Germida, J.J.

Walley, F.L.

Wijesinghe, M.A.K.

English

eCommons@USASK

Root-associated bacterial communities in flax (Linum usitatissimum) and their response to arbuscular mycorrhizal (AM) inoculation

University of Saskatchewan Research Archive

Root- associated Bacterial Communities inFlax (Linum usitatissimum) and Their Response to Arbuscular Mycorrhizal (AM) InoculationM.A.K. Wijesinghe, J.J. Germida & F.L. WalleyDepartment of Soil SciencesUniversity of Saskatchewan   SK   CanadaSustainable Agriculture for Increasing World Population (http://www.fewresources.org)Plant-Associated Microbes in Sustainable Agriculture• More attention to Endophytes- Superior plant growth promotion- Subset of rhizosphere microbes- Simple - Great potential to aid sustainable agriculture (source: Ye et al., 2014)Flax as a Healthy Food•  A functional food  - Poly unsaturated fatty acids (~75%)- Alpha linolenic acid (ALA) (~ 57%) - Dietary fibre and lignans- Health benefits• Lead producer• Mainly grown inCanadian prairies• More flax from SKFlax plant is sensitive to chemical fertilizationGrowth Chamber ExperimentObjectives1. Endophytic and rhizo-Bacterial communities associated with flax2.      Impact of AM inoculation on bacterial communitiesBrief Methodology• Three different soils- Central Butte- Allen- Kelvington• Two rates of AM inoculation- Control (C)- Recommended rate (X) - 4 times recommended rate (4X)Brief Methodology (cont.)Sampling of Rhizosphere Soil and Roots Surface Sterilization of RootsDNA Extraction Amplicon PreparationIllumina SequencingData AnalysisPERMANOVA Results for Bacterial Community Composition Associated with FlaxSource d.f. F value P valueHabitat (Rhizosphere/ Endosphere) 1 326.43927 0.0002*Soil 2 9.97255 0.0006*AMF inoculation 2 0.42485   0.7556Habitat X Soil 2 10.74913 0.0006*Habitat X AMF inoculation 2 0.35087 0.8168Soil X AMF inoculation 4 0.8691 0.9978Bacterial Community Composition of Flax inRhizosphere and Root EndosphereStress 2 (16.96 %)Stress 1  (50.2%)F-Root-Cb - contF-Root-Cb - XF-Root-Cb - 4XF-Root-Al - contF-Root-Al - XF-Root-Al - 4XF-Root-Kel - contF-Root-Kel- XF-Root-Kel - 4XF-Rhiz-Cb - contF-Rhiz-Cb - XF-Rhiz-Cb - 4XF-Rhiz-Al - contF-Rhiz-Al - XF-Rhiz-Al - 4XF-Rhiz-Kel - contF-Rhiz-Kel- XF-Rhiz-Kel - 4XDistribution of Bacterial Genera among Rhizosphere and Root Endosphere in FlaxRhizosphere213262Endosphere41Relative Distribution of Dominant Bacterial Phyla associated with Flax Rhizosphere00.10.20.30.40.50.60.70.80.91Cont X 4X Cont X 4X Cont X 4XRelative Distribution of Bacterial PhylaTreatmentsVerrucomicrobiaProteobacteriaPlanctomycetesNitrospiraeGemmatimonadetesFirmicutesChloroflexiBacteroidetesArmatimonadetesActinobacteriaAcidobacteriaRhizosphere Rhizosphere RhizosphereAllen Central Butte KelvingtonRelative Distribution of Dominant Bacterial Phyla associated with Flax Rhizosphere and Endosphere00.10.20.30.40.50.60.70.80.91Cont X 4X Cont X 4X Cont X 4X Cont X 4X Cont X 4X Cont X 4XRelative Distribution of Bacterial PhylaTreatmentsVerrucomicrobiaProteobacteriaPlanctomycetesNitrospiraeGemmatimonadetesFirmicutesChloroflexiBacteroidetesArmatimonadetesActinobacteriaAcidobacteriaRhizosphere Roots Roots RootsRhizosphere RhizosphereAllen Central Butte KelvingtonRelative Distribution of Dominant ProteobacterialOrders associated with Flax Rhizosphere00.10.20.30.40.50.60.70.80.91Cont  X 4x Cont  X 4x Cont  X 4xRelative Distribution of ProteobacterialOrdersTreatmentsOtherXanthomonadalesPseudomonadalesLegionellalesEnterobacterialesAlteromonadalesMyxococcalesRhodocyclalesNitrosomonadalesMethylophilalesBurkholderialesSphingomonadalesRickettsialesRhodospirillalesRhodobacteralesRhizobialesCaulobacteralesRhizosphereRhizosphere RhizosphereCentral Butte KelvingtonAllenRelative distribution of Dominant Proteobacterial Orders associated with Flax Rhizozphere and Endosphere00.10.20.30.40.50.60.70.80.91Cont  X 4x Cont  X 4x Cont  X 4x Cont  X 4x Cont  X 4x Cont  X 4xRelative Distribution of ProteobacterialOrdersTreatmentsOtherXanthomonadalesPseudomonadalesLegionellalesEnterobacterialesAlteromonadalesMyxococcalesRhodocyclalesNitrosomonadalesMethylophilalesBurkholderialesSphingomonadalesRickettsialesRhodospirillalesRhodobacteralesRhizobialesCaulobacteralesRhizosphereRhizosphere RhizosphereRoots Roots RootsCentral Butte KelvingtonAllenDiversity and Richness of Rhizo- and Root Endophytic- BacteriaTreatments Shannon Diversity Chao richness SOBsSoil AM Inoculation Rhizo * Endo * Rhizo * Endo * Rhizo * Endo *C. Butte Control 8.3  AB                4.9 C 1240.8 ABC 575.3 D 499.3 A 152.3 EFGH1X 8.4  AB 3.8 DE 1156.7 ABC 366.7 DEFG 509.3 AB 211.7 DEF4X 8.3  AB 4.75 CD 1119.7 BC 571.2 D 490.6 A 235 CDEAllen Control 8.35  AB 4.5 CDE 1182.1 ABC 370.8 DEF 496 AB 189.3 DEFG1X 8.34  AB 4.8 CD 1212.5 ABC 427.4 D 493.3 ABC 143.7 EFGH4X 8.3  AB 3.8 DE 1224.1 ABC 394.5 DE 496 ABC 172 DEFGHKelvington Control 8.5  A 3.8 DE 1436.2 A 402.3 DE 541 AB 138.7 EFGH1X 8.5  AB 3.7 E 1309.4 AB 349 DEFGH 524.7 BCD 142 EFGH4X 8.6  A 3.5 E 1319.3 AB 457 D 544.3 A 162.7 DEFGHTake Home Message• Flax accommodate 475 and 303 bacterial genera in their rhizosphere and root endosphere, respectively• Root endophytic-bacterial community of flax is a simple and non-random subset of rhizo-bacterial community• Proteobacteria are more dominant among bacterial root endophytes in flax• Commercial AM inoculation dose not affect community composition and diversity of bacteria associated with flax • Endospheric bacteria will be isolated and evaluated for potential benefitsAcknowledgement• Technical support from Flath Ben and Amanda Bruce

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Root-associated bacterial communities in flax (Linum usitatissimum) and their response to arbuscular mycorrhizal (AM) inoculation

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