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Rhizobium daejeonense sp. nov. isolated from a cyanide treatment bioreactor

Zhe-Xue Quan^1,, Hee-Sung Bae^1,2, Jong-Hwan Baek¹, Wen-Feng Chen³, Wan-Taek Im¹ and Sung-Taik Lee¹

¹ Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 373-1 Guseong-dong, Yuseong-gu, Daejeon 305-701, Korea
² Department of Biological Sciences, 331 Life Sciences Building, Louisiana State University, Baton Rouge, LA 70803, USA
³ College of Biological Sciences, China Agricultural University, Beijing 100094, China

Correspondence
Sung-Taik Lee
e_stlee{at}kaist.ac.kr

A polyphasic study was carried out to determine the taxonomic position of two aerobic, cyanide-degrading bacterial strains, designated L61^T and L22, which had been isolated from a bioreactor for the treatment of nickel-complexed cyanide. The two isolates exhibited almost identical taxonomic characteristics. Phylogenetic analysis inferred from comparative 16S rRNA gene sequences indicated that the isolates fall in a sublineage of the genus Rhizobium comprising the type strains of Rhizobium giardinii, Rhizobium radiobacter, Rhizobium rubi, Rhizobium larrymoorei, Rhizobium vitis, Rhizobium undicola, Rhizobium loessense, Rhizobium galegae and Rhizobium huautlense. Cells of the two isolates are Gram-negative, aerobic, motile and non-spore-forming rods (0·6–0·7x1·1–1·3 µm), with peritrichous flagella. The DNA G+C content is 60·1–60·9 mol%. Cellular fatty acids are C_{16 : 0} (2·2–3·3 %), C_{18 : 0} (2·1–3·2 %), C_{19 : 0} cyclo 8c (9·9–16·8 %), C_{20 : 3}6,9,12c (2·7–3·3 %), summed feature 3 (7·2–7·7 %) and summed feature 7 (67·8–73·7 %). The strains formed nodules on a legume plant, Medicago sativa. A nifH gene encoding denitrogenase reductase, the key component of the nitrogenase enzyme complex, was detected in L61^T by PCR amplification by using a nifH-specific primer system. Strains L61^T and L22 were distinguished from the type strains of recognized Rhizobium species in the same sublineage based on low DNA–DNA hybridization values (2–4 %) and/or a 16S rRNA gene sequence similarity value of less than 96 %. Moreover, some phenotypic properties with respect to substrate utilization as a carbon or nitrogen source, antibiotic resistance and growth conditions could be used to discriminate L61^T and L22 from Rhizobium species in the same sublineage. Based on the results obtained in this study, L61^T and L22 are considered to be representatives of a novel species of Rhizobium, for which the name Rhizobium daejeonense sp. nov. is proposed. The type strain is L61^T (=KCTC 12121^T=IAM 15042^T=CCBAU 10050^T).

The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene sequences of strains L61^T and L22 are AY341343 and DQ089696, respectively. The GenBank/EMBL/DDBJ accession number for the partial nifH gene sequence of strain L61^T is AY428644.

A transmission electron micrograph of cells of strain L61^T, a minimum-evolution phylogenetic tree and a table detailing the fatty acid compositions of L61^T, L22 and related Rhizobium species are available as supplementary material in IJSEM Online.

Present address: Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University, Shanghai 200433, China.

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