Lactobacillus kisonensis sp. nov., Lactobacillus otakiensis sp. nov., Lactobacillus rapi sp. nov. and Lactobacillus sunkii sp. nov., heterofermentative species isolated from sunki, a traditional Japanese pickle

1 Yakult Central Institute for Microbiological Research, 1796 Yaho, Kunitachi, Tokyo 186-8650, Japan 2 NODAI Culture Collection Center, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya, Tokyo 156-8502, Japan Correspondence Koichi Watanabe koichi-watanabe{at}yakult.co.jp Thirty six Gram-positive, rod-shaped, non-spore-forming, non-motile bacterial strains were isolated from the non-salted pickle solution used in producing sunki products, a traditional Japanese pickle. The novel strains were discriminated and separated into four groups by amplified fragment length polymorphism profiling, and by analysis based on recA gene sequences. The strains were classified into four species groups belonging to the Lactobacillus buchneri species group, which consists of L. buchneri , Lactobacillus diolivorans , Lactobacillus hilgardii , Lactobacillus kefiri , Lactobacillus parabuchneri and Lactobacillus parakefiri . The phenotypic and genotypic features of the four groups demonstrated that they represented four novel species, for which the names Lactobacillus kisonensis sp. nov. (type strain YIT 11168 T =NRIC 0741 T =JCM 15041 T =DSM 19906 T ), Lactobacillus otakiensis sp. nov. (type strain YIT 11163 T =NRIC 0742 T =JCM 15040 T =DSM 19908 T ), Lactobacillus rapi sp. nov. (type strain YIT 11204 T =NRIC 0743 T =JCM 15042 T =DSM 19907 T ) and Lactobacillus sunkii sp. nov. (type strain YIT 11161 T =NRIC 0744 T =JCM 15039 T =DSM 19904 T ) are proposed. Abbreviations: AFLP, amplified fragment length polymorphism; LAB, lactic acid bacteria The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene sequences of strains L. buchneri YIT 0077 T , L. diolivorans YIT 10368 T , L. hilgardii YIT 0269 T , L. kefiri YIT 0222 T , L. parabuchneri YIT 0272 T , L. parakefiri YIT 10382 T , L. sunkii sp. nov. YIT 11161 T , L. otakiensis sp. nov. YIT 11163 T , YIT 11167, L. kisonensis sp. nov. YIT 11168 T , L. rapi sp. nov. YIT 11204 T , YIT 11442, YIT 11452, YIT 11505, YIT 11510, YIT 11539, YIT 11549, YIT 11557, YIT 11635, YIT 11661 and YIT 11688 are AB429368 –AB429373 and AB366385 –AB366399, respectively, and for the recA gene sequences AB430354 –AB430374, respectively. A table detailing the DNA G+C contents and DNA–DNA relatedness of the novel strains as compared with type strains of species in the L. buchneri group is available with the online version of this paper. Additional phylogenetic trees based on 16S rRNA and RecA gene sequences and constructed with the neighbour-joini