diff --git a/assays/EnzymaticCarbohydrateDigestionAndTLC/README.md b/assays/EnzymaticCarbohydrateDigestionAndTLC/README.md new file mode 100644 index 0000000000000000000000000000000000000000..e69de29bb2d1d6434b8b29ae775ad8c2e48c5391 diff --git a/assays/EnzymaticCarbohydrateDigestionAndTLC/dataset/.gitkeep b/assays/EnzymaticCarbohydrateDigestionAndTLC/dataset/.gitkeep new file mode 100644 index 0000000000000000000000000000000000000000..e69de29bb2d1d6434b8b29ae775ad8c2e48c5391 diff --git a/assays/EnzymaticCarbohydrateDigestionAndTLC/isa.assay.xlsx b/assays/EnzymaticCarbohydrateDigestionAndTLC/isa.assay.xlsx new file mode 100644 index 0000000000000000000000000000000000000000..36979dca791bbb7c1c35361a9de89ed99823bbd8 Binary files /dev/null and b/assays/EnzymaticCarbohydrateDigestionAndTLC/isa.assay.xlsx differ diff --git a/assays/EnzymaticCarbohydrateDigestionAndTLC/protocols/.gitkeep b/assays/EnzymaticCarbohydrateDigestionAndTLC/protocols/.gitkeep new file mode 100644 index 0000000000000000000000000000000000000000..e69de29bb2d1d6434b8b29ae775ad8c2e48c5391 diff --git a/assays/EnzymaticCarbohydrateDigestionAndTLC/protocols/EnzymaticCarbohydrateDigestion.md b/assays/EnzymaticCarbohydrateDigestionAndTLC/protocols/EnzymaticCarbohydrateDigestion.md new file mode 100644 index 0000000000000000000000000000000000000000..7b4ecbf233cc690c26a2de3c1be54b8bbed97cd3 --- /dev/null +++ b/assays/EnzymaticCarbohydrateDigestionAndTLC/protocols/EnzymaticCarbohydrateDigestion.md @@ -0,0 +1,2 @@ +## Enzymatic carbohydrate digestion +Carbohydrate digestion assays were performed using either purified barley GBP (heterologously expressed in *N. benthamiana*) or barley BGLUII33 (available from Megazyme, E-LAMHV). Preparations of the fungal CW and EPS matrix were incubated overnight in sterile Milli-Q water at 65 °C prior to enzymatic digestion. Substrate and enzyme concentrations, buffer compositions, digestion temperature and time are described in the figure captions. Digestion was stopped by denaturing the enzymes at 95 °C for 10 min and the digestion products were stored at -20 °C prior to use. \ No newline at end of file diff --git a/assays/EnzymaticCarbohydrateDigestionAndTLC/protocols/TLC.md b/assays/EnzymaticCarbohydrateDigestionAndTLC/protocols/TLC.md new file mode 100644 index 0000000000000000000000000000000000000000..a700062235e42e7d54290ecb0b663347a9b991d9 --- /dev/null +++ b/assays/EnzymaticCarbohydrateDigestionAndTLC/protocols/TLC.md @@ -0,0 +1,2 @@ +## Thin layer chromatography (TLC) +An aliquot of each sample was subjected to TLC using a silica gel 60 F254 aluminum TLC plate (Merck Millipore, Burlington, USA), using a running buffer containing ethyl acetate/acetic acid/methanol/formic acid/water at a ratio of 8:4:1:1:1 (v/v). D-glucose, laminaribiose β-1-3-(Glc)2, laminaritriose β-1-3-(Glc)3, gentiobiose β-1-6-(Glc)2, and laminaripentaose β-1-3-(Glc)5 at a concentration of 1.5 mg·mL−1 were used as standards (Megazyme, Bray, Ireland). To visualize the glucan fragments, the TLC plate was sprayed with glucan developer solution (45 mg N-naphthol, 4.8 mL H2SO4, 37.2 mL ethanol and 3 mL water) and baked at 95 °C until the glucan bands became visible (approximately 4-5 min). \ No newline at end of file diff --git a/assays/MS-MSAnalysisOfThePull-downProteins/isa.assay.xlsx b/assays/MS-MSAnalysisOfThePull-downProteins/isa.assay.xlsx index 5da3929640b717f566d608a88f5707a5e1de42af..44163a2c0585bc7c95f806f4fc10c0278f5f6c06 100644 Binary files a/assays/MS-MSAnalysisOfThePull-downProteins/isa.assay.xlsx and b/assays/MS-MSAnalysisOfThePull-downProteins/isa.assay.xlsx differ diff --git a/assays/OxidativeBurstAssay/isa.assay.xlsx b/assays/OxidativeBurstAssay/isa.assay.xlsx index 079eac191b001fc238d086b7f8a5b0bdfb056aa3..d541a79ab966c00422257b31faf24c6420cd1f69 100644 Binary files a/assays/OxidativeBurstAssay/isa.assay.xlsx and b/assays/OxidativeBurstAssay/isa.assay.xlsx differ diff --git a/isa.investigation.xlsx b/isa.investigation.xlsx index 1c5b25406e0b0196214bd30d95f23270918796e0..c182d3e5edbd8c67554fa7b660b50bb64ce0dfce 100644 Binary files a/isa.investigation.xlsx and b/isa.investigation.xlsx differ diff --git a/studies/Barley/isa.study.xlsx b/studies/Barley/isa.study.xlsx index dfe27555ffe70166472148c968e6ae60385cfc29..d02d2cfe355ec6fefb9695137fd7c89cbb644bf1 100644 Binary files a/studies/Barley/isa.study.xlsx and b/studies/Barley/isa.study.xlsx differ diff --git a/studies/FungalMaterialGrowthConditionsAndBarleyColonizationAssays/isa.study.xlsx b/studies/FungalMaterialGrowthConditionsAndBarleyColonizationAssays/isa.study.xlsx index 9746ba5b1312c8b45bb946138b741920ac29cf1e..c6c19e65eb862ae646b4a80b7b296fb77b3b7186 100644 Binary files a/studies/FungalMaterialGrowthConditionsAndBarleyColonizationAssays/isa.study.xlsx and b/studies/FungalMaterialGrowthConditionsAndBarleyColonizationAssays/isa.study.xlsx differ diff --git a/studies/PlantMaterialAndGrowthConditions/isa.study.xlsx b/studies/PlantMaterialAndGrowthConditions/isa.study.xlsx index 5e88e58d545a8b38164e29ebc0987f28c78db3ef..844fbcdb8a80d8e76cff3906aee4b3b44d18d985 100644 Binary files a/studies/PlantMaterialAndGrowthConditions/isa.study.xlsx and b/studies/PlantMaterialAndGrowthConditions/isa.study.xlsx differ