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For Peer Review Bergey?s Manual of Systematics of Archaea and Bacteria Tepidamorphus Journal: Bergey’s Manual of Systematics of Archaea and Bacteria Manuscript ID gbm01455.R1 Wiley - Manuscript type:For Genus Peer Paper Review Date Submitted by the Author: n/a Complete List of Authors: Albuquerque, Luciana; Center for Neuroscience and Cell Biology, University of Coimbra, Biotechnology Rainey, Fred; University of Alaska Anchorage, Biological Sciences da Costa, Milton; Center for Neuroscience and Cell Biology, University of Coimbra, Biotechnology Alphaproteobacteria, Rhodobiaceae, Slightly Thermophilic, Aerobic, Keywords: Budding John Wiley & Sons Page 1 of 26 Bergey?s Manual of Systematics of Archaea and Bacteria 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 For Peer Review 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 16S RNA gene sequence neighbor joining tree demonstration the position of the genus Tepidamorphus within the radiation of genera of the order Rhizobiales. The scale bar represents one nucleotide substitution 46 per 100 nucleotides. Numbers at branching points represent bootstrap values from 1000 replications. The 47 tree was rooted using the sequence of Roseospirillum parvum. 48 49 246x291mm (300 x 300 DPI) 50 51 52 53 54 55 56 57 58 59 60 John Wiley & Sons Bergey?s Manual of Systematics of Archaea and Bacteria Page 2 of 26 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 For Peer Review 20 21 22 23 24 25 26 27 28 29 30 31 Morphology of type strain of Tepidamorphus gemmatus in liquid cultures. Differential interference 32 microscopy showing overall features and budding (a, b). Transmission electron microscopy showing irregular 33 shaped rod- and large ovoid-shaped cells, clear intracellular inclusions may represent polyhydroxyalkanoate 34 (PHA) deposits (c). Aggregates of rod-shaped cells viewed by negative staining (d). Rod shaped cell with 35 and elongated hyphal–like bud (e). Two attached cells with several flagella (f). Rod-shaped cells producing 36 long rod-shaped or ovoid-shaped buds (g, h). *Reprinted with permission from Albuquerque et al. (2010a), Systematic and Applied Microbiology, Elsevier. 37 38 39 254x190mm (96 x 96 DPI) 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 John Wiley & Sons Page 3 of 26 Bergey?s Manual of Systematics of Archaea and Bacteria 1 2 3 1 Proteobacteria / Alphaproteobacteria / Rhizobiales / Rhodobiaceae 4 5 2 6 7 3 Tepidamorphus 8 9 10 4 11 VP 12 5 Albuquerque, Rainey, Pena, Tiago, Veríssimo, Nobre and da Costa 2010b, 1447 13 14 6 (Effective publication: Albuquerque, Rainey, Pena, Tiago, Veríssimo, Nobre and da 15 16 7 Costa 2010a, 65) 17 18 8 19 For Peer Review 20 9 Luciana Albuquerque, Center for Neuroscience and Cell Biology, University of 21 22 23 10 Coimbra, Coimbra, Portugal 24 25 11 Fred A. Rainey, Department of Biological Sciences, University of Alaska Anchorage, 26 27 12 Anchorage, AK, USA 28 29 13 Milton S. da Costa, Center for Neuroscience and Cell Biology, University of Coimbra, 30 31 14 Coimbra, Portugal 32 33 15 34 35 36 16 Te.pi.da.mor’phus. L. masc. adj. tepidus, moderately warm, lukewarm, tepid; Gr. masc. 37 38 17 adj. amorphos, without form, shapeless; N.L. masc. n. Tepidamorphus, an organism 39 40 18 without a distinctive morphology that grows at warm temperatures. 41 42 19 43 44 20 Abstract: 45 46 21 Irregular rod-shaped cells, 0.5–2.0 µm in width and 1.0–1.5 µm in length. Motile. Cells 47 48 with long rod-shaped structures; multiplies by budding. Endospores are not observed. 49 22 50 51 23 Stain Gram-negative. Colonies are nonpigmented. Slightly thermophilic. 52 53 24 Chemoorganotrophic. Strictly aerobic. Bacteriochlorophyll a and puf genes are not 54 55 25 present. Oxidase and catalase positive. Thiosulfate is not oxidized to sulfate. Sugars, 56 57 58 59 60 John Wiley & Sons Bergey?s Manual of Systematics of Archaea and Bacteria Page 4 of 26 1 2 3 26 organic acids and amino acids are used as carbon and energy sources. Major respiratory 4 5 27 quinone is ubiquinone 10 (U-10). Major polar lipids are phosphatidylcholine (PC), 6 7 28 phosphatidylethanolamine (PE), phosphatidylglycerol (PG), diphosphatidylglycerol 8 9 29 (DPG) and phosphatidylmonomethylethanolamine (PME). Major fatty acids are 10 11 30 primarily saturated and monounsaturated straight-chained. Isolated from hydrothermal 12 13 14 31 areas. 15 16 32 DNA G+C content (mol%): 67 (HPLC). 17 18 33 Type species: Tepidamorphus gemmatus Albuquerque, Rainey, Pena, Tiago, 19 For Peer Review 20 34 Veríssimo, Nobre and da Costa 2010b, 1447VP. (Effective publication: Albuquerque, 21 22 35 Rainey, Pena, Tiago, Veríssimo, Nobre and da Costa 2010a, 65). 23 24 36 25 26 27 37 Keywords: 28 29 38 Alphaproteobacteria, Rhodobiaceae, Slightly Thermophilic, Aerobic, Budding 30 31 39 32 33 40 Irregular rod-shaped cells, 0.5–2.0 µm in width and 1.0–1.5 µm in length. Motile. 34 35 41 Cells with long rod-shaped structures; multiplies by budding. Endospores are not 36 37 42 observed. Stain Gramnegative. Colonies are nonpigmented. Slightly thermophilic. 38 39 Chemoorganotrophic. Strictly aerobic. Bacteriochlorophyll a and puf genes are not 40 43 41 42 44 present. Oxidase and catalase positive. Thiosulfate is not oxidized to sulfate. Sugars, 43 44 45 organic acids and amino acids are used as carbon and energy sources. Major 45 46 46 respiratory quinone is ubiquinone 10 (U-10). Major polar lipids are 47 48 47 phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylglycerol 49 50 48 (PG), diphosphatidylglycerol (DPG) and phosphatidylmonomethylethanolamine 51 52 49 (PME). Major fatty acids are primarily saturated and monounsaturated straight- 53 54 55 50 chained. Isolated from hydrothermal areas. 56 57 58 59 60 John Wiley & Sons Page 5 of 26 Bergey?s Manual of Systematics of Archaea and Bacteria 1 2 3 51 DNA G+C content (mol%): 67 (HPLC). 4 5 52 Type species: Tepidamorphus gemmatus Albuquerque, Rainey, Pena, Tiago, 6 7 53 Veríssimo, Nobre and da Costa 2010b, 1447VP. (Effective publication: Albuquerque, 8 9 54 Rainey, Pena, Tiago, Veríssimo, Nobre and da Costa 2010a, 65). 10 11 55 Number of validated species: 1. 12 13 14 56 Family classification: The genus Tepidamorphus is classified whithin the family 15 16 57 Rhodobiaceae (fbm00172). 17 18 58 19 For Peer Review 20 59 Further descriptive information 21 22 23 60 24 25 61 Phylogeny 26 27 62 The genus Tepidamorphus comprises, at this time, one species named T. gemmatus, 28 29 T 63 represented by two strains, CB-27A and CB-26A. 16S rRNA gene sequence-based 30 31 32 64 comparisons show T. gemmatus to fall within the radiation of the order Rhizobiales 33 34 65 (obm00071) (Figure 1). The Tepidamorphus lineage clusters with a clade comprising 35 36 66 the genera Microbaculum, Lutibaculum, and Butyratibacter, sharing 94-96% similarity 37 38 67 with species of these genera (Kumar et al., 2012; Su et al., 2017; Wang et al., 2017). 39 40 68 These similarity values as well as other characteristics support the genus status of the 41 42 69 Tepidamorphus lineage. Genera peripheral to this clade include Afifella, Bauldia, 43 44 45 70 Dichotomicrobium (gbm00817), Methyloceanibacter, Methyloligella, and Rhodobium 46 47 71 (gbm00849) (Albuquerque et al., 2014; Doronina et al., 2013; Hirsch and Hoffmann 48 49 72 1989; Srinivas et al., 2007; Takeuchi et al., 2014; Urdiain et al., 2008; Yee et al., 2010). 50 51 73 The genus Rhodobium is the type genus of the family Rhodobiaceae (fbm00172) 52 53 74 (Garrity et al., 2005). Although the cluster of the genera listed above is not supported by 54 55 75 bootstrap analyses, for the purpose of this outline we include the genus Tepidamorphus 56 57 58 59 60 John Wiley & Sons Bergey?s Manual of Systematics of Archaea and Bacteria Page 6 of 26 1 2 3 76 within the family Rhodobiaceae and await further analysis of the phylogeny of the 4 5 77 Alphaproteobacteria (cbm00041) based on whole genome sequence comparisons. 6 7 78 8 9 79 Cell morphology and colony characteristics 10 11 80 Tepidamorphus gemmatus forms translucent nonpigmented colonies and the cells are 12 13 14 81 irregular rod-shaped, 0.5–2.0 µm in width by 1.0–1.5 µm in length with ovoid- and long 15 16 82 rod-shaped budding structures (Figure 2a, 2b) and is motile by means of one or more 17 18 83 flagella (Albuquerque et al., 2010a). Negative staining electron-microscopy of 19 For Peer Review 20 84 exponential phase cultures grown in liquid medium shows that the cells have irregular 21 22 85 morphologies (Figure 2c), with frequent branching and aggregation (Figure 2d). Some 23 24 86 branches originate from a lateral position of the cells while others are apical. However, 25 26 27 87 many of these branches resemble elongated buds (Figure 2e, 2f, 2g, 2h). A dark electron 28 29 88 dense band is frequently seen at the junction of separation of the cells. Electron dense 30 31 89 circular structures appear to correspond to polyphosphate granules. Lipid like 32 33 90 inclusions, corresponding to polyhydroxyalkanoate (PHA), are very common and these 34 35 91 frequently surround a small electron-dense granule. Carotenoid pigments are not 36 37 92 detected. 38 39 40 93 41 42 94 Nutrition and growth conditions 43 44 95 This slightly thermophilic species has an optimum growth temperature of about 45– 45 46 96 50ºC and the temperature range for growth is between 30ºC and 50ºC.
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