Cd-hyperaccumulator Solanum Nigrum Research Articles

Effect of ammonium sulfate combined with aqueous bio-chelator on Cd uptake by Cd-hyperaccumulator Solanum nigrum L.

The efficacy of using plants to phytoremediate heavy metal (HM) contaminated soils can be improved using soil amendments. These amendments may both increase plant biomasses and HMs uptake. We aimed to determine the composite effect of ammonium sulfate ((NH4)2SO4) combined with the application of an aqueous stem-extracted bio-chelator (Bidens tripartita L) on the plant biomasses and cadmium (Cd) phytoextraction by Solanum nigrum L. The constant (NH4)2SO4 application mode plus bio-chelator additives collectively enhanced the shoot Cd extraction ability owing to the increased plant biomass and shoot Cd concentration by S. nigrum. The shoot Cd extraction and the soil Cd decreased concentration confirmed the optimal Cd phytoextraction pattern in K8 and K9 treatments (co-application of (NH4)2SO4 and twofold/threefold bio-chelators). Accordingly, Cd contamination risk in the soil (2 mg kg−1) could be completely eradicated (<0.2 mg kg−1) after three rounds of phytoremediation by S.nigrum based on K8 and K9 treatments through calculating soil Cd depletion. The microorganism counts and enzyme activities in rhizosphere soils at treatments with the combined soil additives apparently advanced. In general, co-application mode of (NH4)2SO4 and aqueous bio-chelator was likely to be a perfect substitute for conventional scavenger agents on account of its environmental friendliness and cost saving for field Cd contamination phytoremediation by S. nigrum.

Plant growth promoting endophyte promotes cadmium accumulation in Solanum nigrum L. by regulating plant homeostasis

The homeostasis regulating mechanism of endophyte enhancing cadmium (Cd) extraction by hyperaccumulator is poorly understood. Here, an endophyte strain E3 that belonged to Pseudomonas was screened from Cd hyperaccumulator Solanum nigrum L., which significantly improved the Cd phytoextraction efficiency of S. nigrum by 40.26%. The content and translocation factor of nutrient elements indicated that endophyte might regulate Cd accumulation by affecting the uptake and transport of magnesium and iron in S. nigrum. Gene transcriptional expression profile further revealed that SnMGT, SnIRT1, and SnIRT2, etc., were the key genes involved in the regulation of S. nigrum elements uptake by endophyte. However, changes in elemental homeostasis did not negatively affect plant growth. Endophyte inoculation promoted plant growth by fortifying photosynthesis as well as recruiting specific bacteria in S. nigrum endosphere, e.g., Pseudonocardiaceae, Halomonas. Notably, PICRUSt2 analysis and biochemical characterization jointly suggested that endophyte regulated starch degradation in S. nigrum leaves to maintain photosynthetic balance. Our results demonstrated that microecological characteristics of hyperaccumulator could be reshaped by endophyte, also the homeostasis regulation in endophyte enhanced hyperaccumulator Cd phytoextraction was significant.

Proteomic analysis reveals differential responsive mechanisms in Solanum nigrum exposed to low and high dose of cadmium

Cadmium (Cd) contamination is becoming a widespread environmental problem. However, the differential responsive mechanisms of Cd hyperaccumulator Solanum nigrum to low or high dose of Cd are not well documented. In this study, phenotypic and physiological analysis firstly suggested that the seedlings of S. nigrum showed slight leaf chlorosis symptoms under 25 μM Cd and severe inhibition on growth and photosynthesis under 100 μM Cd. Further proteomic analysis identified 105 differentially expressed proteins (DEPs) in the Cd-treated leaves. Under low dose of Cd stress, 47 DEPs are mainly involved in primary metabolic processes, while under high dose of Cd stress, 92 DEPs are mainly involved in photosynthesis, energy metabolism, production of phytochelatin and reactive oxygen species (ROS). Protein-protein interaction (PPI) network analysis of DEPs support above differential responses in the leaves of S. nigrum to low and high dose of Cd treatments. This work provides the differential responsive mechanisms in S. nigrum to low and high dose of Cd, and the theoretical foundation for the application of hyperaccumulating plants in the phytoremediation of Cd-contaminated soils.

Attenuating Effect of an Extract of Cd-Hyperaccumulator Solanum nigrum on the Growth and Physio-chemical Changes of Datura innoxia Under Cd Stress

PurposeThe use of plant extracts obtained from plants that are highly tolerant to heavy metal toxicity has been beneficial in improving the growth of plants grown under metal toxicity conditions. A lab experiment was performed to elucidate the alleviating role of foliar applied cadmium (Cd)-hyperaccumulator Solanum nigrum (S. nigrum) extract on Datura innoxia (D. innoxia) plants grown under Cd stress (0, 50, and 100 mg Cd kg-1 soil).MethodsGrowth parameters, photosynthetic pigment content, osmo-metabolic compounds, reduced glutathione and phytochelatins content, oxidative damage, and lignin content and its related enzyme (cell wall-bound peroxidase, POX) were determined.ResultsApart from the foliar application response of S. nigrum leaf extract (SNE) in either Cd exposed or non-Cd exposed plants, growth parameters of D. innoxia plant grown under both Cd concentrations (50 and 100 mg Cd kg-1 soil) in terms of root and shoot fresh, dry weight, length, and leaf area were noticeably diminished by 29 and 51%, 29 and 54%, 35 and 70%, 40 and 53%, 30 and 69%, 40 and 60%, and 11 and 23%, respectively, compared with untreated control plants. Foliar delivered SNE secured the photosynthetic pigment, free amino acids, soluble proteins, and soluble sugar content. Additionally, it lessened the adverse effects of Cd stress on D. innoxia plants by curtailing the content of hydrogen peroxide (H2O2) and malondialdehyde (MDA) by 28 and 27%, and 21 and 23%, respectively, compared with the plants subjected to 50 and 100 mg Cd kg-1 only. The findings herein indicated that the plant water extract and their interactions in the investigated Cd rates significantly augmented phenolics, alkaloids, reduced glutathione and phytochelatins content. Cell wall stiffening in D. innoxia indicated that lignin content and POX were significantly higher in plants exposed to 100 mg Cd kg-1 soil displaying increase values of 275 and 300%, respectively, against non-Cd treated control. The magnitude of increment imposed by Cd stress was lessened by using SNE that reflects on adequate cell growth advocated by limited lignification, in terms of lignin content, and downregulated POX activity. Owing to SNE application, root and leaves Cd contents were efficiently reduced reflecting apparent plant liveliness compared with the SNE non-treated Cd-stressed plants.ConclusionsThe outcomes of this study designate that foliar application of the Cd-hyperaccumulator S. nigrum leaf extract can be counted as an unconventional and innovative approach in the alleviation of Cd stress and can be employed as integrated practice when Cd-contaminated regions were exploited for sustainable agriculture of the multipurpose plants.

Cosmopolitan cadmium hyperaccumulator Solanum nigrum: Exploring cadmium uptake, transport and physiological mechanisms of accumulation in different ecotypes as a way of enhancing its hyperaccumulative capacity

The non-essential element cadmium (Cd) is one of the most problematic priority soil pollutants due to multitude of pollution sources, mobility in the environment and high toxicity to all living organisms. This strongly limits also the number and occurrence of species – Cd hyperaccumulators to be used for soil phytoremediation. However, efficient Cd hyperaccumulator Solanum nigrum L. appeared to commonly occur worldwide as a representative of Solanum nigrum complex of a great taxonomic diversity. This led to the idea that the search among different ecotypes of Solanum nigrum L. may result in the identifying the most efficient Cd hyperaccumulator without applying to soil any additional measures such as chemical ligands. In this first pioneering comparative study, three randomly selected ecotypes of S. nigrum L. ssp. nigrum from Shenyang (SY) and Hanzhong (HZ) in China, and Kyoto (KY) in Japan were used in pot experiments at soil treatments from 0 to 50 mg Cd kg−1. The Cd accumulation capacity appeared to represent KY > HZ > SY range, KY ecotype accumulating up to 73%, and HZ ecotype up to 67% bigger total Cd load than SY ecotype. At Cd content in soil up to 10 mg kg−1, no significant effect on the all ecotype biomass, photosynthetic activities, contents of first line defense antioxidant enzymes (CAT, SOD, GPX), and scavenging antioxidants ASA, GSH, was observed. At Cd in soil>10 mg kg−1all these parameters showed decreasing, and cell damage indicator MDA increasing trend, however total accumulated Cd load further increased up to 30 mg kg Cd in soil in all ecotypes in the same KY > HZ > SY sequence. The study proved the great potential of enhancing Cd accumulation capacity of S. nigrum species by selecting the most efficient ecotypes among commonly occurring representatives of S. nigrum complex worldwide. Moreover, these first comparative experiments convinced that the cosmopolitan character and great variety of species/subspecies belonging to Solanum nigrum complex all over the world opens the new area for successful soil phytoremediation with the use of the most appropriate eco/genotypes of S. nigtum as a tool for the best Cd-contaminated soil management practice.

Comparative study on different organic acids for promoting Solanum nigrum L. hyperaccumulation of Cd and Pb from the contaminated soil

Organic acids-assisted phytoremediation is a promising strategy to remove metal pollutants from the soil. However, few reports have focused on the mechanisms of organic acids promoting the uptake of heavy metals by hyperaccumulators. In this study, 5 types of organic acids, namely polybasic carboxylic acids, acidic amino acids, acidic plant growth regulators, phosphoric and gluconic acids, were comprehensively investigated the effects on the solubility of Cd and Pb in the soil along with their uptake by Cd hyperaccumulator Solanum nigrum L. The results indicated that the addition of Hydroxyethylidene-1,1-diphosphonic acid (HEDP) and d-Gluconic acid (D-GA) effectively extracted the most of acid-extractable and some of reducible and oxidizable fractions of Cd and Pb in the soil, with the extraction rates of 64.8% and 34.4% for total Cd and 53.6% and 30.0% for total Pb, respectively. HEDP and D-GA significantly increased the accumulations of Cd (57.1% and 35.0%) and Pb (43.4% and 31.9%) by S. nigrum without the inhibition of its biomass, making the great removal efficiencies of Cd (1.35% and 1.16%) and Pb (0.039% and 0.036%) from the soil. The enhanced phytoremediation efficiency of S. nigrum was due to the increase of the extractable Cd and Pb in the rhizosphere but little changes of soil pH and enzyme activities (catalase and urease). Among all of organic acids, HEDP may be an alternative to EDTA because of its characteristics of environmental friendliness and high efficiency.

X-ray absorption spectroscopy evidence of sulfur-bound cadmium in the Cd-hyperaccumulator Solanum nigrum and the non-accumulator Solanum melongena.

It has been proposed that non-protein thiols and organic acids play a major role in cadmium phytoavailability and distribution in plants. In the Cd-hyperaccumulator Solanum nigrum and non-accumulator Solanum melongena, the role of these organic ligands in the accumulation and detoxification mechanisms of Cd are debated. In this study, we used X-ray absorption spectroscopy to investigate Cd speciation in these plants (roots, stem, leaves) and in the soils used for their culture to unravel the plants responses to Cd exposure. The results show that Cd in the 100mgkg-1 Cd-doped clayey loam soil is sorbed onto iron oxyhydroxides. In both S.nigrum and S.melongena, Cd in roots and fresh leaves is mainly bound to thiol ligands, with a small contribution of inorganic S ligands in S.nigrum leaves. We interpret the Cd binding to sulfur ligands as detoxification mechanisms, possibly involving the sequestration of Cd complexed with glutathione or phytochelatins in the plant vacuoles. In the stems, results show an increase binding of Cd to -O ligands (>50% for S.nigrum). We suggest that Cd is partly complexed by organic acids for transportation in the sap.

Small structures with big impact: Multi-walled carbon nanotubes enhanced remediation efficiency in hyperaccumulator Solanum nigrum L. under cadmium and arsenic stress

With the fast development of nanotechnology, nanomaterials are being increasingly applied for the remediation of contaminated soils. However, few researches have been reported on the complex interactions of carbon nanotubes with heavy metal (loid)s in phytoremediation. Here, we conduct a pot experiment to investigate the effects of multi-walled carbon nanotubes (MWCNTs) on the plant growth and behavior of heavy metal (loid)s in hyperaccumulator-soil system. Cd hyperaccumulator Solanum nigrum L. (S. nigrum) were cultivated in Cadmium (Cd) and Arsenic (As) contaminated soils amended with MWCNTs at 100, 500, and 1000 mg kg−1 for 60 days, respectively. The application of MWCNTs increased the shoot length and plant dry biomass by 5.56%∼25.13% and 5.23%∼27.97%. Whereas, root and leaf growth were inhibited in 1000 mg kg−1 MWCNTs treatments. Meanwhile, MWCNTs at 500 mg kg−1 significantly enhanced the accumulation of heavy metal (loid)s in S. nigrum（18.29% for Cd and 32.47% for As）and alleviated co-contamination induced toxicity, by motivating plant growth, stimulating antioxidant enzymatic activities, and increasing micronutrient content (p < 0.05). The bio-concentration factor of As was decreased (15.31–28.08%) under MWCNTs application, which plays an important role in the alleviation of phytotoxicity. Besides, bioavailable Cd and As were reduced in rhizosphere soils, and the most significant reduction (16.29% for Cd and 8.19% for As) were shown in 500 mg kg−1 MWCNTs treatment. These findings demonstrate that suitable concentration of MWCNTs can enhance remediation efficiency. Our study gives a strong evidence to promote the phytoremediation for co-contaminated soils by using nanomaterials.

Phytoremediation of cadmium-polluted soil assisted by D-gluconate-enhanced Enterobacter cloacae colonization in the Solanum nigrum L. rhizosphere

Microbe-assisted phytoremediation for Cd-polluted soil is being regarded increasingly. However, the availability of microbes that can collaborate with Cd-hyperaccumulators effectively has become one of bottlenecks restricting the remediation efficiency. A siderophore-producing bacterium (Y16; Enterobacter cloacae) isolated from the rhizospheric soil of Cd-hyperaccumulator Solanum nigrum L. was identified by 16S rRNA gene sequencing and biochemical analysis, and then used for analyzing microbial chemotaxis, carbon source utilization, and insoluble P/Cd mobilization capacities. Besides, a soil-pot trial was performed to underlie the phytoremediation mechanism of Cd-polluted soil assisted by D-gluconate-enhanced Enterobacter cloacae colonization (DEYC) in the Solanum nigrum L. rhizosphere. Results displayed that D-gluconate was an effective chemoattractant and carbon source strengthening Y16 colonization, and Y16 exhibited strong abilities to mobilize insoluble P/Cd in shake flask by extracellular acidification (p < 0.05). In the soil-pot trial, DEYC observably enhanced soil Cd phytoextraction by Solanum nigrum L., and increased microbial diversity according to alpha- and beta-diversity analysis (p < 0.05). Taxonomic distribution and co-occurrence network analysis suggested that DEYC increased relative abundances of dominant microbial taxa associated with soil acidification (Acidobacteria-6), indoleacetic acid secretion (Ensifer adhaerens), soil fertility improvement (Flavisolibacter, Bdellovibrio bacteriovorus, and Candidatus nitrososphaera), and insoluble Cd mobilization (Massilia timonae) at different classification levels. Importantly, COGs analysis further shown that DEYC aroused the up-regulation of key genes related to chemotactic motility, carbon fixation, TCA cycle, and propanoate metabolism. These results indicated that DEYC drove the rhizospheric enrichment of pivotal microbial taxa directly or indirectly involved in soil Cd mobilization, meanwhile distinctly promoted plant growth for accumulating more mobilizable Cd. Therefore, Y16 could be used as bio-inoculants for assisting phytoremediation of Cd-polluted soil.

Indole-3-acetic acid promotes cadmium (Cd) accumulation in a Cd hyperaccumulator and a non-hyperaccumulator by different physiological responses

To study the effects of indole-3-acetic acid (IAA) on cadmium (Cd) accumulation and the physiological responses of the Cd hyperaccumulator Solanum nigrum and non-hyperaccumulator Solanum melongena, a pot experiment was conducted in soil containing 2mgkg-1 Cd in which different concentrations of IAA (0, 10, 20, or 40mgL-1) were sprayed on plant leaves. The results showed that Cd accumulation in shoots of S. nigrum was significantly increased by 30% after the addition of 10mgL-1 IAA under 2mgkg-1 Cd stress compared to that in the control, but shoot Cd accumulation showed no significant change in S. melongena after this IAA treatment. Additionally, the growth and the proline content in the two species were significantly increased by 20mgL-1 IAA. The activities of peroxidase and catalase in leaves of S. nigrum and the activity of superoxide dismutase (SOD) in S. melongena were significantly increased and their malondialdehyde content was significantly decreased compared to those in the control. The root activity of S. nigrum was significantly improved after 10 and 20mgL-1 IAA treatments, but no significant difference was observed in S. melongena. The correlation analysis results showed that the Cd concentration in leaves of S. nigrum was significantly and positively correlated with the carotenoid and proline contents, and there was also a significant positive correlation between the Cd concentration and SOD activity in leaves of S. melongena. Therefore, S. nigrum is an ideal plant for the phytoextraction of Cd-contaminated soil assisted by IAA. IAA promotes Cd accumulation in plant shoots by enhancing the accumulation of carotenoids and proline in S. nigrum and maintaining a high leaf SOD activity in S. melongena.

Complete genome analysis of Serratia marcescens RSC-14: A plant growth-promoting bacterium that alleviates cadmium stress in host plants.

Serratia marcescens RSC-14 is a Gram-negative bacterium that was previously isolated from the surface-sterilized roots of the Cd-hyperaccumulator Solanum nigrum. The strain stimulates plant growth and alleviates Cd stress in host plants. To investigate the genetic basis for these traits, the complete genome of RSC-14 was obtained by single-molecule real-time sequencing. The genome of S. marcescens RSC-14 comprised a 5.12-Mbp-long circular chromosome containing 4,593 predicted protein-coding genes, 22 rRNA genes, 88 tRNA genes, and 41 pseudogenes. It contained genes with potential functions in plant growth promotion, including genes involved in indole-3-acetic acid (IAA) biosynthesis, acetoin synthesis, and phosphate solubilization. Moreover, annotation using NCBI and Rapid Annotation using Subsystem Technology identified several genes that encode antioxidant enzymes as well as genes involved in antioxidant production, supporting the observed resistance towards heavy metals, such as Cd. The presence of IAA pathway-related genes and oxidative stress-responsive enzyme genes may explain the plant growth-promoting potential and Cd tolerance, respectively. This is the first report of a complete genome sequence of Cd-tolerant S. marcescens and its plant growth promotion pathway. The whole-genome analysis of this strain clarified the genetic basis underlying its phenotypic and biochemical characteristics, underpinning the beneficial interactions between RSC-14 and plants.

Host plant growth promotion and cadmium detoxification in Solanum nigrum, mediated by endophytic fungi

Current investigation conducted to evaluate the associated fungal endophyte interactions of a Cd hyper-accumulator Solanum nigrum Korean ecotype under varying concentrations of Cd. Two indole-3-acetic acid (IAA) producing fungal strains, RSF-4L and RSF-6L, isolated from the leaves of S. nigrum, were initially screened for Cd tolerance and accumulation potential. In terms of dry biomass production, the strain RSF-6L showed higher tolerance and accumulation capacity for Cd toxicity in comparison to RSF-4L. Therefore, RSF-6L was applied in vivo to S. nigrum and grown for six weeks under Cd concentrations of 0, 10, and 30mgKg−1 of dry sand. The effect of fungal inoculation assessed by plant physiological responses, endogenous biochemical regulations, and Cd profile in different tissues. Significant increase were observed in plant growth attributes such as shoot length, root length, dry biomass, leaf area, and chlorophyll contents in inoculated RSF-6L plants in comparison to non-inoculated plants with or without Cd contamination. RSF-6L inoculation decreased uptake of Cd in roots and above ground parts, as evidenced by a low bio-concentration factor (BCF) and improved tolerance index (TI). However, Cd concentration in the leaves remained the same for inoculated and non-inoculated plants under Cd spiking. Fungal inoculation protected the host plants, as evidenced by low peroxidase (POD) and polyphenol peroxidase (PPO) activities and high catalase (CAT) activity. Application of appropriate fungal inoculation that can improve tolerance mechanisms of hyper-accumulators and reduce Cd uptake can be recommended for phyto-stabilisation/immobilisation of heavy metals in crop fields.

Fractionation of Stable Cadmium Isotopes in the Cadmium Tolerant Ricinus communis and Hyperaccumulator Solanum nigrum.

Cadmium (Cd) isotopes provide new insights into Cd uptake, transport and storage mechanisms in plants. Therefore, the present study adopted the Cd-tolerant Ricinus communis and Cd-hyperaccumulator Solanum nigrum, which were cultured under controlled conditions in a nutrient solution with variable Cd supply, to test the isotopic fractionation of Cd during plant uptake. The Cd isotope compositions of nutrient solutions and organs of the plants were measured by multiple collector inductively coupled plasma mass spectrometry (MC-ICPMS). The mass balance of Cd isotope yields isotope fractionations between plant and Cd source (δ114/110Cdorgans-solution) of −0.70‰ to −0.22‰ in Ricinus communis and −0.51‰ to −0.33‰ in Solanum nigrum. Moreover, Cd isotope fractionation during Cd transport from stem to leaf differs between the Cd-tolerant and -hyperaccumulator species. Based on these results, the processes (diffusion, adsorption, uptake or complexation), which may induce Cd isotope fractionation in plants, have been discussed. Overall, the present study indicates potential applications of Cd isotopes for investigating plant physiology.

Effect of Funneliformis mosseae on the growth, cadmium accumulation and antioxidant activities of Solanum nigrum

Arbuscular mycorrhizal (AM) fungi can promote plant growth and resistance to heavy metal (HM), affect HM bioavailability and uptake by plants, and thus have been considered as the most prominent symbiotic fungi for contribution to phytoremediation. A greenhouse pot experiment was performed to assess the effects of Funneliformis mosseae BGC XJ02 (Fm) on the plant growth, P acquisition, Cd accumulation, phytochelatins (PCs) production and antioxidant activities of Cd-hyperaccumulator Solanum nigrum in different Cd-added soils (0, 5, 10, 20 and 40mgCdkg−1soil). Fm associated with S. nigrum very well, and mycorrhizal colonizations were not influenced by Cd addition. Fm inoculation significantly increased P acquisition, plant growth, Cd concentration and Cd uptake in both shoots and roots of S. nigrum at all Cd levels. The improvement of catalase (CAT), ascorbate peroxidase (APX) and peroxidase (POD) activities in the leaves of mycorrhizal plants suggested that Fm colonization helped S. nigrum to relieve oxidative damage to biomolecules in Cd-contaminated soil. Moreover, glutathione reductase (GR) activities in the leaves of mycorrhizal plants were elevated (from 17% to 99%) in lower Cd levels (0, 5 and 10mgCdkg−1 soil), but reduced in higher Cd levels (20 and 40mgCdkg−1 soil). The obvious decrease of malonaldehyde (MDA) content in the leaves of mycorrhizal plants displayed that Fm symbiosis significantly enhanced antioxidant activities. In conclusion, Fm symbiosis alleviated Cd phytotoxicity to S. nigrum through the improvement of P acquisition and activities of CAT, POD, APX in mycorrhizal plants, and then improved plant growth and phytoremediation efficiency for Cd-contaminated soil.

Methyl jasmonate alleviates cadmium toxicity in Solanum nigrum by regulating metal uptake and antioxidative capacity

The growth of the Cd-hyperaccumulator Solanum nigrum L. and its physiological responses to a short-term (7 d) Cd stress and to exogenous methyl jasmonate (MeJA) were investigated. Compared with the leaves of S. nigrum, the roots were more liable to Cd and showed a significantly decreased dry mass and increased malondialdehyde content. Cd accumulation in the shoots and roots of S. nigrum were proportional to the Cd concentration in the hydroponic solution. The application of a low concentration of MeJA (0.01 μM) significantly reduced the translocation/accumulation of Cd in both the shoots and roots compared with a 40 mg dm−3 Cd treatment only. Moreover, 40 mg dm−3 Cd significantly decreased the activity of leaf superoxide dismutase, but 0.01 μM MeJA restored it. MeJA also enhanced the activity of catalase in the leaves but showed no significant effect on peroxidase activity. The content of both endogenous jasmonic acid (JA) and MeJA in the leaves of S. nigrum increased with the increase of exogenous MeJA concentration.

Enhancement of arbuscular mycorrhizal fungus (Glomus versiforme) on the growth and Cd uptake by Cd-hyperaccumulator Solanum nigrum

Arbuscular mycorrhizal fungus (AMF) can enhance plant growth and resistance to toxicity produced by heavy metals (HMs), affect the bioavailability of HMs in soil and the uptake of HMs by plants, and thus has been emerged as the most prominent symbiotic fungus for contribution to phytoremediation. A greenhouse pot experiment was conducted to assess the effect of Glomus versiforme BGC GD01C (Gv) on the growth and Cd accumulation of Cd-hyperaccumulator Solanum nigrum in different Cd-added soils (0, 25, 50, 100mgCdkg−1 soil). Mycorrhizal colonization rates were generally high (from 71% to 82%) in Gv-inoculated treatments at all Cd levels. Gv colonization enhanced soil acid phosphatase activity, and hence elevated P acquisition and growth of S. nigrum at all Cd levels. Moreover, the presence of Gv significantly increased DTPA-extractable (phytoavailable) Cd concentrations in 25 and 50mgCdkg−1 soils, but did not affect phytoavailable Cd in 100mgCdkg−1 soil. Similarly, inoculation with Gv significantly increased Cd concentrations of S. nigrum in 25 and 50mgCdkg−1 soils, but decreased Cd concentrations of the plants in 100mgCdkg−1 soil. Overall, inoculation with Gv greatly improved the total Cd uptakes in all plant tissues at all Cd levels. The present results indicated that S. nigrum associated with Gv effectively improved the Cd uptake by plant and would be a new strategy in microbe-assisted phytoremediation for Cd-contaminated soils.

Interaction of Cd-hyperaccumulator Solanum nigrum L. and functional endophyte Pseudomonas sp. Lk9 on soil heavy metals uptake

Bioaugmentation is a promising method for assisting phytoextraction of heavy metals from contaminated soil, and the development of bioaugmentation-assisted phytoextraction requires the understanding of the mechanism involved in the interaction between plants and inocula. In this study, a pot study was conducted to evaluate the effect of bacterial endophyte Pseudomonas sp. Lk9 which can produce biosurfactants, siderophores and organic acids on the growth and metal uptake of Cd-hyperaccumulator Solanum nigrum L. growing in multi-metal-contaminated soil. The results revealed that Lk9 inoculation could improve soil Fe and P mineral nutrition supplies, enhance soil heavy metal availability, and affect host-mediated low-molecular-weight organic acids secretion, thereby significantly increasing S. nigrum shoot dry biomass by 14% and the total of Cd by 46.6%, Zn by 16.4% and Cu by 16.0% accumulated in aerial parts, compared to those of non-inoculated control. The assessment of phytoextraction showed that Lk9 inoculation elevated the bioaccumulation factor of Cd (28.9%) and phytoextraction rates of all metals (17.4%, 48.6% and 104.6% for Cd, Zn and Cu, respectively), while the translocation factors had negligible difference between Lk9 inoculation (3.30, 0.50 and 0.40 for Cd, Zn and Cu, respectively) and non-inoculated control (2.95, 0.53 and 0.42 for Cd, Zn and Cu, respectively). It was also found that the symbiotic association between S. nigrum and Lk9 significantly increased the soil microbial biomass C by 39.2% and acid phosphatase activity by 28.6% compared to those in S. nigrum without Lk9. This study would provide a new insight into the bioaugmentation-assisted phytoextraction of heavy metal-contaminated soils.

Diversity of endophytic bacterial populations associated with Cd-hyperaccumulator plant Solanum nigrum L. grown in mine tailings

Despite the broad application of molecular approaches for the analysis of bacterial communities in a wide range of natural habitats, little information is available on the diversity of bacterial endophytes in the hyperaccumulators growing in heavy-metal contaminated sites. In this study, the diversity of bacterial endophytes associated with Cd-hyperaccumulator Solanum nigrum L. growing in mine tailing was analyzed by 16S rDNA clone library technique. In total, 197 individual sequences comprising 40 different Operational Taxonomic Units (OTUs) were used to assess the diversity of bacterial endophytes. Phylogenetic analysis revealed that the majority of clones were affiliated with Proteobacteria (61.93%; 19 OTUs), which included α (41.12%; 11 OTUs), β (6.09%; 4 OTUs) and γ (14.72%; 4 OTUs) subclasses. Other clones belonged to Bacteroidetes (12.18%; 5 OTUs), Firmicutes (2.03%; 2 OTUs), Actinobacteria (18.78%; 10 OTUs) and uncultured bacteria (5.08%; 4 OTUs). The dominant genera were Sphingomonas and Pseudomonas, which accounted for 24.37% and 12.18% of total clones respectively. These findings suggested that the diversity of endophytic bacterial populations was abundant in S. nigrum L., and these bacterial endophytes should be targeted for future research to determine their functional role, if any, in heavy metal stress reduction.

Analysis and characterization of cultivable heavy metal-resistant bacterial endophytes isolated from Cd-hyperaccumulator Solanum nigrum L. and their potential use for phytoremediation

This study investigates the heavy metal-resistant bacterial endophytes of Cd-hyperaccumulator Solanum nigrum L. grown on a mine tailing by using cultivation-dependent technique. Thirty Cd-tolerant bacterial endophytes were isolated from roots, stems, and leaves of S. nigrum L. and classified by amplified ribosomal DNA-restriction analysis into 18 different types. Phylogenetic analysis based on 16S rDNA sequences showed that these isolates belonged to four groups: Actinobacteria (43%), Proteobacteria (23%), Bacteroidetes (27%) and Firmicutes (7%). All the isolates were then characterized for their plant growth promoting traits as well as their resistances to different heavy metals; and the actual plant growth promotion and colonization ability were also assessed. Four isolates were re-introduced into S. nigrum L. under Cd stress and resulted in Cd phytotoxicity decrease, as dry weights of roots increased from 55% to 143% and dry weights of above-ground from 64% to 100% compared to the uninoculated ones. The total Cd accumulation of inoculated plants increased from 66% to 135% (roots) and from 22% to 64% (above-ground) compared to the uninoculated ones. Our research suggests that bacterial endophytes are a most promising resource and may be the excellent candidates of bio-inoculants for enhancing the phytoremediation efficiency.

Involvement of auxin and nitric oxide in plant Cd-stress responses

Cadmium (Cd) toxicity inhibited the seedling growth while inducing the occurrences of lateral roots (LR) and adventitious roots (AR). Further study indicated that auxin and nitric oxide (NO) are involved in the processes. In this study, we chose model plant Arabidopsis thaliana and Cd-hyperaccumulator Solanum nigrum as material to examine the involvement of Cd-induced auxin redistribution in NO accumulation in plants and the effect of NO on Cd accumulation. For this aim, the histochemical staining, NO fluorescence probe (DAF-2DA) detections combined with the pharmacological study were used in this study. By using DR5:GUS staining analysis combined with NO fluorescence probe (DAF-2DA) detection, we found that Cd-induced NO accumulation is at least partly due to auxin redistribution in plants exposure to Cd. Supplementation with SNP donor S-nitrosoglutathione (GSNO) increased the number of LR and AR. In contrast, NO-scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethyl imidazoline-1-oxyl-3-oxide (cPTIO) reversed the effects of NO on modulating root system architecture and Cd accumulation. These results suggest that manipulation of the NO level is an effective approach to improve Cd tolerance in plants by modulating the development of LR and AR, and provide insights into novel strategies for phytoremediation.