Isolation And Classification Of 2 Novel Species Of Lactic Acid Bacteria From Northeast Pickle And The Intestinal Tract Of Honey Bees

Northeast pickle and the gut of honey bees are rich in lactic acid bacteria,lactic acid bacteria are the main flora in the fermentation process of pickled cabbage,and play a major role in the production of pickled cabbage;The gut flora of bees consist of specific symbiotic bacteria,and lactic acid bacteria have an important effect on the health of bees.Therefore,it is significant to understand the diversity of lactic acid bacteria for the process of northeast pickle and the health of bees.In this study,lactic acid bacteria in northeast pickle and bee gut were identified and analyzed by phe S gene sequence and 16S r RNA gene sequence.We isloated 39 strains of lactic acid bacteria from northeast pickle and isolated 66 strains of lactic acid bacteria from the intestine of bees,the total number of strains was 105 of lactic acid bacteria.These 105 strains of lactic acid bacteria were distributed in 2 genera,including Lactobacillus and Pediococcus,belonging to 17 known species and 3 unknown species.The known species isolated from northeast pickle were:L.plantarum subsp.plantarum,L.xiangfangensis,L.brevis,L.koreensis,Pediococcus parvulus,L.buchneri,L.parabuchneri,L.coryniformis,L.paracasei,L.nenjiangensis,among them,L.coryniformis was the dominant strain in northeast pickle.The known species isolated from the bee gut were:L.plantarum subsp.plantarum,L.brevis,L.apinorum,L.sakei,L.kullabergensis,L.parabuchneri,L.helsingborgensis,L.apis,L.kunkeei,Pediococcus acidilactici,L.kunkeei was the dominant strain in the bee gut.strains 395-6.2~T and 1206-1~Twere two unknown species which isolated from northeast pickle,and one unknown specie was isolated from bee gut as strain F027-1-2.16S r RNA gene sequence analysis showed that the strain 395-6.2~T was phylogenetically related to L.farciminis,L.formosensis,L.futsaii,L.crustorum,L.nuruki,L.heilongjiangensis,L.musae,L.nantensis,L.mindensis,L.kimchiensis,L.zhachilii,L.alimentarius,L.bobalius,L.kimchii and L.paralimentarius.Strain 395-6.2~T exhibited 95.7-99.4%16S r RNA gene sequence similarities,85.0-94.0%phe S gene sequence similarities,94.2-98.0%rpo A gene sequence similarities to type strains of phylogenetically related species.ANI and d DDH values between strain 395-6.2~T and type strains of phylogenetically related species were 77.9-87.1%and22.5-33.5%,respectively.Based upon the data obtained in the present study,a novel species was proposed and the type strain was 395-6.2~T.The result of 16S r RNA gene sequence analysis indicated that strains 1206-1~T and F027-1-2 were distantly related to the type strains of L.sharpeae,L.hulanensis,L.songhuajiangensis,L.pantheris,L.hailandensis,L.camelliae,L.jixianensis,L.nasuensis,L.baoqingensis,L.manihotivorans,L.porcinae,L.brantae and L.saniviri.Strain1206-1~T exhibited 94.2-96.4%16S r RNA gene sequence similarities,69.5-83.3%phe S gene sequence similarities,73.1-90.3%rpo A gene sequence similarities to type strains of phylogenetically related species.Strains 1206-1~T and F027-1-2 were aligned by MEGAX,the sequences were consistent,and Blast was performed on NCBI.Strains 1206-1~T and F027-1-2 had identical 16S r RNA,phe S and rpo A gene sequences,and their phylogenetic positions were consistent.16S r RNA gene sequence similarities were less than 96.4%,it was shown that strains1206-1~Tand F027-1-2 were the same new lactic acid bacteria.Taxonomic identification of new species 395-6.2~T and 1206-1~T(F027-1-2).This study showed that lactic acid bacteria had diversity in northeast pickle and bee gut,and the new lactic acid bacteria were isolated have expanded the classification system of lactic acid bacteria,It provided new resources and ideas for the application of lactic acid bacteria in northeast pickle,the health research of bees and the development of lactic acid bacteria resources.