A rod-shaped, motile, facultatively anaerobic and moderately halotolerant plant-growth-promoting actinobacterial strain, designated NCCP-11 T , was isolated from paddy grains. To delineate its taxonomic position, the strain was subjected to a polyphasic characterization. Cells of strain NCCP-11 T grew at 10-37 6C (optimum 28-32 6C), at pH 6-9 (optimum pH 7) and in 0-12 % (w/v) NaCl (optimum 1-2 %) in broth medium. Based on 16S rRNA gene sequence analysis, strain NCCP-11 T showed highest similarity to the type strains of Cellulomonas hominis (98.99 %) and Cellulomonas denverensis (98.09 %) and less than 97 % with other closely related taxa. The chemotaxonomic data [major menaquinone: MK-9(H4); cell-wall peptidoglycan: type A4b; major fatty acids: anteiso-C 15 : 0 , C 16 : 0 , C 14 : 0 and anteiso-C 17 : 0 ; major polar lipids: diphosphatidylglycerol, phosphatidylinositol, phosphatidylinositolmannosides and two unknown polar lipids] also supported the affiliation of strain NCCP-11 T to the genus Cellulomonas. The level of DNA-DNA relatedness between strain NCCP-11 T and the two type strains mentioned above was less than 42.7 %. On the basis of DNA-DNA relatedness, physiological and biochemical characteristics and phylogenetic position, strain NCCP-11 T can be differentiated from species of the genus Cellulomonas with validly published names and thus represents a novel species, for which the name Cellulomonas pakistanensis sp. nov. is proposed. The type strain is NCCP-11 T (5DSM 24792 T 5JCM 18755 T 5KCTC 19798 T ).

The taxonomic position of a Gram-stain positive and heavy metal tolerant bacterium, designated strain NCCP-662(T), was investigated by polyphasic characterisation. Cells of strain NCCP-662(T) were observed to be rod to filamentous shaped, motile and strictly aerobic, and to grow at 10-50 °C (optimum 30-37 °C) and at pH range of 6-10 (optimum pH 7-8). The strain was found to be able to tolerate 0-12 % NaCl (w/v) and heavy metals (Cr 1200 ppm, Pb 1800 ppm and Cu 1200 ppm) in tryptic soya agar medium. The phylogenetic analysis based on the 16S rRNA gene sequence of strain NCCP-662(T) showed that it belongs to the genus Bacillus and showed high sequence similarity (98.2 and 98.0 %, respectively) with the type strains of Bacillus niabensis 4T19(T) and Bacillus halosaccharovorans E33(T). The chemotaxonomic data showed that the major quinone is MK-7; the predominant cellular fatty acids are anteiso-C15 :0, iso-C14:0, iso-C16:0 and C16:0 and iso-C15:0; the major polar lipids are diphosphatidylglycerol, phosphatidylglycerol along with several unidentified glycolipids, phospholipids and polar lipids. The DNA G+C content was determined to be 36.9 mol%. These data also support the affiliation of strain NCCP-662(T) with the genus Bacillus. The level of DNA-DNA relatedness between strain NCCP-662(T) and B. niabensis JCM 16399(T) was 20.5 ± 0.5 %. On the basis of physiological and biochemical characteristics, phylogenetic analyses and DNA-DNA hybridization data, strain NCCP-662(T) can be clearly differentiated from the validly named Bacillus species and thus represents a new species, for which the name Bacillus malikii sp. nov. is proposed with the type strain NCCP-662(T) (= LMG 28369(T) = DSM 29005(T) = JCM 30192(T)).

A Gram-negative, non-spore-forming, non-pigmented, strictly aerobic and non-motile short rod bacterium, designated NCCP-1258, was isolated from Cholistan desert soil, Bahawalpur, Pakistan. Growth of strain NCCP-1258 was observed at pH range 6.5-9.5 (optimum 7.5-8.5) and temperature range 20-45 °C (optimum 40 °C), and it tolerated 0-2 % NaCl (optimum 0.5 %, w/v). Phylogenetic analysis based on 16S rRNA gene sequence comparison revealed that strain NCCP-1258 belongs to genus Microvirga and is most closely related to Microvirga lotononidis (98.0 %), Microvirga vignae (97.4 %), Microvirga lupini (97.2 %), Microvirga zambiensis (97.2 %) and Microvirga flocculans (97.1 %). Analysis of the concatenated sequences of four housekeeping gene loci (dnaK, gyrB, recA and rpoB) also confirmed the placement of strain NCCP-1258 within the genus Microvirga. DNA-DNA relatedness values of NCCP-1258 with above-mentioned type strains were less than 42 %. The DNA G+C content of strain NCCP-1258 was 64.3 mol%. Chemotaxonomic data (predominant menaquinone system was Q-10; major fatty acids were C, C ω7c and C cyclo ω8c; the polar lipid profile contained diphosphatidylglycerol, phosphatidylcholine, phosphatidyl dimethyl ethanolamine and phosphatidyl ethanolamine) also supported the affiliation of strain NCCP-1258 to the genus Microvirga. On the basis of physiological and biochemical characteristics, phylogenetic analyses and DNA-DNA relatedness, strain NCCP-1258 can be distinguished from the closely related taxa and thus represents a novel species of the genus Microvirga, for which the name Microvirga pakistanensis sp. nov. is proposed with the type strain NCCP-1258 (=CGMCC 1.15074 = KCTC 42496).

The taxonomic position of a Gram-stain negative, moderately halophilic bacterium, designated NCCP-934(T), was investigated using polyphasic taxonomic approach. The strain NCCP-934(T) was isolated from rhizosphere of a plant (Saccharum spontaneum, family Poaceae) growing in salt mines area in the Karak district of Khyber Pakhtunkhwa Province, Pakistan. Cells of strain NCCP-934(T) are rod shaped and motile. The bacterium is strictly aerobic, can grow at a temperature range of 10-40 °C (optimum at 30-33 °C) and in a pH range of 6.0-10.5 (optimum pH 7.0-9.0). The strain can tolerate 1-30 % (w/v) NaCl (optimal growth occurs in the presence of approximately 3-9 % NaCl). The phylogenetic analysis based on the 16S rRNA gene sequence, showed that strain NCCP-934(T) belongs to the genus Kushneria with the highest sequence similarity to K. marisflavi SW32(T) (98.9 %), K. indalinina CG2.1(T) (98.7 %), K. avicenniae MW2a(T) (98.4 %) and less than 97 % similarity with other related species (94.7 % with the type species of the genus, K. aurantia A10(T)). DNA-DNA relatedness between strain NCCP-934(T) and the type strains of the closely related species was lower than 18 %. The chemotaxonomic data (major respiratory quinone, Q9; predominant fatty acids, C18:1 ω7c and C16:0 followed by C12:0 3-OH and Summed features 3 (C16:1 ω7c/iso-C15:0 2-OH); major polar lipids, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylmonomethylethanol, phosphatidylserine, phosphatidylinositol and three polar lipid of unknown structure) supported the affiliation of strain NCCP-934(T) within the genus Kushneria. The DNA G+C content of strain NCCP-934(T) was 59.2 mol%. On the basis of phenotypic, chemotaxonomic and phylogenetic data, strain NCCP-934(T) can be distinguished from the closely related taxa and thus represents a novel species in the genus Kushneria, for which the name Kushneria pakistanensis sp. nov. is proposed, with the type strain NCCP-934(T) (=LMG 28525(T) = KCTC 42082(T) = JCM 18802(T)).