Phenotypic Characteristics Combined with Genomic Analysis to Identify Different Colony Morphology Bacillus velezensis ACCC 19742

doi:10.13560/j.cnki.biotech.bull.1985.2019-0750

Abstract

Abstract: During the period of regular transfer and preservation of bacteria in Agricultural Culture Collection of China（ACCC）,it was found that Bacillus amyloliquefaciens ACCC 19742 showed two different colonies on the same medium,named these two different colonies as 19742-1 and 19742-2. The morphology,physiology,biochemistry and genome analysis were used to identify ACCC 19742,and to further determine the taxonomic status of the strain. Firstly,the strains were isolated and purified. Secondly,16S rRNA and gyrB genes were amplified and sequenced. Phylogenetic tree was constructed by MEGA 7.0 software. Physiological and biochemical characteristics of the strains were analyzed by API 20NE,BIOLOG and fatty acid. The ANI and DDH values of the strains were analyzed by whole genome analysis. Only the gluconate assimilation reaction in 2 strains was different according to API 20NE detection,the main fatty acid components were consistent,only the percentage was slightly different. Based on the phylogenetic analysis,16S rRNA gene similarity of the two strains was 100%,while the gyrB gene similarity was 99.4%. The ANI value of the two strains was 99.95% and the DDH value was 99.62%. Combining with genetic and phenotypic characteristics,it was confirmed that the two strains were derived from different morphological variants of the same strain,not caused by pollution. Meanwhile,ANI and DDH values of 19742 and Bacillus velezensis NRRL_B 41580^Twere the highest（97% and 77%,respectively）. Evolutionary analysis of 16S rRNA and gyrB also indicated that the strain belonged to the taxonomic status Bacillus velezensis,but not B. amyloliquefaciens. This provides a certain reference value for the preservation of the strain.

Key words: Bacillus amyloliquefaciens, morphovar, whole genome sequencing, Bacillus velezensis

GUO He-bao, WANG Xing, HE Shan-wen, ZHANG Xiao-xia. Phenotypic Characteristics Combined with Genomic Analysis to Identify Different Colony Morphology Bacillus velezensis ACCC 19742[J]. Biotechnology Bulletin, 2020, 36(2): 142-148.

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