Ying Ma

Plant growth promoting rhizobacteria and endophytes accelerate phytoremediation of metalliferous soils

Technogenic activities (industrial-plastic, textiles, microelectronics, wood preservatives; mining-mine refuse, tailings, smelting; agrochemicals-chemical fertilizers, farm yard manure, pesticides; aerosols-pyrometallurgical and automobile exhausts; biosolids-sewage sludge, domestic waste; fly ash-coal combustion products) are the primary sources of heavy metal contamination and pollution in the environment in addition to geogenic sources. During the last two decades, bioremediation has emerged as a potential tool to clean up the metal-contaminated/polluted environment. Exclusively derived processes by plants alone (phytoremediation) are time-consuming. Further, high levels of pollutants pose toxicity to the remediating plants. This situation could be ameliorated and accelerated by exploring the partnership of plant-microbe, which would improve the plant growth by facilitating the sequestration of toxic heavy metals. Plants can bioconcentrate (phytoextraction) as well as bioimmobilize or inactivate (phytostabilization) toxic heavy metals through in situ rhizospheric processes. The mobility and bioavailability of heavy metal in the soil, particularly at the rhizosphere where root uptake or exclusion takes place, are critical factors that affect phytoextraction and phytostabilization. Developing new methods for either enhancing (phytoextraction) or reducing the bioavailability of metal contaminants in the rhizosphere (phytostabilization) as well as improving plant establishment, growth, and health could significantly speed up the process of bioremediation techniques. In this review, we have highlighted the role of plant growth promoting rhizo- and/or endophytic bacteria in accelerating phytoremediation derived benefits in extensive tables and elaborate schematic sketches.

Inoculation of plant growth promoting bacterium Achromobacter xylosoxidans strain Ax10 for the improvement of copper phytoextraction by Brassica juncea

In this study, a copper-resistant plant growth promoting bacterial (PGPB) strain Ax10 was isolated from a Cu mine soil to assess its plant growth promotion and copper uptake in Brassica juncea. The strain Ax10 tolerated concentrations up to 600 mg CuL(-1) on a Luria-Bertani (LB) agar medium and utilized 1-aminocyclopropane-1-carboxylic acid (ACC) as a sole N source in DF salts minimal medium. The strain Ax10 was characterized as Achromobacter xylosoxidans based on its 16S rDNA sequence homology (99%). The bacterium A. xylosoxidans Ax10 has also exhibited the capability of producing indole acetic acid (IAA) (6.4 microg mL(-1)), and solubilizing inorganic phosphate (89.6 microg mL(-1)) in specific culture media. In pot experiments, inoculation of A. xylosoxidans Ax10 significantly increased the root length, shoot length, fresh weight and dry weight of B. juncea plants compared to the control. This effect can be attributed to the utilization of ACC, production of IAA and solubilization of phosphate. Furthermore, A. xylosoxidans Ax10 inoculation significantly improved Cu uptake by B. juncea. Owing to its wide action spectrum, the Cu-resistant A. xylosoxidans Ax10 could serve as an effective metal sequestering and growth promoting bioinoculant for plants in Cu-stressed soil. The present study has provided a new insight into the phytoremediation of Cu-contaminated soil.

Improvement of plant growth and nickel uptake by nickel resistant-plant-growth promoting bacteria

In this study, among a collection of Ni-resistant bacterial strains isolated from the rhizosphere of Alyssum serpyllifolium and Phleum phleoides grown on serpentine soil, five plant growth-promoting bacteria (PGPB) were selected based on their ability to utilize 1-aminocyclopropane-1-carboxylate (ACC) as the sole N source and promote seedling growth. All of the strains tested positive for indole-3-acetic acid (IAA) production and phosphate solubilization. In addition, four of the strains exhibited significant levels of siderophores production. Further, the efficiency of PGPB in enhancing Ni solubilization in soils was analyzed. Compared with control treatment, inoculation of PGPB strains significantly increased the concentrations of bioavailable Ni. Furthermore, a pot experiment was conducted to elucidate the effects of inoculating Ni-resistant PGPB on the plant growth and the uptake of Ni by Brassica juncea and B. oxyrrhina in soil contaminated with 450 mg kg(-1) Ni. Psychrobacter sp. SRA2 significantly increased the fresh (351%) and dry biomass (285%) of the B. juncea test plants (p<0.05), whereas Psychrobacter sp. SRA1 and Bacillus cereus SRA10 significantly increased the accumulation of Ni in the root and shoot tissues of B. juncea compared with non-inoculated controls. This result indicates that the strains SRA1 and SRA10 facilitated the release of Ni from the non-soluble phases in the soil, thus enhancing the availability of Ni to plants. A significant increase, greater than that of the control, was also noted for growth parameters of the B. oxyrrhina test plants when the seeds were treated with strain SRA2. This effect can be attributed to the utilization of ACC, solubilization of phosphate and production of IAA. The results of the study revealed that the inoculation of Ni mobilizing strains Psychrobacter sp. SRA1 and B. cereus SRA10 increases the efficiency of phytoextraction directly by enhancing the metal accumulation in plant tissues and the efficient PGPB, Psychrobacter sp. SRA2 increases indirectly by promoting the growth of B. juncea and B. oxyrrhina.

Inoculation of endophytic bacteria on host and non-host plants--effects on plant growth and Ni uptake.

Among a collection of Ni resistant endophytes isolated from the tissues of Alyssum serpyllifolium, four plant growth promoting endophytic bacteria (PGPE) were selected based on their ability to promote seedling growth in roll towel assay. Further, the PGPE screened showed the potential to produce plant growth promoting (PGP) substances and plant polymer hydrolyzing enzymes. These isolates were further screened for their PGP activity on A. serpyllifolium and Brassica juncea under Ni stress using a phytagar assay. None of the four isolates produced any disease symptoms in either plant. Further, strain A3R3 induced a maximum increase in biomass and Ni content of plants. Based on the PGP potential in phytagar assay, strain A3R3 was chosen for studying its PGP effect on A. serpyllifolium and B. juncea in Ni contaminated soil. Inoculation with A3R3 significantly increased the biomass (B. juncea) and Ni content (A. serpyllifolium) of plants grown in Ni contaminated soil. The strain also showed high level of colonization in tissue interior of both plants. By 16S rRNA gene sequencing analysis, A3R3 was identified as Pseudomonas sp. Successful colonization and subsequent PGP potentiality of Pseudomonas sp. A3R3 indicate that the inoculation with PGPE might have significant potential to improve heavy metal phytoremediation.

Isolation and characterization of Ni mobilizing PGPB from serpentine soils and their potential in promoting plant growth and Ni accumulation by Brassica spp.

The study was undertaken to assess the effects of Ni mobilizing bacteria on the plant growth and the uptake of Ni by Brassica juncea and Brassica oxyrrhina. Among a collection of Ni resistant bacterial strains isolated from the non-rhizosphere and rhizosphere soils of Alyssum serpyllifolium and Astragalus incanus at a serpentine site in Bragança, north-east of Portugal, nine strains were selected based on their ability to solubilize Ni in soil. Further assessment on plant growth-promoting parameters revealed the intrinsic ability of the Ni mobilizing strains to produce indole-3-acetic acid (IAA), siderophores, utilize 1-aminocyclopropane-1-carboxylic acid (ACC) as the sole N source and solubilize insoluble phosphate. All of the strains tested positive for IAA production and phosphate solubilization. In addition, all the strains, except SRS5 exhibited significant levels of siderophore production. Besides, five isolates showed positive for ACC deaminase activity. In pot experiments, inoculation of plants with Ni mobilizing strains increased the biomass of both B. juncea and B. oxyrrhina. Among the strains, Pseudomonas sp. SRI2, Psychrobacter sp. SRS8 and Bacillus sp. SN9 showed maximum increase in the biomass of the test plants. In addition, the strain SN9 significantly increased the Ni concentration in the root and shoot tissues of B. juncea and B. oxyrrhina. Further, a significantly positive correlation was observed between the bacterial Ni mobilization in soil and the total Ni uptake in both plant species. The findings, therefore, revealed that inoculation of Ni mobilizing plant growth-promoting bacterial strain SN9 increases the efficiency of phytoextraction directly by enhancing Ni accumulation in plant tissues and indirectly by promoting the shoot and root biomass of B. juncea and B. oxyrrhina.

Photocatalytic activity of TiO2 films grown on different substrates.

Titanium dioxide films were prepared on glass, indium-tin oxide (ITO) glass and p-type monocrystalline silicon and studied for the photocatalytic degradation of rhodamine B in an aqueous medium. Raman, AFM, and XPS spectroscopic investigations of these films indicated that microstructure of titanium oxide films were greatly affected by the substrate materials. Rutile was confirmed to be easily formed on the surface of ITO glass, and TiO2 tended to grow as closely packed particles that were elongated strips with an average size of 20 nm, and had lovely contrast with the perfectly round particles grown on p-type monocrystalline silicon. Charge transfer between the film and silicon substrate was verified by surface photovoltage spectra. This may be the real reason why the films grown on ITO glass and silicon substrates exhibit higher photocatalytic reactivity than the film on glass substrate. Moreover, the different surface properties also seem to be responsible for the different activity.

Beneficial role of bacterial endophytes in heavy metal phytoremediation

Phytoremediation is an emerging technology that uses plants and their associated microbes to clean up pollutants from the soil, water and air. In recent years, phytoremediation assisted by bacterial endophytes has been highly recommended for cleaning up of metal polluted soils since endophytic bacteria can alleviate metal toxicity in plant through their own metal resistance system and facilitate plant growth under metal stress. Endophytic bacteria improve plant growth in metal polluted soils in two different ways: 1) directly by producing plant growth beneficial substances including solubilization/transformation of mineral nutrients (phosphate, nitrogen and potassium), production of phytohormones, siderophores and specific enzymes; and 2) indirectly through controlling plant pathogens or by inducing a systemic resistance of plants against pathogens. Besides, they also alter metal accumulation capacity in plants by excreting metal immobilizing extracellular polymeric substances, as well as metal mobilizing organic acids and biosurfactants. The present work aims to review the progress of recent research on the isolation, identification and diversity of metal resistant endophytic bacteria and illustrate various mechanisms responsible for plant growth promotion and heavy metal detoxification/phytoaccumulation/translocation in plants.

Characterization of metal-resistant plant-growth promoting Bacillus weihenstephanensis isolated from serpentine soil in Portugal

A metal‐resistant bacterial strain SM3 isolated from a serpentine soil in the north‐east of Portugal was characterized as Bacillus weihenstephanensis based on the morphological and biochemical characteristics and on the comparative analysis of the partial 16S ribosomal DNA sequence. Bacillus weihenstephanensis SM3 showed a high degree of resistance to nickel (1500 mg l–1), copper (500 mg l–1) and zinc (700 mg l–1) and also to antibiotics (ampicillin, penicillin, kanamycin and streptomycin). Strain SM3 has also exhibited the capability of solubilizing phosphate and producing indole‐3‐acetic acid (IAA) both in the absence and in the presence of metals (Ni, Cu and Zn). A pot experiment was conducted to elucidate the effects of strain SM3 on plant growth and uptake of Ni, Cu or Zn by Helianthus annuus. Inoculation with strain SM3 increased the shoot and root biomass of H. annuus grown in both non‐contaminated and contaminated soil. Furthermore, strain SM3 increased the accumulation of Cu and Zn in the root and shoot systems. A batch experiment was also conducted to assess the metal mobilization potential of strain SM3 in soil. Inoculation with this strain increased the concentrations of water soluble Ni, Cu and Zn in soil. Metal solubilization by this bacterial strain may be an important process to promote the uptake of heavy metals by plants. This study elucidates the multifarious role of strain SM3 in plant growth promotion and its metal mobilizing potential. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

The hyperaccumulator Sedum plumbizincicola harbors metal-resistant endophytic bacteria that improve its phytoextraction capacity in multi-metal contaminated soil.

Endophyte-assisted phytoremediation has recently been suggested as a successful approach for ecological restoration of metal contaminated soils, however little information is available on the influence of endophytic bacteria on the phytoextraction capacity of metal hyperaccumulating plants in multi-metal polluted soils. The aims of our study were to isolate and characterize metal-resistant and 1-aminocyclopropane-1-carboxylate (ACC) utilizing endophytic bacteria from tissues of the newly discovered Zn/Cd hyperaccumulator Sedum plumbizincicola and to examine if these endophytic bacterial strains could improve the efficiency of phytoextraction of multi-metal contaminated soils. Among a collection of 42 metal resistant bacterial strains isolated from the tissues of S.xa0plumbizincicola grown on Pb/Zn mine tailings, five plant growth promoting endophytic bacterial strains (PGPE) were selected due to their ability to promote plant growth and to utilize ACC as the sole nitrogen source. The five isolates were identified as Bacillus pumilus E2S2, Bacillus sp. E1S2, Bacillus sp. E4S1, Achromobacter sp. E4L5 and Stenotrophomonas sp. E1L and subsequent testing revealed that they all exhibited traits associated with plant growth promotion, such as production of indole-3-acetic acid and siderophores and solubilization of phosphorus. These five strains showed high resistance to heavy metals (Cd, Zn and Pb) and various antibiotics. Further, inoculation of these ACC utilizing strains significantly increased the concentrations of water extractable Cd and Zn in soil. Moreover, a pot experiment was conducted to elucidate the effects of inoculating metal-resistant ACC utilizing strains on the growth of S.xa0plumbizincicola and its uptake of Cd, Zn and Pb in multi-metal contaminated soils. Out of the five strains, B.xa0pumilus E2S2 significantly increased root (146%) and shoot (17%) length, fresh (37%) and dry biomass (32%) of S.xa0plumbizincicola as well as plant Cd uptake (43%), whereas Bacillus sp. E1S2 significantly enhanced the accumulation of Zn (18%) in plants compared with non-inoculated controls. The inoculated strains also showed high levels of colonization in rhizosphere and plant tissues. Results demonstrate the potential to improve phytoextraction of soils contaminated with multiple heavy metals by inoculating metal hyperaccumulating plants with their own selected functional endophytic bacterial strains.

Chemically responsive luminescent switching in transparent flexible self-supporting [EuW10O36]9−-agarose nanocomposite thin films

Europium-containing polyoxometalates (Eu-POMs) are widely used for the fabrication of hybrid inorganic-organic luminescent materials. A few efforts have been devoted to develop active Eu-POM-based luminescent sensors and switches. In this study, highly transparent flexible self-supporting decatungsteuropate(EuW10)-agarose thin films were successfully fabricated by a facile hydrogel casting technique. It was identified that strong interactions between agarose and EuW10 by hydrogen bonds at the hydroxyl sites and densely-packed 3D network structure of agarose in the gel state account for the homogenous distribution of EuW10 and good mechanical properties of the nanocomposite films. More importantly, the obtained thin films displayed strong red emission of Eu(III) ion, and the luminescence of these thin films was sensitive to the acid and base gases. When the thin films were exposed to HCl gas, their luminescence was sharply decreased, while the luminescence was recovered upon subsequently exposing the films to NH3 gas. Such process could be repeated many times and a portable switch based on these thin films was proposed.