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 9 – Native Legume RNB 1600 - 1720 Authors: Rashmita Parihar 1 , Neetu Poonar 1 , Nisha Tak 1 , Alkesh Tak 1 , Indu Sankhla 1 , Anil Tripathi 2 , Ravi Tiwari 3 , Euan K. James 4 , Janet I. Sprent 5 and H.S.Gehlot 1 1 Department of Botany, Jai Narain Vyas University, Jodhpur, 342001, India 2 School of Biotechnolgy, Banaras Hindu University, Varanasi, 221005, India 3 Centre for Rhizobium Studies, Murdoch University, Murdoch, 6150, W Australia 4 James Hutton Institute, EPI Division, Invergowrie, Dundee DD2 5DA, UK 5 Division of Plant Sciences, University of Dundee at JHI, Invergowrie, Dundee DD2 5DA, UK Presentation Title: Burkholderia phymatum isolated from root nodules of two species of Indigofera from alkaline soils of the Indian Thar desert Presentation Time: 1600 – 1620 It is long established that many species in the large Mimosoid genus Mimosa are nodulated by Beta-rhizobia (which so far include strains of Cupriavidus and Burkholderia), and that in its major centre of diversification in central Brazil there has probably been co-evolution between Mimosa and its Burkholderia symbionts (Bontemps et al. 2010; dos Reis Junior et al. 2010). Recently there has also been increasing evidence of nodulation and nitrogen fixation by Burkholderia in symbiosis with legumes outside the genus Mimosa. These include species of Cyclopia (Podalyrieae) and Rhynchosia (Phaseoleae) native to South Africa, and the agriculturally important legume Phaseolus vulgaris has also been shown to harbor B. phymatum as a symbiont in Morocco. In the present study we have isolated four strains (IL24, IL26, IC12 and IC14) of Burkholderia from root nodules of two Indian endemic species from the Papilionoid genus Indigofera (I. cordifolia and I. linnaei) growing in alkaline soils in the semi-arid Thar Desert in Western Rajasthan. Based on 16S rRNA gene sequences the strains are very close to those isolated from Mimosa spp., P. vulgaris and Indigofera suffruticosa. An ARDRA pattern obtained using AluI and Sau3AI, as well as an RPO1-based RAPD profile, showed that isolates IL24, IL26, IC12 and IC14 from Indigofera were genetically identical to five isolates of Burkholderia from M. pudica (MP17, MP20, MP21, MP22 and MP23). Moreover, phenotypic characteristics (eg. colony morphology, pH range, NaCl tolerance, antibiotic sensitivity and C-utilization patterns) were also similar. These data are discussed in terms of the legume hosts and their various ecological niches within India, including such characteristics as climate and soils. Bontemps et al. 2010. Burkholderia species are ancient symbionts of legumes. Mol. Ecol. 19:44-52. dos Reis Junior et al. 2010. Nodulation and nitrogen fixation by Mimosa spp. in the Cerrado and Caatinga biomes of Brazil. New Phytol 186:934-946. Acknowledgement: Authors acknowledge the financial support from ATSE Crawford Training Fund; Centre for Rhizobium Studies, Murdoch University and DBT, Govt. of India (BT/PR11461/AGR/21/270/2008). 60 2011
17 th International Congress on Nitrogen Fixation Fremantle, Western Australia 27 November – 1 December 2011 Session Details: Tuesday 29 November 2011 Concurrent Session 9 – Native Legume RNB 1600 - 1720 Authors: Robert Walker 1,2 , Graham O‟Hara 2 , Lesley Mutch 1 , Lambert Bräu 2 & Elizabeth Watkin 1 1 School of Biomedical Sciences, Curtin University, Bentley, 6845, Western Australia. 2 Centre for Rhizobium Studies, Murdoch University, Murdoch, 6150, Western Australia. Presentation Title: The phylogenetic diversity of australian Burkholderia root nodule bacteria Presentation Time: 1620 – 1640 Recently, members of the beta-proteobacteria were described as novel root nodule bacteria (coined betarhizobia) with many species represented by the genus Burkholderia and some Cupriavidus. Endosymbionts isolated from nodules of Mimosa spp. are predominantly Burkholderia spp. and this appears to be a preferential association however, Burkholderia spp. have also been isolated from Papillionoideae and other members of the Mimosoideae subfamily of legumes. Beta-rhizobia from Australia have been isolated from Acacia spp. and from members of the tribes Kennediinae and the endemic Mirbeliaea, however in glasshouse trials these isolates form ineffective nodules on many other Australian legume species. This study examined twelve authenticated Burkholderia sp. isolated from both native and invasive legumes from New South Wales, Western Australia, the Northern Territory, and two South African isolates from Rhynchosia ferulifolia. The type strains Burkholderia phymatum STM815 and Cupriavidus taiwanensis LMG19424 were also included. The phylogenetic diversity of these isolates was explored by examining the partial sequences from coding regions of the 16S rRNA, recA, dnaK and atpD genes. To determine the phylogenetic distribution within Australian isolates only, a concatenated neighbour-joining tree was constructed and most isolates fell into geographical clades. When including non- Australian isolates, Australian Burkholderia root nodule bacteria appear to form a distinct monophyletic group, separate to the Burkholderia cepacia complex (BCC) and from non-Australian isolates. Nodulation and nitrogen fixation genes were also partially sequenced and strongly align to nod and nif genes from Australian Bradyrhizobium spp. however, very few sequence data exist for the nodulation and nitrogen fixation genes of Australian Burkholderia root nodule bacteria. Australian root nodule Burkholderia may have evolved from a distinct monophyletic lineage from other beta-rhizobia allowing them to nodulate Australian legumes with varying effectiveness providing a competitive advantage over other soil saprophytes however, Australian beta-rhizobia may not be useful as effective nitrogen fixing endosymbionts. 61 2011
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IN INOCULANTS Nodulaid - 17th International Nitrogen Fixation ...

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