[Blocking Effects of Foliar Conditioners on Cadmium, Arsenic, and Lead Accumulation in Wheat Grain in Compound-contaminated Farmland].

Heavy metal contamination of soil has become a hot issue of social concern due to its impact on the safety of agricultural products in recent years. Wheat is one of the most dominant staple food crops worldwide and has become a major source of toxic metals in human diets. Foliar application was considered to be a more efficient and economical method of heavy metal remediation. Field experiments were carried out in Cd-, As-, and Pb-contaminated farmland soils. The effects of foliar conditioners on the accumulation of Cd, As, and Pb in wheat grains were investigated after being sprayed with Zn (0.2% ZnSO4), Mg (0.4% MgSO4), and Mn (0.2% MnSO4) separately and in combination. Thus, the effective foliar conditioners were selected to block the accumulation of Cd, As, and Pb in wheat grains grown in combined heavy metal-contaminated farmland in north China. The results showed that, compared with that in the control, the Cd, As, and Pb contents in wheat grains of the Zn+Mg+Mn foliar treatment were significantly decreased by 18.96%, 23.87%, and 51.31%, respectively, and TFgrain/straw decreased by 14.62%, 27.73%, and 47.70%, respectively. Thus, spraying the compound foliar conditioner of Zn+Mg+Mn could effectively reduce heavy metal accumulation in wheat grains through inhibition translocation of those metals from stem leaves to grain. In addition, the results indicated that Cd and As were mainly distributed at the central endosperm (34.08%-37.08%), whereas Pb was primarily distributed at the pericarp and seed coat (27.78%) of the wheat grain. Compared with that in the control, spraying the compound foliar conditioner of Zn+Mg+Mn extremely decreased Cd and As accumulation in the aleurone layer of the wheat grain by 81.10% and 82.24%, respectively. Except for the pericarp, seed coat, and central endosperm layers, the Pb content in each grain layer was dramatically decreased by 42.85% to 91.15%. There was only a significant negative correlation between heavy metal content and Zn content in the aleurone layer (P2) of wheat flour. In summary, the accumulation of Cd, As, and Pb in wheat grains, especially in the aleurone layer, could be effectively reduced by foliar conditioner application at the jointing, booting, and early filling stages of wheat, separately. Furthermore, besides the foliar treatment, removing wheat bran to reduce Cd contamination in wheat grains is highly recommended to ensure the safe production of wheat.

[Distribution and Transport of Cadmium and Arsenic in Different Aboveground Parts of Wheat After Flowering].

To investigate the effects of leaves and stems on the accumulation and transport of cadmium(Cd) and arsenic(As) in wheat shoots after flowering, a field experiment was conducted in a typical Cd and As co-contaminated agricultural land to explore the distribution and translocation of Cd and As in the different parts of two wheat cultivars after flowering. The results showed that Cd was mainly distributed in the nodes of two varieties, and the translocation factors of Cd from internode 3 to node 2, from internode 2 to node 1, and from sheath 1 to node 1 were markedly higher than those of other aboveground parts during the grain-filling stage. However, Cd was mainly distributed in the leaves, and the translocation factors of Cd from sheath to leaf and from node 1 to rachis was significantly higher than those of other parts at the mature stage. In addition, the transport capacity of Cd from glume to rachis and from rachis to grain in JM22 was significantly lower than that in SN28, which significantly reduced Cd concentrations in the rachis, glume, and grain of JM22 by 22.3%, 40.8%, and 44.4%, respectively. Meanwhile, As was mainly distributed in the wheat leaves from the grain-filling stage to the mature stage, and As concentrations in the glume and grain of JM22 were 25.8% and 33.3% lower than those of SN28, respectively. Additionally, the translocation factors of As from the sheath to the node were significantly 438% and 190% higher than that from leaf to sheath and from node to internode during the whole grain filling stage and mature stage. Moreover, the translocation factors of As from glumes to grains and from rachis to grains in JM22 were 40.6% and 44.4% lower than that in SN28, respectively. In summary, flag leaf, node 1, and the rachis had regulated Cd transport and accumulation in wheat grains, whereas leaf 3, flag leaf, node 1, the glumes, and the rachis were mainly responsible for As transport and accumulation in wheat grains.

A dirigent protein complex directs lignin polymerization and assembly of the root diffusion barrier.

Functionally similar to the tight junctions present in animal guts, plant roots have evolved a lignified Casparian strip as an extracellular diffusion barrier in the endodermis to seal the root apoplast and maintain nutrient homeostasis. How this diffusion barrier is structured has been partially defined, but its lignin polymerization and assembly steps remain elusive. Here, we characterize a family of dirigent proteins (DPs) essential for both the localized polymerization of lignin required for Casparian strip biogenesis in the cell wall and for attachment of the strip to the plasma membrane to seal the apoplast. We reveal a Casparian strip lignification mechanism that requires cooperation between DPs and the Schengen pathway. Furthermore, we demonstrate that DPs directly mediate lignin polymerization as part of this mechanism.

Transcriptomic mechanisms of reduced PM2.5-Pb retention in the leaves of the low-Pb-accumulation genotype of Chinese cabbage.

Atmospheric fine particulate matter (PM2.5) mainly contributes to Pb accumulation in the edible leaves of Chinese cabbage in North China. It was found that a low-Pb-accumulation (LPA) genotype of Chinese cabbage contained less Pb in leaves than high-Pb-accumulation (HPA) genotype exposed to PM2.5-Pb. However, there are no data on the transcriptional regulatory mechanisms of foliar PM2.5-Pb uptake by Chinese cabbage. The present study investigated the retention of PM2.5-Pb in foliar apoplast and symplasm and the underlying molecular mechanisms of reduced Pb in LPA leaves. It appeared more Pb in apoplast and less Pb in symplasm of LPA leaves, whereas the pattern was opposite in HPA. There were 2646 and 3095 differentially expressed genes (DEGs) in LPA and HPA leaves under PM2.5-Pb stress with clearly genotype-specific function, respectively. Furthermore, mRNA levels of XTH16 regulating cell wall thickening, PME2 and PME6 involved in cell wall remodification were significantly expressed in LPA, but not in HPA. Meanwhile, foliar PM2.5-Pb stress downregulated expression of ZIP1, YSL1, and CNGC3 responsible for Pb influx to cell, and upregulated expression of ABCG36 regulated Pb efflux from symplasm in LPA leaves. These results improve our understanding to the mechanisms underlying foliar Pb uptake from PM2.5-Pb at transcriptomic level.

[Characteristics and Health Risk Assessment of Cadmium, Lead, and Arsenic Accumulation in Leafy Vegetables Planted in a Greenhouse].

In recent years, dust pollution has occurred frequently in spring and haze or fog in autumn and winter. The inhalable particulate matters in the atmosphere, especially PM2.5, loaded in heavy metals such as cadmium, lead, and arsenic, are easily taken up by leafy vegetables and accumulate in the edible parts. It is not clear whether the accumulation of heavy metals in the edible parts of leafy vegetables in greenhouses is also affected by atmospheric deposition. Therefore, a field experiment was conducted to explore characteristics and health risk assessment of cadmium, lead, and arsenic accumulation in leafy vegetables planted in a greenhouse using six types of common leafy vegetables (spinach, leaf lettuce, lettuce, pakchoi, Chrysanthemum coronarium, and fennel) in the Beijing-Tianjin-Hebei region. The results showed that C. coronarium, pakchoi, and spinach are the leafy vegetables with a low accumulation of Cd, Pb, and As, respectively. Fennel is the leafy vegetable with a low accumulation of Cd and Pb. In the greenhouse, Pb concentrations in PM2.5 were 42.6 and 8.4 times of Cd and As, respectively. Moreover, PM2.5-Pb contributed on average 36.5% to the edible parts of six kinds of leafy vegetables, which indicated that the Cd, Pb, and As accumulated in leafy vegetables were mainly derived from the soil. Meanwhile, the concentrations of Cd, Pb, and As in the edible parts of vegetables did not exceed the safety limitations of three heavy metals (GB 2762-2017), and Pb accumulation in leafy vegetables does not pose a health risk to humans. However, Cd in the leafy vegetables could threaten the health of adults and children, except for the intake of fennel. Conversely, As in the C. coronarium could threaten the health of adults and children.

Mechanism of Pb accumulation in Chinese cabbage leaves: Stomata and trichomes regulate foliar uptake of Pb in atmospheric PM2.5.

Chinese cabbage (Brassica rapa ssp. pekinensis) is one of the most popular and frequently consumed leafy vegetables. It was found that atmospheric PM2.5-Pb contributes to Pb accumulation in the edible leaves of Chinese cabbage via stomata in North China during haze seasons with high concentrations of fine particulate matter in autumn and winter. However, it is unclear whether both stomata and trichomes co-regulate foliar transfer of PM2.5-Pb from atmospheric deposition to the leaf of Chinese cabbage genotypes with trichomes. Field and hydroponic experiments were conducted to investigate the effects of foliar uptake of PM2.5-Pb on Pb accumulation in leaves using two genotypes of Chinese cabbage, one without trichomes and one with trichomes. It was verified that open stoma is a prominent pathway of foliar PM2.5-Pb transfer in the short-term exposure for 6 h, contributing 74.5% of Pb accumulation in leaves, whereas Pb concentrations in the leaves of with-trichome genotype in the rosette stage were 6.52- and 1.04-fold higher than that of without-trichome genotype in greenhouse and open field, respectively, which suggests that stomata and trichomes co-regulate foliar Pb uptake of from atmospheric PM2.5. Moreover, subcellular Pb in the leaves was distributed in the following order of cytoplasm (53.8%) > cell wall (38.5%)> organelle (7.8%), as confirmed through high-resolution secondary ion mass spectrometry (NanoSIMS). The Leadmium™ Green AM dye manifested that Pb in PM2.5 entered cellular space of trichomes and accumulated in the basal compartment, enhancing foliar Pb uptake in the edible leaves of cabbage. The results of these experiments are evidence that both stomata and trichomes are important pathways in the regulation of foliar Pb uptake and translocation in Chinese cabbage.

Bioimaging of Pb by LA-ICP-MS and Pb isotopic compositions reveal distributions and origins of Pb in wheat grain.

Atmospheric heavy metal deposition in agroecosystems has increased recently, especially in northern China, which poses serious risks to crop safety and human health via food chain. Wheat grains can accumulate high levels of Pb even when wheat is planted in soils with low levels of Pb. However, the influence of atmospheric deposition on the accumulation and distribution of Pb in wheat grain is still unclear. A field survey was conducted in three districts (A: a district with industrial and traffic pollution; B: a district with traffic pollution; and C: an unpolluted district) in Hebei Province, North China. The grain of wheat cultivated in district A accumulated more Pb from soil and atmospheric deposition than those in other districts, and the bran from district A contained 3.50 and 2.04 times more Pb than those from districts B and C, respectively. The Pb distribution pattern in wheat grain detected by laser ablation inductively coupled mass spectrometry (LA-ICP-MS) was characterized by accumulation mostly in the pericarp and seed coat rather than in the crease, embryo and endosperm. Furthermore, Pb isotopic data showed that airborne Pb was the major source (>50%) of Pb in wheat grain. Interestingly, average contributions of Pb from atmospheric deposition to white flour (78.22%) were higher than its contributions to bran (56.27%). In addition, wheat flag leaves were exposed to PbSO4 at the booting stage, and much greater Pb accumulation (0.33-0.48 mg/kg) was observed in exposed wheat grain than in the control (P < 0.05), PbSO4 constituted most (82.80-100%) of the Pb in the wheat grain. In summary, the results confirmed the efficient foliar Pb uptake and transfer from atmospheric deposition into wheat grain. It would be a new sight for understanding the contribution of airborne Pb to Pb accumulation in wheat grains.

Contribution of PM2.5-Pb in atmospheric fallout to Pb accumulation in Chinese cabbage leaves via stomata.

Foliar uptake of Pb is especially important when Chinese cabbage (Brassica rapa spp. pekinensis), having a large leaf surface area, is cultivated in North China during seasons with heavy haze. However, the mechanisms of foliar Pb uptake via stomata by Chinese cabbage exposed to atmospheric fallout are unclear. A field experiment was conducted to explore the impacts of Pb in particulate matter with sizes ≤ 2.5 µm (PM2.5-Pb) from atmospheric fallout to Pb accumulation in cabbage leaves through stomata. Cabbage varieties with low-Pb-accumulation (LPA) and high-Pb-accumulation (HPA) were examined using inductively coupled plasma-mass spectrometry and scanning electron microscopy/energy-dispersive X-ray analysis. The 206Pb/207Pb and 208Pb/207Pb ratios of PM2.5, plants, and soil demonstrated that the major source of Pb in cabbage leaves was PM2.5. The average width and length of the stomatal apertures were 7.14 and 15.61 µm for LPA cabbage and 8.10 and 16.64 µm for HPA cabbage, which are large enough for PM2.5-Pb to enter the leaves. The HPA cabbage had significantly higher stomatal width-to-length ratios than the LPA cabbage, indicating that the former trapped much more PM2.5-Pb and accumulated more Pb. These results clarify the contributions of the stomatal characteristics to PM2.5-Pb accumulation in the edible parts of Chinese cabbage.

[Characteristics of Cd, As, and Pb in Soil and Wheat Grains and Health Risk Assessment of Grain-Cd/As/Pb on the Field Scale].

In recent years, heavy metal pollution in farmlands has become increasingly serious because of human activities such as metal smelting, sewage irrigation, and road traffic in China. A field survey was conducted to investigate characteristics of Cd, As, and Pb in soil and wheat grains and assess the health risk of grain-Cd/As/Pb to humans on the fields scale. The farmland was influenced by smelter and sewage irrigation in the attitude and by road traffic in the horizon. The results showed that in farmland soil with moderate pollution levels, Cd, As, and Pb concentrations in soil samples all exceeded the risk screening values of farmland soil (GB 15618-2018), and the exceeding rates were 100%, 100%, and 36.7% respectively; the exceeding rates of Cd and Pb concentrations in wheat grains were 76.7% and 13.3%, respectively (GB 2762-2017). Distance from smelter, river of sewage irrigation, and road had no significant effect on Cd, As, and Pb concentrations in soil but had a significant effect on Cd and As concentrations in wheat grains, with the median Cd and As concentrations of the closest group being 14.9% and 41.8%, respectively, higher than the highest group (P<0.05). The Pb concentrations in soil and wheat grains were influenced by road traffic; the median Pb concentrations of the closest group were 78.9% and 471%, respectively, higher than the highest group (P<0.05). Cd and As in wheat grains have carcinogenic risks (Ri>1×10-4), RCd > RAs, Rchildren > Radult, while Pb poses no health risks in this farmland.

[Mechanisms of copper uptake by submerged plant Hydrilla verticillata ( L. F. ) royle and Myriophyllum spicatum L].

Cultivation experiments, including short-term copper uptake kinetics, uptake kinetics of Cu by different subcellular fractions and efflux of Cu, were carried out to investigate mechanisms of copper uptake by two species of submerged macrophyte, Hydrilla verticillata (L. f. ) Royle and Myriophyllum spicatum L. The results showed that: (1) Both H. verticillata and M. spicatum showed high influx rates for copper ions with no statistically significant differences [ Vmax (DW) is 2 micromol. (g min) - 1] ; (2) Both leaf and stem Cu concentrations of two strains of plants showed sharp increase with the external medium, and the equilibrium was reached for about 12 h after being exposed to solution with 2 micromol L-1 Cu. Leaves of these two plants accumulated much higher concentrations of Cu than stems. Furthermore, Cu in leaves concentrated predominantly in cell walls ( > 60% ) , followed by soluble and organell fractions; (3) Copper accumulated in cell wall of H. verticillata leaf was considerably higher than that of M. spicatum, and Cu concentrated in intracellular fractions of leaves showed no statistically significant difference between these two plants. Moreover, ability of Cu accumulation and efflux by H. verticillata was higher than that by M. spicatum.

Arsenic accumulation and translocation in the submerged macrophyte Hydrilla verticillata (L.f.) Royle.

Worldwide contamination of arsenic in aquatic systems requires the development of a cost-effective, in situ phytoremediation technology. Hydrilla verticillata (L.f.) Royle, a submerged macrophyte widely distributed throughout the world, has the potential to effectively remove heavy metals from water. In order to understand the potential of H. verticillata for As phytofiltration and its impacts on As cycling in the water system, we investigated As accumulation, speciation and translocation in H. verticillata plants. Plant shoots showed a significant accumulation of As, with a maximum of >700 µg g(-1) dry weight (DW) after exposure to 20 µM arsenate [As(V)] or arsenite [As(III)] for 4d, with no significant differences between the As(V) and As(III) treatments (P>0.05). In addition, results of an in planta transport experiment showed that, after exposure of root and shoot to 2µM As(V) and As(III) for 4d, the bioconcentration factor (BCF) in roots for As(V) was almost twofold than that of As(III). Higher As BCFs in roots compared to shoots was also observed. Arsenic accumulated primarily in the cell walls of root cells (>73% of the total As in roots) and in the soluble parts of leaves (>60% of the total As in leaves). Regardless of the form of As supplied [As(III) or As(V)], As(V) was the dominant form in roots and As(III) was the dominant form in leaves. Further, basipetal translocation of As in this plant (≥17%) was markedly higher than acropetal translocation (≤3%). Because of accumulation of As in the shoot and immobilization of As below ground in roots, H. verticillata is a potential As phytofiltrator for bioremediation.

[Effect of competitive interference on the biosorption of lead (II) by Hydrilla verticallata].

Using submerged aquatic plant is a cheap and clean technique to remediate heavy metals in wastewaters. Batch experiments were conducted to assess the biosorption characteristics of Pb (II) ions by fresh tissues of Hydrilla verticallata. The biosorption of Pb(II) was examined for single, binary and ternary solutions at different initial concentrations and different pH values. The experimental results showed that the biosorption capacity increased with increasing pH from 2.0 to 6.0. The biosorption value reached 44.65 mg/g when initial lead concentration was 250 mg/L and pH = 5. Both Cu (II) and Zn(II) ions were found to have an adverse effect on the biosorption of Pb(II) for binary and ternary solutions. In Pb-Cu binary metal solution, when both initial concentrations of lead and copper ions were 250 mg/L, the biosorption capacity for lead ions was decreased to 49.29% of that in single lead ion solution. The biosorption equilibrium data for the Pb-Cu binary metal solution fitted the Langmuir competitive model well (R2 = 0.966). The theoretical q(max) value (58.02 mg/g) was in excellent consistent with that obtained experimentally, and the average relative error between calculated q(e) and experimental q(e) values was only 15.6%. Comparison between biosorption of Pb (II) and Cu (II) by H. verticallata in the binary solution could lead to the conclusion that H. verticallata has no preference of Pb(II) over Cu(II).

Copper uptake and translocation in a submerged aquatic plant Hydrilla verticillata (L.f.) Royle.

A comprehensive understanding of the uptake, tolerance and transport of heavy metals in the wetland system through aquatic plants will be essential for the development of phytoremediation technologies. Copper accumulation and translocation of a submersed macrophyte Hydrilla verticillata (L.f.) Royle were investigated. Plant shoots showed a significant accumulation of Cu with a maximum of 30830 mg Cu kg⁻¹ dry weight after exposed to 4000 µg L⁻¹ Cu for 4d. Both roots and shoots can directly take up Cu from solution and Cu mainly accumulated in cell wall fractions. Moreover, H. verticillata predominantly accumulated Cu through shoots from the aqueous solutions because of the higher weights and bioaccumulation factors of shoots than those of roots. Acropetal translocation of Cu in the plant is higher than the basipetal translocation, which implies that upward translocation of Cu is mainly via the xylem and downward translocation is mainly through the phloem. These findings contribute to the application of submerged aquatic plants to copper removal from moderately contaminated waters.