Unveiling mechanisms of silicon-mediated resistance to chromium stress in rice using a newly-developed hierarchical system.

Silicon (Si) has been well-known to enhance plant resistance to heavy-metal stress. However, the mechanisms by which silicon mitigates heavy-metal stress in plants are not clear. In particular, information regarding the role of Si in mediating resistance to heavy-metal stress at a single cell level is still lacking. Here, we developed a hierarchical system comprising the plant, protoplast, and suspension cell subsystems to investigate the mechanisms by which silicon helps to alleviate the toxic effects of trivalent chromium [Cr(III)] in rice. Our results showed that in whole-plant subsystem silicon reduced shoot Cr(III) concentration, effectively alleviating Cr(III) stress in seedlings and causing changes in antioxidant enzyme activities similar to those observed at lower Cr(III) concentrations without silicon added. However, in protoplast subsystem lacking the cell wall, no silicon deposition occurred, leading to insignificant changes in cell survival or antioxidation processes under Cr(III) stress. Conversely, in suspension cell subsystem, silicon supplementation substantially improved cell survival and changes in antioxidant enzyme activities under Cr(III) stress. This is due to the fact that >95% of silicon was on the cell wall, reducing Cr(III) concentration in cells by 7.7%-10.4%. Collectively, the results suggested that the silicon deposited on the cell wall hindered Cr(III) bio-uptake, which consequently delayed Cr(III)-induced changes in antioxidant enzyme activities. This research emphasizes the significance of cell walls in Si-alleviated heavy-metal stress and deepens our understanding of silicon functioning in plants. Furthermore, the hierarchical system has great potential for application in studying the functioning of other elements in plant cell walls.

Multi-imaging platform for rhizosphere studies: Phosphorus and oxygen fluxes.

Rhizosphere is a soil volume of high spatio-temporal heterogeneity and intensive plant-soil-microbial interactions, for which visualization and process quantification is of highest scientific and applied relevance, but still very challenging. A novel methodology for quick assessment of two-dimensional distribution of available phosphorus (P) in rhizosphere was suggested, tested, and development up to the application platform. Available P was firstly trapped by an in-situ diffusive gradients in thin-films (DGT) sampler with precipitated zirconia as the binding gel, and subsequently, the loaded gel was analyzed with an optimized colorimetric imaging densitometry (CID). The imaging platform was established linking: i) DGT, ii) planar optode, and iii) soil zymography techniques to simultaneously determine available P, oxygen, and acid phosphatase in rhizosphere at sub-millimeter spatial scales. The DGT identified available P level in rice rhizosphere were spatially overlapping to the localized redox hotspots and phosphatase activity. The spatial relationship between available P and acid phosphatase activity was dependent on root development. The root radial oxygen loss (ROL) remained active during the experimental observations (2-3 days), while a flux of available P of 10 pg cm-2 s-1 was visualized within 2-3 mm of roots, confirming the correlative response of rice roots to oxygen secretion and P uptake. Summarizing, the established imaging platform is suitable to capture spatial heterogeneity and temporal dynamics of root activities, nutrient bioavailability, ROL and enzyme activities in rhizosphere.

Probability of cultivating Se-rich maize in Se-poor farmland based on intensive field sampling and artificial neural network modelling.

Selenium (Se) is a necessary micronutrient for humans, and its supplementation from crop grains is important to address the ubiquitous Se deficiency in people worldwide. Se uptake by crops largely depend on soil bioavailable Se rather than soil total Se content, which provides possibilities to explore the Se-rich crops in Se-poor area. Here, the possibility of cultivating Se-rich maize grains in Se-poor farmland was tested based on intensive field sampling and mathematical modelling. Sampling was conducted at county scale, and a total of 7779 topsoil samples and 109 maize samples with paired rhizosphere soils samples were collected. Results showed that although the soil Se content in the study county from southwestern China was at a low level (0.01-2.75 mg kg-1), 54.1% of the maize grain samples satisfied the standard for Se-rich products (0.02-0.30 mg kg-1). Soil organic matter, iron oxide, and phosphorus levels were correlated negatively with Se bioconcentration factor (BCF) of maize grain. Compared with the multivariate linear regression model, the artificial neural network (ANN) model was more accurate and reliable in predicting maize Se BCF. Prediction using the ANN model showed that 22.7% of the county's farmland was suitable for cultivating naturally Se-rich maize, which increased 21.3% growing areas than that from cultivation based on simply soil total Se. This study provided a new methodological framework for natural Se-rich maize production and verified the probability of cultivating naturally Se-rich maize in Se-poor farmland.

Selenium Increased Arsenic Accumulation by Upregulating the Expression of Genes Responsible for Arsenic Reduction, Translocation, and Sequestration in Arsenic Hyperaccumulator Pteris vittata.

Selenate enhances arsenic (As) accumulation in As-hyperaccumulator Pteris vittata, but the associated molecular mechanisms are unclear. Here, we investigated the mechanisms of selenate-induced arsenic accumulation by exposing P. vittata to 50 µM arsenate (AsV50) and 1.25 (Se1.25) or 5 µM (Se5) selenate in hydroponics. After 2 weeks, plant biomass, plant As and Se contents, As speciation in plant and growth media, and important genes related to As detoxification in P. vittata were determined. These genes included P transporters PvPht1;3 and PvPht1;4 (AsV uptake), arsenate reductases PvHAC1 and PvHAC2 (AsV reduction), and arsenite (AsIII) antiporters PvACR3 and PvACR3;2 (AsIII translocation) in the roots, and AsIII antiporters PvACR3;1 and PvACR3;3 (AsIII sequestration) in the fronds. The results show that Se1.25 was more effective than Se5 in increasing As accumulation in both P. vittata roots and fronds, which increased by 27 and 153% to 353 and 506 mg kg-1. The As speciation analyses show that selenate increased the AsIII levels in P. vittata, with 124-282% more AsIII being translocated into the fronds. The qPCR analyses indicate that Se1.25 upregulated the gene expression of PvHAC1 by 1.2-fold, and PvACR3 and PvACR3;2 by 1.0- to 2.5-fold in the roots, and PvACR3;1 and PvACR3;3 by 0.6- to 1.1-fold in the fronds under AsV50 treatment. Though arsenate enhanced gene expression of P transporters PvPht1;3 and PvPht1;4, selenate had little effect. Our results indicate that selenate effectively increased As accumulation in P. vittata, mostly by increasing reduction of AsV to AsIII in the roots, AsIII translocation from the roots to fronds, and AsIII sequestration into the vacuoles in the fronds. The results suggest that selenate may be used to enhance phytoremediation of As-contaminated soils using P. vittata.

Total and available metal concentrations in soils from six long-term fertilization sites across China.

Approximately 19% of agricultural soils in China are contaminated by heavy metals. However, the effects of agricultural management practices on soil contamination are not well understood. Taking advantage of six long-term (23-34 years) field sites across China, this study examined the effects of different agricultural fertilization treatments, including control (no fertilization), inorganic nitrogen, phosphorus and potassium fertilization (NPK), manure fertilization (M), and NPK plus manure fertilization (NPKM), on the total and available metal concentrations in soils. The results showed that after 23-34 years of fertilization, the M and NPKM treatments significantly increased the total concentration of cadmium (Cd), copper (Cu), and zinc (Zn) in soils compared with the concentrations measured for the control and NPK treatments. In contrast, the fertilization treatments had almost no influence on soil lead (Pb) and nickel (Ni) concentrations. The results of analysis via diffusive gradients in thin films demonstrated that long-term sheep or cattle manure fertilization increased the available metals, especially Cd, Cu, and Zn, but long-term swine manure application decreased the available metals, except for Cu and Zn, in soils. Further analysis revealed that the manure source, soil pH level, and biogeochemical properties of metals affected the availability of Cd, Cu, Pb, Zn, and Ni in soils. Collectively, organic fertilizers had the potential to reduce metal uptake by crops, but caution should be taken to reduce metal concentrations in manure.