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.