A Hybrid Approach Enabling Large-Scale Glycomic Analysis of Post-Golgi Vesicles Reveals a Transport Route for Polysaccharides.

The plant endomembrane system facilitates the transport of polysaccharides, associated enzymes, and glycoproteins through its dynamic pathways. Although enzymes involved in cell wall biosynthesis have been identified, little is known about the endomembrane-based transport of glycan components. This is partially attributed to technical challenges in biochemically determining polysaccharide cargo in specific vesicles. Here, we introduce a hybrid approach addressing this limitation. By combining vesicle isolation with a large-scale carbohydrate antibody arraying technique, we charted an initial large-scale map describing the glycome profile of the SYNTAXIN OF PLANTS61 (SYP61) trans-Golgi network compartment in Arabidopsis (Arabidopsis thaliana). A library of antibodies recognizing specific noncellulosic carbohydrate epitopes allowed us to identify a range of diverse glycans, including pectins, xyloglucans (XyGs), and arabinogalactan proteins in isolated vesicles. Changes in XyG- and pectin-specific epitopes in the cell wall of an Arabidopsis SYP61 mutant corroborate our findings. Our data provide evidence that SYP61 vesicles are involved in the transport and deposition of structural polysaccharides and glycoproteins. Adaptation of our methodology can enable studies characterizing the glycome profiles of various vesicle populations in plant and animal systems and their respective roles in glycan transport defined by subcellular markers, developmental stages, or environmental stimuli.

pH dependence of arsenic speciation in paddy soils: The role of distinct methanotrophs.

Arsenic (As) is a priority environmental pollutant in paddy field. The coupling of arsenate (As(V)) reduction with anaerobic methane (CH4) oxidation was recently demonstrated in paddy soils and has been suggested to serve as a critical driver for As transformation and mobilization. However, whether As(V)-dependent CH4 oxidation is driven by distinct methanotrophs under different pH conditions remains unclear. Here, we investigated the response of As(V)-dependent CH4 oxidation to pH shifts (pH 5.5-8.0) by employing isotopically labelled CH4. Furthermore, the underlying mechanisms were also investigated in well-controlled anoxic soil suspension incubations. Our results showed that As(V)-dependent CH4 oxidation is highly sensitive to pH changes (1.6-6.8 times variation of arsenite formation). A short-term (0-10 d) pH shift from near-neutral pH to acidic conditions (i.e., pH 5.5, -85% arsenite formation) had an inhibitory effect on As(V)-dependent CH4 oxidation. However, prolonged acidic conditions (i.e., >15 d) had no significant influence on As(V)-dependent CH4 oxidation. The microbial analyses indicated that As reduction in paddies can be driven by anaerobic CH4 oxidation archaea (ANME) and methanotrophs. And, methanotrophs may serve as a critical driver for As(V)-dependent CH4 oxidation. Moreover, type I methanotrophs Methylobacter were more active in oxidizing CH4 than type II methanotrophs Methylocystis when the pH ≥ 6.5. However, Methylocystis had a higher tolerance to soil acidification than Methylobacter. This study illustrates that As(V)-dependent CH4 oxidation could be dominated by distinct methanotrophs along with pH shifts, which eventually enhances As release in paddy soils.

Influence of elemental sulfur on cadmium bioavailability, microbial community in paddy soil and Cd accumulation in rice plants.

Cadmium (Cd) is highly toxic to living organisms and the contamination of Cd in paddy soil in China has received much attention. In the present study, by conducting pot experiment, the influence of S fertilizer (S0) on rice growth, iron plaque formation, Cd accumulation in rice plants and bacterial community in rice rhizosphere soil was investigated. The biomass of rice plants was significantly increased by S0 addition (19.5-73.6%). The addition of S0 increased the formation of iron plaque by 24.3-45.8%, meanwhile the amount of Cd sequestered on iron plaque increased. In soil treated with 5 mg/kg Cd, addition of 0.2 g/kg S0 decreased the diffusive gradients in thin films (DGT) extractable Cd by 60.0%. The application of S0 significantly decreased the concentration of Cd in rice grain by 12.1% (0.1 g/kg) and 36.6% (0.2 g/kg) respectively. The addition of S0 significantly increased the ratio of Acidobacteria, Bacteroidetes in rice rhizosphere soil. Meanwhile, the ratio of Planctomycetes and Chloroflexi decreased. The results indicated that promoting Fe- and S-reducing and residue decomposition bacterial in the rhizosphere by S0 may be one biological reason for reducing Cd risk in the soil-rice system.

Regulation mechanisms of humic acid on Pb stress in tea plant (Camellia sinensis L.).

Though the interaction between humic acid (HA) and heavy metals has been widely reported, the effects of HA on the toxicity of heavy metals to plants are still in debate. In this study, the regulation mechanisms of HA on Pb stress in tea plant (Camellia sinensis L.) was investigated through hydroponic experiments, and the experimental results were explained by using transmission electron microscope (TEM), scanning transmission X-ray microscopes (STXM) and isobaric tags for relative and absolute quantitation (iTRAQ) differential proteomics. Significant alleviation of Pb stress was found with HA coexistence. TEM results showed that HA greatly mitigated the damage of cells caused by Pb stress. Compared with sole Pb treatment, the addition of HA increased the contents of pectin and pectic acid in the cell wall by 10.5% and 30.5%, while arabinose (Ara) and galactose (Gal) decreased by 20.5% and 15.9%, respectively, which were beneficial for increasing Pb adsorption capacity of the cell wall and promoting cell elongation. Moreover, iTRAQ differential proteomics analysis proved that HA strengthened the antioxidant system, promoted the synthesis of cell wall, and stabilized protein and sulfur-containing substance metabolism in molecular level. Notably, the concentration of calcium (Ca) in the cell wall of HA coexistence treatment was 47.4% higher than Pb treatment. STXM results also indicated that the distribution of Ca in the cell wall was restored with the presence of HA. This might promote the formation of the egg-box model, thus alleviating Pb stress in cells. Our results reveal the regulation mechanisms of HA on Pb detoxification in plants and provide useful information for improving the safety of agricultural products.

Influence of Humic Acid on Pb Uptake and Accumulation in Tea Plants.

A hydroponic experiment combined with synchronous radiation X-ray fluorescence (SRXRF) analysis was designed to understand the influence of humic acid (HA) in tea plants under lead stress. The results showed that the quantitative relationship (QR) between humic acid and Pb is an important factor affecting the regulation of humic acid with respect to the accumulation of Pb in tea plants. Besides, excess humic acid might stimulate the accumulation of Pb in the root cell wall and transfer to the shoot organs through undifferentiated casparian band structure. This study could provide a theoretical basis for the scientific evaluation of the effect of humic acid on tea uptake and the accumulation of Pb and the practical application of humic acid in reducing Pb pollution in the field.

Effects of ferrous sulfate amendment and water management on rice growth and metal(loid) accumulation in arsenic and lead co-contaminated soil.

Arsenic (As) and lead (Pb) commonly co-exist with high concentrations in paddy soil mainly due to human activities in south of China. This study investigates the effect of ferrous sulfate (FeSO4) amendment and water management on rice growth and arsenic (As) and lead (Pb) accumulation in rice plants. A paddy soil co-contaminated with As and Pb was chosen for the pot experiment with three FeSO4 levels (0, 0.25, and 1%, on a dry weight basis) and two water managements (flooded, non-flooded). The concentrations of As and Pb in iron plaques and rice plants were determined. Application of FeSO4 and non-flooded conditions significantly accelerated the growth of rice plants. With the addition of FeSO4, iron plaques were significantly promoted and most of the As and Pb were sequestered in the iron plaques. The addition of 0.25% FeSO4 and non-flooded conditions did not significantly change the accumulation of As and Pb in rice grains. The practice also significantly decreased the translocation factor (TF) of As and Pb from roots to above-ground parts which might have been aided by the reduction of As and Pb availability in soil, the preventing effect of rice roots, and the formation of more reduced glutathione (GSH). Flooded conditions decreased the Pb concentration in rice plants, but increased As accumulation. Moreover, rice grew thin and weak and even died under flooded conditions. Overall, an appropriate FeSO4 dose and non-flooded conditions might be feasible for rice cultivation, especially addressing the As issue in the co-contaminated soil. However, further detailed studies to decrease the accumulation of Pb in edible parts and the field application in As and Pb co-contaminated soil are recommended.

Mechanism study of sulfur fertilization mediating copper translocation and biotransformation in rice (Oryza sativa L.) plants.

Metabolism of sulfur (S) is suggested to be an important factor for the homeostasis and detoxification of Cu in plants. We investigated the effects of S fertilizers (S0, Na2SO4) on Cu translocation and biotransformation in rice plants by using multiple synchrotron-based techniques. Fertilization of S increased the biomass and yield of rice plants, as well as the translocation factor of Cu from root to shoot and shoot to grain, resulting in enhanced Cu in grain. Sulfur K-edge X-ray near edge structure (XANES) analysis showed that fertilization of S increased the concentration of glutathione in different rice tissues, especially in rice stem and leaf. Copper K-edge XANES results indicated that a much higher proportion of Cu (I) species existed in rice grain than husk and leaf, which was further confirmed by soft X-ray scanning transmission microscopy results. Sulfur increased the proportion of Cu (I) species in rice grain, husk and leaf, suggesting the inducing of Cu (II) reduction in rice tissues by S fertilization. These results suggested that fertilization of S in paddy soils increased the accumulation of Cu in rice grain, possibly due to the reduction of Cu (II) to Cu (I) by enhancing glutathione synthesis and increasing the translocation of Cu from shoot to grain.

Effect of heavy-metal-resistant bacteria on enhanced metal uptake and translocation of the Cu-tolerant plant, Elsholtzia splendens.

A hydroponics trial was employed to study the effects of Pseudomonas putida CZ1 (CZ1), a heavy-metal-resistant bacterial strain isolated from the rhizosphere of Elsholtzia splendens (E. splendens), on the uptake and translocation of copper (Cu) in E. splendens. Significant promotion of plant growth coupled with the obvious plant-growth-promoting (PGP) characters of the bacteria suggested that CZ1 would be a plant-growth-promoting rhizobacterium (PGPR) to E. splendens under Cu stress condition. The results of inductively coupled plasma optical emission spectrometry (ICP-OES) showed that CZ1 increased the concentration of Cu in the shoots (up to 211.6% compared to non-inoculation treatment) and translocation factor (TF) (from 0.56 to 1.83%) of those exposed to Cu. The distribution of Cu in root cross section measured by synchrotron-based X-ray fluorescence microscopy (SRXRF) indicated that CZ1 promoted the transport of Cu from cortex to xylem in roots, which contributed to the accumulation of Cu in shoots. Furthermore, CZ1 improved the uptake of nutrient elements by plants to oppose to the toxicity of Cu. In summary, P. putida CZ1 acted as a PGPR in resistance to Cu and promoted the accumulation and translocation of Cu from root to shoot by element redistribution in plant root; hence, CZ1 is a promising assistance to phytoremediation.

Translocation and biotransformation of CuO nanoparticles in rice (Oryza sativa L.) plants.

Metal-based nanoparticles (MNPs) may be translocated and biochemically modified in vivo, which may influence the fate of MNPs in the environment. Here, synchrotron-based techniques were used to investigate the behavior of CuO NPs in rice plants exposed to 100 mg/L CuO NPs for 14 days. Micro X-ray fluorescence (µ-XRF) and micro X-ray absorption near edge structure (µ-XANES) analysis revealed that CuO NPs moved into the root epidermis, exodermis, and cortex, and they ultimately reached the endodermis but could not easily pass the Casparian strip; however, the formation of lateral roots provided a potential pathway for MNPs to enter the stele. Moreover, bulk-XANES data showed that CuO NPs were transported from the roots to the leaves, and that Cu (II) combined with cysteine, citrate, and phosphate ligands and was even reduced to Cu (I). CuO NPs and Cu-citrate were observed in the root cells using soft X-ray scanning transmission microscopy (STXM).

MITIGATION OF Cu(II) PHYTOTOXICITY TO RICE (ORYZA SATIVA) IN THE PRESENCE OF TiO2 AND CeO2 NANOPARTICLES COMBINED WITH HUMIC ACID.

Engineered nanoparticles (NPs) and natural organic matter (NOM) in the environment may interact with background contaminants such as heavy metals and modify their bioavailability and toxicity. In the present study, the combined influences of 2 common NPs (TiO2 and CeO2 ) and humic acid (HA; as a model NOM) on Cu(II) phytotoxicity to rice were investigated by a 3-d root elongation assay performed on filter paper media. The results showed that the adsorption coefficients of bare TiO2 and CeO2 NPs (100 mg/L) toward Cu(2+) are 2.65 and 4.37, respectively, at an initial concentration of 10 mg/L, suggesting that Cu(II) could be strongly adsorbed by NPs, whereas HA-coated TiO2 and CeO2 NPs further enhanced the adsorption coefficients to 4.37 and 6.85, respectively. In addition, compared with Cu-alone treatment, the addition of bare TiO2 and CeO2 NPs (1000 mg/L) increased the length of rice root by 32.5% and 39.0%, respectively; however, the presence of HA-coated TiO2 and CeO2 NPs increased the root length by 90.2% and 100.1%, respectively, which indicated that the mitigation effect of HA-coated NPs on Cu(II) phytotoxicity was more visible than that of bare NPs. The results demonstrated that coexistence of NPs and HA significantly alleviated Cu(II) phytotoxicity as a result of a decrease in bioavailable soluble Cu(II) concentration, which contributes to an understanding of the potential behavior of NPs in the environment.

[Research advances in uptake, translocation, accumulation and detoxification of Pb in plants].

Contamination of soils by lead (Pb) is of widespread occurrence because of the industrialization, urbanization, mining, and many other anthropogenic activities. It is urgent and necessary for scientists to uncover the mechanisms of uptake, translocation, accumulation and detoxification of Pb in plants for the following two reasons. First, it helps target and regulate the key process of Pb uptake by crops and vegetables and minimize the threat of Pb introduction to the food chain. Second, it helps cultivate Pb hyperaccumulating plants that can absorb and sequester excessive amounts from contaminated soils in their biomass without incurring damage to basic metabolic functions. The purpose of this review was to summarize the research advances in uptake, translocation and accumulation of Pb in plants and address the mechanisms by which plants or plant systems detoxify Pb. The further researches on the foliar uptake, the interactions between soil components and plant cell wall, as well as the integrated technologies for phytoremediation of Pb-contaminated soils were prospected.

[Influence of sulfur on the speciation transformation and phyto-availability of heavy metals in soil: a review].

The biogeochemical cycle of sulfur can directly affect the speciation transformation of heavy metals in soils and their accumulation in plants. The toxicity of heavy metals in plants can be alleviated by their complexation with sulfur compounds like phytochelatins or metallothiones, consisting of the major mechanisms of phytodetoxification. Sulfur deficiency is becoming one of the limiting factors that influence Chinese agricultural production. Although the applications of sulfur fertilizer in soil have received much attention in recent years, the interaction mechanism of heavy metal with sulfur metabolism has not been studied extensively. In this paper, we reviewed current research advance on the impact of sulfur on the speciation transformation of soil heavy metals and their accumulation in plants, discussed the effect of sulfur on the detoxification mechanism of heavy metal in plants and provided further research prospective in this field.

Tracing intracellular localization and chemical forms of copper in Elsholtzia splendens with cluster analysis.

Copper (Cu) accumulation in soils is becoming a major environmental problem. Elsholtzia splendens, a Cu-tolerant plant growing in Cu mine areas, can accumulate high levels of Cu. This study develops a systematic method of detecting the intracellular localization and different chemical forms of Cu to comprehensively understand the mechanisms involved in Cu tolerance and detoxification of E. splendens. Various chemical forms of Cu were absorbed by E. splendens, the highest proportion of which was stored in its cell walls (68%) and vacuoles (42%). Most of the Cu in the roots, stems and leaves were extracted by 2% hydrate acetic acid (HAc) in controls. The distribution characteristics of Cu, particularly in the roots, were closely related to the Cu tolerance of E. splendens. In addition, the binding capacity of Cu to soluble salts or proteins decreased and its binding capacity to insoluble oxalate increased with the increasing Cu supply. These results indicated that Cu ions were integrated with compounds of low bioavailable, such as undissolved phosphate or oxalate, which contributed to the metal tolerance and detoxification of E. splendens. Cluster analysis, based on Cu abundance levels, reveals that correlations exist between the chemical forms of Cu and their intracellular locations, which in turn indicates a relationship between their migration activity and toxicity.

Risk assessment of heavy metals pollution in agricultural soils of siling reservoir watershed in Zhejiang Province, China.

Presence of heavy metals in agriculture soils above the permissible limit poses threats to public health. In this study, concentrations of seven metals were determined in agricultural soils from Yuhang county, Zhejiang, China. Multivariate statistical approaches were used to study the variation of metals in soils during summer and winter seasons. Contamination of soils was evaluated on the basis of enrichment factor (EF), geoaccumulation index (I(geo)), contamination factor (C(f)), and degree of contamination (C(deg)). Heavy metal concentrations were observed higher in winter as compared to summer season. Cr and Cd revealed random distribution with diverse correlations in both seasons. Principal component analysis and cluster analysis showed significant anthropogenic intrusions of Zn, Cd, Pb, Cr, and Cu in the soils. Enrichment factor revealed significant enrichment (EF > 5) of Zn, Cd, and Pb, whereas geoaccumulation index and contamination factor exhibited moderate to high contamination for Zn, Cr, Cd, and Pb. In light of the studied parameters, permissible limit to very high degree of contamination (C(deg) > 16) was observed in both seasons.

Cr(VI) reduction by a potent novel alkaliphilic halotolerant strain Pseudochrobactrum saccharolyticum LY10.

A novel Cr(VI)-reducing strain, Pseudochrobactrum saccharolyticum LY10, was isolated and characterized for its high Cr(VI)-reducing ability. Strain LY10 had typical characteristics of alkali-tolerance and halotolerance. Kinetic analysis indicated that the maximum reduction rate was achieved under optimum conditions with initial pH 8.3, 20gL(-1) NaCl, 55mgL(-1) Cr(VI), and 1.47×10(9)cellsmL(-1) of cell concentration. Further mechanism studies verified that the removal of Cr(VI) was mainly achieved by a metabolism-dependent bioreduction process. Strain LY10 accumulated chromium both in and around the cells, with cell walls acting as the major binding sites for chromium. X-ray absorption near-edge structure (XANES) analysis further confirmed that the chromium immobilized by the cells was in the Cr(III) state. In the present study, Pseudochrobactrum saccharolyticum was, for the first time, reported to be a Cr(VI)-reducing bacteria. Results from this research would provide a potential candidate for bioremediation of Cr(VI)-contaminated environments, especially alkaline and saline milieus with Cr(VI) at low-to-mid concentrations.

Effect of bamboo biochar on pentachlorophenol leachability and bioavailability in agricultural soil.

In this study, bamboo biochar (BBC) was proposed as a practical carbonaceous material for the in situ remediation of organic pollutants. Column leaching experiments were used to evaluate the leachability of pentachlorophenol (PCP) in soil amended with BBC (0, 1, 2, and 5%; w/w). After leaching, the residual bioavailability of PCP in the soil column was determined by chemical extraction methods (methanol and distilled water). The results showed that compared to the control, the addition of 5% BBC decreased the cumulative leach-loss content of PCP by 42%, and also reduced PCP concentrations by 56 and 65% in methanol and distilled water extracts, respectively. Moreover, the linear leaching curves for the 10-h leaching time indicated that the rapid release process may be dependent on the continuous diffusion and partition mechanism. Regression analyses showed a significant (p<0.001) relationship between the BBC percentage and cumulative content of leached PCP, in addition, a significant relationship between BBC percentage and the chemical extraction of PCP with correlation coefficients (R(2)) greater than 0.9. Therefore, adding BBC to soil could prevent PCP further contamination such as bioavailability, surface or groundwater contamination through leaching.

[Characteristics of distribution and chemical forms of Pb in tea plant varieties].

A hydroponics experiment combined with subcellular fractionation and sequential extraction was conducted to study the Pb concentration in different organs of two tea plant varieties (Longjing 43 and Yingshuang) and the Pb subcellular distribution and chemical forms in the roots of the varieties. Under Pb stress, the root system of the two varieties had different features in morphology. With the increasing concentration of Pb in culture solution, the Pb concentration in Longjing 43 young leaves increased, but that in Yingshuang' s had no significant variation. A marked difference was observed in the Pb subcellular distribution and its chemical forms in roots between the two varieties under Pb stress. In Longjing 43 roots, all subcellular fractions except soluble ones had a lower Pb concentration at low Pb stress, and all the subcellular fractions except cell wall ones had a higher Pb concentration at higher Pb stress, compared with those in Yingshuang's. In Longjing 43 roots, the HAc-extractable Pb occupied the greatest proportion, followed by NaCl-extractable Pb, HCl- and H2O- extractable Pb, and ethanol-extractable Pb; while in Yingshuang's, NaCl-extractable Pb had the greatest proportion, followed by HAc-extractable Pb, HCl- and H2O-extractable Pb, and ethanol-extractable Pb. Based on these findings, tea plant variety Yingshuang was likely to possess a higher tolerance to Pb than Longjing 43 did.