About Frankia Literature Review Suzanna Kruger Biology 523 Microbial Ecology Dr. Anna-Louise Reysenbach 3 June 2003

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  • About Frankia Literature Review Suzanna Kruger Biology 523 Microbial Ecology Dr. Anna-Louise Reysenbach 3 June 2003
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  • Overview Frankia taxonomy Host species Role in nitrogen cycle Methods of isolation Anatomy of nodule formation Anatomy and Metabolism Ecological relationships
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  • Frankia Taxonomy Filamentous, High GC, Gram-Positive Bacteria Group III Actinomycete Nitrogen-fixing symbiont of plants (Madigan, 2003) http://www.msu.edu/~batzli/whats_frankia.html http://helios.bto.ed.ac.uk/bto/microbes/microbes.htm#Microorganisms% 20and%20the%20"Tree%20of%20Life"
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  • Host Species Hosts are non- leguminous, woody angiosperms 8 Families, 23 genera including: Alnus (Betulacaceae) Ceanothus (Rhamnaceae) Rubus (Rosaceae) http://www.msu.edu/~batzli/graphics/alder_forest.jpg Red Alder Forest, Washington
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  • Role in Nitrogen Cycle (Madigan, 2003) Nitrogen fixation: N 2 + 8H+ + 8e - + 16 ATP = 2NH 3 + H 2 + 16ADP + 16 Pi Catalyzed by nitrogenase Reasons for interest in Frankia are related to the Nitrogen Cycle: Forestry management Land reclamation Soil improvment
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  • Methods of Isolation and Culture of Frankia ISOLATION 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.
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  • Identification of Frankia strains 11th 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)
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  • Anatomy of Nodule Formation Berg, 1999: Nodule development is under the plants controlgreat variety of nodule morphology across species Three steps to invasion of host by Frankia : Infection threads with interfacial matrix, formation of cytoplasmic bridges (filamentous) Vegetative hyphae (filamentous) Symbiotic vesicles - where N 2 - fixation occurs (multicellular and spherical) http://helios.bto.ed.ac.uk/bto/microbes/nitrogen.htm
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  • Anatomy and Metabolism: O 2 and Nitrogenase Silvester et al., 1999 Frankia grows best at atmospheric pO 2. 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: Miscroscopy High concentrations of mitochondria around the base of the symbiotic vesicles may protect nitrogenase. Tjepkema et al., 1999 Dastica has a similar morphology to legumes, which have a variable diffusion barrier to O 2. Does Dastica ? Methods: Nitrogenase activity was measured via C 2 H 2 reduction and the evolution of CO 2. Results: Nitrogenase activity was not restored to predecline values by increasing external pO 2. Furthermore, calculations of O 2 uptake by mitochondria are not sufficient to reduce the diffusion of O 2 past the mitochondria H A : Nitrogenase activity is compartmenalized in multiple vesicles so that if activity is decreased in one, it is increased in another.
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  • Ecological Relationships Clawson et al., 1999 What 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, 1999 Does 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.
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  • Relationship to Mycorrhizal co-Symbionts Cervantes, 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: Cortinariaceae Hygrophoraceae Russulaceae Boletaceae Fungi Imperfecti Presence 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.
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  • References Berg, 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. 77:1351-1357 Cervantes, 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. 77:1358-1366 Tisa, 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 pO 2. Canadian Journal of Botany. 77:1367-1372.