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).

Lactobacillus kosoi Chiou et al. 2018 and Lactobacillus micheneri McFrederick et al. 2018 are closely related, and they share 100 % 16S rRNA gene sequence similarity, 99.6 % pheS gene sequence similarity, 100 % rpoA gene sequence similarity, 97.3 % average nucleotide identity (ANI) value and 76.6 % in silico DNA–DNA hybridization (isDDH) value, indicating that they represent the same species. Fatty acid methyl esters (FAME) analysis and phenotypic characterization also indicated that L. kosoi and L. micheneri are very similar. We propose L. kosoi Chiou et al. 2018 as a later heterotypic synonym of L. micheneri McFrederick et al. 2018. The taxonomic position of Lactobacillus plantarum subsp. argentoratensis in the L. plantarum group was re-examined using a polyphasic approach, including sequence analyses of 16S rRNA, pheS, rpoA and recA genes, average nucleotide identity analysis, in silico DNA–DNA hybridization, fatty acid methyl ester analysis and phenotypic characterization. Results of 16S rRNA gene sequence analysis indicated that L. plantarum subsp. argentoratensis was closely related to L. plantarum subsp. plantarum , L. pentosus and L. paraplantarum in the L. plantarum group, sharing 99.6–99.7 % 16S rRNA gene sequence similarities. Results of pheS, rpoA and recA gene sequence analyses indicated that L. plantarum subsp. argentoratensis was most closely related to L. plantarum subsp. plantarum , having 91.8 % pheS gene sequence similarity, 98.9 % rpoA gene sequence similarity and 93.1 % recA gene sequence similarity. L. plantarum subsp. argentoratensis DSM 16365T shared 95.6 % ANI value and 62.9 % isDDH value with L. plantarum subsp. plantarum ATCC 14917T. The low isDDH value confirmed that L. plantarum subsp. argentoratensis and L. plantarum subsp. plantarum represent two different species, rather than two different subspecies in the L. plantarum group. On the basis of the data from polyphasic characterization obtained in the present study and in previous studies, L. plantarum subsp. argentoratensis is elevated to the species level and represents a novel species of the genus Lactobacillus , for which the name Lactobacillus argentoratensis sp. nov. is proposed and the type strain is DKO 22T (=CIP 108320T=DSM 16365T=JCM 16169T). Two novel Gram-stain-positive bacterial strains, designated 1206-1T and F027-1-2, were isolated from traditional pickle in Heilongjiang Province, PR China, and from the intestinal tract of a honey bee (Apis mellifera) in Hubei Province, PR China, respectively. The two bacteria were characterized by a polyphasic approach, including 16S rRNA gene sequence analysis, pheS gene sequence analysis, rpoA gene sequence analysis, fatty acid methyl ester analysis, average nucleotide identity analysis, in silico DNA–DNA hybridization analysis and an analysis of phenotypic features. The results of 16S rRNA gene sequence analysis indicated that strains 1206-1T and F027-1-2 were distantly related to Lactobacillus sharpeae , Lactobacillus hulanensis , Lactobacillus songhuajiangensis , Lactobacillus pantheris , Lactobacillus thailandensis , Lactobacillus camelliae , Lactobacillus jixianensis , Lactobacillus nasuensis , Lactobacillus baoqingensis , Lactobacillus manihotivorans and Lactobacillus porcinae . Strain 1206-1T exhibited 94.2–96.4 % 16S rRNA gene sequence similarities, 69.5–83.3 % pheS gene sequence similarities and 73.1–90.3 % rpoA gene sequence similarities to type strains of phylogenetically related species. ANI and isDDH values between strain 1206-1T and the type strains of phylogenetically related species were 52.7–73.7 % and 21.1–30.1 %, respectively. On the basis of the data obtained in the present study, a novel species, Lactobacillus zhaodongensis sp. nov. is proposed and the type strain is 1206-1T (=CCM 8981T=CCTCC AB 2019200T=LMG 31620T).

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).

Thirty-one rhizobial strains isolated from nodules of legumes native of Xinjiang, China, were characterized. These strains were classified as belonging to the genus Rhizobium based on amplified 16S rDNA restriction analysis (ARDRA). The strains were distinguished from recognized Rhizobium species using analysis of 16S-23S rDNA intergenic spacers (IGS-RFLP), SDS-PAGE analysis of whole proteins and BOX-PCR; the test strains always formed a distinct cluster with patterns that were quite different from those of the reference rhizobial strains used. According to the phylogenetic analysis based on the 16S rRNA gene, the test strains belonged to the genus Rhizobium, with Rhizobium tropici, Rhizobium rhizogenes and Rhizobium lusitanum as the closest related species, with 99.6, 99.2 and 99.4 % sequence similarities, respectively, between the type strains of the three Rhizobium species and strain CCBAU 83401T .Phylogenetic analyses of the representative strains using IGS and atpD, recA and glnII genes all confirmed the phylogenetic arrangements obtained using the 16S rRNA gene. The DNA-DNA relatedness values between strain CCBAU 83401 T and strains CCBAU 83364, CCBAU 83345and CCBAU 83523 ranged from 80.8 to 100 %, showing that they belong to the same species. The DNA-DNA relatedness between strain CCBAU 83401 T and R. tropici IIB CIAT 899 T , R. tropici IIA CFN 299, R. rhizogenes LMG 150 T and R. lusitanum P1-7 T were 26.9, 27.7, 38.2 and 22.6 %, respectively, clearly indicating that strain CCBAU 83401 T represents a novel species.Phenotypic characterization of four representative strains, CCBAU 83401 T , CCBAU 83364, CCBAU 83345 and CCBAU 83523, showed several distinctive features that differentiated them from closely related species. The 31 strains had identical nodD and nifH genes, which were very similar to those of the bean-nodulating R. lusitanum, Devosia neptuniae and R. tropici IIB. Based upon these results, the strains from this study are considered to represent a novel species, for which the name Rhizobium multihospitium sp. nov. is proposed. The DNA G+C content ranged from 65.3 to 66.0 mol% (T m ). The type strain is CCBAU 83401 T (5LMG 23946 T 5HAMBI 2975 T ), which nodulates Robinia pseudoacacia, but not Leucaena leucocephala, Phaseolus vulgaris, Pisum sativum or Medicago sativa.

A Gram-stain-positive bacterial strain, S4-3 T , was isolated from traditional pickle in Heilongjiang Province, China. The bacterium was characterized by a polyphasic approach, including 16S rRNA gene sequence analysis, pheS gene sequence analysis, rpoA gene sequence analysis, dnaK gene sequence analysis, fatty acid methyl ester (FAME) analysis, determination of DNA G+C content, DNA-DNA hybridization and an analysis of phenotypic features. Strain S4-3 T showed 97.9-98.7 % 16S rRNA gene sequence similarities, 84.4-94.1 % pheS gene sequence similarities and 94.4-96.9 % rpoA gene sequence similarities to the type strains of Lactobacillus nantensis, Lactobacillus mindensis, Lactobacillus crustorum, Lactobacillus futsaii, Lactobacillus farciminis and Lactobacillus kimchiensis. dnaK gene sequence similarities between S4-3 T and Lactobacillus nantensis LMG 23510 T , Lactobacillus mindensis LMG 21932 T , Lactobacillus crustorum LMG 23699 T , Lactobacillus futsaii JCM 17355 T and Lactobacillus farciminis LMG 9200 T were 95.4, 91.5, 90.4, 91.7 and 93.1 %, respectively. Based upon the data obtained in the present study, a novel species, Lactobacillus heilongjiangensis sp. nov., is proposed and the type strain is S4-3 T (5LMG 26166 T 5NCIMB 14701 T ).Pickle is rich in lactic acid bacteria (LAB). In recent years, many novel LAB species and subspecies have been isolated from pickle, such as Lactobacillus senmaizukei (Hiraga et al., et al., 2012). In the present study, a Gram-stain-positive bacterial strain, S4-3 T , was isolated from traditional pickle in Heilongjiang Province, China. The bacterium was characterized by a polyphasic approach. The strains used in this study are listed in Table 1. All strains were incubated aerobically at 30 u C on mMRS medium (pH 6.2-6.4) consisting of 0.5 % peptone, 0.5 % meat extract, 1 % tryptone, 0.5 % yeast extract, 1 % glucose, 0.1 % Tween 80, 0.2 % K 2 HPO 4 . 3H 2 O, 0.5 % sodium acetate anhydrous, 0.2 % diammonium hydrogen citrate, 0.025 % MnSO 4 . H 2 O and 0.01 % MgSO 4 . 7H 2 O.

Fourteen Gram-stain-positive bacterial strains were isolated from Chinese traditional pickle and yogurt. The strains were characterized using a polyphasic taxonomic approach, including 16S rRNA gene sequence analysis, pheS gene sequence analysis, rpoA gene sequence analysis, fatty acid methyl ester analysis, determination of DNA G+C content, determination of average nucleotide identity (ANI), in silico DNA-DNA hybridization (isDDH) and an analysis of phenotypic features. The data demonstrated that the 14 strains represented ten novel species belonging to the genus Lactobacillus, strains 73-4 T , 247-3 T , 143-4(a) T , 33-1 T , 143-6 T , 247-4 T , 17-4 T , 143-1 T , 735-2 T and M1530-1 T were designated as the type strains. Strains 73-4 T and 247-3 T were phylogenetically related to the type strains of Lactobacillus camelliae and Lactobacillus jixianensis, having 97.0-98.9 % 16S rRNA gene sequence similarities, 83.9-87.2 % pheS gene sequence similarities and 86.8-93.3 % rpoA gene sequence similarities. Strains 143-4(a) T and 33-1 T were phylogenetically related to the type strains of Lactobacillus rhamnosus, Lactobacillus paracasei and Lactobacillus casei, having 93.6-96.5 % 16S rRNA gene sequence similarities, 73.9-77.2 % pheS gene sequence similarities and 76.1-77.6 % rpoA gene sequence similarities. Strains 143-6 T , 247-4 T , 17-4 T and 143-1 T were phylogenetically related to the type strains of Lactobacillus concavus, Lactobacillus dextrinicus and Lactobacillus bayanensis, exhibiting 95.5-99.9 % 16S rRNA gene sequence similarities, 76.5-83.1 % pheS gene sequence similarities and 83.6-98.3 % rpoA gene sequence similarities. Strain 735-2 T was phylogenetically related to the type strains of Lactobacillus zhaoyuanensis, Lactobacillus jiayinensis and Lactobacillus coryniformis, having 98.2-99.1 % 16S rRNA gene sequence similarities, 82.8-84.1 % pheS gene sequence similarities and 93.0-93.9 % rpoA gene sequence similarities. Strain M1530-1 T was phylogenetically related to the type strains of Lactobacillus suantsaiihabitans and Lactobacillus brevis, having 99.5 and 99.0 % 16S rRNA gene sequence similarities, 90.3 and 81.7 % pheS gene sequence similarities and 97.7 and 91.1 % rpoA gene sequence similarities. The ANI and isDDH values between strains 73-4 T , 247-3 T , 143-4(a) T , 33-1 T , 143-6 T , 247-4 T , 17-4 T , 143-1 T , 735-2 T , M1530-1 T and type strains of phylogenetically related species were less than 86.8 % and 33.9 % respectively, confirming that they represent ten novel species within the genus Lactobacillus. Based upon the data of polyphasic characterization obtained in the present study, ten novel species,

Three Gram-stain-positive bacterial strains, 11050T, 7-19T and 11102T, were isolated from traditional pickle and sourdough in Heilongjiang Province, China. These bacteria were characterized by a polyphasic approach, including 16S rRNA gene sequence analysis, pheS gene sequence analysis, rpoA gene sequence analysis, dnaK gene sequence analysis, fatty acid methyl ester (FAME) analysis, determination of DNA G+C content, DNA–DNA hybridization and an analysis of phenotypic features. Strain 11050T belonged to the Lactobacillus plantarum species group and shared 98.0–98.4 % 16S rRNA gene sequence similarities and 84.7–88.9 % dnaK gene sequence similarities with type strains of Lactobacillus plantarum subsp. plantarum , Lactobacillus plantarum subsp. argentoratensis , Lactobacillus pentosus , Lactobacillus paraplantarum , Lactobacillus fabifermentans and Lactobacillus xiangfangensis and had 75.9–80.7 % pheS gene sequence similarities and 90.7–92.5 % rpoA gene sequence similarities with Lactobacillus plantarum subsp. plantarum LMG 6907T, Lactobacillus plantarum subsp. argentoratensis LMG 9205, Lactobacillus pentosus LMG 10755T, Lactobacillus paraplantarum LMG 16673T, Lactobacillus fabifermentans LMG 24284T and Lactobacillus xiangfangensis 3.1.1T, respectively. Strain 7-19T was phylogenetically related to Lactobacillus thailandensis , Lactobacillus pantheris and Lactobacillus sharpeae , having 94.1–96.7 % 16S rRNA gene sequence similarities, 71.5–82.3 % pheS gene sequence similarities and 71.2–83.4 % rpoA gene sequence similarities with type strains of Lactobacillus thailandensis , Lactobacillus pantheris and Lactobacillus sharpeae , respectively. Strain 11102T was phylogenetically related to Lactobacillus oligofermentans , Lactobacillus suebicus , Lactobacillus vaccinostercus and Lactobacillus hokkaidonensis . Strain 11102T had 99.2 % 16S rRNA gene sequence similarity, 81.3 % pheS gene sequence similarity and 96.1 % rpoA gene sequence similarity with Lactobacillus oligofermentans LMG 22743T, respectively. Strain 11102T shared 96.0–96.8 % 16S rRNA gene sequence similarities, 73.3–81.0 % pheS gene sequence similarities and 74.6–76.9 % rpoA gene sequence similarities with type strains of Lactobacillus suebicus , Lactobacillus vaccinostercus and Lactobacillus hokkaidonensis , respectively. Based upon the data from polyphasic characterization obtained in the present study, three novel species, Lactobacillus mudanjiangensis sp. nov., Lactobacillus songhuajiangensis sp. nov. and Lactobacillus nenjiangensis sp. nov., are proposed and the type strains are 11050T ( = LMG 27194T = CCUG 62991T), 7-19T ( = LMG 27191T = NCIMB 14832T = CCUG 62990T) and 11102T ( = LMG 27192T = NCIMB 14833T), respectively.