Microfluidics and Metabolomics Reveal Symbiotic Bacterial–Fungal Interactions Between Mortierella elongata and Burkholderia Include Metabolite Exchange

We identified two poplar (Populus sp.)-associated microbes, the fungus, Mortierella elongata strain AG77, and the bacterium, Burkholderia strain BT03, that mutually promote each other’s growth. Using culture assays in concert with a novel microfluidic device to generate time-lapse videos, we found growth specific media differing in pH and pre-conditioned by microbial growth led to increased fungal and bacterial growth rates. Coupling microfluidics and comparative metabolomics data results indicated that observed microbial growth stimulation involves metabolic exchange during two ordered events. The first is an emission of fungal metabolites, including organic acids used or modified by bacteria. A second signal of unknown nature is produced by bacteria which increases fungal growth rates. We find this symbiosis is initiated in part by metabolic exchange involving fungal organic acids.

[1]  Claire E. Stanley,et al.  Combining microfluidics and RNA-sequencing to assess the inducible defensome of a mushroom against nematodes , 2019, BMC Genomics.

[2]  Joseph M. Martel-Foley,et al.  Rapid Isolation and Concentration of Pathogenic Fungi Using Inertial Focusing on a Chip-Based Platform , 2019, Front. Cell. Infect. Microbiol..

[3]  Claire E. Stanley,et al.  Bidirectional Propagation of Signals and Nutrients in Fungal Networks via Specialized Hyphae , 2019, Current Biology.

[4]  S. Hacquard,et al.  Bacterial-fungal interactions: ecology, mechanisms and challenges. , 2018, FEMS microbiology reviews.

[5]  Hui Sun,et al.  Bacterial endosymbionts influence host sexuality and reveal reproductive genes of early divergent fungi , 2017, Nature Communications.

[6]  C. Schadt,et al.  Comparative genomics of Mortierella elongata and its bacterial endosymbiont Mycoavidus cysteinexigens , 2017, Environmental microbiology.

[7]  A. Aharoni,et al.  Live imaging of root–bacteria interactions in a microfluidics setup , 2017, Proceedings of the National Academy of Sciences.

[8]  S. Henry,et al.  Lipid metabolic changes in an early divergent fungus govern the establishment of a mutualistic symbiosis with endobacteria , 2016, Proceedings of the National Academy of Sciences.

[9]  P. Dorrestein,et al.  Role of secondary metabolites in the interaction between Pseudomonas fluorescens and soil microorganisms under iron-limited conditions. , 2016, FEMS microbiology ecology.

[10]  H. Ohta,et al.  Mycoavidus cysteinexigens gen. nov., sp. nov., an endohyphal bacterium isolated from a soil isolate of the fungus Mortierella elongata. , 2016, International journal of systematic and evolutionary microbiology.

[11]  I. Nookaew,et al.  Two Poplar-Associated Bacterial Isolates Induce Additive Favorable Responses in a Constructed Plant-Microbiome System , 2016, Front. Plant Sci..

[12]  M. Newman,et al.  Fungal Innate Immunity Induced by Bacterial Microbe-Associated Molecular Patterns (MAMPs) , 2016, G3: Genes, Genomes, Genetics.

[13]  Gayathri Ilangumaran,et al.  Inter-organismal signaling and management of the phytomicrobiome , 2015, Front. Plant Sci..

[14]  P. Bagnaresi,et al.  Symbiosis with an endobacterium increases the fitness of a mycorrhizal fungus, raising its bioenergetic potential , 2015, The ISME Journal.

[15]  D. Zühlke,et al.  Molecular mechanisms underlying the close association between soil Burkholderia and fungi , 2015, The ISME Journal.

[16]  Ruth D Gates,et al.  The coral core microbiome identifies rare bacterial taxa as ubiquitous endosymbionts , 2015, The ISME Journal.

[17]  L J Millet,et al.  Modular microfluidics for point-of-care protein purifications. , 2015, Lab on a chip.

[18]  M. Hattori,et al.  Draft Genome Sequence of the Betaproteobacterial Endosymbiont Associated with the Fungus Mortierella elongata FMR23-6 , 2014, Genome Announcements.

[19]  D. Weston,et al.  Newly identified helper bacteria stimulate ectomycorrhizal formation in Populus , 2014, Front. Plant Sci..

[20]  Claire E. Stanley,et al.  Probing bacterial-fungal interactions at the single cell level. , 2014, Integrative biology : quantitative biosciences from nano to macro.

[21]  N. Moran,et al.  The tiniest tiny genomes. , 2014, Annual review of microbiology.

[22]  C. Schadt,et al.  Plant host and soil origin influence fungal and bacterial assemblages in the roots of woody plants , 2014, Molecular ecology.

[23]  Yves Moné,et al.  The oxidative environment: a mediator of interspecies communication that drives symbiosis evolution , 2014, Proceedings of the Royal Society B: Biological Sciences.

[24]  Shaoli Wang,et al.  The Endosymbiont Hamiltonella Increases the Growth Rate of Its Host Bemisia tabaci during Periods of Nutritional Stress , 2014, PloS one.

[25]  B. Wolfe,et al.  Towards an Ecosystem Approach to Cheese Microbiology. , 2013, Microbiology spectrum.

[26]  C. Schadt,et al.  A Multifactor Analysis of Fungal and Bacterial Community Structure in the Root Microbiome of Mature Populus deltoides Trees , 2013, PloS one.

[27]  J. Caplan,et al.  Transcriptomics of the Rice Blast Fungus Magnaporthe oryzae in Response to the Bacterial Antagonist Lysobacter enzymogenes Reveals Candidate Fungal Defense Response Genes , 2013, PloS one.

[28]  U. Paszkowski,et al.  Polyphony in the rhizosphere: presymbiotic communication in arbuscular mycorrhizal symbiosis. , 2013, Current opinion in plant biology.

[29]  D. Beebe,et al.  Low-Volume Toolbox for the Discovery of Immunosuppressive Fungal Secondary Metabolites , 2013, PLoS pathogens.

[30]  M. Packirisamy,et al.  TipChip: a modular, MEMS-based platform for experimentation and phenotyping of tip-growing cells. , 2013, The Plant journal : for cell and molecular biology.

[31]  G. Bending,et al.  Increased hyphal branching and growth of ectomycorrhizal fungus Lactarius rufus by the helper bacterium Paenibacillus sp , 2013, Mycorrhiza.

[32]  Jerry M. Parks,et al.  Down-regulation of the caffeic acid O-methyltransferase gene in switchgrass reveals a novel monolignol analog , 2012, Biotechnology for Biofuels.

[33]  T. Pawlowska,et al.  EVOLUTIONARY STABILITY IN A 400‐MILLION‐YEAR‐OLD HERITABLE FACULTATIVE MUTUALISM , 2012, Evolution; international journal of organic evolution.

[34]  J. Liao,et al.  Combined inactivation of the Clostridium cellulolyticum lactate and malate dehydrogenase genes substantially increases ethanol yield from cellulose and switchgrass fermentations , 2012, Biotechnology for Biofuels.

[35]  A. Deveau,et al.  Bacterial-Fungal Interactions: Hyphens between Agricultural, Clinical, Environmental, and Food Microbiologists , 2011, Microbiology and Molecular Reviews.

[36]  S. Cruveiller,et al.  The genome of the obligate endobacterium of an AM fungus reveals an interphylum network of nutritional interactions , 2011, The ISME Journal.

[37]  W. Boer,et al.  Dual transcriptional profiling of a bacterial/fungal confrontation: Collimonas fungivorans versus Aspergillus niger , 2011, The ISME Journal.

[38]  S. Tringe,et al.  Metagenomic Discovery of Biomass-Degrading Genes and Genomes from Cow Rumen , 2011, Science.

[39]  K. Yamaguchi,et al.  Detection of betaproteobacteria inside the mycelium of the fungus Mortierella elongata. , 2010, Microbes and environments.

[40]  Dan V. Nicolau,et al.  Microfluidics structures for probing the dynamic behaviour of filamentous fungi , 2010 .

[41]  T. Pawlowska,et al.  Molecular evolution in bacterial endosymbionts of fungi. , 2010, Molecular biology and evolution.

[42]  G. Muyzer,et al.  Application of bacteria as self-healing agent for the development of sustainable concrete , 2010 .

[43]  Wolfgang Schmidt-Heck,et al.  Intimate bacterial–fungal interaction triggers biosynthesis of archetypal polyketides in Aspergillus nidulans , 2009, Proceedings of the National Academy of Sciences.

[44]  Qing Nie,et al.  Robust Spatial Sensing of Mating Pheromone Gradients by Yeast Cells , 2008, PloS one.

[45]  T. Coenye,et al.  Diversity and occurrence of Burkholderia spp. in the natural environment. , 2008, FEMS microbiology reviews.

[46]  P. Frey-Klett,et al.  The mycorrhiza helper bacteria revisited. , 2007, The New phytologist.

[47]  J. Pierrat,et al.  The mycorrhiza helper Pseudomonas fluorescens BBc6R8 has a specific priming effect on the growth, morphology and gene expression of the ectomycorrhizal fungus Laccaria bicolor S238N. , 2007, The New phytologist.

[48]  K. L. Hanson,et al.  Fungi use efficient algorithms for the exploration of microfluidic networks. , 2006, Small.

[49]  J. Guillamón,et al.  Real-Time Quantitative PCR (QPCR) and Reverse Transcription-QPCR for Detection and Enumeration of Total Yeasts in Wine , 2006, Applied and Environmental Microbiology.

[50]  N. Gerardo,et al.  Symbiosis and Insect Diversification: an Ancient Symbiont of Sap-Feeding Insects from the Bacterial Phylum Bacteroidetes , 2005, Applied and Environmental Microbiology.

[51]  J. Wernegreen,et al.  Genome evolution in bacterial endosymbionts of insects , 2002, Nature Reviews Genetics.

[52]  R. Kolter,et al.  Pseudomonas-Candida Interactions: An Ecological Role for Virulence Factors , 2002, Science.

[53]  R. Ibrahim,et al.  Aldonic Acids: A Novel Family of nod Gene Inducers of Mesorhizobium loti, Rhizobium lupini, and Sinorhizobium meliloti , 1998 .

[54]  M. Selosse,et al.  Variations in symbiotic efficiency, phenotypic characters and ploidy level among different isolates of the ectomycorrhizal basidiomycete Laccaria bicolor strain S 238 , 1996 .

[55]  C. Bandi,et al.  An obligately endosymbiotic mycorrhizal fungus itself harbors obligately intracellular bacteria , 1996, Applied and environmental microbiology.

[56]  M. Gessner,et al.  Extraction and quantification of ergosterol as a measure of fungal biomass in leaf litter , 1991, Microbial Ecology.

[57]  H A Isack,et al.  Honeyguides and Honey Gatherers: Interspecific Communication in a Symbiotic Relationship , 1989, Science.

[58]  A. Kondorosi,et al.  Rhizobium fix genes mediate at least two communication steps in symbiotic nodule development , 1988, The Journal of cell biology.

[59]  A P Trinci,et al.  A kinetic study of the growth of Aspergillus nidulans and other fungi. , 1969, Journal of general microbiology.

[60]  D. E. Contois Kinetics of bacterial growth: relationship between population density and specific growth rate of continuous cultures. , 1959, Journal of general microbiology.

[61]  F. Martin,et al.  Pairwise Transcriptomic Analysis of the Interactions Between the Ectomycorrhizal Fungus Laccaria bicolor S238N and Three Beneficial, Neutral and Antagonistic Soil Bacteria , 2014, Microbial Ecology.

[62]  P. Frey-Klett,et al.  Mycorrhiza Helper Bacteria , 2008 .

[63]  U. Nehls,et al.  Sugar for my honey: carbohydrate partitioning in ectomycorrhizal symbiosis. , 2007, Phytochemistry.

[64]  Robert Saftner,et al.  Relationship Between Host Acidification and Virulence of Penicillium spp. on Apple and Citrus Fruit. , 2004, Phytopathology.

[65]  Michael D. Abràmoff,et al.  Image processing with ImageJ , 2004 .

[66]  Andy F. S. Taylor,et al.  Mycelial growth and substrate acidification of ectomycorrhizal fungi in response to different minerals. , 2004, FEMS microbiology ecology.

[67]  Peter Vandamme,et al.  'Candidatus glomeribacter gigasporarum' gen. nov., sp. nov., an endosymbiont of arbuscular mycorrhizal fungi. , 2003, International journal of systematic and evolutionary microbiology.

[68]  B. Lang,et al.  Mitochondrial evolution. , 1999, Science.

[69]  B. Lang,et al.  Mitochondrial Evolution , 1999 .

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