Pararobbsia silviterrae gen. nov., sp. nov., isolated from forest soil and reclassification of Burkholderia alpina as Pararobbsia alpina comb. nov.

A novel Gram-stain-negative, aerobic, non-spore-forming, non-motile and rod-shaped bacterial strain, DHC34T, was isolated from forest soil of Dinghushan Biosphere Reserve, Guangdong Province, China (112° 31' E 23° 10' N). It grew optimally on R2A medium at 28 °C, at pH 6.0-7.0 and in the presence of 0-1 % (w/v) NaCl. Strain DHC34T was closely related to Burkholderia alpina LMG 28138T (98.5 % 16S rRNA gene sequence similarity). 16S rRNA gene sequence analysis showed that strain DHC34T formed a clade with B. alpina LMG 28138T, which is next to but branched deeply with Robbsia andropogonis ICMP 2807T. The phylogenetic relationships among these three strains were also supported with the phylogram based on concatenated partial gyrB, recA and trpB gene sequences. The phylogenomic tree generated with the UBCG tool showed that strains DHC34T and R. andropogonis ICMP 2807T were in a different clade. The DNA-DNA relatedness values between strain DHC34T and B. alpina LMG 28138T and R. andropogonis ICMP 2807T were much lower than 70 %. Strain DHC34T contained ubiquinone 8 as the major respiratory quinone. Its major fatty acids were C16 : 0, C17 : 0 cyclo and C19 : 0 cyclo ω8c. The DNA G+C content of strain DHC34T was 64.2 mol%. The major polar lipids were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, three unidentified aminophospholipids, four unidentified phospholipids, one unidentified aminolipid and a polar lipid. The phenotypic, phylogenetic, genotypic and chemotaxonomic data showed that strain DHC34T represents a novel species of a new genus in the family Burkholderiaceae, for which the name Pararobbsia silviterrae gen. nov., sp. nov. is proposed. The type strain of Pararobbsia silviterrae is DHC34T (=KCTC 42628T=LMG 28845T). On the basis of the current data, Burkholderia alpina is renamed as Pararobbsia alpina comb. nov.

Paraburkholderia phosphatilytica sp. nov., a phosphate-solubilizing bacterium isolated from forest soil.

A novel Gram-stain-negative, aerobic, non-spore-forming, non-motile and rod-shaped bacterial strain, 7QSK02T, was isolated from forest soil of Dinghushan Biosphere Reserve, Guangdong Province, China. It grew at 12-37 °C, at pH 4.0-7.5 and in the presence of 0-1.0 % (w/v) NaCl on R2A agar medium, with optimum growth at 28 °C, pH 5.5 and 0 % NaCl. Strain 7QSK02T was closely related to members of the genus Paraburkholderia: P. acidipaludis NBRC 101816T (98.1 % 16S rRNA gene sequence similarity), P. piptadeniae STM 7183T (97.6 %), P. kururiensis JCM 10599T (97.3 %), P. caballeronis TNe-841T (97.3 %) and P. diazotrophica JPY461T (97.1 %). 16S rRNA gene sequence analysis showed that strain 7QSK02T and two closely strains, P. kururiensis JCM 10599T and P. caballeronis TNe-841T, formed a clade within the genus Paraburkholderia, but was clearly separated from the established species. The genomic G+C content of strain 7QSK02T was 64.9 mol% based on total genome calculations. The average nucleotide identity and digital DNA-DNA hybridization value for the complete genomes were 79.2-81.5 and 23.2-24.9 % between strain 7QSK02T and its closely related species listed above. Strain 7QSK02T contained ubiquinone 8 as the major respiratory quinone. Major fatty acids were C16 : 0, C17 : 0 cyclo and C19 : 0 cyclo ω8c. The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylmethylethanolamine, one unidentified aminophospholipid, aminolipid and polar lipid. The phenotypic, chemotaxonomic and phylogenetic properties, and genome analysis suggest that strain 7QSK02T represents a novel species of the genus Paraburkholderia, for which the name Paraburkholderia phosphatilytica sp. nov. is proposed. The type strain is 7QSK02T (=GDMCC 1.1283T=CGMCC 1.15470T=KCTC 62473T).

Paraburkholderia acidiphila sp. nov., Paraburkholderia acidisoli sp. nov. and Burkholderia guangdongensis sp. nov., isolated from forest soil, and reclassification of Burkholderia ultramafica as Paraburkholderia ultramafica comb. nov.

Three Gram-stain-negative, aerobic, motile and rod-shaped bacterial strains, 7Q-K02T, DHF22T and DHOM02T, were isolated from forest soil sampled at Dinghushan Biosphere Reserve, Guangdong Province, China. Strains 7Q-K02T, DHF22T and DHOM02T grew at 4-37, 4-42 and 12-37 °C, pH 3.0-8.5, 3.5-8.5 and 5.0-8.0, and in the presence of 0-3.0, 0-3.5 and 0-2.5 % (w/v) NaCl; with optima at 28-33, 28 and 28-33 °C, pH 3.5-6.5, 4.0-5.5 and 6.5-7.0, and 0-1.5, 0-1.5 and 0.5-1.5 % (w/v) NaCl, respectively. Strains 7Q-K02T and DHF22T have the highest 16S rRNA gene sequence similarities of 99.0 and 98.0 % to Paraburkholderia sacchari LMG 19450T and 97.7 % between themselves, while strain DHOM02T shares the highest similarity of 98.4 % to 'Burkholderia rinojensis' A396T followed by 98.3 % to Burkholderia plantarii ATCC 43733T. In the 16S rRNA gene sequence phylogram, strain 7Q-K02T formed a sister branch with Paraburkholderia sacchari, Paraburkholderia oxyphila and Paraburkholderia paradisi, and strain DHF22T was separated from all other species within the genus Paraburkholderia, while strain DHOM02T formed a separated clade with members of the genus Burkholderia. The DNA G+C contents of strains 7Q-K02T, DHF22T and DHOM02T wwe 64.3, 65.4 and 66.6 %, respectively. Digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) values of strains 7Q-K02T, DHF22T and closely related Paraburkholderia strains were in the ranges of 25.5-43.7 % and 81.5-91.3 %, respectively. While dDDH and ANI values between strain DHOM02T and Burkholderia strains with genome sequence data were in the ranges of 22.4-31.0 % and 78.2-86.1 %, respectively. These three strains have the same major respiratory quinone: ubiquinone-8. Strains 7Q-K02T, DHF22T and DHOM02T have C16 : 0, C17 : 0 cyclo, C19 : 0 cyclo ω8c and summed feature 8 (C18 : 1 ω7c/C18 : 1 ω6c) as their major fatty acid compositions. The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. On the basis of phenotypic, phylogenetic, genomic analyses and chemotaxonomic data, strains 7Q-K02T and DHF22T represent two novel species of the genus Paraburkholderia, for which the names Paraburkholderia acidiphila sp. nov. (type strain 7Q-K02T=CGMCC 1.15433T=KCTC 62472T=LMG 29209T) and Paraburkholderia acidisoli sp. nov. (type strain DHF22T=GDMCC 1.1448T=LMG 30262T) are proposed, while strain DHOM02T represents a novel species in the genus Burkholderia, for which the name Burkholderia guangdongensis sp. nov. (type strain DHOM02T=KCTC 42625T=LMG 28843T) is proposed. We also propose to transfer Burkholderia ultramafica to the genus Paraburkholderia as Paraburkholderia ultramafica comb. nov. based mainly on the results of phylogenomic analysis.

Paraburkholderia dinghuensis sp. nov., isolated from soil.

A novel Gram-stain-negative, aerobic, non-spore-forming, non-motile and rod-shaped bacterial strain, designated DHOA04T, was isolated from a forest soil sample collected at Dinghushan Biosphere Reserve, Guangdong Province, PR China (112° 31' E 23° 10' N). It grew optimally at 28-33 °C and pH 6.5-7.0. Strain DHOA04T contained Q-8 as the major respiratory quinone. Its main fatty acids were C16 : 0, C17 : 0cyclo, summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c) and summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c). The DNA G+C content of DHOA04T was 63.0 mol%, which is in the range of the genus Paraburkholderia. The average nucleotide identity and digital DNA-DNA hybridization values for the complete genomes were 81.6-83.0 and 25.5-27.0 % between strain DHOA04T and five closely related type strains. The major polar lipids were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol and two unidentified aminophospholipids. On the basis of 16S rRNA gene sequence analysis, the strain was found to be closely related to members of the genus Paraburkholderia, but clearly separated from the established species. Phylogenetic analysis based on the 16S rRNA gene sequences using the maximum-likelihood algorithm indicated that strain DHOA04T was most closely related to Paraburkholderia ferrariae NBRC 106233T. The phenotypic, chemotaxonomic and phylogenetic data, and genome analysis showed that strain DHOA04T represents a novel species of the genus Paraburkholderia, for which the name Paraburkholderia dinghuensis sp. nov. is proposed. The type strain is DHOA04T (=KCTC 42627T=LMG 28839T).

Chitinophaga flava sp. nov., isolated from monsoon evergreen broad-leaved forest soil.

A Gram-stain-negative, aerobic, non-motile strain, K3CV102501T, was isolated from a soil sample collected from the monsoon evergreen broad-leaved forest of Dinghushan Biosphere Reserve located in Guangdong Province, PR China. The primal colony of strain K3CV102501T was very similar to the fruiting body of myxobacteria on the original isolation plates. Young cultures of strain K3CV102501T contained long (2-4×0.4-0.5 µm) filamentous cells and divided into rod shapes (0.7-1.0×0.6-0.8 µm) after 4 days of incubation at 28 °C. Strain K3CV102501T grew at pH 6.0-9.5 (optimum, pH 6.5-7.5) and 7-42 °C (optimum, 28-35 °C). Phylogenetic analysis based on its 16S rRNA gene sequence showed that strain K3CV102501T belonged to the genus Chitinophagaand showed the highest similarity to C.hitinophaga jiangningensis JCM 19354T (96.9 %). The DNA G+C content of the type strain was 46.6 mol%. The major fatty acids (>10 %) were iso-C15 : 0, C16 : 1ω5c and iso-C17 : 0 3-OH. The major polar lipids were phosphatidylethanolamine and an unidentified aminolipid. Menaquinone-7 was the predominant quinone. The phenotypic, chemotaxonomic and phylogenetic data clearly showed that strain K3CV102501T represents a novel species of the genus Chitinophaga, for which the name Chitinophaga flava sp. nov. is proposed. The type strain is K3CV102501T (=KCTC 62435T=GDMCC 1.1325T).

Trinickia dinghuensis sp. nov. and Trinickia fusca sp. nov., isolated from forest soil.

Two Gram-stain-negative, aerobic, non-spore forming and rod-shaped bacterial strains, designated DHOM06T and 7MK8-2T, were isolated from forest soil sampled at Dinghushan Biosphere Reserve, Guangdong Province, PR China. Strain DHOM06T grew at 12-37 °C (optimum, 28-33 °C), pH 4.5-7.5 (pH 5.5) and in the presence of 0-0.5 % NaCl (w/v); while strain 7MK8-2T grew at 12-42 °C (28-33 °C), pH 4.0-8.5 (pH 4.5-5.5) and in the presence of 0-1.0 % NaCl (w/v). Strains DHOM06T and 7MK8-2T each has a 16S rRNA gene sequence similarity of 97.1-98.9 % as well as 97.4-97.9 % to Trinickia strains, respectively. In the 16S rRNA gene sequence phylogram, both strains and all five currently described Trinickia species formed a clade but they were all distinct from each other. The average nucleotide identity and digital DNA-DNA hybridization values for strains DHOM06T and 7MK8-2T and all Trinickia species were in the range of 77.4-82.6 % and 21.7-26.2 %, respectively. The DNA G+C content of DHOM06T and 7MK8-2T was 63.2 and 63.5 mol%, respectively, based on total genome calculations. These two strains contained ubiquinone 8 as the major respiratory quinone and C16 : 0, C17 : 0cyclo, C19 : 0cyclo ω8c, summed feature 3 (C16 : 1ω7c/C16 : 1ω6c) and summed feature 8 (C18 : 1ω7c/C18 : 1ω6c) as the major cellular fatty acids. The major polar lipids of DHOM06T and 7MK8-2T were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. On the basis of phenotypic, chemotaxonomic, phylogenetic and genomic analysis data, strains DHOM06T and 7MK8-2T represent two novel species of the genus Trinickia, for which the names Trinickia dinghuensis sp. nov. (type strain DHOM06T=GDMCC 1.1280T=LMG 30259T) and Trinickia fusca sp. nov. (type strain 7MK8-2T=GDMCC 1.1449T=KCTC 62469T) are proposed.

Chitinophaga varians sp. nov., isolated from forest soil.

A Gram-stain-negative, aerobic, non-motile bacterium, designated strain 10-7W-9003T, was isolated from the forest soil of Limushan National Forest Park, south-east China (19° 10' 42″ N, 109° 44' 45″ E). Strain 10-7W-9003T showed a shape change during the course of culture from long filamentous cells (5-10×0.4-0.5 µm) at 5-36 h, to rod shaped (1.0-1.5×0.5-0.7 µm) with inoculation after 2 days. It grew optimally at 28-30 °C and pH 6.5-7.5. On the basis of 16S rRNA gene sequence analysis, it belongs to the genus Chitinophaga and is most closely related to Chitinophaga eiseniae KACC 13774T and Chitinophaga qingshengii JCM 30026T, with 16S rRNA gene sequences similarities of 98.8 and 98.3 %, respectively. However, the DNA-DNA hybridization study showed that strain 10-7W-9003T shared relatively low relatedness values with KACC 13774T (21.8 %) and JCM 30026T (20.4 %), respectively. The major fatty acids (>10 %) were iso-C15 : 0, C16 : 1ω5c and iso-C17 : 0 3-OH. The genomic DNA G+C content was 50.7 mol%. It contained MK-7 as the major quinone. The phenotypic, chemotaxonomic and phylogenetic data clearly showed that strain 10-7W-9003T represents a novel species of the genus Chitinophaga, for which the name Chitinophaga varians sp. nov. is proposed. The type strain is 10-7W-9003T (=GDMCC 1.1252T=KACC 19415T=KCTC 52926T).

Dyella caseinilytica sp. nov., Dyella flava sp. nov. and Dyella mobilis sp. nov., isolated from forest soil.

Three strains, DHOB09T, DHOC52T and DHON07T, were isolated from the forest soil of Dinghushan Biosphere Reserve, Guangdong Province, PR China. They were all Gram-stain-negative, aerobic, rod-shaped cells. The ranges (optimum) for the temperature, pH and NaCl concentration for growth of DHOB09T, DHOC52T and DHON07T were 10-42 (25-28) °C, pH 5.5-9.0 (7.0-7.5) and 0-4.0 (0-0.5) % (w/v); 10-42 (28) °C, pH 4.0-7.0 (4.5-6.5) and 0-2.0 (0) % (w/v) and 10-37 (25-28) °C, pH 4.0-7.5 (5.5-6.0) and 0-2.5 (0)  % (w/v), respectively. Phylogenetic analysis based on 16S rRNA gene sequences showed that DHOB09T, DHOC52T and DHON07T formed a phyletic cluster with seven species of the genus Dyella within the major clade of Dyella with sequence similarities ranged from 96.9 to 98.6 %. This indicated that the three strains may represent three novel species of the genus Dyella. This result was also strongly supported by the concatenated analysis of partial gyrB, lepA and recA gene sequences. DNA-DNA hybridization between strains DHON07T and DHOB09T, as well as DHON07T and Dyella koreensis BB4T was much lower than 70 %. The G+C content of strains DHOB09T, DHOC52T and DHON07T were 59.4, 60.7 and 59.5 %, respectively. The major fatty acids of the three strains were iso-C15 : 0, iso-C16 : 0 and iso-C17 : 0 and the predominant respiratory lipoquinone was ubiquinone-8. All of the physiological, phylogenetic and chemotaxonomic data showed that strains DHOB09T, DHOC52T and DHON07T are distinctive from each other and from all species of the genus Dyellawith validly published names. Therefore, we suggest that they represent three novel species of the genus, for which the names Dyella caseinilytica sp. nov. (type strain DHOB09T=CGMCC 1.15434T=LMG 29202T), Dyella flava sp. nov. (type strain DHOC52T=NBRC 111979T=KCTC 52128T) and Dyella mobilis sp. nov. (type strain DHON07T=CGMCC 1.15400T=NBRC 111475T) are proposed.

Puia dinghuensis gen. nov., sp. nov., isolated from monsoon evergreen broad-leaved forest soil.

A Gram-stain-negative, rod-shaped, aerobic, non-motile bacterial strain, 4GSH07T, was originally isolated from the monsoon evergreen broad-leaved forest soil of Dinghushan Biosphere Reserve, Guangdong Province, PR China (112° 31' E 23° 10' N). It grew optimally at 28-33 °C and pH 4.0. The 16S rRNA gene sequence analysis showed that strain 4GSH07T had the highest sequence similarity of 94.0 % to Parasegetibacter terrae JCM 19942T, and formed an independent lineage separable from other described genera of the family Chitinophagaceae. The main fatty acids (>5 %) were iso-C15 : 0, iso-C17 : 0 3-OH, C16 : 0 3-OH, summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c) and C15 : 0 2-OH. The organism contained MK-7 as the predominant respiratory quinone, and the total DNA G+C content was 50.3 mol%. The phenotypic, chemotaxonomic and phylogenetic data showed consistently that strain 4GSH07T represents a novel species of a novel genus of the family Chitinophagaceae, for which the name Puia dinghuensis gen. nov., sp. nov., is proposed, with 4GSH07T (=CGMCC 1.15448T=LMG 29214T) as the type strain.

Dyella acidisoli sp. nov., D. flagellata sp. nov. and D. nitratireducens sp. nov., isolated from forest soil.

Bacterial strains 4M-Z03T, 4M-K16T and DHG59T were isolated from forest soil samples collected from the Dinghushan Biosphere Reserve, Guangdong Province, PR China (112° 31' E 23° 10' N). The three strains grew well at 28 °C, pH 5.0-6.0 on R2A medium. On the basis of 16S rRNA gene sequence analysis, the three strains, together with Dyella humi DHG40T, formed a distinct phyletic clade within the genus Dyella, and the sequence similarities between any strains of the clade ranged from 97.8 to 98.5 %. Sequence analysis of concatenated partial gyrB, lepA and recA gene sequences also strongly suggested that the three strains represented three novel species of the genus Dyella. The respiratory lipoquinone of the three strains was ubiquinone-8, and their DNA G+C content was 58.2-59.0 mol%. The fatty acid profiles differed substantially among these three strains, although they had two common major fatty acids, iso-C15 : 0 and iso-C17 : 1ω9c. The DNA-DNA relatedness among the three strains and the type strains of the closest species of the genus Dyella examined was lower than 50 %. The results of genotypic and phenotypic characterization presented above demonstrate that the three strains examined represent three novel species of the genus Dyella, for which the names Dyella acidisoli sp. nov. (type strain 4M-Z03T=NBRC 111980T=KCTC 52131T), Dyella flagellata sp. nov. (type strain 4M-K16T=NBRC 111981T=KCTC 52130T) and Dyella nitratireducens sp. nov. (type strain DHG59T=NBRC 111472T=LMG 29201T=CGMCC 1.15439T) are proposed.

Aliidongia dinghuensis gen. nov., sp. nov., a poly-ß-hydroxybutyrate-producing bacterium isolated from Pinus massoniana forest soil.

A Gram-stain-negative, aerobic, motile, rod-shaped bacterium, designated 7M-Z19T, was isolated from a soil sample collected from a Pinus massoniana forest of Dinghushan Biosphere Reserve, Guangdong Province, PR China. Strain 7M-Z19T grew at pH 4.5-7.5 (optimum pH 6.0-6.5), 10 to 37 °C (optimum 28 °C) and NaCl concentration up to 2.0 % (optimum 0 %, w/v). iso-C17 : 0, C18 : 1ω7c and C19 : 0ω8c cyclo were the major fatty acids (>10 %) while ubiquinone-10 was the only respiratory quinone detected in 7M-Z19T. The polar lipids of the strain consisted of phosphatidylethanolamine, phosphatidyldimethylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, six unidentified aminophospholipids, three unidentified phospholipids, six unidentified lipids and a glycolipid. The DNA G+C content was 65.8 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the isolate formed a distinct lineage with Dongia mobilis and Dongia rigui within the family Rhodospirillaceae, but with a low sequence similarity of 92.7 and 92.0 %, respectively. On the basis of phylogenetic, phenotypic, physiological and chemotaxonomic distinctiveness, strain 7M-Z19T should be placed in the family Rhodospirillaceae as a representative of a novel genus and species, for which the name Aliidongia dinghuensis gen. nov., sp. nov., is proposed. The type strain of the type species is 7M-Z19T (=NBRC 112240T=KCTC 52134T=CGMCC 1.15725T).

Dinghuibacter silviterrae gen. nov., sp. nov., isolated from forest soil.

A novel Gram-stain negative, non-motile, rod-shaped, aerobic bacterial strain, designated DHOA34T, was isolated from forest soil of Dinghushan Biosphere Reserve, Guangdong Province, China. Comparative 16S rRNA gene sequence analysis showed that it exhibited highest similarity with Flavisolibacter ginsengiterrae Gsoil 492T and Flavitalea populi HY-50RT, at 90.89 and 90.83%, respectively. In the neighbour-joining phylogenetic tree based on 16S rRNA gene sequences, DHOA34T formed an independent lineage within the family Chitinophagaceae but was distinct from all recognized species and genera of the family. The major cellular fatty acids of DHOA34T included iso-C15:0, anteiso-C15:0, iso-C17:0 3-OH and summed feature 3 (C16:1ω6c and/or C16:1ω7c). The DNA G+C content was 51.6 mol% and the predominant quinone was menaquinone 7 (MK-7). Flexirubin pigments were produced. The phenotypic, chemotaxonomic and phylogenetic data demonstrate consistently that strain DHOA34T represents a novel species of a new genus in the family Chitinophagaceae, for which the name Dinghuibacter silviterrae gen. nov., sp. nov. is proposed. The type strain of Dinghuibacter silviterrae is DHOA34T (=CGMCC 1.15023T=KCTC 42632T).

Paraburkholderiapallidirosea sp. nov., isolated from a monsoon evergreen broad-leaved forest soil.

A Gram-stain-negative, rod-shaped, aerobic and motile bacterial strain, DHOK13T, was isolated from the forest soils of Dinghushan Biosphere Reserve, Guangdong Province, PR China (112° 31' E, 23° 10' N). It grew optimally at 28-33 °C and pH 7.0-7.5. The main fatty acids were C16 : 0, C17 : 0 cyclo, C19 : 0 cycloω8c, summed feature 2 (C12 : 0 aldehyde and/or unknown 10.9525) and summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c). The organism contained ubiquinone Q-8 as the predominant isoprenoid quinone. The total DNA G+C content of strain DHOK13T was 62.0 mol%. Phylogenetic analysis of the 16S rRNA gene, as well as the sequence of the partial housekeeping genes, gyrB and recA, showed consistently that strain DHOK13T formed an independent cluster with Paraburkholderia phenazinium LMG 2247T. DNA-DNA hybridization studies showed relatively low relatedness values (39 %) of strain DHOK13T with P. phenazinium LMG 2247T. The phenotypic, chemotaxonomic and phylogenetic data showed that strain DHOK13T represents a novel species of the genus Paraburkholderia for which the name Paraburkholderia pallidirosea sp. nov. is proposed. The type strain is DHOK13T (=KCTC 42626T=LMG 28846T).

Dyella humi sp. nov., isolated from forest soil.

A Gram-stain-negative, aerobic, motile, rod-shaped, yellow-pigmented bacterium, designated as DHG40T, was isolated from a soil sample collected from the forest of Dinghushan Biosphere Reserve, Guangdong Province, China. Strain DHG40T grew at pH 4.0-8.0 and 10-37 °C (optimum at pH 6.0-7.0 and 25-28 °C). NaCl inhibited growth at concentrations above 2.5 % (w/v). Phylogenetic analysis based on 16S rRNA gene sequences revealed that the isolate fell within the cluster of the genus Dyella. Strain DHG40T was closely related to Dyella ginsengisoli Gsoil 3046T (97.6 % 16S rRNA gene sequence similarity), Dyella marensis CS5-B2T (97.5 %), Dyella koreensis BB4T (97.4 %) and Dyella jejuensis JP1T (97.4 %). The DNA-DNA relatedness values between strain DHG40T and its phylogenetically closest relatives were all below 40 %. The DNA G+C content was 60.3 mol%. In addition, iso-C15 : 0, iso-C16 : 0, iso-C17 : 0 and iso-C17 : 1ω9c were the major fatty acids (>10 %) and ubiquinone-8 was the respiratory quinone. The major polar lipids were phosphatidylethanolamine, phosphatidylmethylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, an unidentified aminolipid and an unidentified phospholipid. On the basis of phylogenetic, phenotypic, physiological and chemotaxonomic distinctiveness, strain DHG40T should be placed in the genus Dyella as a representative of a novel species, for which the name Dyella humi sp. nov. is proposed. The type strain is DHG40T (=KCTC 42629T=LMG 28842T).

Edaphobacter dinghuensis sp. nov., an acidobacterium isolated from lower subtropical forest soil.

An aerobic bacterium, designated DHF9T, was isolated from a soil sample collected from the lower subtropical forest of Dinghushan Biosphere Reserve, Guangdong Province, PR China. Cells were Gram-stain-negative, non-motile, short rods that multiplied by binary division. Strain DHF9T was an obligately acidophilic, mesophilic bacterium capable of growth at pH 3.5-5.5 (optimum pH 4.0) and at 10-33 °C (optimum 28-33 °C). Growth was inhibited at NaCl concentrations above 2.0 % (w/v). The major fatty acids were iso-C15 : 0, C16 : 0 and C16 : 1ω7c. The polar lipids consist of phosphatidylethanolamine, two unidentified aminolipids, two unidentified phospholipids, two unidentified polar lipids and an unidentified glycolipid. The DNA G+C content was 57.7 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the strain belongs to the genus Edaphobacter in subdivision 1 of the phylum Acidobacteria, with the highest 16S rRNA gene sequence similarity of 97.0 % to Edaphobacter modestus Jbg-1T. Based on phylogenetic, chemotaxonomic and physiological analyses, it is proposed that strain DHF9T represents a novel species of the genus Edaphobacter, named Edaphobacter dinghuensis sp. nov. The type strain is DHF9T ( = DSM 29920T = CGMCC 1.12997T).

Acidipila dinghuensis sp. nov., an acidobacterium isolated from forest soil.

An aerobic, chemoheterotrophic, non-motile, capsule-forming bacterium designated DHOF10T was isolated from a soil sample collected from the forest of Dinghushan Biosphere Reserve, Guangdong Province, PR China. Strain DHOF10T was able to grow at pH 3.5-8.0 (optimum pH 4.0-4.5) and at 10-37 °C (optimum 28-37 °C). NaCl tolerance was up to 1.0 % (w/v). Major fatty acids consisted of iso-C15 : 0, C18 : 1ω9c and C16 : 1ω7c. The quinone was MK-8 and the DNA G+C content was 56.3 mol%. The polar lipids consisted of phosphatidylethanolamine, an unidentified aminolipid, an unidentified phospholipid, two unidentified aminophospholipids and two unidentified polar lipids. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the isolate was a member of genus Acidipila of the phylum Acidobacteria, with the highest 16S rRNA gene sequence similarity of 97.3 % to Acidipila rosea AP8T. On the basis of phylogenetic, phenotypic, physiological and chemotaxonomic distinctiveness, strain DHOF10T represents a novel species of the genus Acidipila, for which the name Acidipila dinghuensis sp. nov. is proposed. The type strain is DHOF10T ( = CGMCC 1.13007T = KCTC 42631T).

Chitinophaga dinghuensis sp. nov., isolated from soil.

A Gram-reaction-negative, aerobic, non-motile bacterial strain, DHOC24T, was isolated from the forest soil of Dinghushan Biosphere Reserve, Guangdong Province, PR China. Strain DHOC24T underwent a shape change during the course of culture from long filamentous cells (10-30 × 0.4-0.5 µm) at 2 days to coccobacilli (0.5-1.0 × 0.7-1.0 µm) at 15 days after inoculation. It grew optimally at 28-33 °C and pH 6.5-7.5. The major quinone of strainDHOC24T was MK-7, the main fatty acids were iso-C15 : 0, C16 : 1ω5c and iso-C17 : 0 3-OH and the DNA G+C content was 43.1 mol%. On the basis of 16S rRNA gene sequence analysis, the strain was found to be affiliated with members of the genus Chitinophaga, but was clearly separated from established species of the genus. Strain DHOC24T was most closely related to Chitinophaga jiangningensis JN53T (98.3 % 16S rRNA gene sequence similarity) and Chitinophaga terrae KP01T (97.9 %). DNA-DNA hybridization study showed relatively low relatedness values (32.1 %) of strain DHOC24T with C. jiangningensis JN53T. The phenotypic, chemotaxonomic and phylogenetic data showed that strain DHOC24T represents a novel species of the genus Chitinophaga, for which the name Chitinophaga dinghuensis sp. nov. is proposed. The type strain is DHOC24T ( = CGMCC 1.12995T = DSM 29821T).

A simple and efficient protocol for transient transformation of sliced grape berries.

Grape is an economically important crop but recalcitrant to Agrobacterium-mediated genetic transformation and in vitro regeneration. Here, we have developed a protocol for transient transformation of grapes by investigating the effects of explant pre-culture and duration of vacuum infiltration on transformation efficiency. Using sliced grape berries of "Shine-Muscat" (Vitis labrusca × Vitis vinifera) between the end of fruit expansion phase and the mature stage as explants, we firstly compared the effect of pre-culture explants into a susceptible state (incubation on Murashige and Skoog (MS) agar plate in the dark at 25 ± 1 °C for 48 h) with no pre-culture and then tested different vacuum infiltration times on transformation efficiency using ß-glucuronidase (GUS) reporter system. Pre-culture increased the susceptibility of explants to the agrobacteria infection and increased transient transformation efficiency as assessed by histochemical GUS activity, with intense blue coloration compared with the faint staining observed in the non-susceptible explants. Using a Circulating Water Vacuum Pump system to facilitate agrobacteria entry into berry cells, we tested vacuum durations of 5, 10, and 15 min and observed that transformation efficiency increased with vacuum duration of infiltration. These results were confirmed by relative gene expression of GUS transgene as assessed by RT-qPCR and GUS activity assay. To further confirm the usefulness of our protocol, we transiently transformed grape berries with the hydrogen peroxide sensor gene VvHPCA3, and this was confirmed by gene expression analysis as well as increased sensitivity of the explants to hydrogen peroxide treatment. Overall, this study has resulted in a simple but efficient transient transformation protocol for grape berries and would be a valuable tool for the rapid testing of gene function and the study of key regulatory networks in this important crop.

The complete chloroplast genome of Rhododendron shanii W.P. Fang (Ericaceae), a endemic plant from the Southern Dabie Mountains of China.

Rhododendron shanii W.P. Fang 1983 (Ericaceae) is woody plant naturally distributed in the southwest of Anhui, China. The complete chloroplast genome sequence of R. shanii was generated by whole-genome next-generation sequencing data and assembled based on three Rhododendron species chloroplast genome. The complete chloroplast genome sequence of R. shanii was 204,170 bp and divided into four distinct regions: small single-copy region (2615 bp), large single-copy region (107,189 bp), and a pair of inverted repeat regions (47,183 bp). The genome annotation displayed 150 genes, including 95 protein-coding genes, 47 tRNA genes, and eight rRNA genes. Phylogenetic analysis with the Ericaceae reported chloroplast genomes revealed that R. shanii is sister to the clade comprising R. delavayi, R. griersonianum and R. platypodum.