title: Soil suppressiveness to fusarium disease: shifts in root microbiome associated with reduction of pathogen root colonization.
abstract: Soil suppressiveness to Fusarium disease was induced by incubating sandy soil with debris of wild rocket (WR; Diplotaxis tenuifolia) under field conditions. We studied microbial dynamics in the roots of cucumber seedlings following transplantation into WR-amended or nonamended soil, as influenced by inoculation with Fusarium oxysporum f. sp. radicis-cucumerinum. Disease symptoms initiated in nonamended soil 6 days after inoculation, compared with 14 days in WR-amended soil. Root infection by F. oxysporum f. sp. radicis-cucumerinum was quantified using real-time polymerase chain reaction (PCR). Target numbers were similar 3 days after inoculation for both WR-amended and nonamended soils, and were significantly lower (66%) 6 days after inoculation and transplanting into the suppressive (WR-amended) soil. This decrease in root colonization was correlated with a reduction in disease (60%) 21 days after inoculation and transplanting into the suppressive soil. Fungal community composition on cucumber roots was assessed using mass sequencing of fungal internal transcribed spacer gene fragments. Sequences related to F. oxysporum, Fusarium sp. 14005, Chaetomium sp. 15003, and an unclassified Ascomycota composed 96% of the total fungal sequences in all samples. The relative abundances of these major groups were highly affected by root inoculation with F. oxysporum f. sp. radicis-cucumerinum, with a 10-fold increase in F. oxysporum sequences, but were not affected by the WR amendment. Quantitative analysis and mass-sequencing methods indicated a qualitative shift in the root's bacterial community composition in suppressive soil, rather than a change in bacterial numbers. A sharp reduction in the size and root dominance of the Massilia population in suppressive soil was accompanied by a significant increase in the relative abundance of specific populations; namely, Rhizobium, Bacillus, Paenibacillus, and Streptomyces spp. Composition of the Streptomyces community shifted significantly, as determined by PCR denaturing gradient gel electrophoresis, resulting in an increase in the dominance of a specific population in suppressive soils after only 3 days. This shift was related mainly to the increase in Streptomyces humidus, a group previously described as antagonistic to phytopathogenic fungi. Thus, suitable soil amendment resulted in a shift in the root's bacterial communities, and infection by a virulent pathogen was contained by the root microbiome, leading to a reduced disease rate.
authors.affiliation: Department of Plant Pathology and Microbiology, The Hebrew University of Jerusalem, The Robert H. Smith Faculty of Agriculture, Food, and Environment, Rehovot 76100, Israel.
authors.forename: Eyal
authors.initials: E
authors.lastname: Klein
authors.forename: Maya
authors.initials: M
authors.lastname: Ofek
authors.forename: Jaacov
authors.initials: J
authors.lastname: Katan
authors.forename: Dror
authors.initials: D
authors.lastname: Minz
authors.forename: Abraham
authors.initials: A
authors.lastname: Gamliel
mesh_terms: D001203:Ascomycota / Q000235:genetics / Q000302:isolation & purification; D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification; D061046:Biological Control Agents; D019607:Brassicaceae / Q000737:chemistry; D018553:Cucumis sativus / Q000254:growth & development / Q000382:microbiology*; D016172:DNA Fingerprinting; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000737:chemistry / Q000235:genetics; D005670:Fusarium / Q000254:growth & development / Q000472:pathogenicity*; D054884:Host-Pathogen Interactions; D064307:Microbiota; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000382:microbiology*; D018515:Plant Leaves / Q000737:chemistry / Q000382:microbiology; D018517:Plant Roots / Q000382:microbiology; D018547:Plant Stems / Q000737:chemistry; D012331:RNA, Fungal / Q000737:chemistry / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D036226:Seedlings / Q000382:microbiology; D017422:Sequence Analysis, DNA; D012988:Soil Microbiology; D013302:Streptomyces / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D061046:Biological Control Agents; D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer; D012331:RNA, Fungal; D012336:RNA, Ribosomal, 16S