Biofilm-overproducing Bacillus amyloliquefaciens P29ΔsinR decreases Pb availability and uptake in lettuce in Pb-polluted soil.

In this study, one mutant strain P29ΔsinR with increased biofilm production was constructed from a biofilm-producing Bacillus amyloliquefaciens strain P29. Then, the effect of strain P29 and its biofilm-overproducing mutant strain P29ΔsinR on Pb availability and accumulation in lettuce and the associated mechanisms were characterized in the Pb-contaminated soil. The live strains P29 and P29ΔsinR increased the dry masses of roots and edible tissues by 31-74% compared to the controls. The live strains P29 and P29ΔsinR reduced the Pb uptake in the roots by 36-52% and edible tissues by 24-43%, Pb bioconcentration factor by 36-52%, and rhizosphere soil available Pb content by 12-25%, respectively, compared to the controls. The live strains P29 and P29ΔsinR increased the pH, proportion of biofilm-producing bacteria by 46-154%, contents of polysaccharides by 99-139% and proteins by 32-57%, and gene relative abundances of epsC by 7.1-10.2-fold, tasA by 10.3-10.8-fold, and sipW by 6.5-26.1-fold, which were associated with biofilm formation and Pb adsorption in the rhizosphere soils, respectively, compared to the controls. Furthermore, the mutant strain P29ΔsinR showed higher ability to reduce Pb availability and uptake in lettuce and increase the pH, proportion of biofilm-producing bacteria, polysaccharide and protein contents, and relative abundances of these genes. These results showed that the biofilm-overproducing strain P29ΔsinR induced lower Pb availability and accumulation in the vegetable and more biofilm-producing bacteria, polysaccharide and protein production, and Pb-immobilizing related gene abundances in the Pb-contaminated soil.

Biofilm-overproducing Bacillus subtilis B12ΔYwcc decreases Cd uptake in Chinese cabbage through increasing Cd-immobilizing related gene abundance and root surface colonization.

Biofilm-producing bacteria can decrease Cd uptake in vegetables, but mechanisms underlying this effect are poorly characterized. In this study, two mutant strains B12ΔYwcc and B12ΔSlrR were constructed from a biofilm-producing Bacillus subtilis strain B12. Then, the impacts of strain B12 and its high biofilm-producing mutant strain B12ΔYwcc and low biofilm-producing mutant strain B12ΔSlrR on Cd availability and uptake in Chinese cabbage and the related mechanisms were investigated in the Cd-polluted soil. Strain B12 and its mutants B12ΔYwcc and B12ΔSlrR increased the dry biomasses of edible tissues by 54%-130% compared with the controls. Strain B12 and its mutant B12ΔYwcc reduced the soil available Cd content by 36%-50% and root and edible tissue Cd contents by 23%-50% compared with the controls. Furthermore, the mutant strain B12ΔYwcc reduced the edible tissue Cd content by 40% and increased the polysaccharide content by 23%, invertase activity by 139%, and gene copies of the cumA by 4.5-fold, epsA by 7.1-fold, and cadA by 4.3-fold, which were involved in Cd adsorption in the rhizosphere soils, respectively, compared with strain B12. The polysaccharide content and cumA, epsA, and cadA gene copy numbers showed significantly reverse correlations with the available Cd content. Notably, the mutant strain B12ΔYwcc showed better ability to colonize the vegetable root surface than strain B12. These findings demonstrated that the biofilm-overproducing mutant strain B12ΔYwcc increased the polysaccharide production and Cd-immobilizing related cumA, epsA, and cadA gene copies, resulting in lower Cd availability and accumulation in Chinese cabbage in the Cd-polluted soil.

A Combination of Genomics, Transcriptomics, and Genetics Provides Insights into the Mineral Weathering Phenotype of Pseudomonas azotoformans F77.

Silicate mineral weathering (dissolution) plays important roles in soil formation and global biogeochemical cycling. In this study, a combination of genomics, transcriptomics, and genetics was used to identify the molecular basis of mineral weathering activity and acid tolerance in Pseudomonas azotoformans F77. Biotite was chosen as a silicate mineral to investigate mineral weathering. The genome of strain F77 was sequenced, and the genes significantly upregulated when grown in the presence of biotite included mineral weathering-related genes associated with gluconic acid metabolism, flagellar assembly, and pilus biosynthesis and acid tolerance-related genes associated with neutralizing component production, reducing power, and proton efflux. The biotite-weathering behaviors of strain F77 and its mutants that were created by deleting the tkt, tal, and gntP genes, which are involved in gluconic acid metabolism, and the potF, nuoF, and gdtO genes, which are involved in acid tolerance, were determined. The Fe and Al concentrations in the strain F77-inoculated medium increased 2.2- to 13.7-fold compared to the controls. The cell numbers of strain F77 increased over time, while the pH values in the medium ranged from 3.75 to 3.90 between 20 and 36 h of incubation. The release of Al and Fe was significantly reduced in the F77 Δtal, F77 ΔgntP, F77 ΔpotF, and F77 ΔnuoF mutants. Bacterial growth was significantly reduced in the presence of biotite in the F77 ΔpotF and F77 ΔnuoF mutants. Our results demonstrated the acid tolerance of strain F77 and suggested that multiple genes and metabolic pathways in strain F77 are involved in biotite weathering and acid tolerance during the mineral weathering process. IMPORTANCE Acid production and tolerance play important roles in effective and persistent mineral weathering in bacteria, although the molecular mechanisms governing acid production and acid tolerance in bacteria have not been fully elucidated. In this study, the molecular mechanisms underlying biotite (as a silicate mineral) weathering (dissolution) and acid tolerance of P. azotoformans F77 were characterized using genomics, transcriptomics, and genetics analyses. Our results showed that the genes and metabolic pathways for gluconic acid metabolism, flagellar assembly, and pilus biosynthesis may play important roles in mineral weathering by strain F77. Notably, the genes associated with neutralizing component production, reducing power, and proton efflux may be related to acid tolerance in strain F77. The expression of these acid production- and acid tolerance-related genes was observed to be increased by biotite in strain F77. Our findings may help to elucidate the molecular mechanisms governing mineral weathering and, especially, acid tolerance in mineral-weathering bacteria.

Interactions between Biotite and the Mineral-Weathering Bacterium Pseudomonas azotoformans F77.

In this study, the mineral-weathering bacterium Pseudomonas azotoformans F77, which was isolated from the soil of a debris flow area, was evaluated for its weathering activity under direct contact with biotite or without contact. Then, biotite-weathering behaviors of strain F77, mutants that had been created by deleting the gcd and adh genes (which are involved in gluconic acid metabolism and pilus formation, respectively), and the double mutant F77ΔgcdΔadh were compared. The relative gene expression levels of F77 and its mutants F77Δgcd and F77Δadh were also analyzed in the presence of biotite. Direct contact with biotite increased Fe and Al release from the mineral in the presence of F77. All strains had similar abilities to release Fe and Al from the mineral except for F77Δgcd and F77Δadh Mobilized Fe and Al concentrations were decreased by up to 72, 26, and 87% in the presence of F77Δgcd, F77Δadh, and F77ΔgcdΔadh, respectively, compared to levels observed in the presence of F77 during the mineral-weathering process. Gluconic acid production was decreased for F77Δgcd and F77ΔgcdΔadh, while decreased cell attachment on the mineral surface was observed for F77Δadh, compared to findings for F77. The F77 genes involved in pilus formation and gluconic acid metabolism showed increased expression levels in the presence of biotite. The results of this study showed important roles for the genes involved in gluconic acid metabolism and pilus formation in mineral weathering by F77 and demonstrated the distinctive effect of these genes on mineral weathering by F77.IMPORTANCE Bacteria play important roles in mineral weathering and soil formation, although the molecular mechanisms underlying the interactions between bacteria and silicate minerals are poorly understood. In this study, the interactions between biotite and the highly effective mineral-weathering bacterium P. azotoformans F77 were characterized. Our results showed that the genes involved in gluconic acid metabolism and pilus formation play important roles in mineral weathering by F77. The presence of biotite could promote the expression of these genes in F77, and a distinctive effect of these genes on mineral weathering by F77 was observed in this study. Our results provide new knowledge and promote better understanding regarding the interaction between silicate minerals and mineral-weathering bacteria, as well as the molecular mechanisms involved in these processes.

Combined biochar and metal-immobilizing bacteria reduces edible tissue metal uptake in vegetables by increasing amorphous Fe oxides and abundance of Fe- and Mn-oxidising Leptothrix species.

In this study, a highly effective combined biochar and metal-immobilizing bacteria (Bacillus megaterium H3 and Serratia liquefaciens CL-1) (BHC) was characterized for its effects on solution Pb and Cd immobilization and edible tissue biomass and Pb and Cd accumulation in Chinese cabbages and radishes and the mechanisms involved in metal-polluted soils. In the metal-containing solution treated with BHC, the Pb and Cd concentrations decreased, while the pH and cell numbers of strains H3 and CL-1 increased over time. BHC significantly increased the edible tissue dry weight by 17-34% and reduced the edible tissue Pb (0.32-0.46 mg kg-1) and Cd (0.16 mg kg-1) contents of the vegetables by 24-45%. In the vegetable rhizosphere soils, BHC significantly decreased the acid-soluble Pb (1.81-2.21 mg kg-1) and Cd (0.40-0.48 mg kg-1) contents by 26-47% and increased the reducible Pb (18.2-18.8 mg kg-1) and Cd (0.38-0.39 mg kg-1) contents by 10-111%; while BHC also significantly increased the pH, urease activity by 115-169%, amorphous Fe oxides content by 12-19%, and relative abundance of gene copy numbers of Fe- and Mn-oxidising Leptothrix species by 28-73% compared with the controls. These results suggested that BHC decreased edible tissue metal uptake of the vegetables by increasing pH, urease activity, amorphous Fe oxides, and Leptothrix species abundance in polluted soil. These results may provide an effective and eco-friendly way for metal remediation and reducing metal uptake in vegetables by using combined biochar and metal-immobilizing bacteria in polluted soils.

Metal-immobilizing and urease-producing bacteria increase the biomass and reduce metal accumulation in potato tubers under field conditions.

In this study, the effect of two metal-immobilizing bacterial strains, Serratia liquefaciens CL-1 and Bacillus thuringiensis X30, on the availability of Cd and Pb and the metal accumulation in potato tubers, as well as the underlying mechanisms in metal-contaminated soils were characterized. Moreover, the impacts of the strains on metal immobilization, pH, and NH4+ concentration in metal-contaminated soil solutions were evaluated. Strains CL-1 and X30 increased tuber dry weight by 46% and 40%, reduced tuber Cd and Pb contents by 68-83% and 42-47%, and decreased the Cd and Pb translocation factors by 61-70% and 30-34%, respectively, compared to the controls. Strains CL-1 and X30 decreased the available Cd and Pb contents by 52-67% and 30-44% and increased the NH4+ content by 55% and 31%, pH, urease activity by 70% and 41%, and relative abundance of ureC gene copies by 37% and 20% in the rhizosphere soils, respectively, compared with the controls. Reduced Cd and Pb concentrations and increased pH and NH4+ concentration were found in the bacteria-inoculated soil solution compared to the controls. These results suggested that the strains reduced tuber metal uptake through decreasing the metal availability and increasing the pH, ureC gene relative abundance and urease activity as well as decreasing the metal translocation from the leaves to tubers. These results may provide an effective metal-immobilizing bacteria (especially strain CL-1)-enhanced approach to reduce metal uptake of potato tubers in metal-polluted soils.

Distinct biotite-weathering activities of Arthrobacter pascens F74 under different contact conditions.

Bacteria play important roles in mineral weathering and soil formation. However, little is known regarding the interactions between biotite and Arthrobacter strains. In this study, the mineral-mineral activities of the Arthrobacter pascens F74 isolated from a weathered rock surface were evaluated for its weathering behavior under direct contact and no contact with biotite. No contact was obtained by using dialysis bags. When directly in contact with biotite, Al and Fe concentrations increased by 9- to 47-fold compared with the controls in the presence of strain F74. Furthermore, strain F74 increased mobilized Al by 106% to 175% and Fe by 29% to 123% under direct contact than under no contact conditions. During biotite dissolution, significantly higher cell numbers and lower pH in the culture medium were observed in the presence of strain F74 under direct contact conditions than under no contact conditions. Significantly higher gluconic acid concentration and glucose dehydrogenase activity were found under direct contact conditions than under no contact and no biotite conditions. Scanning electron microscopy analysis showed cell adhesion on the biotite surface. These results demonstrated that strain F74 behaved differently with respect to biotite-weathering effectiveness and mechanisms under different contact conditions. The results also suggested that direct contact between biotite and strain F74 was important for the production of gluconic acid, cell adhesion on the mineral surface, and the mineral dissolution of the strain.

Metal-immobilizing Serratia liquefaciens CL-1 and Bacillus thuringiensis X30 increase biomass and reduce heavy metal accumulation of radish under field conditions.

In this study, metal-tolerant bacteria Serratia liquefaciens CL-1 and Bacillus thuringiensis X30 were compared for their Cd and Pb immobilization in solution and impacts on biomass and Cd and Pb uptake in a radish in metal-contaminated soils under field conditions. Strains CL-1 and X30 significantly reduced water-soluble Cd and Pb concentrations (45-67%) and increased the pH in solution compared to the controls. These strains significantly increased the biomass (25-99%) and decreased edible tissue Cd and Pb uptake in the radish (37-81%) and DTPA-extractable Cd and Pb contents (18-44%) of the rhizosphere soil compared to the un-inoculated controls. Strain CL-1 had higher potential to reduce edible tissue Cd and Pb uptake in the radish and DTPA-extractable Cd content than strain X30. Also, these strains significantly increased Cd translocation factor and strain CL-1 also significantly increased Pb translocation factor of the radish. Furthermore, strain CL-1 significantly increased the ratio of small soil aggregates (< 0.25 mm and 0.25-0.50 mm) of the rhizosphere soil. The results showed that these strains reduced the edible tissue Cd and Pb uptake through decreasing Cd and Pb availability in the soil and increasing Cd or Pb translocation from the roots to the leaves of the radish. The results also suggested the bacteria-related differences in reduced heavy metal uptake in the radish and the mechanisms involved under field conditions.

Increased biomass and quality and reduced heavy metal accumulation of edible tissues of vegetables in the presence of Cd-tolerant and immobilizing Bacillus megaterium H3.

A Cd-resistant and immobilizing Bacillus megaterium H3 was characterized for its impact on the biomass and quality and heavy metal uptake of edible tissues of two vegetables (Brassica campestris L. var. Aijiaohuang and Brassica rapa L. var. Shanghaiqing) grown in heavy metal-polluted soil. The impact of strain H3 on the soil quality was also evaluated. The increase in the edible tissue biomass and the contents of soluble proteins and vitamin C of the vegetables inoculated with strain H3 ranged from 18% to 33%, 17% to 31%, and 15% to 19%, respectively, compared with the controls. Strain H3 significantly decreased the edible tissue Cd and Pb contents of the two greens (41-80%), DTPA-extractable Cd content (35-47%) of the rhizosphere soils, and Cd and Pb translocation factors (25-56%) of the greens compared with the controls. Moreover, strain H3 significantly increased the organic matter content (17-21%) and invertase activity (13-14%) of the rhizosphere soils compared with the controls. Our results demonstrated the increased edible tissue biomass and quality, decreased Cd and Pb uptake of the edible tissues, and improved soil quality in the presence of strain H3. The results also suggested an effective bacterial-enhanced technique for decreased metal uptake of greens and improved vegetable and soil qualities in the metal-contaminated soils.

Impact of poxB, pta, and ackA genes on mineral-weathering of Enterobacter cloacae S71.

In this study, biotite weathering behaviors were compared between mineral-weathering bacteria Enterobacter cloacae S71, mutant strains created by the deletion of poxB, pta, and ackA genes involved in acetate formation, and their complemented strains. Compared to strain S71, a decrease in bacterial growth was observed during the early and middle stages for the mutant ΔpoxB and at the middle and later stages for the mutants Δpta and ΔackA. Dissolved Al and Fe concentrations were lower during the early stage for strain ΔpoxB, at the early or middle stage for strain Δpta, and at the middle and later stages and throughout the weathering process for strain ΔackA, compared to strain S71. Acetate production was depressed during the early stage for strain ΔpoxB, at the early and middle stages for strain Δpta, and throughout the weathering process for strain ΔackA. Overall, the ackA gene exhibited a larger impact on dissolved Fe and acetate concentrations than both the poxB and pta genes. Reduced bacterial growth and lower dissolved Al, Fe, and acetate concentrations recovered by the complemented strains. These results show that strain S71 promoted mineral weathering through the production of acetic acid with distinctive impacts by the genes involved in acetate.

The abundance and mineral-weathering effectiveness of Bacillus strains in the altered rocks and the soil.

In this study, we collected different levels of altered rocks of a rocky mountain and the adjacent soil and characterized the abundance and weathering effectiveness of Bacillus strains. Based on qPCR and culture-dependent approaches, the gene copies or the numbers of Bacillus strains were significantly higher in the soil than in the altered rocks, while the ratio of the gene copies or the numbers of Bacillus strains to those of total bacteria was higher in the less altered rock, followed by the more altered rock and the soil. The relative abundance of the highly active Al-solubilizing Bacillus strains was higher in the more altered rock, followed by the less altered rock and the soil. Among the Al-solubilizing Bacillus species, 30-36% of them were different between the altered rocks and the soil, however, similar Al-solubilizing Bacillus species were found in the less altered rocks and the more altered ones. The results showed the alteration-related changes in the abundance and mineral weathering effectiveness of Bacillus strains and suggested the ecological adaptation of the mineral-weathering Bacillus populations and their role in mineral weathering in the rock and soil environments.

Location-Related Differences in Weathering Behaviors and Populations of Culturable Rock-Weathering Bacteria Along a Hillside of a Rock Mountain.

Bacteria play important roles in rock weathering, elemental cycling, and soil formation. However, little is known about the weathering potential and population of bacteria inhabiting surfaces of rocks. In this study, we isolated bacteria from the top, middle, and bottom rock samples along a hillside of a rock (trachyte) mountain as well as adjacent soils and characterized rock-weathering behaviors and populations of the bacteria. Per gram of rock or surface soil, 106-107 colony forming units were obtained and total 192 bacteria were isolated. Laboratory rock dissolution experiments indicated that the proportions of the highly effective Fe (ranging from 67 to 92 %), Al (ranging from 40 to 48 %), and Cu (ranging from 54 to 81 %) solubilizers were significantly higher in the top rock and soil samples, while the proportion of the highly effective Si (56 %) solubilizers was significantly higher in the middle rock samples. Furthermore, 78, 96, and 6 % of bacteria from the top rocks, soils, and middle rocks, respectively, significantly acidified the culture medium (pH < 4.0) in the rock dissolution process. Most rock-weathering bacteria (79 %) from the rocks were different to those from the soils and most of them (species level) have not been previously reported. Furthermore, location-specific rock-weathering bacterial populations were found and Bacillus species were the most (66 %) frequently isolated rock-weathering bacteria in the rocks based on cultivation methods. Notably, the top rocks and soils had the highest and lowest diversity of rock-weathering bacterial populations, respectively. The results suggested location-related differences in element (Si, Al, Fe, and Cu) releasing effectiveness and communities of rock-weathering bacteria along the hillside of the rock mountain.

Cd immobilization and reduced tissue Cd accumulation of rice (Oryza sativa wuyun-23) in the presence of heavy metal-resistant bacteria.

Two metal-resistant Bacillus megaterium H3 and Neorhizobium huautlense T1-17 were investigated for their immobilization of Cd in solution and tissue Cd accumulation of rice (Oryza sativa wuyun-23) in the Cd-contaminated soil. Strains H3 and T1-17 decreased 79-96% of water-soluble Cd in solution and increased grain biomass in the high Cd-contaminated soil. Inoculation with H3 and T1-17 significantly decreased the root (ranging from 25% to 58%), above-ground tissue (ranging from 13% to 34%), and polished rice (ranging from 45% to 72%) Cd contents as well as Cd bioconcentration factor of the rice compared to the controls. Furthermore, H3 and T1-17 significantly reduced the exchangeable Cd content of the rhizosphere soils compared with the controls. Notably, strain T1-17 had significantly higher ability to reduce Cd bioconcentration factor and polished rice Cd uptake than strain H3. The results demonstrated that H3 and T1-17 decreased the tissue (especially polished rice) Cd uptake by decreasing Cd availability in soil and Cd bioconcentration factor and the effect on the reduced polished rice Cd uptake was dependent on the strains. The results may provide an effective synergistic bioremediation of Cd-contaminated soils in the bacteria and rice plants and bacterial-assisted safe production of rice in Cd-contaminated soils.

Change in mineral weathering behaviors of a bacterium Chitinophaga jiangningensis JN53 under different nutrition conditions.

Bacteria play important roles in mineral weathering and soil formation. However, little is known about the nutrition-related changes in mineral weathering potential and pattern of bacteria. In this study, mineral weathering behaviors of a novel mineral-weathering bacterium Chitinophaga jiangningensis JN53 were characterized in the presence of three contrasting biotite or potassium feldspar-added media. C. jiangningensis JN53 increased more Fe release from the minerals in the M-BHm (nutrition-poor medium) than in the SSKM (nutrition-rich medium) and BHm (nutrition-moderate medium), while C. jiangningensis JN53 released more Al from the minerals and Si from biotite in the SSKM. Similar Si release from potassium feldspar by C. jiangningensis JN53 was observed in the SSKM, BHm, and M-BHm. K releasing ability of C. jiangningensis JN53 was significantly higher in the biotite-added M-BHm. Highest and lowest growth of C. jiangningensis JN53 was observed in the SSKM and M-BHm, respectively. In the presence of the minerals, C. jiangningensis JN53 mainly produced gluconic acid in the SSKM and acetic acid in the BHm and M-BHm. C. jiangningensis JN53 also produced large amount of succinic acid in the biotite-added SSKM and oxalic acid in the potassium feldspar-added M-BHm. The results showed the growth, production of organic acids, and mineral weathering ability of C. jiangningensis JN53 in the three contrasting nutrition conditions. The results also suggested the change in the mineral weathering behaviors of C. jiangningensis JN53 under different levels of nutrition conditions.

Distinct Mineral Weathering Behaviors of the Novel Mineral-Weathering Strains Rhizobium yantingense H66 and Rhizobium etli CFN42.

UNLABELLED: Bacteria play important roles in mineral weathering, soil formation, and element cycling. However, little is known about the interaction between silicate minerals and rhizobia. In this study, Rhizobium yantingense H66 (a novel mineral-weathering rhizobium) and Rhizobium etli CFN42 were compared with respect to potash feldspar weathering, mineral surface adsorption, and metabolic activity during the mineral weathering process. Strain H66 showed significantly higher Si, Al, and K mobilization from the mineral and higher ratios of cell numbers on the mineral surface to total cell numbers than strain CFN42. Although the two strains produced gluconic acid, strain H66 also produced acetic, malic, and succinic acids during mineral weathering in low- and high-glucose media. Notably, higher Si, Al, and K releases, higher ratios of cell numbers on the mineral surface to total cell numbers, and a higher production of organic acids by strain H66 were observed in the low-glucose medium than in the high-glucose medium. Scanning electron microscope analyses of the mineral surfaces and redundancy analysis showed stronger positive correlations between the mineral surface cell adsorption and mineral weathering, indicated by the dissolved Al and K concentrations. The results showed that the two rhizobia behaved differently with respect to mineral weathering. The results suggested that Rhizobium yantingense H66 promoted potash feldspar weathering through increased adsorption of cells to the mineral surface and through differences in glucose metabolism at low and high nutrient concentrations, especially at low nutrient concentrations. IMPORTANCE: This study reported the potash feldspar weathering, the cell adsorption capacity of the mineral surfaces, and the metabolic differences between the novel mineral-weathering Rhizobium yantingense H66 and Rhizobium etli CFN42 under different nutritional conditions. The results showed that Rhizobium yantingense H66 had a greater ability to weather the mineral in low- and high-glucose media, especially in the low-glucose medium. Furthermore, Rhizobium yantingense H66 promoted mineral weathering through the increased adsorption of cells to the mineral surface and through increased organic acid production. Our results allow us to better comprehend the roles of different rhizobia in silicate mineral weathering, element cycling, and soil formation in various soil environments, providing more insight into the geomicrobial contributions of rhizobia to these processes.

Saccharibacillus qingshengii sp. nov., isolated from a lead-cadmium tailing.

A Gram-stain-positive, strictly aerobic strain, H6T, was isolated from a soil sample of lead-cadmium tailing in Qixia district, Nanjing (China). Cells of the strain are rod-shaped and colonies on LB agar are red. Strain H6T has subpolar and polar flagella and the optimal condition for growth is 30 °C, with 1 % (w/v) NaCl and at pH 7.0. Based on the 16S rRNA gene sequences, phylogenetic analysis showed that strain H6T was closely related to the genus Saccharibacillus, and the closest relatives were Saccharibacillus deserti WLJ055T (99.0 % 16S rRNA gene sequence similarity), Saccharibacillus kuerlensis HR1T (97.0 %) and Saccharibacillus sacchari GR21T (96.4 %). The DNA-DNA relatedness value between strain H6T and S. deserti WLJ055T was 55.0 %. The major polar lipids of strain H6T were diphosphatidylglycerol, phosphatidylglycerol, phosphoglycolipid and three unknown glycolipids. The DNA G+C content was 58.4 mol% and MK-7 was the major isoprenoid quinone. The major fatty acids were anteiso-C15 : 0 and C16 : 0. meso-Diaminopimelic acid was detected in the peptidoglycan. Based on the phylogenetic, biochemical and chemotaxonomic data, strain H6T represents a novel species of the genus Saccharibacillus, for which the name Saccharibacillus qingshengii sp. nov., is proposed. The type strain is H6T (=CCTCC AB 2016001T=JCM 31172T).

Increased growth and root Cu accumulation of Sorghum sudanense by endophytic Enterobacter sp. K3-2: Implications for Sorghum sudanense biomass production and phytostabilization.

Endophytic bacterial strain K3-2 was isolated from the roots of Sorghum sudanense (an bioenergy plant) grown in a Cu mine wasteland soils and characterized. Strain K3-2 was identified as Enterobacter sp. based on 16S rRNA gene sequence analysis. Strain K3-2 exhibited Cu resistance and produced 1-aminocyclopropane-1-carboxylate (ACC) deaminase, indole-3-acetic acid (IAA), siderophores, and arginine decarboxylase. Pot experiments showed that strain K3-2 significantly increased the dry weight and root Cu accumulation of Sorghum sudanense grown in the Cu mine wasteland soils. Furthermore, increase in total Cu uptake (ranging from 49% to 95%) of the bacterial inoculated-Sorghum sudanense was observed compared to the control. Notably, most of Cu (83-86%) was accumulated in the roots of Sorghum sudanense. Furthermore, inoculation with strain K3-2 was found to significantly increase Cu bioconcentration factors and the proportions of IAA- and siderophore-producing bacteria in the root interiors and rhizosphere soils of Sorghum sudanense compared with the control. Significant decrease in the available Cu content was also observed in the rhizosphere soils of the bacterial-inoculated Sorghum sudanense. The results suggest that the endophytic bacterial strain K3-2 may be exploited for promoting Sorghum sudanense biomass production and Cu phytostabilization in the Cu mining wasteland soils.

Paenibacillus qingshengii sp. nov., isolated from a lead-zinc tailing.

A novel bacterial strain, S1-9(T), was isolated from a lead-zinc tailing in Nanjing, Jiangsu Province, China. Cells of strain S1-9(T) were Gram-stain-negative, ellipsoidal endospore-forming, aerobic rods and motile by means of peritrichous flagella. On the basis of 16S rRNA gene sequence analysis, strain S1-9(T) was shown to belong to the genus Paenibacillus and the closest phylogenetic relatives were Paenibacillus glucanolyticus DSM 5162(T) (96.8% similarity), Paenibacillus lautus NRRL NRS-666(T) (96.5%) and Paenibacillus lactis MB 1871(T) (95.4%). The predominant menaquinone was MK-7. The major cellular fatty acids were anteiso-C15:0 and iso-C16:0. The polar lipid profile contained phosphatidylglycerol, phosphatidylethanolamine, diphosphatidylglycerol, two unknown phospholipids and two unknown lipids. The total DNA G+C content of strain S1-9(T) was 49.9 mol%. Based on the low levels of DNA-DNA relatedness (ranging from 21.8 to 48.4%) to the type strains of the above species of the genus Paenibacillus and unique phenotypic characteristics, strain S1-9(T) is considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus qingshengii sp. nov. is proposed. The type strain is S1-9(T) ( = CCTCC AB 2014290(T) = JCM 30613(T)).

Rhizobium yantingense sp. nov., a mineral-weathering bacterium.

A Gram-stain-negative, rod-shaped bacterial strain, H66(T), was isolated from the surfaces of weathered rock (purple siltstone) found in Yanting, Sichuan Province, PR China. Cells of strain H66(T) were motile with peritrichous flagella. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain H66(T) belongs to the genus Rhizobium. It is closely related to Rhizobium huautlense SO2(T) (98.1 %), Rhizobium alkalisoli CCBAU 01393(T) (98.0 %) and Rhizobium cellulosilyticum ALA10B2(T) (98.0 %). Analysis of the housekeeping genes, recA, glnII and atpD, showed low levels of sequence similarity (<92.0 %) between strain H66(T) and other recognized species of the genus Rhizobium. The predominant components of the cellular fatty acids were summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c) and C16 : 0. The G+C content of strain H66(T) was 60.3 mol%. Strain H66(T) is suggested to be a novel species of the genus Rhizobium based on the low levels of DNA-DNA relatedness (ranging from 14.3 % to 40.0 %) with type strains of species of the genus Rhizobium and on its unique phenotypic characteristics. The namehttp://dx.doi.org/10.1601/nm.1279Rhizobium yantingense sp. nov. is proposed for this novel species. The type strain is H66(T) ( = CCTCC AB 2014007(T) = LMG 28229(T)).

Chitinophaga longshanensis sp. nov., a mineral-weathering bacterium isolated from weathered rock.

A Gram-stain-negative, aerobic, yellow-pigmented, non-motile, non-spore-forming, rod-shaped bacterial strain, Z29(T), was isolated from the surface of weathered rock (potassic trachyte) from Nanjing, Jiangsu Province, PR China. Phylogenetic analysis based on 16S rRNA gene sequences suggested that strain Z29(T) belongs to the genus Chitinophaga in the family Chitinophagaceae. Levels of 16S rRNA gene sequence similarity between strain Z29(T) and the type strains of recognized species of the genus Chitinophaga ranged from 92.7 to 98.2 %. The main fatty acids of strain Z29(T) were iso-C15 : 0, C16 : 1ω5c and iso-C17 : 0 3-OH. It also contained menaquinone 7 (MK-7) as the respiratory quinone and homospermidine as the main polyamine. The polar lipid profile contained phosphatidylethanolamine, unknown aminolipids, unknown phospholipids and unknown lipids. The total DNA G+C content of strain Z29(T) was 51.3 mol%. Phenotypic properties and chemotaxonomic data supported the affiliation of strain Z29(T) with the genus Chitinophaga. The low level of DNA-DNA relatedness (ranging from 14.6 to 29.8 %) to the type strains of other species of the genus Chitinophaga and differential phenotypic properties demonstrated that strain Z29(T) represents a novel species of the genus Chitinophaga, for which the name Chitinophaga longshanensis sp. nov. is proposed. The type strain is Z29(T) ( = CCTCC AB 2014066(T) = LMG 28237(T)).