A Gram-stain-negative bacterial strain, 10-17T, was isolated from traditional sourdough in Heilongjiang Province, China. The bacterium was characterized by a polyphasic approach, including 16S rRNA gene sequence analysis, RNA polymerase β subunit (rpoB) gene sequence analysis, DNA gyrase (gyrB) gene sequence analysis, initiation translation factor 2 (infB) gene sequence analysis, ATP synthase β subunit (atpD) gene sequence analysis, fatty acid methyl ester analysis, determination of DNA G+C content, DNA–DNA hybridization and an analysis of phenotypic features. Strain 10-17T was phylogenetically related to Enterobacter hormaechei CIP 103441T, Enterobacter cancerogenus LMG 2693T, Enterobacter asburiae JCM 6051T, Enterobacter mori LMG 25706T, Enterobacter ludwigii EN-119T and Leclercia adecarboxylata LMG 2803T, having 99.5 %, 99.3 %, 98.7 %, 98.5 %, 98.4 % and 98.4 % 16S rRNA gene sequence similarity, respectively. On the basis of polyphasic characterization data obtained in the present study, a novel species, Enterobacter xiangfangensis sp. nov., is proposed and the type strain is 10-17T ( = LMG 27195T = NCIMB 14836T = CCUG 62994T). Enterobacter sacchari Zhu et al. 2013 was reclassified as Kosakonia sacchari comb. nov. on the basis of 16S rRNA, rpoB, gyrB, infB and atpD gene sequence analysis and the type strain is strain SP1T( = CGMCC 1.12102T = LMG 26783T).
Six strains were isolated from root nodules of Vicia faba grown in Nanchang, Yifeng, Taihu, Huaibei, Bengbu and Lujiang, in the middle and lower reaches of the Yangtze River. According to phylogenetic analyses of 16S rRNA gene, atpD and recA sequences, these strains belong to the genus Rhizobium, with Rhizobium etli and Rhizobium leguminosarum as the closest related species. CCBAU 33202 T , a representative of these novel isolates, showed sequence similarity to its closest relatives R. etli CFN 42 T and R. leguminosarum USDA 2370 T of 99.5 and 99.1 % for the 16S rRNA gene, 91.9 and 91.9 % for atpD and 90.3 and 93.2 % for recA. The strains from this study could also be differentiated from R. etli CFN 42 T and R. leguminosarum USDA 2370T by 16S-IGS RFLP and SDS-PAGE of whole-cell proteins, fatty acid profiles and several phenotypic characteristics. DNA-DNA hybridization yielded relatedness of 19 and 14-43 %, respectively, with R. etli CFN 42 T and strains representing different biovars of R. leguminosarum.All data obtained in this study showed that these V. faba isolates belong to a novel species, for which the name Rhizobium fabae sp. nov. is proposed. The type strain, CCBAU 33202 T (5LMG 23997 T 5JCM 14381 T ), was isolated from Nanchang.It is well known that Vicia faba and other species of Vicia, Pisum, Lathyrus and Lens form nodules with Rhizobium leguminosarum bv. viciae, one of the three biovars of R. leguminosarum (Jordan 1984). Earlier studies revealed that rhizobia isolated from nodules of V. faba in different geographical locations were diverse in their chromosomal background and plasmid content, and rhizobial strains of V. faba were distinguishable from rhizobial strains of Pisum sativum, especially in the nodulation genotypes (van Berkum et al., 1995;Laguerre et al., 2003;Young et al., 2003;Mutch et al., 2003;Mutch & Young, 2004). However, no species other than Rhizobium leguminosarum bv. viciae has been reported to nodulate V. faba. In our previous study, wide diversity was revealed amongst rhizobia associated with V. faba in Chinese fields (Tian et al., 2007). Six strains isolated from six sites in the middle and lower reaches of the Yangtze River formed a distinct group belonging to Rhizobium based upon analysis of BOX-A1R PCR, amplified fragment length polymorphism analysis, amplified 16S rDNA restriction analysis (ARDRA), 16S rRNA gene phylogeny and DNA-DNA hybridization (Tian et al., 2007). In the present study, a polyphasic approach was used to clarify the taxonomic affiliation of this Rhizobium group.Strains CCBAU 33202 T and CCBAU 33201, isolated from Nanchang and Yifeng in Jiangxi province, and CCBAU 23123, CCBAU 23122, CCBAU 23127 and CCBAU 23132, isolated from Taihu, Huaibei, Bengbu and Lujiang in Anhui province, all isolated from V. faba, were used in this study. Standard procedures and YMA medium (Vincent, 1970) were used to isolate and cultivate the strains. DNA samples were prepared as described by Terefework et al. (2001).Previously, these six strains have been grouped in the same ARDR...
Currently, Lactococcus lactis contains four subspecies: L. lactis subsp. lactis , L. lactis subsp. hordniae , L. lactis subsp. cremoris and L. lactis subsp. tructae . In the study of Pérez et al., these four subspecies could be clearly divided into two groups based on recA sequence analysis: L. lactis subsp. lactis and L. lactis subsp. hordniae ; L. lactis subsp. cremoris and L. lactis subsp. tructae . The two groups had a relatively low DNA–DNA hybridization value (about 60 %). In the present study, the taxonomic position of L. lactis subsp. cremoris and L. lactis subsp. tructae was re-examined based on sequence analyses of 16S rRNA, rpoB, recA and pheS genes, average nucleotide identity (ANI) values and digital DNA–DNA hybridization (dDDH) values. The result of 16S rRNA gene sequence analysis indicated that L. lactis subsp. cremoris NCDO 607T and L. lactis subsp. tructae L105T were phylogenetically related to the type strains of L. lactis subsp. hordniae , L. lactis subsp. lactis , Lactococcus taiwanensis , Lactococcus kimchii , Lactococcus allomyrinae , Lactococcus protaetiae, Lactococcus hircilactis , Lactococcus fujiensis and Lactococcus nasutitermitis . The 16S rRNA gene, rpoB, recA, pheS and concatenated rpoB, recA and pheS sequence similarities, ANI values, and dDDH values between the type strains of L. lactis subsp. cremoris , L. lactis subsp. tructae and phylogenetically related species were 93.5–99.4 %, 83.3–97.6 %, 80.6–92.4 %, 79.7–92.7 %, 83.5–94.3 %, 72.4–86.9 % and 21.4–32.5 %, respectively. Lower than 95–96 % ANI values and lower than 70 % dDDH values confirmed that the type strains of L. lactis subsp. cremoris and L. lactis subsp. tructae represent a novel species in the genus Lactococcus . Because L. lactis subsp. cremoris was proposed and validated before L. lactis subsp. tructae , L. lactis subsp. cremoris is elevated to the species level as Lactococcus cremoris sp. nov. and L. lactis subsp. tructae is transferred to L. cremoris as L. cremoris subsp. tructae comb. nov. The type strain of L. cremoris sp. nov. is NCDO 607T (=ATCC 19257T=DSM 20069T=JCM 16167T=LMG 6897T=NBRC 100676T). The type strain of L. cremoris subsp. tructae comb. nov. is L105T (=NBRC 110453T=DSM 21502T=JCM 31125T=LMG 24662T).
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