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By Erick Breathwaite

By Erick Breathwaite. Identification of Gunnera genes encoding components of the common sym pathway potentially responsible for symbiosis between Gunnera manicata and Nostoc Puntiforme. Soil Microbes. http://permaculturetokyo.blogspot.com/2009/02/rhizobium-symbiosis-with-woody-plants.html.

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By Erick Breathwaite

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  1. By Erick Breathwaite Identification of Gunnera genes encoding components of the common sym pathway potentially responsible for symbiosis between Gunneramanicataand NostocPuntiforme

  2. Soil Microbes http://permaculturetokyo.blogspot.com/2009/02/rhizobium-symbiosis-with-woody-plants.html

  3. Gunnera-Nostoc symbiosis webdev.one365.com/.../dynamic/Image/gunnera.jpg microbiology.ucdavis.edu/.../pics/hormogonia.jpg

  4. Gunnera-Nostoc symbiosis papillae Hormogonia • microbiology.ucdavis.edu/.../pics/hormogonia.jpg

  5. The Question • The genetic basis for Gunnera/cyanobacteria symbiosis remains unknown therefore… • We will look specifically for potential genes in Gunnera plants that are responsible for establishing and maintaining symbiosis between the cyanobacteria, NostocPunctiformeand Gunnera plants.

  6. Our Hypothesis • Previous experiments have observed that there are commonalities observed in the signaling components (Sym pathway) and modulation of host cell responses in both arbuscularmycorhizal (AM) and root-nodule (RN) symbioses • Therefore, we hypothesize that the genes that are responsible for the communication between the cyanobacteria and Gunnera may also use components of the common sym pathway/communication mechanisms

  7. Preliminary Steps • Created normalized cDNA library using mRNA isolated from glands • About 3000 cDNA clones from the library was sequenced from the 5’ end using the traditional Sanger method and 454-pyrosequencing technology was also used to identify and characterize the cDNAlibrary http://cdna-library.com/ http://www.medigenomix.de/pics/seq/GS20_sequencing.jpg

  8. Experimental procedure • We will utilize programs such as NCBI BLAST to identify homologous genes in other plants. Biobike will be used to compare and further analyze unique sequences and identify genes of interest. • Of particular interest in this experiment will be genes that are similar to components of the common Sym pathway such as LRR receptor gene, cation channel genes, lecuine rich receptor-like kinase genes, calcium and calmodulin-dependent kinase genes, the CYCLOPS gene and nodulation specific transcription factors. • Design gene specific primers for each gene and use qRT-PCR to compare their expression levels in glands vs. normal stem tissue and potentially see upregulation of genes in gland tissue www.people.vcu.edu/.../BioBIKE/highwheeler.gif

  9. Potential results • Determine if specific genes in the common sym pathway genes are used for Gunnera to recognize and internalize Nostoccyanobacteria as we hypothesized • If not, we intend to investigate if other groups of genes known to be involved in plant-pathogen interactions are necessary for Gunnera to recognize and internalize Nostoccyanobacteria. (Jonathan's experiment)

  10. Applications of results • By understandingthe mechanism and genes responsible for the Gunnera-Nostoc symbiosis, we can possibly suggest a means of establishing useful associationsbetween nitrogen-fixing cyanobacteria and crop plants eliminating the need for environmentally harmful fertilizers. http://www.ecofriend.org/archives/200707/5/?sort=desc

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