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 /_publication/Chandrasekar[[:space:]]et[[:space:]]al.[[:space:]]-[[:space:]]2022[[:space:]]-[[:space:]]Fungi[[:space:]]hijack[[:space:]]a[[:space:]]ubiquitous[[:space:]]plant[[:space:]]apoplastic[[:space:]]endoglu.pdf filter=lfs diff=lfs merge=lfs -text
+_publication/koac114-suppl_data/tpc.21.00790_Supplemental[[:space:]]Data.pdf filter=lfs diff=lfs merge=lfs -text

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README.md

+# Fungi hijack a ubiquitous plant apoplastic endoglucanase to release a ROS scavenging β-glucan decasaccharide to subvert immune responses
+
+## Original Publication
+
+Balakumaran Chandrasekar, Alan Wanke, Stephan Wawra, Pia Saake, Lisa Mahdi, Nyasha Charura, Miriam Neidert, Gereon Poschmann, Milena Malisic, Meik Thiele, Kai Stühler, Murali Dama, Markus Pauly, Alga Zuccaro, Fungi hijack a ubiquitous plant apoplastic endoglucanase to release a ROS scavenging β-glucan decasaccharide to subvert immune responses, The Plant Cell, Volume 34, Issue 7, July 2022, Pages 2765–2784, https://doi.org/10.1093/plcell/koac114
+
+## Abstract
+
+Plant pathogenic and beneficial fungi have evolved several strategies to evade immunity and cope with host-derived hydrolytic enzymes and oxidative stress in the apoplast, the extracellular space of plant tissues. Fungal hyphae are surrounded by an inner insoluble cell wall layer and an outer soluble extracellular polysaccharide (EPS) matrix. Here, we show by proteomics and glycomics that these two layers have distinct protein and carbohydrate signatures, and hence likely have different biological functions. The barley (*Hordeum vulgare*) β-1,3-endoglucanase *Hv*BGLUII, which belongs to the widely distributed apoplastic glycoside hydrolase 17 family (GH17), releases a conserved β-1,3;1,6-glucan decasaccharide (β-GD) from the EPS matrices of fungi with different lifestyles and taxonomic positions. This low molecular weight β-GD does not activate plant immunity, is resilient to further enzymatic hydrolysis by β-1,3-endoglucanases due to the presence of three β-1,6-linked glucose branches and can scavenge reactive oxygen species. Exogenous application of β-GD leads to enhanced fungal colonization in barley, confirming its role in the fungal counter-defensive strategy to subvert host immunity. Our data highlight the hitherto undescribed capacity of this often-overlooked EPS matrix from plant-associated fungi to act as an outer protective barrier important for fungal accommodation within the hostile environment at the apoplastic plant–microbe interface.
+
+<img src="./_publication/koac114f6.jpeg" style="width:400px; margin: auto; padding: 30px 0px;">
+
+## License
+
+This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
+
+## Data availability
+
+***Hordeum vulgare* BGLU2**: P15737 (UniProt), HORVU.MOREX.r3.3HG0319100.1 (MorexV3_pseudomolecules_assembly, EnsemblePlants).
+
+[def]: oac114f6.jpe
\ No newline at end of file

_publication/README.md

+# Supplementary Data Information
+## Supplemental Figure S1. EPS matrix isolated from S. indica grown in CM medium using a cryogelation approach.
+
+## Supplemental Figure S2. Proteome analysis of S. indica EPS matrix, CW, and culture filtrate.
+
+## Supplemental Figure S3. Expression analysis of selected genes with WSC domains in S. indica during plant colonization at 3, 7, and 14 dpi.
+
+## Supplemental Figure S4. Glycosyl linkage analysis of Si EPS and Si CW.
+
+## Supplemental Figure S5. Analysis of oligosaccharides released from the EPS or protein-free CW of S. indica by the action of β-1,3-glucanases.
+
+## Supplemental Figure S6. Glycosyl linkage and MALDI-TOF analysis of the glucan fraction (β-GD) released from the EPS matrix of S. indica.
+
+## Supplemental Figure S7. Purification of the β-GD fragment for 1H NMR analysis.
+
+## Supplemental Figure S8. Mechanically released fragments from S. indica EPS matrix and CW layer do not exhibit ROS scavenging activity.
+
+## Supplemental Figure S9. Chitohexaose-triggered ROS accumulation is decreased by S. indica β-GD treatment in a concentration-dependent manner.
+
+## Supplemental Figure S10. Purification of native β-GD and immunogenic characterization.
+
+## Supplemental Figure S11. Detoxification of apoplastic ROS by S. indica β-GD is independent of elicitor treatment and plant species.
+
+## Supplemental Figure S12. Serendipita indica β-GD treatment does not trigger cytosolic calcium influx in A. thaliana seedlings.
+
+## Supplemental Figure S13. Digestion of laminariheptaose with HvBGLUII enhances ROS production in barley roots.
+
+## Supplemental Figure S14. Glycan controls are not degraded by hydrogen peroxide during Fenton reaction.
+
+## Supplemental Figure S15. Glycosyl linkage analysis of B. sorokiniana EPS matrix and CW.
+
+## Supplemental Figure S16. Analysis of oligosaccharides released from the EPS of B. sorokiniana by the action of Trichoderma harzianum lysing enzymes.
+
+## Supplemental Figure S17. The β-GD released from the EPS matrix of B. sorokiniana consists of a seven unit β-1,3-linked glucan backbone substituted with three β-1,6-glucosyl residues.
+
+## Supplemental Figure S18. Mechanically released fragments from B. sorokiniana EPS matrix or CW do not scavenge ROS.
+
+## Supplemental Data Set S1. Total proteins detected in the proteome analysis of the EPS matrix, CW, and culture filtrate isolated from S. indica grown in CM, TSB, or YPD liquid media.
+
+## Supplemental Data Set S2. Relative abundance of signal peptide (SP) containing proteins detected in the EPS matrix, CW, and culture filtrate isolated from the S. indica under three culture mediums: CM, TSB, and YPD. LFQ intensities of the proteins were used to calculate their relative abundance.
+
+## Supplemental Data Set S3. Statistics, enrichment score, Benjamini–Hochberg corrected P-value.
+
+## Supplemental Data Set S4. Glycosyl sugar residues detected in the total ion chromatogram of EPS matrix isolated from S. indica, n = 4 independent biological replicates.
+
+## Supplemental Data Set S5. Glycosyl sugar residues detected in the total ion chromatogram of alcohol insoluble residue (cell wall) isolated from S. indica, n = 4 independent biological replicates.
+
+## Supplemental Data Set S6. List of Barley CAZymes detected in the root apoplastic fluids (AFs) of barley after mock, S. indica or B. sorokiniana treatments.
+
+## Supplemental Data Set S7. Glycosyl sugar residues detected in the total ion chromatogram of alcohol insoluble residue (cell wall) isolated from B. sorokiniana, n = 3 independent biological replicates.
+
+## Supplemental Data Set S8. Glycosyl sugar residues detected in the total ion chromatogram of EPS matrix isolated from B. sorokiniana, n = 3 independent biological replicates.
+
+## Supplemental Data Set S9. List of Barley CAZymes differentially regulated during B. sorokiniana infection.
+
+## Supplemental Data Set S10. Summary of statistical analyses.
+
+https://academic.oup.com/plcell/article/34/7/2765/6571157#supplementary-data
\ No newline at end of file