Rhizobium cremeum sp. nov., isolated from sewage and capable of acquisition of heavy metal and aromatic compounds resistance genes.

Two strains of Rhizobia isolated from sewage collected from the Chinese Baijiu distillery were characterized using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strains W15T and W16 were grouped as a separate clade closely related to Rhizobium daejeonense L61T (98.6%). Multilocus sequence analysis (MLSA) with three housekeeping genes (recA, glnII and rpoA) also showed that strains W15T and W16 belonged to the genus Rhizobium. Average nucleotide identity and digital DNA-DNA hybridization values between genome sequences of strain W15T and the closely related species ranged from 77.0% to 87.8% and from 23.9% to 30.9%. The DNA G + C content of strain W15T was 61.6 mol%. Strain W15T contained Q-10 as the major ubiquinone and the dominant fatty acids were summed feature 8 (C 18:1ω7c and/or C 18:1ω6c; 73.1%) and C18:0 (7.6%). The main polar lipids are phosphatidylcholine, phosphatidylmethylethanolamine, phosphatidylethanolamine and phosphatidylglycerol. On the basis of the evidences presented in this study, strains W15T and W16 represents a novel species of the genus Rhizobium, for which the name Rhizobium cremeum sp. nov. is proposed. The type strain is W15T (= CGMCC 1.18731T = KACC 22344T).

Lowered Cd toxicity, uptake and expression of metal transporter genes in maize plant by ACC deaminase-producing bacteria Achromobacter sp.

In this study, an ACC deaminase-producing bacterial strain Achromobacter sp. A1 was isolated from maize rhizosphere soil, characterized and evaluated for the effects on cadmium (Cd) immobilization in solution/rhizosphere, physiological characteristics and the tissue Cd contents in maize and the molecular mechanisms involved by hydroponic and pot experiments. ACC deaminase activity of strain A1 was significantly enhanced by Cd addition and Cd concentration decreased (55.54-63.62%) in solution supplemented with various Cd concentrations. Strain A1 significantly increased the maize dry weights (30.77-105%) and chlorophyll content (7.46-14.46%), decreased MDA content (25.16-36.87%) and ethylene production (20.93-35.86%) in hydroponic experiment. Strain A1 significantly reduced the above-ground tissue Cd uptake by 12.64-33.68% and 42-48% in hydroponic and pot experiments, reduced the DTPA-extractable Cd content and elevated invertase, urease and catalase activity in rhizosphere soils. In addition, the expression levels of Cd transporter genes HMA3 and Nramp5 were significantly reduced in root and shoot after strain A1 inoculation. These results indicate that strain A1 has great potential for application as a novel and environmentally friendly inoculant to immobilize Cd and reduce maize Cd uptake in Cd-contaminated environments, and will improve the understanding of the relative molecular mechanisms underlying the response to strain A1 in maize plant.

Responses of rhizosphere bacterial communities, their functions and their network interactions to Cd stress under phytostabilization by Miscanthus spp.

Miscanthus has good tolerance to heavy metals (HMs) and has received increasing attention in studies of HM-contaminated soil remediation. In this study, four Miscanthus cultivars (M. lutarioriparius Xiangnadi NO4, M. sinensis Xiangmang NO1, M. lutarioriparius × M. sinensis hybrid Xiangzamang NO1, and M. floridulus Wujiemang NO1) that grow in China were studied. Their tolerance and enrichment abilities in soils containing 50 mg kg-1 cadmium (Cd) and the structure and function of their rhizosphere bacterial communities during the remediation process were analyzed. The results exhibiting a tolerance index (TI) higher than 75 in roots and the aboveground parts (TI > 60, indicating highly tolerant plants) indicated that all four Miscanthus cultivars were tolerant to high Cd concentrations. Moreover, Cd was mainly enriched in roots, the translocation ability from roots to aboveground parts was weak, and the four cultivars exhibited phytostabilization ability in Cd-contaminated soils. High-throughput sequencing (HTS) analysis showed that the Miscanthus rhizosphere bacterial community comprised 33 phyla and 446 genera, including plant growth-promoting rhizobacteria (PGPRs), such as Bacillus, Sphingomonas, and Mesorhizobium. The addition of Cd affected the Miscanthus rhizosphere bacterial community and reduced community diversity. Phylogenetic molecular ecological networks (pMENs) indicated that Cd addition reduced interactions between Miscanthus rhizosphere bacteria and thereby led to a simpler network structure, increased the number of negative-correlation links, enhanced the competition between rhizosphere bacterial species, reduced the number of key bacteria, and changed the composition of those bacteria. PICRUSt functional predictive analysis indicated that Cd stress reduced soil bacterial functions in the Miscanthus rhizosphere. The results of this study provide a basis for the remediation of Cd-contaminated soils by Miscanthus and provide a reference for the subsequent regulation of Miscanthus remediation efficiency by PGPRs or key bacteria.

Massilia atriviolacea sp. nov., a dark purple-pigmented bacterium isolated from soil.

A bacterial strain, designated SODT, with Gram-stain-negative and motile rod-shaped cells, was isolated from soil in Hefei, PR China, and was characterized using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain SODT belonged to the genus Massilia and showed the highest similarities to Massilia violaceinigra B2T (99.3 %), followed by Massilia glaciei B448-2T (98.7 %), Massilia eurypsychrophila CGMCC 1.12828T (98.6 %) and Rugamonas rubra CCM3730T (97.8 %). Average nucleotide identity and digital DNA-DNA hybridization values between genome sequences of strain SODT and the closely related species ranged from 77.1 to 89.3% and from 22.2 to 34.7 %. The DNA G+C content of strain SODT was 65.4 mol%. Strain SODT contained Q-8 as the major ubiquinone and the dominant fatty acids were summed feature 3 (C16 : 1ω7c and/or C15 : 0iso 2-OH; 58.5 %), C16 : 0 (26.8 %) and C18 : 1ω7c (5.0 %). The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol. On the basis of the evidence presented in this study, strain SODT represents a novel species of the genus Massilia, for which the name Massiliaatriviolacea sp. nov. is proposed. The type strain is SODT (=KCTC 62720T=LMG 30840T).

Paenibacillus zeisoli sp. nov., isolated from maize-cultivated soil artificially contaminated with cadmium.

A Gram-stain-negative, rod-shaped, motile bacterial strain, designated 3-5-3T, was isolated from maize-cultivated soil artificially contaminated with cadmium, in Nanyang, Henan Province, China. Strain 3-5-3T was oxidase- and catalase-positive. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain 3-5-3T was affiliated with the genus Paenibacillus and most closely related to Paenibacillus anaericanus MH2T (96.5 % similarity). The average nucleotide identity and digital DNA-DNA hybridization values between 3-5-3T and the closely related species ranged 69.4-84.5 % and 18.1-18.4 %. The genomic G+C content was 53.8 mol%. Anteiso-C15 : 0 was the major fatty acid and MK-7 was the only menaquinone. The diamino acid in the cell-wall peptidoglycan contains meso-diaminopimelic acid. The polar lipid profile contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylmethylethanolamine, two unidentified glycolipids, two unidentified aminolipids, one unidentified phospholipid, one unidentified phosphoaminolipid and six unidentified lipids. On the basis of the results obtained in this study, strain 3-5-3T is considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacilluszei soli sp. nov. is proposed. The type strain is 3-5-3T (=CGMCC 1.13686T=KCTC 33998T).

Bacillus acidinfaciens sp. nov., isolated from farmland soil.

A Gram-stain-positive, rod-shaped, motile bacterial strain, designated 3-2-2T, was isolated from field topsoil collected from a western suburb of Nanyang city, Henan province, China. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain 3-2-2T was a member of the genus Bacillus and most closely related to Bacillus fortis R-6514T (98.9 % similarity), Bacillus terrae RA9T (98.0 %) and Bacillus fordii R-7190T (97.7 %). A draft genome sequence determined for strain 3-2-2T revealed a DNA G+C content of 42.2 mol%. The average nucleotide identity and digital DNA-DNA hybridization values between 3-2-2T and the closely related Bacillus species ranged 79.4-84.2 % and 23.4-24.6 %. The major fatty acids of strain 3-2-2T were iso-C15 : 0, anteiso-C15 : 0, iso-C14 : 0 and iso-C16 : 0. The major isoprenoid quinone was MK-7. meso-Diaminopimelic acid was detected in the peptidoglycan. The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, an unidentified phospholipid and an unidentified lipid. The results of phylogenetic analyses, in silico genomic comparisons, and chemotaxonomic and phenotypic analyses clearly indicated that strain 3-2-2T represents a novel species within the genus Bacillus, for which the name Bacillus acidinfaciens sp. nov. is proposed. The type strain is 3-2-2T (=CGMCC 1.13685T=LMG 30839T).

Complete genome sequence of Caulobacter flavus RHGG3T, a type species of the genus Caulobacter with plant growth-promoting traits and heavy metal resistance.

Caulobacter flavus RHGG3T, a novel type species in the genus Caulobacter, originally isolated from rhizosphere soil of watermelon (Citrullus lanatus), has the ability to improve the growth of watermelon seedling and tolerate heavy metals. In vitro, C. flavus RHGG3T was able to solubilize phosphate (80.56 mg L-1), produce indole-3-acetic acid (IAA) (11.58 mg L-1) and was resistant to multiple heavy metals (copper, zinc, cadmium, cobalt and lead). Inoculating watermelon with this strain increased shoot and root length by 22.1% and 43.7%, respectively, and the total number of lateral roots by 55.9% compared to non-inoculated watermelon. In this study, we present the complete genome sequence of C. flavus RHGG3T, which was comprised of a single circular chromosome of 5,659,202 bp with a G + C content of 69.25%. An annotation analysis revealed that the C. flavus RHGG3T genome contained 5172 coding DNA sequences, 9 rRNA and 55 tRNA genes. Genes related to plant growth promotion (PGP), such as those associated with phosphate solubilization, nitrogen fixation, IAA, phenazine, volatile compounds, spermidine and cobalamin synthesis, were found in the C. flavus RHGG3T genome. Some genes responsible for heavy metal tolerance were also identified. The genome sequence of strain RHGG3T reported here provides new insight into the molecular mechanisms underlying the promotion of plant growth and the resistance to heavy metals in C. flavus. This study will be valuable for further exploration of the biotechnological applications of strain RHGG3T in agriculture.

Leucobacter triazinivorans sp. nov., a s-triazine herbicide prometryn-degrading bacterium isolated from sludge.

A Gram-stain-positive, rod-shaped, non-motile bacterial strain, designated JW-1T, was isolated from activated sludge collected from the outlet of an aeration tank in a prometryn-manufacturing plant, located in Binzhou City, Shandong province, PR China. Phylogenetic analysis, based on 16S rRNA gene sequences, indicated that strain JW-1T belongs to the genus Leucobacter and its closest neighbours are 'Leucobacter kyeonggiensis' F3-P9 (98.95 % similarity), Leucobacter celer subsp. astrifaciens CBX151T (98.62 %), Leucobacter celer subsp. celer NAL101T (98.53 %), Leucobacter chromiiresistens JG31T (97.86 %) and Leucobacter chironomi DSM 19883T (97.37 %). DNA-DNA hybridization values with the above strains were <55 %. The DNA G+C content of strain JW-1T was 72.6 mol%. The major fatty acids of strain JW-1T were iso-C16 : 0, anteiso-C15 : 0, anteiso-C17 : 0 and iso-C15 : 0. The predominant polar lipids were diphosphatidylglycerol, phosphatidylglycerol and glycolipid. The predominant menaquinone was MK-11. The cell wall amino acids were 2,4-diaminobutyric acid, alanine, glutamic acid, glycine and threonine. Based on the molecular and chemotaxonomic data, as well as the physiological and biochemical characteristics, strain JW-1T is considered to represent a novel species of the genus Leucobacter, for which the name Leucobacter triazinivorans is proposed. The type strain is JW-1T (=DSM 105188T=LMG 30083T).

Massilia buxea sp. nov., isolated from a rock surface.

A Gram-stain-negative, rod-shaped and motile bacterial strain, designated A9T, was isolated from the surface of rock collected from the shore of Nvshan lake in Mingguang, Anhui province, China. Phylogenetic analysis based on 16S rDNA sequence data showed that strain A9T was affiliated with the genus Massilia and showed the highest sequence similarities to Massilia plicata KCTC 12344T (98.8 %) and Massilia lurida CGMCC 1.10822T (97.9 %). The major fatty acids (>5 %) were summed feature 3 (C16 : 1ω7c and/or C15 : 0 iso 2-OH), C16 : 0 and C18 : 1ω7c. Strain A9T contained Q-8 as the predominant ubiquinone and diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and an unidentified aminophospholipid as the predominant polar lipids. The DNA G+C content was 69.9 mol%. Mean DNA-DNA relatedness values between strain A9T and its closest phylogenetic relatives, M. plicata KCTC 12344T and M. lurida CGMCC 1.10822T, were 38.8 % and 23.23 %, respectively. On the basis of the results obtained in this study, strain A9T is considered to represent a novel species of the genus Massilia, for which the name Massilia buxea sp. nov. is proposed. The type strain is A9T (=DSM 103547T=CGMCC 1.15931T=KCTC 52429T).

Mucilaginibacter rubeus sp. nov., isolated from rhizosphere soil.

A Gram-stain-negative, aerobic, non-motile, rod-shaped and non-spore-forming bacterium, designated EF23T, was isolated from rhizosphere soil of watermelon. Growth of strain EF23T was observed at 10-37 °C, at pH 5.0-9.0 and in the presence of 0-0.5 % (w/v) NaCl. Strain EF23T contained menaquinone 7 (MK-7) as the major isoprenoid quinone, and summed feature 3 (C16:1ω7c and/or iso-C15 : 0 2-OH), iso-C15 : 0, C16 : 0 and iso-C17 : 0 3-OH as the major fatty acids. Phosphatidylethanolamine was identified as the major polar lipid. The genomic DNA G+C content of strain EF23T was 43.7 mol%. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain EF23T was most closely related to Mucilaginibacter gossypii Gh-67T (98.9 % similarity) and Mucilaginibacter gossypiicola Gh-48T (97.6 %). DNA-DNA relatedness values between strain EF23T and M. gossypii KCTC 22380T and M. gossypiicola KCTC 22379T were 31.6 and 53.7 %. On the basis of the evidence presented in this polyphasic taxonomic study, strain EF23T is considered to represent a novel species of the genus Mucilaginibacter, for which the name Mucilaginibacter rubeus sp. nov. is proposed. The type strain is EF23T (=CGMCC 1.15913T=KCTC 52516T).

Caulobacter rhizosphaerae sp. nov., a stalked bacterium isolated from rhizosphere soil.

The Gram-reaction-negative, aerobic, white- to pale-yellow-coloured and rod-shaped bacterium with a single polar flagellum or a stalk, designated strain 7F14T, was isolated from rhizosphere soil of cultivated watermelon (Citrullus lanatus) collected from Hefei, China. Growth of strain 7F14T was observed at pH 6.0-9.0, 10-30 °C and in the presence of 0-1 % (w/v) NaCl. Cells were catalase-negative and oxidase-positive. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain 7F14T formed a phyletic lineage within the genus Caulobacter of the family Caulobacteraceae and showed the highest 16S rRNA gene sequence similarities to Caulobacter henricii ATCC 15253T (98.66 %), Caulobacter segnis ATCC 21756T (98.27 %), Caulobacter vibrioides CB51T (97.92 %) and Caulobacter flavus RHGG3T (97.44 %). The G+C content of the genomic DNA was 68.6 mol%. Strain 7F14T contained Q-10 as the sole ubiquinone and 11-methyl C18 : 1ω7c, C18 : 1ω7c, C16 : 0 and summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH) as the major fatty acids. The polar lipids profile consisted of phosphatidylglycerol, an unknown phosphoglycolipid, five unknown glycolipids, an unknown phospholipid and three unknown lipids. DNA-DNA relatedness values to the most closely related type strains Caulobacter henricii DSM 4730T and Caulobacter segnis DSM 7131T were 26.0 and 19.7 %, respectively. Based on unique phenotypic traits, and phylogenetic, chemotaxonomic and DNA-DNA hybridization results, strain 7F14T should be classified as a representative of a novel species of the genus Caulobacter, for which the name Caulobacter rhizosphaerae sp. nov. is proposed. The type strain is 7F14T (=CGMCC 1.15915T=KCTC 52515T).

Complete genome sequence of a novel chlorpyrifos degrading bacterium, Cupriavidus nantongensis X1.

Cupriavidus nantongensis X1 is a chlorpyrifos degrading bacterium, which was isolated from sludge collected at the drain outlet of a chlorpyrifos manufacture plant. It is the first time to report the complete genome sequence of C. nantongensis species, which has been reported as a novel species of Cupriavidus genus. It could provide further pathway information in chlorpyrifos degradation.

Cupriavidus nantongensis sp. nov., a novel chlorpyrifos-degrading bacterium isolated from sludge.

A Gram-stain-negative, aerobic, coccoid to small rod-shaped bacterium, designated X1T, was isolated from sludge collected from the vicinity of a pesticide manufacturer in Nantong, Jiangsu Province, China. Based on 16S rRNA gene sequence analysis, strain X1T belonged to the genus Cupriavidus, and was most closely related to Cupriavidus taiwanensis LMG 19424T (99.1 % 16S rRNA gene sequence similarity) and Cupriavidus alkaliphilus LMG 26294T (98.9 %). Strain X1T showed 16S rRNA gene sequence similarities of 97.2-98.2 % with other species of the genus Cupriavidus. The major cellular fatty acids of strain X1T were C16 : 0, C16 : 1ω7c and/or iso-C15 : 0 2-OH (summed feature 3), C18 : 1ω7c and C17 : 0 cyclo, and the major respiratory quinone was ubiquinone Q-8. The major polar lipids of strain X1T were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, aminophospholipid, phospholipid and hydroxyphosphatidylethanolamine. The DNA G+C content was 66.6 mol%. The DNA-DNA relatedness values of strain X1T with the five reference strains C. taiwanensis LMG 19424T, C. alkaliphilus LMG 26294T, Cupriavidus necator LMG 8453T, Cupriavidus gilardii LMG 5886T and 'Cupriavidus yeoncheonense' KCTC 42053 were lower than 70 %. The results obtained from phylogenetic analysis, phenotypic characterization and DNA-DNA hybridization indicated that strain X1T should be proposed to represent a novel species of the genus Cupriavidus, for which the name Cupriavidus nantongensis sp. nov. is proposed. The type strain is X1T (=KCTC 42909T=LMG 29218T).

Increased plant growth and copper uptake of host and non-host plants by metal-resistant and plant growth-promoting endophytic bacteria.

The effects of inoculation with two metal-resistant and plant growth-promoting endophytic bacteria (Burkholderia sp. GL12 and Bacillus megaterium JL35) were evaluated on the plant growth and Cu uptake in their host Elsholtzia splendens and non-host Brassica napus plants grown in natural Cu-contaminated soil. The two strains showed a high level of ACC deaminase activities. In pot experiments, inoculation with strain GL12 significantly increased root and above-ground tissue dry weights of both plants, consequently increasing the total Cu uptake of E. splendens and Brassica napus by 132% and 48.2% respectively. Inoculation with strain JL35 was found to significantly increase not only the biomass of B. napus, consequently increasing the total Cu uptake of B. napus by 31.3%, but Cu concentration of E. splendens for above-ground tissues by 318% and roots by 69.7%, consequently increasing the total Cu uptake of E. splendens by 223%. The two strains could colonize the rhizosphere soils and root interiors of both plants. Notably, strain JL35 could colonize the shoot tissues and significantly increase the translocation factors and bioaccumulation factors of E. splendens. These results suggested that Burkholderia sp. GL12 and B. megaterium JL35 were valuable bacterial resource which had the potential in improving the efficiency of Cu phytoextraction by E. splendens and B. napus in a natural Cu-contaminated soil.

Caulobacter flavus sp. nov., a stalked bacterium isolated from rhizosphere soil.

A Gram-stain-negative, aerobic, yellow-pigmented and rod-shaped bacterium with a single polar flagellum or a stalk, designated strain RHGG3T, was isolated from rhizosphere soil of cultivated watermelon (Citrullus lanatus) collected from Hefei, China. Optimal growth of strain RHGG3T was observed at pH 7.0 and 28-30 °C. Cells were catalase-positive and oxidase-negative. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain RHGG3T belonged to the genus Caulobacter and showed the highest 16S rRNA gene sequence similarities to Caulobacter segnis ATCC 21756T (98.6 %), Caulobacter vibrioides CB51T (98.3 %) and Caulobacter henricii ATCC 15253T (97.2 %). The G+C content of the genomic DNA was 70 mol%. Strain RHGG3T contained Q-10 as the sole ubiquinone and the major fatty acids (>8 %) were 11-methyl C18 : 1ω7c, C18 : 1ω7c, C16 : 0, C15 : 0 and summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH). The polar lipids were various unknown glycolipids, phosphatidylglycerol and phosphoglycolipids. DNA-DNA relatedness of strain RHGG3T to type strains of the most closely related species (Caulobacter segnis ATCC 21756T, Caulobacter vibrioides DSM 4738 and Caulobacter henricii ATCC 15253T) was 32.4-40.9 %. Based on polyphasic taxonomy analysis (phylogenetic, unique phenotypic traits, chemotaxonomic and DNA-DNA hybridizations), strain RHGG3T represents a novel species of the genus Caulobacter, for which the name Caulobacter flavus sp. nov. is proposed. The type strain is RHGG3T ( = CGMCC 1.15093T = KCTC 42581T = JCM 30763T).

The role of wheat germ agglutinin in the attachment of Pseudomonas sp. WS32 to wheat root.

Wheat germ agglutinin (WGA), which is secreted on the surface of wheat root, has been defined as a protein that reversibly and non-enzymatically binds to specific carbohydrates. However, little attention has been paid to the function of WGA in the attachment of bacteria to their host plants. The aim of this study was to investigate the role of WGA in the attachment of Pseudomonas sp. WS32 to wheat roots. Wheat roots were initially treated with double-distilled water, WGA-H (WGA solution that was heated at 100°C for 15 min) and WGA, independently. Subsequently, the roots were co-incubated with cell solutions (109 cells/ml). A dilution plate method using a solid nutrient medium was employed to determine the adsorption of WS32 to wheat roots. WGA was labeled with fluorescein isothiocyanate and detected using the fluorescent in situ hybridization (FISH) technique. The number of adsorptive WS32 cells on wheat roots was significantly increased when the wheat roots were pretreated with WGA, compared with the control treatment (p = 0.01). However, WGA-H failed to increase the amount of bacterial cells that attached to the wheat roots because of the loss of its physiological activity. The FISH assay also revealed that more cells adhered to WGA-treated wheat roots than to control or WGA-H-treated roots. The results indicated that WGA can mediate Pseudomonas strain WS32's adherence to wheat seedling roots. The findings of this study provide a better understanding of the processes involved in plant-microbe interactions.

Breeding and characterization of amino acid-analogue-resistant mutants of Arthrospira platensis.

To obtain amino acid-analogue-resistant mutants the wild strain A9 of Arthrospira platensis was mutated by ethylmethane sulfonate (EMS). Mutagenic effects of strain A9 by EMS were studied. The experimental results indicated that the survival rate curve of strain A9 took a typical "exponential shape" with lethal dosage of EMS being 1%. The survival of A9 strain was 13.2% when treated with 0.4% of EMS, and the resistant mutation rates to two amino acid analogues, ρ-fluorophenylalanine (FPA) and L-canavanine sulphate (CS), were greatly increased with the highest rates being at 4.9 × 10(-4) and 3.24 × 10(-4), respectively. By repeated screening, two stable mutants resistant to amino acid analogues, A9f resistant to FPA and A9c resistant to CS, were obtained. Resistances of the two mutants to corresponding amino acid-analogues were both significantly increased. Compared with their parent strain A9, A9f appeared larger than A9 performance in filament diameter, spiral diameter, spiral pitch, filament length and spiral number, and A9c showed much longer length and spiral pitch than those of the initial strain. Analysis results on amino acids compositions and contents showed that both two mutants accumulated quite higher concentration of amino acids in cells. The two mutants might be excellent high amino acids producing strain. By this means two useful mutants with stable genetic makers for further genetic study of A. platensis were obtained, which laid a good foundation for further study on the transformation of A. platensis.

Isolation and characterization of plant growth-promoting rhizobacteria from wheat roots by wheat germ agglutinin labeled with fluorescein isothiocyanate.

Thirty-two isolates were obtained from wheat rhizosphere by wheat germ agglutinin (WGA) labeled with fluorescein isothiocyanate (FITC). Most isolates were able to produce indole acetic acid (65.6%) and siderophores (59.3%), as well as exhibited phosphate solubilization (96.8%). Fourteen isolates displayed three plant growth-promoting traits. Among these strains, two phosphate-dissolving ones, WS29 and WS31, were evaluated for their beneficial effects on the early growth of wheat (Triticum aestivum Wan33). Strain WS29 and WS31 significantly promoted the development of lateral roots by 34.9% and 27.6%, as well as increased the root dry weight by 25.0% and 25.6%, respectively, compared to those of the control. Based on 16S rRNA gene sequence comparisons and phylogenetic positions, both isolates were determined to belong to the genus Bacillus. The proportion of isolates showing the properties of plant growth-promoting rhizobacteria (PGPR) was higher than in previous reports. The efficiency of the isolation of PGPR strains was also greatly increased by WGA labeled with FITC. The present study indicated that WGA could be used as an effective tool for isolating PGPR strains with high affinity to host plants from wheat roots. The proposed approach could facilitate research on biofertilizers or biocontrol agents.

Promotion of growth and Cu accumulation of bio-energy crop (Zea mays) by bacteria: implications for energy plant biomass production and phytoremediation.

Three metal-resistant and plant growth-promoting bacteria (Burkholderia sp. GL12, Bacillus megaterium JL35 and Sphingomonas sp. YM22) were evaluated for their potential to solubilize Cu(2) (OH)(2)CO(3) in solution culture and their plant growth promotion and Cu uptake in maize (Zea mays, an energy crop) grown in a natural highly Cu-contaminated soil. The impacts of the bacteria on the Cu availability and the bacterial community in rhizosphere soils of maize were also investigated. Inductively coupled-plasma optical emission spectrometer analysis showed variable amounts of water-soluble Cu (ranging from 20.5 to 227 mgL(-1)) released by the bacteria from Cu(2) (OH)(2)CO(3) in solution culture. Inoculation with the bacteria was found to significantly increase root (ranging from 48% to 83%) and above-ground tissue (ranging from 33% to 56%) dry weights of maize compared to the uninoculated controls. Increases in Cu contents of roots and above-ground tissues varied from 69% to 107% and from 16% to 86% in the bacterial-inoculated plants compared to the uninoculated controls, respectively. Inoculation with the bacteria was also found to significantly increase the water-extractive Cu concentrations (ranging from 63 to 94%) in the rhizosphere soils of the maize plants compared to the uninoculated controls in pot experiments. Denaturing gradient gel electrophoresis and sequence analyses showed that the bacteria could colonize the rhizosphere soils and significantly change the bacterial community compositions in the rhizosphere soils. These results suggest that the metal-resistant and plant growth-promoting bacteria may be exploited for promoting the maize (energy crop) biomass production and Cu phytoremediation in a natural highly Cu-contaminated soil.

Genetic diversity and characterization of heavy metal-resistant-endophytic bacteria from two copper-tolerant plant species on copper mine wasteland.

The diversity of endophytic bacteria from the copper-tolerant species Elsholtzia splendens and Commelina communis was evaluated by using cultivation and cultivation-independent techniques. Phylogenetic analysis based on 16S rDNA sequences showed that the Cu-resistant isolates belonged to three major phylogenetic groups: Firmicutes, Actinobacteria and Proteobacteria. Bacillus and Acinetobacter were predominant among the Cu-resistant isolates. Sequence analysis from the 16S rDNA clone libraries of the two plant roots revealed sequences of Alpha-, Beta-, Gamma-Proteobacteria, Firmicutes,Actinobacteria and Bacteroidetes. Gammaproteobacteria was predominant in the two 16S rDNA clone libraries. Thirty-two endophytic bacteria were isolated and characterized with respect to heavy metal resistance and production of plant growth-promoting factors. In experiments involving rape plants grown in vermiculite containing 4 mg kg(-1) of Cu, inoculation with the isolates was found to increase the dry weights of roots (ranging from 132% to 155%) and above-ground tissues (ranging from 71% to 83%) compared to the uninoculated control. Increase in above-ground tissue Cu content varied from 63% to 125% in inoculated-rape plants cultivated in Cu-contaminated substrate compared to the uninoculated control.