteins (e.g. collagens), or modified sugars (e.g. chitin). Plants, however, have come up with a particular solution, based on the polymerisation of lipid‐like precursors, giving rise to cutin and suberin. Suberin is a structural lipophilic polyester of fatty acids, glycerol and some aromatics found in cell walls of phellem, endodermis, exodermis, wound tissues, abscission zones, bundle sheath and other tissues. It deposits as a hydrophobic layer between the (ligno)cellulosic primary cell wall and plasma membrane. Suberin is highly protective against biotic and abiotic stresses, shows great developmental plasticity and its chemically recalcitrant nature might assist the sequestration of atmospheric carbon by plants. The aim of this review is to integrate the rapidly accelerating genetic and cell biological discoveries of recent years with the important chemical and structural contributions obtained from very diverse organisms and tissue layers. We critically discuss the order and localisation of the enzymatic machinery synthesising the presumed substrates for export and apoplastic polymerisation. We attempt to explain observed suberin linkages by diverse enzyme activities and discuss the spatiotemporal relationship of suberin with lignin and ferulates, necessary to produce a functional suberised cell wall.

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Visualizing polymeric components that define distinct root barriers across plant lineages

Cited by 22 publications

References 28 publications

Knockout of mitogen-activated protein kinase 3 causes barley root resistance against Fusarium graminearum

Knockout of mitogen-activated protein kinase 3 causes barley root resistance against Fusarium graminearum

Knockout of MITOGEN-ACTIVATED PROTEIN KINASE 3 causes barley root resistance against Fusarium graminearum

The making of suberin

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