Decreasing arsenic accumulation in rice by overexpressing OsNIP1;1 and OsNIP3;3 through disrupting arsenite radial transport in roots
TL;DR: Overexpression ofOsNIP1;1 or OsNIP3;3 provides a route for arsenite to leak out of the stele, thus restricting arsenite loading into the xylem, and is effective in reducing arsenic accumulation in rice grain.
Abstract: Rice is a major dietary source of the toxic metalloid arsenic. Reducing arsenic accumulation in rice grain is important for food safety. We generated transgenic rice overexpressing two aquaporin genes, OsNIP1;1 and OsNIP3;3, under the control of a maize ubiquitin promoter or the rice OsLsi1 promoter, and tested the effect on arsenite uptake and translocation. OsNIP1;1 and OsNIP3;3 were highly permeable to arsenite in Xenopus oocyte assays. Both transporters were localized at the plasma membrane. Knockout of either gene had little effect on arsenite uptake or translocation. Overexpression of OsNIP1;1 or OsNIP3;3 in rice did not affect arsenite uptake but decreased root-to-shoot translocation of arsenite and shoot arsenic concentration markedly. The overexpressed OsNIP1;1 and OsNIP3;3 proteins were localized in all root cells without polarity. Expression of OsNIP1;1 driven by the OsLsi1 promoter produced similar effects. When grown in two arsenic-contaminated paddy soils, overexpressing lines contained significantly lower arsenic concentration in rice grain than the wild-type without compromising plant growth or the accumulation of essential nutrients. Overexpression of OsNIP1;1 or OsNIP3;3 provides a route for arsenite to leak out of the stele, thus restricting arsenite loading into the xylem. This strategy is effective in reducing arsenic accumulation in rice grain.
Citations
More filters
TL;DR: In this paper, the authors reviewed recent progress in understanding the biogeochemical processes controlling As and Cd bioavailability in paddy soil, the mechanisms of their uptake, translocation and detoxification in rice plants, and strategies to reduce their accumulation in rice grain.
Abstract: Arsenic (As) and cadmium (Cd) are two toxic elements that have a relatively high risk of transfer from paddy soil to rice grain. Rice is a major dietary source of these two elements for populations consuming rice as a staple food. Reducing their accumulation in rice grain is important for food safety and human health. We review recent progress in understanding the biogeochemical processes controlling As and Cd bioavailability in paddy soil, the mechanisms of their uptake, translocation and detoxification in rice plants, and strategies to reduce their accumulation in rice grain. Similarities and differences between the two elements are emphasized. Some knowledge gaps are also identified. The concentrations of As and Cd in rice grain vary by three orders of magnitude, depending on the bioavailability of the two elements in soil, rice genotype and growing conditions. The redox potential in paddy soil has a profound but opposite effect on As and Cd bioavailability, whereas soil pH affects Cd bioavailability more than As bioavailability. A number of key genes involved in As and Cd uptake, translocation, sequestration, and detoxification in rice have been characterized. Allelic variations of several genes underlying the variations in Cd accumulation have been identified, but more remains to be elucidated, especially for As. Two types of strategies can be used to reduce As and Cd accumulation, reducing their bioavailability in soil or their uptake and translocation in rice. Reducing the accumulation of both As and Cd in rice simultaneously remains a great challenge.
272 citations
TL;DR: It is shown that OsNRAMP1 was able to transport Cd and manganese (Mn) when expressed in yeast, but did not transport Fe or As, and contributes significantly to the uptake of Mn and Cd in rice.
Abstract: Rice is a major dietary source of the toxic metal, cadmium (Cd). Previous studies reported that the rice transporter, OsNRAMP1, (Natural resistance-associated macrophage protein 1) could transport iron (Fe), Cd and arsenic (As) in heterologous yeast assays. However, the in planta function of OsNRAMP1 remains unknown. Here, we showed that OsNRAMP1 was able to transport Cd and manganese (Mn) when expressed in yeast, but did not transport Fe or As. OsNRAMP1 was mainly expressed in roots and leaves and encoded a plasma membrane-localized protein. OsNRAMP1 expression was induced by Cd treatment and Fe deficiency. Immunostaining showed that OsNRAMP1 was localized in all root cells, except the central vasculature, and in leaf mesophyll cells. The knockout of OsNRAMP1 resulted in significant decreases in root uptake of Cd and Mn and their accumulation in rice shoots and grains, and increased sensitivity to Mn deficiency. The knockout of OsNRAMP1 had smaller effects on Cd and Mn uptake than knockout of OsNRAMP5, while knockout of both genes resulted in large decreases in the uptake of the two metals. Taken together, OsNRAMP1 contributes significantly to the uptake of Mn and Cd in rice, and the functions of OsNRAMP1 and OsNRAMP5 are similar but not redundant.
134 citations
TL;DR: Manipulation of some transporters for these elements, especially those localized in the node for allocation of mineral elements to the grain, has been successful in generating rice with higher density and bioavailability of essential elements, but less toxic elements.
97 citations
TL;DR: In this paper, the effects of SiO2 NPs on inhibiting arsenic uptake into rice using individual rice cells were investigated for the first time, and the results indicated that the addition of siO2NPs could enhance the proportion of live cells by weakening oxidative stress upon As exposure.
Abstract: Our previous studies indicated that the foliar application of silica nanoparticles (SiO2 NPs) could obviously reduce arsenic (As) accumulation in rice. However, the mechanism underlying this effect at the single-cell level has not been reported. In this study, we investigated for the first time the effects of SiO2 NPs on inhibiting As uptake into rice using individual rice cells. The results indicated that the addition of SiO2 NPs could enhance the proportion of live cells by weakening oxidative stress upon As exposure. Compared to the treatment of cells with As only, treatment with SiO2 NPs could maintain the integrity of the cell, increase the thickness of the cell wall (77.4%) and the ratio of As in the pectin (19.6%). In addition, the pectin content, cation exchange capacity (CEC) and pectin methylesterase (PME) activity were also increased in the SiO2 NPs-pretreated cells, leading to a decreased degree of pectin methylesterification and an improved mechanical force of the cell walls. Furthermore, in the SiO2 NPs-pretreated rice cells, the expression of the OsLis1 and OsLis2 genes was lower, whereas the expression of the OsNIP1;1 and OsNIP3;3 genes was higher than that of the As-only group. This finding provides new insights into the mechanism of how the addition of SiO2 NPs inhibits As uptake into rice at the single-cell level and lays the foundation for its application in As-contaminated paddy soil.
93 citations
TL;DR: The abundance of proteins involved in photosynthesis, energy metabolism, transport, signaling and ROS homeostasis were found higher in As + Se than in As alone exposure, which resulted in reduced As accumulation and toxicity.
86 citations
References
More filters
TL;DR: The protocol presented here has underpinned much of the in vivo results highlighting the dynamic nature of the plant secretory pathway and is a relatively fast technique to assess expression of genes of interest.
Abstract: Expression and tracking of fluorescent fusion proteins has revolutionized our understanding of basic concepts in cell biology. The protocol presented here has underpinned much of the in vivo results highlighting the dynamic nature of the plant secretory pathway. Transient transformation of tobacco leaf epidermal cells is a relatively fast technique to assess expression of genes of interest. These cells can be used to generate stable plant lines using a more time-consuming, cell culture technique. Transient expression takes from 2 to 4 days whereas stable lines are generated after approximately 2 to 4 months.
1,437 citations
TL;DR: The identification of a silicon transporter provides both an insight into the silicon uptake system in plants, and a new strategy for producing crops with high resistance to multiple stresses by genetic modification of the root's silicon uptake capacity.
Abstract: Silicon is beneficial to plant growth and helps plants to overcome abiotic and biotic stresses by preventing lodging (falling over) and increasing resistance to pests and diseases, as well as other stresses. Silicon is essential for high and sustainable production of rice, but the molecular mechanism responsible for the uptake of silicon is unknown. Here we describe the Low silicon rice 1 (Lsi1) gene, which controls silicon accumulation in rice, a typical silicon-accumulating plant. This gene belongs to the aquaporin family and is constitutively expressed in the roots. Lsi1 is localized on the plasma membrane of the distal side of both exodermis and endodermis cells, where casparian strips are located. Suppression of Lsi1 expression resulted in reduced silicon uptake. Furthermore, expression of Lsi1 in Xenopus oocytes showed transport activity for silicon only. The identification of a silicon transporter provides both an insight into the silicon uptake system in plants, and a new strategy for producing crops with high resistance to multiple stresses by genetic modification of the root's silicon uptake capacity.
1,398 citations
TL;DR: It is reported that two different types of transporters mediate transport of arsenite, the predominant form of arsenic in paddy soil, from the external medium to the xylem, which explains why rice is efficient in arsenic accumulation.
Abstract: Arsenic poisoning affects millions of people worldwide. Human arsenic intake from rice consumption can be substantial because rice is particularly efficient in assimilating arsenic from paddy soils, although the mechanism has not been elucidated. Here we report that two different types of transporters mediate transport of arsenite, the predominant form of arsenic in paddy soil, from the external medium to the xylem. Transporters belonging to the NIP subfamily of aquaporins in rice are permeable to arsenite but not to arsenate. Mutation in OsNIP2;1 (Lsi1, a silicon influx transporter) significantly decreases arsenite uptake. Furthermore, in the rice mutants defective in the silicon efflux transporter Lsi2, arsenite transport to the xylem and accumulation in shoots and grain decreased greatly. Mutation in Lsi2 had a much greater impact on arsenic accumulation in shoots and grain in field-grown rice than Lsi1. Arsenite transport in rice roots therefore shares the same highly efficient pathway as silicon, which explains why rice is efficient in arsenic accumulation. Our results provide insight into the uptake mechanism of arsenite in rice and strategies for reducing arsenic accumulation in grain for enhanced food safety.
1,181 citations
TL;DR: A range of mitigation methods, from agronomic measures and plant breeding to genetic modification, may be employed to reduce As uptake by food crops.
Abstract: Arsenic (As) is an environmental and food chain contaminant. Excessive accumulation of As, particularly inorganic arsenic (As(i)), in rice (Oryza sativa) poses a potential health risk to populations with high rice consumption. Rice is efficient at As accumulation owing to flooded paddy cultivation that leads to arsenite mobilization, and the inadvertent yet efficient uptake of arsenite through the silicon transport pathway. Iron, phosphorus, sulfur, and silicon interact strongly with As during its route from soil to plants. Plants take up arsenate through the phosphate transporters, and arsenite and undissociated methylated As species through the nodulin 26-like intrinsic (NIP) aquaporin channels. Arsenate is readily reduced to arsenite in planta, which is detoxified by complexation with thiol-rich peptides such as phytochelatins and/or vacuolar sequestration. A range of mitigation methods, from agronomic measures and plant breeding to genetic modification, may be employed to reduce As uptake by food crops.
1,025 citations
TL;DR: Hyperaccumulation appears to involve enhanced arsenate uptake, decreased arsenite-thiol complexation and arsenite efflux to the external medium, greatly enhanced xylem translocation of arsenite, and vacuolar sequestration of arsenites in fronds.
Abstract: Arsenic (As) is an element that is nonessential for and toxic to plants. Arsenic contamination in the environment occurs in many regions, and, depending on environmental factors, its accumulation in food crops may pose a health risk to humans.Recent progress in understanding the mechanisms of As uptake and metabolism in plants is reviewed here. Arsenate is taken up by phosphate transporters. A number of the aquaporin nodulin26-like intrinsic proteins (NIPs) are able to transport arsenite,the predominant form of As in reducing environments. In rice (Oryza sativa), arsenite uptake shares the highly efficient silicon (Si) pathway of entry to root cells and efflux towards the xylem. In root cells arsenate is rapidly reduced to arsenite, which is effluxed to the external medium, complexed by thiol peptides or translocated to shoots. One type of arsenate reductase has been identified, but its in planta functions remain to be investigated. Some fern species in the Pteridaceae family are able to hyperaccumulate As in above-ground tissues. Hyperaccumulation appears to involve enhanced arsenate uptake, decreased arsenite-thiol complexation and arsenite efflux to the external medium, greatly enhanced xylem translocation of arsenite, and vacuolar sequestration of arsenite in fronds. Current knowledge gaps and future research directions are also identified.
1,012 citations