IN INOCULANTS Nodulaid - 17th International Nitrogen Fixation ...

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17 th International Congress on Nitrogen Fixation Fremantle, Western Australia 27 November – 1 December 2011 Session Details: Tuesday 29 November 2011 Concurrent Session 8 – Plant Symbiotic Genes 1600 – 1720 Authors: Matthew Bellgard1 , Wayne Reeve2 , Huaan Yang3 , Jingjuan Zhang4 , Roberto Barrero1 and John Howieson2, 4 1 Centre for Comparative Genomics, Murdoch University, South Street, Murdoch 6150, Australia. 2 Centre for Rhizobium Studies, School of Biological Sciences and Biotechnology, Murdoch University, South Street, Murdoch 6150, Australia. 3 Department of Agriculture and Food, Western Australia (DAFWA), Baron Hay Court, South Perth, 6151, Western Australia. 4 Crop and Plant Research Institute, Murdoch University Presentation Title: Interpreting sub-optimal nitrogen fixation in Lupinus angustifolius utilising the genome sequence of both symbiotic partners. Presentation Time: 1600 – 1620 WSM1417, WSM1253 and WSM471 are nodule bacteria that infect Lupinus angustifolius (NLL narrow-leaf lupin) and which have different nodulation and nitrogen fixation profiles. These three strains were sequenced within the JGI GEBA project through the US Department of Energy. To understand the genetic control of the relationship between these three micro-symbionts and their legume host we required both the lupin genome sequence, and transcriptome information. We thus sequenced NLL cv Tanjil using a WGS approach and achieved a 30 fold coverage of the whole genome. We obtained 31 billion base pair high quality sequencing data and are undertaking the lupin genome assembly and annotation with a view to identify candidate genes in nodule formation and function. The transcriptome data was of nodules of three NLL plants inoculated with the three sequenced strains. From the NLL genome sequence we obtained 23,000 contigs, which were assembled into 12,500 scaffolds. The largest contig we have is 149Mb, the N50 size of 13.6 Kb and there is ~80Mb of repeat sequences. Further analysis of the genome sequence data will be provided. 56 2011
17 th International Congress on Nitrogen Fixation Fremantle, Western Australia 27 November – 1 December 2011 Session Details: Tuesday 29 November 2011 Concurrent Session 8 – Plant Symbiotic Genes 1600 – 1720 Authors: Brett Ferguson 1 , Dongxue Li 1 , Meng-Han Lin 1 , Satomi Hayashi 1 , Yu-Hsiang Lin 1 , Dugald Reid 1 , Saeid Mirzaei 1 , Alina Tollenaere 1 , Peter Gresshoff 1 1 Australian Research Council Center of Excellence for Integrative Legume Research, School of Agriculture and Food Sciences, University of Queensland, St. Lucia, Brisbane, QLD, 4072, Australia Presentation Title: Novel components involved in legume nodule development and autoregulation Presentation Time: 1620 – 1640 To discover novel legume components involved in nodule formation (Ferguson et al. 2010), we used nextgeneration sequencing (Illumina) and the available soybean genome (www.phytozome.net/soybean) to identify genes that are differentially expressed in the zone of root hair emergence following rhizobia-inoculation. Legumes regulate the number of nodules they form via the Autoregulation Of Nodulation (AON). Following rhizobia-inoculation, AON commences with the production of a root-derived signal called Q (Reid et al. 2011a). We have identified soybean Q candidate genes encoding CLAVATA3/ESR related (CLE) peptides that exhibit increased expression following rhizobia inoculation or inhibitory nitrate treatment (Reid et al. 2011b). Overexpression of these genes significantly reduces soybean nodule numbers. The nitrate-induced Q peptide appears to act locally, whereas the rhizobia-induced Q peptide is transported to the shoot where it, or a product of its action, is perceived by a LRR receptor kinase called NARK. We used next-generation sequencing to identify soybean components acting downstream of NARK in the leaf. Candidate genes identified can now be used in our recently established bioassay to confirm the presence of Q in various samples and biological extracts (e.g., xylem sap). We also isolated and characterized the phenotype of a soybean line mutated in the homeologous gene of GmNARK, GmCLAVATA1A. Following the perception of Q by NARK in the leaf, a number of factors are produced including a novel signaling compound called the Shoot-Derived Inhibitor (SDI). SDI is predicted to be transported from the shoot to the root where it inhibits continued nodule development. We found SDI to be NARK- and Nod factor-dependent, heat stable, small, and likely not a peptide or RNA molecule (Lin et al. 2010, 2011). Findings regarding our progress in identifying and characterising the abovementioned nodulation factors will be presented. Ferguson BJ, Indrasumunar A, Hayashi S, Lin Y-H, Lin M-H, Reid D, Gresshoff PM (2010) Molecular analysis of legume nodule development and autoregulation. Journal of Integrative Plant Biology 52: 61-76 Lin Y-H, Ferguson BJ, Kereszt A, Gresshoff PM (2010) Suppression of hypernodulation in soybean by a leaf-extracted, NARK- and Nod factor-dependent, low molecular mass fraction. New Phytologist 185: 1074-1086 Lin Y-H, Lin M-H, Gresshoff PM, Ferguson BJ (2011) An efficient petiole-feeding bioassay for introducing aqueous solutions into dicotyledonous plants. Nature Protocols 6: 36-45 Reid DE, Ferguson BJ, Hayashi S, Lin Y-H, Gresshoff PM (2011a) Molecular mechanisms controlling legume autoregulation of nodulation. Annals of Botany 108: 789-795 Reid DE, Ferguson BJ, Gresshoff PM (2011b) Inoculation- and nitrate-induced CLE peptides of soybean control NARK-dependent nodule formation. Molecular Plant-Microbe Interactions 24: 606-618 57 2011
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