Classification Of 13 New Species Of Lactobacillus

Northeast pickle is a traditional fermented food in China,mainly distributed in the northeastern regions of China.It has a long history of making and eating,and naturally fermented sauerkraut is rich in lactic acid bacteria.The naturally fermented yogurt in Tibet has a history of thousands of years and unique nutritional value,and is also rich in lactic acid bacteria.The purpose of this study was to identify 12 potential new lactic acid bacteria isolated from northeast sauerkraut and a new potential lactic acid bacteria isolated from Tibetan yoghurt by using a polyphasic method.New bacteria were characterized by using a polyphasic method,including 16 S r RNA gene sequence analysis,phe S gene sequence analysis,rpo A gene sequence analysis,DNA G+C mol% determination,average nucleotide identity(ANI)analysis,fatty acid methyl ester(FAME)analysis and an analysis of phenotypic features.Strain 54-2~T was phylogenetically related to type strains of Lactobacillus composti,Lactobacillus floricola,Lactobacillus selangorensis,Lactobacillus perolens,Lactobacillus harbinensis and Lactobacillus shenzhenensis,having 90.4-96.5% 16 S r RNA gene sequence similarities,56.7-74.6% phe S gene sequence similarities,57.2-81.6% rpo A gene sequence similarities and 66.26-72.50% ANI values.Strains 54-5~T,143-6~T,247-4~T,17-4~T and 143-1~T were phylogenetically related to type strains of Lactobacillus dextrinicus and Lactobacillus concavus,sharing 94.9-99.9% 16 S r RNA gene sequence similarities;70.1-83.1% phe S gene sequence similarities,81.0-98.3% rpo A gene sequence similarities,and 70.34-81.73% ANI values.Strains 33-7~T,116-2~T,184-8~T,204-8~T,8-1(1)~T,256-3~T and M1575 T were phylogenetically related to type strains of Lactobacillus tucceti,Lactobacillus nodensis,Lactobacillus insicii,Lactobacillus allii,Lactobacillus metriopterae,Lactobacillus terrae,Lactobacillus versmoldensis and Lactobacillus furfuricola,sharing 95.6-100% 16 S r RNA gene sequence similarities,73.1-91.6% phe S gene sequence similarities,78.9-98.2% rpo A gene sequence similarities,and 70.97-89.40% ANI values.The DNA G+C mol% of 54-2~T,54-5~T,143-6~T,247-4~T,17-4~T,143-1~T,33-7~T,116-2~T,184-8~T,204-8~T,8-1(1)~T were 39.96%,40.41%,42.48%,49.01%,41.47%,40.81%,35.56%,36.16%,34.57%,36.52%,36.71%,36.62%,and 39.36% respectively.The major fatty acids of strains 54-2~T,54-5~T,143-6~T,17-4~T and 33-7~T were C16:0 and C18:1?9c.The major fatty acids of strains 247-4~T,143-1~T,116-2~T,8-1(1)~T,M1575 T were C16:0,C18:1?9c and C18:1?6c/ C18:1?7c.The major fatty acids of strains 184-8~T,204-8~T and 256-3~T were C16 : 0,C18 : 1 ?9c,C19 : 1 ?6c/C19 : 0 cyclo ?10c and C18 : 1 ?6c/C18 : 1 ?7c.In this study,13 new lactic acid bacteria were identified by multiple classification methods to determine their taxonomic status,and 13 new lactic acid bacteria species were proposed.The type strains and corresponding names were as follows: 54-2~T(Lactobacillus yilanensis sp.nov.),54-5~T(Lactobacillus bayanensis sp.nov.),143-6~T(Lactobacillus mulanensis sp.nov.),247-4~T(Lactobacillus achengensis sp.nov.),17-4~T(Lactobacillus wuchangensis sp.nov.),143-1~T(Lactobacillus gannanensis sp.nov.),33-7~T(Lactobacillus keshanensis sp.nov.),116-2~T(Lactobacillus kedongensis sp.nov.),184-8~T(Lactobacillus baiquanensis sp.nov.),204-8~T(Lactobacillus jidongensis sp.nov.),8-1(1)~T(Lactobacillus hulinensis sp.nov.),256-3~T(Lactobacillus mishanensis sp.nov.),M1575~T(Lactobacillus zhongbaensis sp.nov.).