Downloaded from www.microbiologyresearch.org by IP: 54.70.40.11 On: Sun, 02 Dec 2018 01:25:48 Bacillus lycopersici sp. nov., isolated from a tomato plant (Solanum lycopersicum L.) Shih-Yao Lin, 1 Asif Hameed, 1 You-Cheng Liu, 1 Cheng-Zhe Wen, 2 Wei-An Lai, 1,3 Yi-Han Hsu 1 and Chiu-Chung Young 1,3 Correspondence Chiu-Chung Young ccyoung@mail.nchu.edu.tw 1 Department of Soil and Environmental Sciences, College of Agriculture and Natural Resources, National Chung Hsing University, Taichung, Taiwan, ROC 2 Bachelor Program of Bio-Industry Management, College of Agriculture and Natural Resources, National Chung Hsing University, Taichung, Taiwan, ROC 3 Agricultural Biotechnology Center, National Chung Hsing University, Taichung, Taiwan, ROC A Gram-stain-positive, rod-shaped, endospore-forming bacterium (designated strain CC-Bw-5 T ) was isolated from chopped tomato stems. The isolate grew at 20–40 8C, pH 6.0–8.0 and tolerated 6 % (w/v) NaCl. The most closely related strains in terms of 16S rRNA gene sequence similarity were Bacillus isabeliae (95.3 %) and Bacillus oleronius (95.3 %). The G+C content of the genomic DNA was 37.2¡3.6 mol%. Strain CC-Bw-5 T was determined to possess C 16 : 0 , iso- C 14 : 0 , iso-C 15 : 0 , anteiso-C 15 : 0 and iso-C 16 : 0 as predominant fatty acids. The polar lipid profile consisted of predominant amounts of diphosphatidylglycerol, and moderate-to-trace amounts of phosphatidylethanolamine, phosphatidylglycerol, two unidentified glycolipids, an unidentified phospholipid and an unidentified phosphoglycolipid. The cell-wall peptidoglycan contained meso- diaminopimelic acid; menaquinone (MK-7) was the predominant respiratory quinone. According to distinct phylogenetic, phenotypic and chemotaxonomic features, strain CC-Bw-5 T is proposed to represent a novel species within the genus Bacillus for which the name Bacillus lycopersici sp. nov. is proposed. The type strain is CC-Bw-5 T (5BCRC 80623 T 5JCM 19140 T ). In 1872, Ferdinand Cohn proposed the genus Bacillus, with Bacillus subtilis as the type species. Most species of the genus represent Gram-stain-positive, rod-shaped, aerobic or facultatively anaerobic bacteria. Typical characteristics of members of the genus Bacillus are low DNA G+C con- tent and containing menaquinone MK-7 as the major respiratory quinone. Due to their capacity to form endo- spores, they can survive long periods of high temperature. Whereas most species of the genus Bacillus prefer neutral pH and low salt concentrations, some can also grow in environments with high alkalinity (Denizci et al., 2010), low pH (Albert et al., 2005) or high salinity (Pappa et al., 2010). The genus Bacillus contains more than two hundred species with validly published names listed on the LPSN website (http://www.bacterio.net/bacillus.html). At the time of writing, three novel species, Bacillus luteus (Subhash et al., 2014), Bacillus panacisoli (Choi & Cha, 2014) and Bacillus solimangrovi (Lee et al., 2014) have been newly described. In this study, we aimed to clarify the taxonomic status of a novel bacterium, designated CC-Bw-5 T , which was isolated from tomato stems. Tomato plant (Solanum lycopersicum L.) was collected from a greenhouse belonging to Taichung senior high school of National Chung Hsing University (248 129 N 1208 689 E), Taichung, Taiwan. The chopped tomato stems were introduced into sterile water and shaken at 30 8C for 30 min. Subsequently, standard serial dilution was carried out and the mixture spread-plated on tryptic soy agar (TSA; Difco). Distinct bacterial strains were isolated, identified and preserved as glycerol suspen- sion (30 %, v/v) at 280 8C for further characterization. In this study, the morphological, biochemical and phyloge- netic characteristics of a novel bacterium were described according to the minimal standards published by Logan et al. (2009) and Tindall et al. (2010), employing the most closely related type strains, Bacillus isabeliae BCRC 17837 T (Albuquerque et al., 2008) and Bacillus oleronius KCTC 3776 T (Heyndrickx et al., 2012) as reference strains in parallel tests. For direct comparative analysis, B. isabeliae BCRC 17837 T was restricted to growth on marine agar (MA; Hi-media), B. oleronius KCTC 3776 T was grown on TSA at 30 8C for 2 days, unless specified otherwise. In order to clarify the taxonomic position of the novel The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain CC-Bw-5 T is KC849424. Two supplementary figures are available with the online Supplementary Material. International Journal of Systematic and Evolutionary Microbiology (2015), 65, 2085–2090 DOI 10.1099/ijs.0.000223 000223 G 2015 IUMS Printed in Great Britain 2085