Endophytic bacteria enhance remediation of tannery effluent in constructed wetlands vegetated with Leptochloa fusca

The use of constructed wetlands (CWs) is a promising approach for the remediation of wastewater. The present study aims to develop a plant–bacteria system within CWs for the efficient remediation of tannery effluent. In a vertical-flow CW vegetated with Leptochloa fusca (Kallar grass), a consortium of three different endophytic bacteria, Pantoea stewartii ASI11, Microbacterium arborescens HU33, and Enterobacter sp. HU38, was used for bioaugmentation. CWs vegetated with only L. fusca had the potential to remediate tannery effluent, but augmentation with endophytic bacteria enhanced the growth of L. fusca while aiding in the removal of both organic and inorganic pollutants from the tannery effluent. Moreover, the bacterial augmentation decreased toxicity in the effluent as well. A higher number of chromium (Cr)-resistant bacteria were isolated from the rhizosphere and endosphere of L. fusca inoculated with the endophytes than from uninoculated plants. Due to promising bioremediation and detoxification potential of L. fusca, it is reported for the first time as a potential candidate to develop effective CWs for the remediation of polluted effluents in combination with pollutant-degrading endophytic bacteria.     

Successful phytoremediation of crude-oil contaminated soil at an oil exploration and production company by plants-bacterial synergism

Phytoremediation is a promising approach for the cleanup of soil contaminated with petroleum hydrocarbons. This study aimed to develop plant-bacterial synergism for the successful remediation of crude oil-contaminated soil. A consortia of three endophytic bacteria was augmented to two grasses, Leptochloa fusca and Brachiaria mutica, grown in oil-contaminated soil (46.8 g oil kg^?1 soil) in the vicinity of an oil exploration and production company. Endophytes augmentation improved plant growth, crude oil degradation, and soil health. Maximum oil degradation (80%) was achieved with B. mutica plants augmented with the endophytes and it was significantly (P < 0.05) higher than the use of plants or bacteria individually. Moreover, endophytes showed more persistence, the abundance and expression of alkB gene in the rhizosphere as well as in the endosphere of the tested plants than in unvegetated soil. A positive relationship (r = 0.70) observed between gene expression and crude oil reduction indicates that catabolic gene expression is important for hydrocarbon mineralization. This investigation showed that the use of endophytes with appropriate plant is an effective strategy for the cleanup of oil-contaminated soil under field conditions.     

Phytoremediation of lead by a wild,non-edible Pb accumulator Coronopus didymus (L.) Brassicaceae

Coronopus didymus was examined in terms of its ability to remediate Pb-contaminated soils. Pot experiments were conducted for 4 and 6 weeks to compare the growth, biomass, photosynthetic efficiency, lead (Pb) uptake, and accumulation by C. didymus plants. The plants grew well having no visible toxic symptoms and 100% survivability, exposed to different Pb-spiked soils 100, 350, 1500, and 2500 mg kg^?1, supplied as lead nitrate. After 4 weeks, root and shoot concentrations reached 1652 and 502 mg Pb kg^?1 DW, while after 6 weeks they increased up to 3091 and 527 mg Pb kg^?1 DW, respectively, at highest Pb concentration. As compared to the 4 week experiments, the plant growth and biomass yield were higher after 6 weeks of Pb exposure. However, the chlorophyll content of leaves decreased but only a slight decline in photosynthetic efficiency was observed on exposure to Pb at both 4 and 6 weeks. The Pb accumulation was higher in roots than in the shoots. The bioconcentration factor of Pb was > 1 in all the plant samples, but the translocation factor was < 1. This suggested C. didymus as a good candidate for phytoremediation of Pb-contaminated soils and can be used for future remediation purposes.     

Effect of Pseudomonas fluorescens RB4 and Bacillus subtilis 189 on the phytoremediation potential of Catharanthus roseus (L.) in Cu and Pb-contaminated soils

The remediation of heavy metal-contaminated soils has become a critical issue due to toxic effects of these metals on living organisms. The current research was conducted to study the effect of Pseudomonas fluorescens RB4 and Bacillus subtilis 189 on the growth and phytoremediation potential of Catharanthus roseus in Cu- and Pb-contaminated soils. The bacterial strains exhibited significantly higher level of water-extractable Pb and Cu in Pb, Cu, and Cu+Pb-contaminated. The P. fluorescens RB4 inoculated plants, produced 102%, 48%, and 45% higher fresh weight (FW) in soils contaminated with Cu, Pb, and both elements, respectively, as compared to un-inoculated control plants. Similarly, B. subtilis 189 inoculated plants produced 108%, 43%, and 114% more FW in the presence of Cu, Pb, and both elements. The plants co-cultivated with both bacteria exhibited 121%, 102%, and 177% higher FW, in Cu, Pb, and both elements contaminated soils, as compared to respective un-inoculated control. Co-cultivation of P. fluorescens RB4, B. subtilis 189, and P. fluorescens RB4 + B. subtilis 189 resulted in higher accumulation of Cu and Pb in shoots of the C. roseus grown in contaminated soils as compared to un-inoculated control. Bacterial treatments also improved the translocation and metal bioconcentration factors. The growth and phytoextraction capability of C. roseus was improved by inoculation of P. fluorescens RB4 and B. subtilis 189.     

Hyperaccumulator Oilcake Manure as an Alternative for Chelate-Induced Phytoremediation of Heavy Metals Contaminated Alluvial Soils

The ability of hyperaccumulator oilcake manure as compared to chelates was investigated by growing Calendula officinalis L for phytoremediation of cadmium and lead contaminated alluvial soil. The combinatorial treatment T₆ [2.5 g kg^?1oilcake manure + 5 mmol kg^?1 EDDS] caused maximum cadmium accumulation in root, shoot and flower up to 5.46, 4.74 and 1.37 mg kg^?1and lead accumulation up to 16.11, 13.44 and 3.17 mg kg^?1, respectively at Naini dump site, Allahabad (S₃). The treatment showed maximum remediation efficiency for Cd (RR = 0.676%) and Pb (RR = 0.202%) at Mumfordganj contaminated site (S₂). However, the above parameters were also observed at par with the treatment T₅ [2.5 g kg^?1oilcake manure +2 g kg^?1 humic acid]. Applied EDDS altered chlorophyll–a, chlorophyll–b, and carotene contents of plants while application of oilcake manure enhanced their contents in plant by 3.73–8.65%, 5.81–17.65%, and 7.04–17.19%, respectively. The authors conclude that Calendula officinalis L has potential to be safely grown in moderately Cd and Pb-contaminated soils and application of hyperaccumulator oilcake manure boosts the photosynthetic pigments of the plant, leading to enhanced clean-up of the cadmium and lead-contaminated soils. Hence, the hyperaccumulator oilcake manure should be preferred over chelates for sustainable phytoremediation through soil-plant rhizospheric process.     

Arbuscular Mycorrhizae Effects on Heavy Metal Uptake by Corn

Maximizing uptake of soil-borne metals into plants is important for successful phytoremediation. Arbuscular mycorrhizae (AM) have been shown previously by our group to sequester metals in the roots of plants and prevent translocation to the shoot. If AM colonization of roots can be reduced, it may be possible to increase metal uptake into plants, thus increasing the efficiency of phytoremediation. The fungicide Benomy® was applied to a Pb-contaminated soil and seeded to corn (Zea mays). Because soil pH affects metal solubility, two pHs were also examined. Colonization of roots by AM was significantly decreased by application of Benomyl to soil, but only at the higher soil pH. Benomyl increased the concentration of several elements, including Pb, in shoots. However, the total Pb content in the shoot decreased due to the reduced shoot weight associated with Benomyl application.

     

Chelator-assisted phytoextraction of arsenic,cadmium and lead by Pteris vittata L. and soil microbial community structure response

Abstract

Using biodegradable chelators to assist in phytoextraction may be an effective approach to enhance the heavy-metal remediation efficiencies of plants. A pot experiment was conducted to investigate the effects of ethylenediamine disuccinic acid (EDDS), citric acid (CA), and oxalic acid (OA) on the growth of the arsenic (As) hyperaccumulator Pteris vittata L., its arsenic (As), cadmium (Cd), and lead (Pb) uptake and accumulation, and soil microbial responses in multi-metal(loid)-contaminated soil. The addition of 2.5-mmol kg^?1 OA (OA-2.5) produced 26.7 and 14.9% more rhizoid and shoot biomass, respectively compared with the control, while EDDS and CA treatments significantly inhibited plant growth. The As accumulation in plants after the OA-2.5 treatment increased by 44.2% and the Cd and Pb accumulation in plants after a 1-mmol kg^?1 EDDS treatment increased by 24.5 and 19.6%, respectively. Soil urease enzyme activities in OA-2.5 treatment were significantly greater than those in the control and other chelator treatments (p?<?0.05). A PCR–denatured gradient gel electrophoresis analysis revealed that with the addition of EDDS, CA and OA enhanced soil microbial diversity. It was concluded that the addition of OA-2.5 was suitable for facilitating phytoremediation of soil As and did not have negative effects on the microbial community.     

Enhanced remediation of chlorpyrifos by ryegrass (Lolium multiflorum) and a chlorpyrifos degrading bacterial endophyte Mezorhizobium sp. HN3

For effective remediation of contaminants, plant-endophyte partnership is a promising field to be explored. Generally endophytic bacteria assist their host plant by withstanding the stress induced by the contaminants. The objective of this study was to explore the suitability of plant-bacterial partnership for chlorpyrifos (CP) remediation using ryegrass and a CP degrading endophyte, Mesorhizobium sp. HN3 which belongs to plant growth promoting rhizobia. The inoculated yfp-tagged Mesorhizobium sp. HN3 efficiently colonized in the rhizosphere, enhanced plant growth and degradation of CP and its metabolite 3,5,6 trichloro-2-pyridinol (TCP). Significantly lower CP residues were observed in the roots and shoots of plants vegetated in inoculated soil which might be attributed to the efficient root colonization of HN3yfp. These results suggest the involvement of Mesorhizobium sp. HN3yfp in CP degradation inside the roots and rhizosphere of plants and further emphasize on the effectiveness of endophytic bacteria in stimulating the remediation of pesticide contaminants. This is the first report which demonstrates the efficacy of bacterial endophyte for degradation of CP residues taken up by the plant and enhanced remediation of chlorpyrifos contaminated soil.     

Effect of<Emphasis Type="Italic">Glomus intraradices</Emphasis> isolated from Pb-contaminated soil on Pb uptake by<Emphasis Type="Italic">Agrostis capillaris</Emphasis> is changed by its cultivation in a metal-free substrate

Development and heavy metal tolerance of two cultivation lineages of the indigenous isolate of arbuscular mycorrhizal fungus (AMF)Glomus intraradices PH5 were compared in a pot experiment in soil from lead (Pb) smelter waste deposits. One lineage was sub-cultured in original Pb-contaminated soil; the second one was maintained for 13 months in an inert substrate (river sand) without Pb stress. The contribution of these cultivation lineages to the Pb uptake and accumulation by the host plantAgrostis capillaris was investigated. The experiment was conducted in a compartmented system where the lateral compartments withAgrostis seedlings were separated from the central pot containing 4-week olderAgrostis plants by a nylon mesh for allowing out-growing of extraradical mycelium (ERM) from the pot. No differences in mycorrhizal colonization, ERM length and viability were observed between the two lineages ofG. intraradices PH5 in the soil of the isolate origin. However, the ability to support plant growth and Pb uptake differed between the lineages and also between the plants in the central pots and the lateral compartments. The growth of the plants in the central pots was positively affected by AMF inoculation. The plants inoculated with the lineage maintained in original soil showed larger shoot biomass and higher shoot P content as compared to the other inoculation treatments. The shoot Pb concentration of these plants was lower when compared to the plants inoculated with the lineage sub-cultured in the inert substrate. However the concentration did not differ from non-mycorrhizal control or from the reference isolateG. intraradices BEG75 from non-contaminated soil. Also shoot Pb contents were similar for all inoculation treatments. The development ofG. intraradices BEG75 in the contaminated soil was very poor; this isolate was not able to initiate colonization of seedlings in lateral compartments. In lateral compartments, growth of seedlings in contaminated soil was inhibited by theG. intraradices PH5 lineage maintained in the inert substrate. Pb translocation from the seedling roots to shoots was increased for plants inoculated with either lineage as compared to the non-mycorrhizal control; however, the increase for the lineage cultivated in the inert substrate was significantly higher in comparison with that maintained in the original soil. After 13 months of cultivation in a metal free substrate, theG. intraradices isolate from Pb contaminated soil did not lose its tolerance to Pb as regards colonization of plant roots and growth of ERM in the soil of its origin. However, its ability to support plant growth and to prevent Pb translocation from the roots to the shoots was decreased.     

Application of endophytic bacteria for the biocontrol of Rhizoctonia solani (Cantharellales: ceratobasidiaceae) damping-off disease in cotton seedlings

The antifungal potentialities of three endophytic bacterial strains, Stenotrophomonas maltophila H8 (Xanthomonadales: Xanthomonadaceae), Pseudomonas aeruginosa H40 (Pseudomonadales: Pseudomonadaceae) and Bacillus subtilis H18 (Bacillales: Bacillaceae) were evaluated against the phytopathogenic fungus Rhizoctonia solani in cotton seedlings under greenhouse conditions. The bacterial strains were applied as a soil drench or talc-based bioformulation in R. solani-infested soil and non-infested soil. Results indicated that the soil drench treatment was more efficient than talc-based bioformulation. A significant increase of seed emergence and seedling survival with a clear reduction of disease severity was achieved with the endophytic bacterial treatments. At the same time, the fresh weight, dry weight, shoot length and root length of the treated plants were markedly enhanced. Moreover, there was an apparent induction of the antioxidant enzymes (peroxidase, polyphenol oxidase and catalase) of the treated seedlings. Gas chromatography–mass spectrometry revealed the presence of various bioactive compounds in the bacterial supernatant. The antagonistic activity of the bacterial strains against R. solani was attributed to their capability to produce a broad spectrum of antifungal compounds in addition to bioactive molecules that can trigger the systemic resistance in the infected seedlings.     

Effect of Medicago sativa L. and compost on organic and inorganic pollutant removal from a mixed contaminated soil and risk assessment using ecotoxicological tests

Several Gentle Remediation Options (GRO), e.g., plant-based options (phytoremediation), singly and combined with soil amendments, can be simultaneously efficient for degrading organic pollutants and either stabilizing or extracting trace elements (TEs). Here, a 5-month greenhouse trial was performed to test the efficiency of Medicago sativa L., singly and combined with a compost addition (30% w/w), to treat soils contaminated by petroleum hydrocarbons (PHC), Co and Pb collected at an auto scrap yard. After 5 months, total soil Pb significantly decreased in the compost-amended soil planted with M. sativa, but not total soil Co. Compost incorporation into the soil promoted PHC degradation, M. sativa growth and survival, and shoot Pb concentrations [3.8 mg kg^?1 dry weight (DW)]. Residual risk assessment after the phytoremediation trial showed a positive effect of compost amendment on plant growth and earthworm development. The O₂ uptake by soil microorganisms was lower in the compost-amended soil, suggesting a decrease in microbial activity. This study underlined the benefits of the phytoremediation option based on M. sativa cultivation and compost amendment for remediating PHC- and Pb-contaminated soils.     

Use of Energy Crop (Ricinus communis L.) for Phytoextraction of Heavy Metals Assisted with Citric Acid

Ricinus communis L. is a bioenergetic crop with high-biomass production and tolerance to cadmium (Cd) and lead (Pb), thus, the plant is a candidate crop for phytoremediation. Pot experiments were performed to study the effects of citric acid in enhancing phytoextraction of Cd/Pb by Ricinus communis L. Citric acid increased Cd and Pb contents in plant shoots in all treatments by about 78% and 18–45%, respectively, at the dosage of 10 mM kg^?1 soil without affecting aboveground biomass production. Addition of citric acid reduced CEC, weakened soil adsorption of heavy metals and activated Cd and Pb in soil solutions. The acid-exchangeable fraction (BCR-1) of Pb remained lower than 7% and significantly increased with citric acid amendment. Respective increases in soil evaluation index induces by 14% and 19% under the Cd1Pb50 and Cd1Pb250 treatments upon addition of citric acid resulted in soil quality improvement. Ricinus communis L. has great potential in citric acid-assisted phytoextraction for Cd and Pb remediation.     

Silicate-Mediated Alleviation of Pb Toxicity in Banana Grown in Pb-Contaminated Soil

Silicate (Si) can enhance plant resistance or tolerance to the toxicity of heavy metals. However, it remains unclear whether Si can ameliorate lead (Pb) toxicity in banana (Musa xparadisiaca) roots. In this study, treatment with 800 mg kg⁻¹ Pb decreased both the shoot and root weight of banana seedlings. The amendment of 800 mg kg⁻¹ Si (sodium metasilicate, Na₂SiO₃·9H₂O) to the Pb-contaminated soil enhanced banana biomass at two growth stages significantly. The amendment of 800 mg kg⁻¹ Si significantly increased soil pH and decreased exchangeable Pb, thus reducing soil Pb availability, while Si addition of 100 mg kg⁻¹ did not influence soil pH. Results from Pb fractionation analysis indicated that more Pb were in the form of carbonate and residual-bound fractions in the Si-amended Pb-contaminated soils. The ratio of Pb-bound carbonate to the total Pb tended to increase with increasing growth stages. Treatment with 100 mg kg⁻¹ Si had smaller effects on Pb forms in the Si-amended soils than that of 800 mg kg⁻¹ Si. Pb treatment decreased the xylem sap greatly, but the addition of Si at both levels increased xylem sap and reduced Pb concentration in xylem sap significantly in the Si-amended Pb treatments. The addition of Si increased the activities of POD, SOD, and CAT in banana roots by 14.2% to 72.1% in the Si-amended Pb treatments. The results suggested that Si-enhanced tolerance to Pb toxicity in banana seedlings was associated with Pb immobilization in the soils, the decrease of Pb transport from roots to shoots, and Si-mediated detoxification of Pb in the plants.     

Ecology of bacterial endophytes associated with wetland plants growing in textile effluent for pollutant-degradation and plant growth-promotion potentials

In this study, 41 culturable endophytic bacteria were isolated from the roots and shoots of three wetland plants, Typha domingensis, Pistia stratiotes and Eichhornia crassipes, and identified by 16S rRNA gene sequencing. Textile effluent-degrading and plant growth-promoting activities of these endophytes were determined. The analysis of endophytic bacterial communities indicated that plant species had a pronounced effect on endophytic bacterial association and maximum endophytes (56.5%) were associated with T. domingensis. These endophytic bacteria mainly belonged to different species of the genera Bacillus (39%), Microbacterium (12%) and Halomonas (12%). Eight of the 41 strains showing maximum efficiency of textile effluent degradation also exhibited plant growth-promoting activities such as production of indole-3-acetic acid and siderophore, presence of 1-amino-cyclopropane-1-carboxylic acid deaminase, and solubilization of inorganic phosphorous. This is the first study describing the diversity and plant-beneficial characteristics of the textile effluent-degrading endophytic bacteria associated with wetland plants. T. domingensis showed better growth in textile effluent and also hosted maximum number of endophytic bacteria in roots and shoots. The interactions between T. domingensis and its associated endophytic bacteria could be exploited to enhance the efficiency of constructed wetlands during the remediation of industrial effluent.     

Enhanced Cd extraction of oilseed rape (Brassica napus) by plant growth-promoting bacteria isolated from Cd hyperaccumulator Sedum alfredii Hance

Four plant growth-promoting bacteria (PGPB) were used as study materials, among them two heavy metal-tolerant rhizosphere strains SrN1 (Arthrobacter sp.) and SrN9 (Bacillus altitudinis) were isolated from rhizosphere soil, while two endophytic strains SaN1 (Bacillus megaterium) and SaMR12 (Sphingomonas) were identified from roots of the cadmium (Cd)/zinc (Zn) hyperaccumulator Sedum alfredii Hance. A pot experiment was carried out to investigate the effects of these PGPB on plant growth and Cd accumulation of oilseed rape (Brassica napus) plants grown on aged Cd-spiked soil. The results showed that the four PGPB significantly boosted oilseed rape shoot biomass production, improved soil and plant analyzer development (SPAD) value, enhanced Cd uptake of plant and Cd translocation to the leaves. By fluorescent in situ hybridization (FISH) and green fluorescent protein (GFP), we demonstrated the studied S. alfredii endophytic bacterium SaMR12 were able to colonize successfully in the B. napus roots. However, all four PGPB could increase seed Cd accumulation. Due to its potential to enhance Cd uptake by the plant and to restrict Cd accumulation in the seeds, SaMR12 was selected as the most promising microbial partner of B. napus when setting up a plant–microbe fortified remediation system.     

Isolation of toxic metal-tolerant bacteria from soil and examination of their bioaugmentation potentiality by pot studies in cadmium- and lead-contaminated soil

Heavy metals, also regarded as toxic metals, are the important environmental pollutants that affect all forms of life. Accumulation of toxic metals in plants results in various biochemical, physiological and structural disturbances, leading to inhibited growth and sometimes plant death. Toxic metal contamination disturbs the soil ecology as well as the agricultural productivity. Several indigenous microbes can withstand the effect of toxic metal and play a vital role in the revival of tarnished soil. In the present study, soil samples were collected from contaminated crop field of Cachar district of Assam, India. Segregation, enumeration and identification of bacteria from soil samples were performed. Among all the tested isolates, very few were able to withstand a high concentration of Cd and Pb in nutrient agar plates. Toxic metal-tolerant bacteria were identified as Pseudomonas aeruginosa and Bacillus cereus. The isolates having a higher tolerance for Cd and Pb were taken into consideration for pot studies. P. aeruginosa strain SN4 and strain SN5 showed significant results at Cd- and Pb-contaminated soil, evidenced by the healthy growth of Oryza sativa seedlings. However, B. cereus strain SN6 showed high tolerance towards Cd and Pb, but pot experimental studies showed adverse effects on seedling germination and shoot growth of O. sativa. P. aeruginosa strains were significantly able to reduce the negative impact of Cd and Pb in the soil, thus finding an alternative in removal, recovery and remediation of toxic metal-contaminated crop field.     

The contribution of endophytic bacteria to Albizia lebbeck-mediated phytoremediation of tannery effluent contaminated soil

Toxicity of chromium often impairs the remediation capacity of plants used in phytoremediation of polluted soils. In this study, we have identified Albizia lebbeck as a prospective chromium hyperaccumulator and examined cultivable diversity of endophytes present in chromium-treated and control saplings. High numbers (22–100%) of endophytic bacteria, isolated from root, stem, and leaf tissues, could tolerate elevated (1–3 mM) concentrations of K₂CrO₇. 16S rRNA gene sequence-based phylogenetic analysis showed that the 118 isolates obtained comprised of 17 operational taxonomic units affiliated with the proteobacterial genera Rhizobium (18%), Marinomonas (1%), Pseudomonas (16%), and Xanthomonas (7%) but also with members of Firmicutes genera, such as Bacillus (35%) and Salinococcus (3%). The novel isolates belonging to Salinococcus and Bacillus could tolerate high K₂CrO₇ concentrations (3 mM) and also showed elevated activity of chromate reductase. In addition, majority (%) of the endophytic isolates also showed production of indole-3-acetic acid. Taken together, our results indicate that the innate endophytic bacterial community assists plants in reducing heavy metal toxicity.     

Assessing the Tolerance of Castor Bean to Cd and Pb for Phytoremediation Purposes

This study evaluated Cd and Pb accumulation by castor bean (Ricinus communis cv. Guarany) plants grown in nutrient solution, aiming to assess the plant’s ability and tolerance to grow in Cd- and Pb-contaminated solutions for phytoremediation purposes. The plants were grown in individual pots containing Hoagland and Arnon’s nutrient solution with increasing concentrations of Cd and Pb. The production of root and shoot dry matter and their contents of Cd, Pb, Ca, Mg, Cu, Fe, Mn, and Zn were evaluated in order to calculate the translocation and bioaccumulation factors, as well as toxicity of Cd and Pb. Cadmium caused severe symptoms of phytotoxicity in the plant’s root and shoot, but no adverse effect was observed for Pb. Castor bean is an appropriate plant to be used as indicator plant for Cd and tolerante for Pb in contaminated solution and it can be potentially used for phytoremediation of contaminated areas.     

Soil amendment application frequency contributes to phytoextraction of lead by sunflower at different nutrient levels

Phytoextraction has been proposed as an alternative remediation technology for soils polluted with heavy metals, but is generally perceived to be too slow. Enhancing accumulation of trace pollutants in harvestable plant tissues is a prerequisite for such technology to be practical. The main aims of this paper were to investigate whether a combination of nutrients and ethylenediaminetetraacetic acid (EDTA) enhanced Pb uptake of sunflower (Helianthus annuus) plants, and if timing of EDTA application altered Pb uptake and environmental persistence. Plants were grown in greenhouse pot experiments. Pb distributions and uptake of the whole plant were studied using chemical and flame atomic absorption spectrometry analyses. Pb mobilization by EDTA appeared to be dose dependent, with more mobilization for the high than the low dose. There were distinct differences in mobilization patterns of various nutrient amendments. EDTA mobilized Pb more in the medium than the highest and lowest nutrient levels. Heterogeneous soil humus components exerted mobilizing and stabilizing effects, so the medium nutrition was most effective for phytoextraction. At low nutrient levels, Pb concentration in the shoot with one low EDTA application was less than two applications to the same total EDTA dosage. So in the poor soil, two applications of EDTA was more effective than once. The half-life of two low EDTA treatment applications was longer than for one application, to the same total dosage. In general, sunflower was suited to phytoremediation of moderately Pb-contaminated soil by phytoextraction.     

茶树叶际选择性富集的内生细菌的鉴定

【目的】茶树(Camelliasinensis)为多年生常绿木本植物,其叶片用于生产茶叶。本论文通过测定茶树叶际内生细菌的菌群组成,比较叶际内生菌与土壤菌群的异同,以鉴定选择性富集于茶树叶际的内生细菌。【方法】本研究采集湖北宜昌邓村茶树,对茶叶表面进行除菌处理,提取茶叶及其内生细菌的总基因组DNA,通过细菌16S核糖体RNA基因(16S rDNA)保守区引物799F和1193R扩增16Sr DNA的V5–V7可变区序列,并通过Illumina二代测序平台对扩增子进行建库测序。【结果】茶叶内生菌群由变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)和极少量的梭杆菌门(Fusobacteria)5个门,共百余属组成,其中,甲基杆菌属(Methylobacterium)、代尔夫特菌属(Delftia)、微杆菌属(Microbacterium)、红球菌属(Rhodococcus)、Aureimonas和鞘氨醇单胞菌属(Sphingomonas)等以较高丰度富集于叶片组织内部。约40%的茶叶叶际内生细菌也在相应的土壤中存在,表明其可能的土壤来源;非土壤来源的茶叶内生细菌达60%,如Aureimonas和Delftia等。【结论】本研究解析了茶树叶际内生细菌的群落组成与结构,分析了内生细菌的可能来源,为以叶际内生菌为靶标,改善茶叶品质、降解农药残留、防御茶叶病虫害的研究提供基础。