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4.3. Description of Bradyrhizobium altum sp. nov. Bradyrhizobium altum (al’ tum. L. neut. adj. altum, high, referring to the high-altitude region where this species was isolated from). Cells are motile, Gram-negative rods and is approximately 1.29 × 0.45 μm in size. Colonies are circular, convex, opaque, cream in colour and < 1 mm in diameteron YMA after 7 daysincubation at 28 ◦ C. The pH range for growth on YMA is 5 to 10 and growth can occur between 15 and 37 ◦ Cand thesestrainscangrowinthe presenceof upto 2.0 % (w/v) NaCl. Reduction of nitrates, β- glucosidase urease, tested positive. It is also positive for the production of L-arabinose, Tween 40, Tween 80, adonitol, N-acetyl-D-glucosamine, D-cellobiose, D-arabitol, D-galactose, D- and L-fructose, m-inositol, D-mannitol, maltose, D-mannose, Draffinose, D-melibiose, L-rhamnose, D-sorbitol, sucrose, xylitol, Dtrehalose, turanose, methyl pyruvate, D-galacturonic acid, acetic acid, malonic acid, propionic acid, D-saccharic acid, quinic acid, sebacic acid, L-asparagine, D- and L-alanine, L-leucine, L-proline, L-ornithine and glycerol. Negative reactions were recorded for glucose fermentation, indole production, β- galactosidase activity and L-arginine dihydrolase. It is also negative for the assimilation of N-acetyl-glucosamine, potassium gluconate, D-maltose, adipic acid, capric acid, malic acid, phenylacetic acid, glycogen, α- cyclodextrin, L-arabinose, i-erythritol, Lthreonine, D- and L-serine, glucose-1-phosphate and glucose-6- phosphate. Strain is resistant to penicillin (10 μg/ml), ampicillin (10 μg/ml), gentamycin (10 μg/ml), chloramphenicol (30 μg/ml) and neomycin (10 μg/ml) and is susceptible to kanamycin (30 μg/ml), erythromycin (30 μg/ml), streptomycin (10 and 25 μg/ml) and tetracycline (30 μg/ml). The type strain Pear77 T (=SARCC 754 = LMG 31407), was isolated from root nodules of Pearsonia obovata collected in the Dullstroom area of the Mpumalanga Province, South Africa. This species forms effective nodules on cowpea (Vigna unguiculata) and siratro (Macroptilium atropurpureum). The DNA G + Ccontent of the type strain is 63.3 mol%. The GenBank/EMBL/DDBJ accession numbers for the genome sequence and gene sequences for this type strain are: whole genome (JACMYP000000000) and 16S rRNA (LR877298), atpD (LN650104), glnII (LN650146), dnaK (LN650125), gyrB (LN650209), rpoB (LN650188) and nodA (LN650251).

Avontuur, Juanita R., Palmer, Marike, Beukes, Chrizelle W., Chan, Wai Y., Tasiya, Taponeswa, Zyl, Elritha van, Coetzee, Martin P.A., Stepkowski, Tomasz, Venter, Stephanus N., Steenkamp, Emma T. (2022): Bradyrhizobium altum sp. nov., Bradyrhizobium oropedii sp. nov. and Bradyrhizobium acaciae sp. nov. from South Africa show locally restricted and pantropical nodA phylogeographic patterns. Molecular Phylogenetics and Evolution (107338) 167: 1-13, DOI: 10.1016/j.ympev.2021.107338

4.2. Description of Bradyrhizobium oropedii sp. nov. Bradyrhizobium oropedii (o.ro.pe’ di.i. Gr. neut. n. oropedion mountain plain; N.L. gen. n. oropedii, of the central Southern African plateau, from where the species was isolated). Cells are motile, Gram-negative, rod shaped and approximately 1.87 × 0.48 μm in size. Growth occurs on YMA at 28 ◦ C after 7 days and colonies are translucent to opaque, circular, convex and cream in colour. Grows at pH values ranging from 5 to 10 and at temperatures ranging from 15 to 35 ◦ C. Isolates are unable to grow in the presence of more than 0.5% NaCl (w/v). Strains tested positive for oxidase, catalase, urease and β- glucosidase activity as well as the production of Tween 40 and Tween 80. Tested positive for the following carbon sources: Dfructose, L-fructose, D-gluconic acid, D-mannitol, α and β- hydroxy butyric acid, p-hydroxy phenylacetic acid, L-alaninamide, L-lactic acid, L-leucine, D-saccharic acid, and urocanic acid. Negative reactions were recorded for indole production and the assimilation of capric acid, adipic acid, malic acid, glycogen, α- D-lactose, N-acetyl-D-galactosamine, α- cyclodextrin, Adonitol, D-cellobiose, D-raffinose, i-erythritol, maltose, lactulose, D-melibiose, sucrose, D-trehalose, turanose, xylitol, L-threonine, uridine, inosine, thymine, glucose-1-phosphate and glucose-6-phosphate, and m-inositol. Resistant to ampicillin (10 μg/ml), chloramphenicol (30 μg/ml), tetracycline (30 μg/ml), penicillin (10 μg/ ml), and susceptible to streptomycin (25 μg/ml). The type strain, Pear76 T (=SARCC 731 = LMG 31408) was isolated from root nodules of Pearsonia obovata in the Dullstroom area of the Mpumalanga Province, South Africa and forms effective nodules on cowpea (Vigna unguiculata) and siratro (Macroptilium atropurpureum). The DNA G + Ccontent of the type strain is 63.5 mol%. GenBank/ EMBL/DDBJ accession numbers for the genome sequence and the gene sequences of this type strain are: whole genome (JACMYO000000000) and 16S rRNA (LR877297), atpD (LN650103), glnII (LN650145), dnaK (LN650124), gyrB (LN650208), rpoB (LN650187) and nodA (LN650250).