Soilborne TOWARDS EXPANDING THE SOUTH AFRICAN RHIZOBIUM CULTURE COLLECTION (SARCC) AS A GENETIC RESOURCE AND ITS APPLICATION IN SUSTAINABLE AGRICULTURE AI Hassen , FL Bopape, K Mashau & DM Sibanyoni Agricultural Research Council Plant Protection Research Institute (ARC-PPRI) Email: hassena@arc.agric.za Joint Forum Biodiversity Information Management & Foundational Information Program, 14 -17 August 2017, Salt Lake Beach Resort, Durban

Presentation Outline 2. Maintenance of the rhizobium culture 1. SARCC- Historical Background 2. Maintenance of the rhizobium culture 3. The SARCC database and prospects for improvement 4. Applications of the SARCC data (Biological Nitrogen Fixation) 5. Concluding summary and recommendations

1. SARCC- Historical Background 1903, Nobbs, field trials with legumes at Ellensburg, Stellenbosch. Did not result in significant research in Nitrogen -fixing systems in South Africa until the late 1950’s. Establishment of Serradella (Ornithopus compressus) and the role of its nitrogen fixing rhizobial symbiont (Sim, 1958). In 1959, Margareta Mes – Rhizobium - legume symbiosis (University of Pretoria, Univ. Stellenbosch). 1965, exploitation of the nodule bacterium legume symbiosis and nodulation status of indigenous legumes & legumes of economic importance in South Africa.

Historical Background cont.. Indigenous Trifolium spp. (T. africanum, T. burchellianum) Rhizobium specificity confirmed among various African clover species; broad spectrum R. leguminosarum biov. trifolii Studies on exotic Trifolium spp, early 1960’s (need to inoculate with R. leguminosarum biov. trifolii). Populations of Sinorhizobium meliloti effective on lucerne (Medicago sativa) naturalized in most areas with suitable pH. Also effective on annual medics Medicago truncata, M. littoralis South African soils rich in indigenous Bradyrhizobium spp. that nodulate Peanuts (Arachis hypogaea) & Cowpea (V. unguiculata)

Historical Background cont.. Lack of B. japonicum strains effective on local soybean (Glycine max L.) cultivars (show specific B. japonicum requirement) Since 1960s, ARC-Plant Protection Research Institute started to expand research on the rhizobium-legume symbiosis in South Africa Became the custodian of a wide range of rhizobium collections that led to the establishment of the SARC, now with the acronym SARCC Several hundreds of rhizobia (>1500 strains) belonging to Rhizobium, Bradyrhizobium, Sinorhizobium & Mesorhizobium species. Routinely maintained (viability, purity, identity).

2. Maintenance of the Rhizobium collections SARCC- an integral part of the SA legume industry. Main source of legume rhizobium inoculant strains. Significant economic and social benefits (commercial & subsistence). Collection activities of the SARCC support research in Biological Nitrogen Fixation (BNF) SARCC cultures obtained from various sources: field collections, international sources (overseas culture collections, universities, client deposits etc.).

Maintenance of the Rhizobium collections P y Cultural characteristics, purity Purity and preliminary ID Growth M o l e c u a r Viability and concentration Species/Genus ID

Preservation of the cultures at SARCC Agar plates/slants/cold rooms Lyophilized (freeze dried) -80 ultra freezer (frozen)

3. The SARCC data base & prospects for improvement

World Federation of Culture Collections (WFCC)

SARCC- on the Global Catalogue of Microorganisms

SARCC- on the global catalogue of microorganisms

SARCC- Home page gcm.wfcc.info/cc/sarcc

4. Applications of the SARCC strains - BNF Schultze and Kondorosi, 1998. Ann Rev Gen; www.cilr.uq.edu.au

Development of rhizobium inoculants (commercial strains) TJ14 SR4 SAC2 Rhizobium rhizogenes 163C UD5 Rhizobium tropici strain LNW10 Rhizobium tropici strain LNP6 XCR6 RAK1 Sinorhizobium meliloti QTX 8-2 Sinorhizobium meliloti Qtx-8-1 RF14 Rhizobium taeanense strain PSB 2-6 XAP2 Mesorhizobium amorphae XEL1 Mesorhizobium sp. SOD33 Azospirillum brasilense REC3 Azospirillum brasilense strain 13-2C Bradyrhizobium elkanii DASA35013 XHX1 WB74 Bradyrhizobium japonicum USDA 110 Bradyrhizobium japonicum DASA38025 XHT1 XBQ5 Sinorhizobium fredii USDA 194 Mesorhizobium loti Bradyrhizobium sp. USDA 4424 Bradyrhizobium sp. AP28 FN566882.1 89 68 98 72 58 56 54 100 90 79 99 60 94 91 81 78 0.1 SAC2 Trifolium sp Hassen et al. 2015 , unpublished

Nodulation of Aspalathus linearis (Rooibos) SARCC-RB16h Mesorhizobium amorphae CCBAU 85061 (EU256436.1) SARCC-RB15b SARCC-RB10g SARCC-RB15i SARCC-RB10a SARCC-RB 16b SARCC-RB15d SARCC-RB19c SARCC-RB15g Mesorhizobium sp. SOD33 (EF125932.1) SARCC-RB15a SARCC-RB16f SARCC-RB16i SARCC-RB16p SARCC-RB16l SARCC-RB16n Mesorhizobium amorphae SEMIA 848 (FJ025122.1) SARCC-RB14g SARCC-RB5e Rhizobium sp. kmd 046 Rhizobium rhizogenes 163C SARCC-RB1d SARCC-RB8c SARCC-RB17a SARCC-RB17b R.tropici strain LNW10 R.tropici LNP7 SARCC-RB1h Rhizobium tropici sp CCBAU 25295 Rhizobium tropici sp LNP6 Bradyrhizobium japonicum sp DASA38025 Bradyrhizobium elkanii DASA35013 SARCC-RB10m Bradyrhizobium japonicum USDA 110 Azospirillum brasilense A154 SARCC-RB14h SARCC-RB17c SARCC-RB14i Burkholderia sp. WSM4206 (HQ698907.1) SARCC-RB1i Burkholderia sp T132 GU191167.1 Burkholderia pyrrocinia sp.H5 SARCC-RB5f SARCC-RB16g SARCC-RB15c SARCC-RB10c SARCC-RB16k Herbasprillum.seropedicae Z67 SARCC-RB10h Herbasprillum lusitanum P6-12 99 69 96 93 85 89 95 77 94 0.02 α-rhizobia β-rhizobia Hassen et al. 2012 Biol. Fert. Soils

5. Concluding summary & recommendations Culture collections with valuable biodiversity data must have a reliable and user-friendly system: in house data management system; home page; online catalogue. Help the collections to manage, disseminate and share the information related to their holdings. Improves the scientific status of the collections; distributions & holdings; utilization of microbial resources & applications in research Proper maintenance of national assets required to provide key services, sustainable crop production and crop protection (Safeguard food security). .

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