A GH81-type β-glucan-binding protein enhances colonization by mutualistic fungi in barley

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flowchart

subgraph Legend
l1(Sample)
l2>Process]
l3(Material)
l4(Data)
end

subgraph studies\NicotianaBenthamiana
direction TB
s1(wild type seeds)---p1>PlantMaterialAndGrowthConditions.md]-->s2(N. benthamiana plants)
end

subgraph studies\HordeumVulgare
direction TB
s3(barley seeds)---p2>PlantMaterialAndGrowthConditions.md]-->s4(barley plants)
end

subgraph studies\CarbohydrateSubstratesForImmunityAndEnzymaticDigestionAssays
direction TB
p3>CarbohydrateSubstratesForImmunityAndEnzymaticDigestionAssays.md]-->s5(carbohydrate substrates)
end

subgraph assays\HeterologousProteinProductionAndPurification
direction TB
a1(codon-optimized GBP1 sequence)---p4>PlasmidConstruction.md]-->a2(pXCScpmv-GBP1-HAStrep)
a3(catalitically inactive GBP1 sequence)---p4-->a4(pXCScpmv-GBP1_E500A-HAStrep)
a2----p6>"`isolation
        sequencing
        _Agrobacterium tumefaciens_ transformation`"]
a4---p5>"`gel electrophoresis
        purification
        _E.coli_ transformation`"]---p6
p6-->a5(Agrobacterium_GBP1)
p6-->a6(Agrobacterium_GBP1_E500A)
a5 & a6---p7>infiltration]
s2----p7
p7---p8>HeterologousProteinProductionAndPurification.md]-->a7(purified proteins)
end

subgraph assays\EnzymaticCarbohydrateDigestionAndTLC
direction TB
a7 & s5---p9>EnzymaticCarbohydrateDigestion.md]-->a8(digestion products)
a8---p10>TLC.md]-->d1("`gr3b_lrg.jpg
                    gr4a_lrg.jpg
                    gr4b_lrg.jpg
                    gr4c_lrg.jpg
                    FigS4A.jpg
                    FigS4B.jpg
                    FigS5A.png`")
end

subgraph assays\MALDI-TOFAnalysis
direction TB
a8---p11>MALDI-TOFAnalysis.md]-->d2("`gr3c_large.jpg
FigS4C.jpg`")
end

subgraph assays\CRISPRCas9-basedMutagenesis
direction TB
s4---p12>CRISPRCas9-basedMutagenesis.md]-->a9(barley mutants)-->d15(gr1b_lrg.jpg)
end

subgraph assays\OxidativeBurstAssay
direction TB
s2 & s4 & a9---p13>PreparationOfThePlantMaterial.md]-->a10(plant material)
a10---p14>ROSBurstAssay.md]-->d3("`gr5_lrg.jpg
                                FigS1A.jpg
                                FigS1B.jpg
                                FigS5B.jpg
                                FigS5C.jpg`")
end

subgraph assays\FungalMaterialGrowthConditionsAndBarleyColonizationAssays
direction TB
s4 & a9---p15>RhizophagusIrregularis.md]-->a11(R. irregularis colonized plants)-->d11(FigS3B.jpg)
s4 & a9---p16>BlumeriaHordei.md]-->a12(B. hordei colonized plants)-->d13(FigS3D.jpg)
s4 & a9---p17>SerendipitaIndicaSerendipitaVermiferaAndBipolarisSorokiniana.md]-->a13(S. indica colonized plants) & a14(S. vermifera colonized plants) & a15(B. sorokiniana colonized plants)
a15-->d12(FigS3C.jpg)
d14(gr6b_lrg.jpg)
end

subgraph assays\qRT-PCR
direction TB
a12 & a13 & a14 & a15---p18>qRT-PCR.md]-->d4("`gr1c_lrg.jpg
gr2_lrg.jpg
gr6d_lrg.jpg`")
end

subgraph assays\StainingForConfocalMicroscopy
direction TB
a13---p19>HordeumVulgare.md]-->d5("`gr6a_lrg.jpg
                                FigS6A.jpg
                                FigS6B.jpg
                                FigS6C.jpg`")
a11---p20>RootStainingOfRIrregularis.md]-->d6(FigS3A.jpg)
end

subgraph assays\ProteinPull-down
direction TB
a16(laminarin)---p21>BiotinylationOfLaminarin.md]-->a17(biotinylated laminarin)
s4 & a17---p22>ProteinPull-downWithBiotinylatedLaminarin.md]-->a18(pull-down proteins)
a18---p23>SDS-PAGE separation]-->d7(FigS1C.jpg)
end

subgraph assays\MS-MSAnalysisOfThePull-downProteins
direction TB
a18---p24>MS-MSAnalysisOfThePull-downProteins.md]-->d8("`gr1a_lrg.jpg
         mmc2.xlsx`")
end

subgraph assays\MultipleSequenceAlignment
d9(gr4a_lrg.jpg)
end

subgraph assays\QuantificationAndStatisticalAnalysis
direction TB
pr25>QuantificationAndStatisticalAnalysis.md]-->d10("`gr6c_lrg.jpg
        mmc3.xlsx`")
end

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