Downloaded from www.microbiologyresearch.org by IP: 54.70.40.11 On: Mon, 05 Nov 2018 14:23:02 Falsirhodobacter halotolerans gen. nov., sp. nov., isolated from dry soils of a solar saltern Y. Subhash, 1 L. Tushar, 1 Ch. Sasikala 2 and Ch. V. Ramana 1 Correspondence Ch. V. Ramana r449@sify.com or sasi449@yahoo.ie 1 Department of Plant Sciences, School of Life Sciences, University of Hyderabad, PO Central University, Hyderabad 500046, India 2 Bacterial Discovery Laboratory, Centre for Environment, Institute of Science and Technology, J.N.T. University Hyderabad, Kukatpally, Hyderabad 500085, India Two bacterial strains (JA744 T and JA745) were isolated from dry soil samples collected from solar salterns at Humma, Odisha, India. Both strains were Gram-stain-negative, catalase- and oxidase- positive, motile rods. Major fatty acids in both strains included C 18 : 1 v7c,C 18 : 0 and C 16 : 0 , while minor amounts of C 10 : 0 3-OH, C 12 : 0 ,C 12 : 0 3-OH, C 14 : 0 and C 16 : 0 were also present. Diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phosphatidylglycerol, an unidentified glycolipid, five unidentified lipids, an unidentified aminolipid and an unidentified phospholipid made up the polar lipids of both strains. Both strains had bacteriohopane derivatives (BHD1,2) and diploptene as major hopanoids. Mean genomic DNA G+C content was 75±1 mol% and the two strains were closely related (mean DNA–DNA hybridization .90 %). Phylogenetic analysis based on the 16S rRNA gene sequence showed that the two strains clustered with species of the genus Rhodobacter belonging to the family Rhodobacteraceae of the class Alphaproteobacteria. The highest sequence similarity was observed with Rhodobacter sphaeroides ATH2.4.1 T (96 %) and other members of the genera Rhodobacter and Pseudorhodobacter (,96 %). However, the two strains were positioned distinctly outside the group formed by the other genera of the family Rhodobacteraceae. Distinct morphological, physiological and genotypic differences from previously described taxa support the classification of these isolates as representatives of a novel species in a new genus, for which the name Falsirhodobacter halotolerans gen. nov., sp. nov. is proposed. The type strain of Falsirhodobacter halotolerans is JA744 T (5KCTC 32158 T 5NBRC 108897 T ). Phototrophy is considered to be an important character in differentiating genera of phototrophs and chemotrophs (Imhoff & Caumette, 2004). Such a distinction is par- ticularly clear for members of the family Rhodobacteraceae, where the phototrophic genera are distinguished from the nearest interspersed chemotrophs. A good example is the transfer of Rhodobacter massiliensis (Greub & Raoult, 2003) to a new genus, as Haematobacter massiliensis (Helsel et al., 2007). In this communication, we report two novel non- phototrophic members of a new genus associated with the family Rhodobacteraceae, isolated from a solar saltern. Strains JA744 T and JA745 were isolated from dry soils of an unused solar saltern at Humma, Odisha, India (GPS positioning of the sample collection site is 19 u 259 N 85 u 049 E), in December 2011. One gram dry soil was serially diluted [10- fold dilution in saline (0.6 % NaCl)] and plated on a medium (pH 7.0) containing (g l 21 ) KH 2 PO 4 (0.2), NH 4 Cl (0.25), KCl (0.5), CaCl 2 . 2H 2 O (0.15), NaCl (1), MgCl 2 . 6H 2 O (0.62), Na 2 SO 4 (2.84), HEPES (2.83), yeast extract (3.0), peptone (3), Casamino acids (0.5), glucose (0.5) and sodium pyruvate (3.0). Three distinct colony morphologies were observed from plates incubated at 30 u C for 3 days. Two pale-yellow colonies were selected for further purification and the purified isolates were designated strains JA744 T and JA445. Both cultures were preserved as glycerol stocks and by lyophilization and stored at 4 u C. Genomic DNA was extracted and purified according to the method of Marmur (1961) and the G+C content of the DNA as determined by reversed-phase HPLC (Mesbah et al., 1989) was 75±1 mol% for both strains JA744 T and JA745. Cell material for 16S rRNA gene sequencing was taken from a colony. DNA was extracted and purified by using a Qiagen genomic DNA extraction kit. Recombinant Taq polymerase (Genei) was used for PCR. The almost- complete 16S rRNA gene sequence was obtained using The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene sequences of strains JA744 T and JA745 are HE662814 and HE662815. Four supplementary figures and a supplementary table are available with the online version of this paper. Abbreviation: BChl, bacteriochlorophyll. International Journal of Systematic and Evolutionary Microbiology (2013), 63, 2132–2137 DOI 10.1099/ijs.0.044107-0 2132 044107 G 2013 IUMS Printed in Great Britain