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About Frankia Literature Review. Suzanna Kruger Biology 523 Microbial Ecology Dr. Anna-Louise Reysenbach 3 June 2003. Overview. Frankia taxonomy Host species Role in nitrogen cycle Methods of isolation Anatomy of nodule formation Anatomy and Metabolism Ecological relationships. - PowerPoint PPT Presentation

Text of About Frankia Literature Review

  • About Frankia Literature ReviewSuzanna KrugerBiology 523 Microbial EcologyDr. Anna-Louise Reysenbach3 June 2003

  • OverviewFrankia taxonomyHost speciesRole in nitrogen cycleMethods of isolationAnatomy of nodule formationAnatomy and MetabolismEcological relationships

  • Frankia TaxonomyFilamentous, High GC, Gram-Positive Bacteria

    Group III ActinomyceteNitrogen-fixing symbiont of plants(Madigan, 2003)http://www.msu.edu/~batzli/whats_frankia.htmlhttp://helios.bto.ed.ac.uk/bto/microbes/microbes.htm#Microorganisms%20and%20the%20"Tree%20of%20Life"

  • Host SpeciesHosts are non-leguminous, woody angiosperms

    8 Families, 23 genera including:Alnus (Betulacaceae)Ceanothus (Rhamnaceae)Rubus (Rosaceae) http://www.msu.edu/~batzli/graphics/alder_forest.jpgRed Alder Forest, Washington

  • Role in Nitrogen Cycle

    (Madigan, 2003)Nitrogen fixation:N2 + 8H+ + 8e- + 16 ATP = 2NH3 + H2 + 16ADP + 16 Pi

    Catalyzed by nitrogenaseReasons for interest inFrankia are related to theNitrogen Cycle:

    Forestry management Land reclamation Soil improvment

  • Methods of Isolation and Culture of FrankiaISOLATION of:First isolated 1978 (Cervantes and Rodrguez-Barrueco, 1992)Sterilization, mechanical disruption, or lytic enzymes (Cervantes and Rodrguez-Barrueco, 1992)CULTURE of:Simple culture media which includes glucose and propionate as a carbon source, casein hydrolysate as a nitrogen source, and vitamins, salts and trace elements. Lipids may also be helpful. (Cervantes and Rodrguez-Barrueco, 1992)WHY:Its not an obligate relationship, so having these tools allows scientists and those interested in forestry management or soil amelioration to inoculate stands of alder or other sp.

  • Identification of Frankia strains11th International Conference on Frankia and Actinorhizal Plants (1998)

    1. 16s rRNA (Ritchie et al., 1999)

    2. PCR + DNA hybridization (Nalin et al., 1999)

    3. PCR + RFLPs (Lumini and Bosco, 1999)

    4. Antibiotic resistance patterns (Tisa et al., 1999)

  • Anatomy of Nodule FormationBerg, 1999:Nodule development is under the plants controlgreat variety of nodule morphology across speciesThree steps to invasion of host by Frankia:Infection threads with interfacial matrix, formation of cytoplasmic bridges (filamentous)Vegetative hyphae (filamentous)Symbiotic vesicles - where N2-fixation occurs (multicellular and spherical)http://helios.bto.ed.ac.uk/bto/microbes/nitrogen.htm

  • Anatomy and Metabolism: O2 and NitrogenaseSilvester et al., 1999Frankia grows best at atmospheric pO2.In most species, symbiotic vesicles develop an envelope to protect nitrogenase.Example: In Alnus, envelope thickness adjusts to ambient thickness.Coriaria and Dastica do not have a multilaminate envelope - how do they protect nitrogenase?Methods: MiscroscopyHigh concentrations of mitochondria around the base of the symbiotic vesicles may protect nitrogenase.Tjepkema et al., 1999Dastica has a similar morphology to legumes, which have a variable diffusion barrier to O2. Does Dastica?Methods: Nitrogenase activity was measured via C2H2 reduction and the evolution of CO2.Results: Nitrogenase activity was not restored to predecline values by increasing external pO2.Furthermore, calculations of O2 uptake by mitochondria are not sufficient to reduce the diffusion of O2 past the mitochondriaHA: Nitrogenase activity is compartmenalized in multiple vesicles so that if activity is decreased in one, it is increased in another.

  • Ecological RelationshipsClawson et al., 1999What are the interactions and relationships that address dominance, competition and distribution among Frankia strains growing in a variety of conditions?

    Methods: Obtained strains and developed clades based on 16s rRNA.

    Results: Diversity in a single alder stand is low; instead it is dominated by a single strain or set of similar strains.

    Further research: Are there different ecotypes associated with different soil conditions?Markham and Chanway, 1999Does past contact reduce the degree of mutualism in the Alnus rubra - Frankia symbiosis?Methods: Three low-elevation and three high-elevation populations of Red Alder were inoculated with Frankia, either a familiar or an unfamiliar strain.Results: Low elevation: High nitrogen in soils. Plants inoculated with the unfamiliar strain grew larger than those with the familiar strain.High elevation: Low nitrogen in soils. Plants inoculated with the familiar strain grew larger.Conclusion: The degree of mutualism depends on environmental conditions and may decrease with time.

  • Relationship to Mycorrhizal co-SymbiontsCervantes, E. and C. Rodrguez-Barrueco. 1992. Relationships between the Mycorrhizal and Actinorhizal Symbioses in Non-legumes, in Methods in Microbiology, Volume 24: Techniques for the Study of Mycorrhiza, Norris J.R., D.J. Read and A.K. Varma, Editors. Academic Press: New York. 417-432.

    Alnus sp. forms symbiotic relationships with fungi also, including:CortinariaceaeHygrophoraceaeRussulaceaeBoletaceaeFungi ImperfectiPresence of Frankia and N-fixing activity may put increased pressure on the plant for the production of ATP.Ectomycorrhizal association as been shown to enhance phosphate uptake for Alnus viridis.

  • ReferencesBerg, R. Howard. 1999. Frankia forms infection threads. Canadian Journal of Botany. 77:1327-1333.Berg, R. Howard. 1999. Cytoplasmic bridge formation in the nodule apex of actinorhizal root nodules. Canadian Journal of Botany. 77:1351-1357Cervantes, E. and C. Rodrguez-Barrueco. 1992. Relationships between the Mycorrhizal and Actinorhizal Symbioses in Non-legumes, in Methods in Microbiology, Volume 24: Techniques for the Study of Mycorrhiza, Norris J.R., D.J. Read and A.K. Varma, Editors. Academic Press: New York. 417-432.Clawson, Michael L., Jeffrey Gawronski and David R. Benson. 1999. Dominance of Frankia strains in stands of Alnus incana subsp. Rugosa and Myrica pensylvanica. Canadian Journal of Botany. 77:1203-1207.Lumini, Erica and Marco Bosco. 1999. Polymerase chain reaction - restriction fragment length polymorphisms for assessing and increasing biodiversity of Frankia culture collections. Canadian Journal of Botany. 77:1261-1269.Madigan, M.T., J.M. Martinko and J. Parker. Brock Biology of Microorganisms, 10th Edition. Prentice-Hall: Upper Saddle River, NJ, 2003.Markham, John H. and Chris P. Chanway. 1999. Does past contact reduce the degree of mutualism in the Alnus rubra - Frankia symbiosis? Canadian Journal of Botany. 77:434-441.Nalin, R., P Normand, P. Simonet and A.M. Domenach. 1999. Polymerase chain reaction and hybridazation on DNA extracted from soil as a tool for Frankia spp. Population distribution studies in soil. Canadian Journal of Botany. 77:1239-1247.Ritchie, Nancy J. and David D. Myrold. 1999. Phylogenetic placement of uncultured Ceanothus microsymbionts using 16s rRNA gene sequences. Canadian Journal of Botany. 77:1208-1213.Silverster, Warwick B., Birgit Langenstein, R. Howard Berg. 1999. Do mitochondria provide the oxygen diffusion barrier in root nodules of Coriaria and Dastica? Canadian Journal of Botany. 77:1358-1366Tisa, Louis S., Matthew Chval, Glenn D. Krumholz and Joel Richards. 1999. Antibiotic resistance patterns of Frankia stains. Canadian Journal of Botany. 77:1257-1260. Tjepkema, John D., Gangyi Du and Christa R. Schwintzer. 1999. Response of respiration and nitrogenase activity in Dastica glomerata (Presl.) Baill. to changes in pO2. Canadian Journal of Botany. 77:1367-1372.