Dissecting the promotional effect of zinc on cadmium translocation from roots to shoots in rice.

It is often expected that Zn decreases Cd accumulation in plants due to competition for the same transporters. Here, we found that increasing Zn supply markedly increased the root-to-shoot translocation of Cd in rice. RNA sequencing showed that high Zn up-regulated expression of genes involved in glutathione biosynthesis and metabolism and the Zn/Cd transporter gene OsHMA2, but down-regulated expression of genes related to Zn uptake. Knockout of the iron or Zn transporter genes OsIRT1, OsIRT2, or OsZIP9 did not affect the Zn promotional effect on Cd translocation. Knockout of the manganese/Cd transporter gene OsNRAMP5 greatly reduced Cd uptake but did not affect the Zn promotional effect. Variation in the tonoplast transporter gene OsHMA3 affected Cd translocation but did not change the Zn promotional effect. Knockout of the Zn/Cd transporter gene OsHMA2 not only decreased Cd and Zn translocation, but also abolished the Zn promotional effect. Increased expression of OsHMA2 under high Zn conditions supports the hypothesis that this transporter participates in the promotional effect of Zn on Cd translocation. The results also show that OsIRT1, OsIRT2, and OsZIP9 made only small contributions to Cd uptake under low Zn conditions but not under high Zn conditions, whereas the dominant role of OsNRAMP5 in Cd uptake diminished under low Zn conditions.

Molecular mechanisms underlying the toxicity and detoxification of trace metals and metalloids in plants.

Plants take up a wide range of trace metals/metalloids (hereinafter referred to as trace metals) from the soil, some of which are essential but become toxic at high concentrations (e.g., Cu, Zn, Ni, Co), while others are non-essential and toxic even at relatively low concentrations (e.g., As, Cd, Cr, Pb, and Hg). Soil contamination of trace metals is an increasing problem worldwide due to intensifying human activities. Trace metal contamination can cause toxicity and growth inhibition in plants, as well as accumulation in the edible parts to levels that threatens food safety and human health. Understanding the mechanisms of trace metal toxicity and how plants respond to trace metal stress is important for improving plant growth and food safety in contaminated soils. The accumulation of excess trace metals in plants can cause oxidative stress, genotoxicity, programmed cell death, and disturbance in multiple physiological processes. Plants have evolved various strategies to detoxify trace metals through cell-wall binding, complexation, vacuolar sequestration, efflux, and translocation. Multiple signal transduction pathways and regulatory responses are involved in plants challenged with trace metal stresses. In this review, we discuss the recent progress in understanding the molecular mechanisms involved in trace metal toxicity, detoxification, and regulation, as well as strategies to enhance plant resistance to trace metal stresses and reduce toxic metal accumulation in food crops.

OsNRAMP5 Is a Major Transporter for Lead Uptake in Rice.

Lead (Pb) is one of the most toxic metals affecting human health globally. Food is an important source of chronic Pb exposure in humans. How Pb is taken up by rice, a staple food for over half of the global population, remains unknown. In the present study, we investigated the role of OsNRAMP5, a member of the NRAMP (Natural Resistance-Associated Macrophage Protein) transporter family, in Pb uptake by rice roots. Heterologous expression of OsNRAMP5 in yeast increased Pb uptake and sensitivity toward Pb. Knockout of OsNRAMP5 in rice by CRISPR/Cas9 gene editing resulted in significant decreases in root uptake of Pb and accumulation in rice shoots. The maximum influx velocity (Vmax) for Pb uptake of the knockout mutants was 70% lower than that of wild-type plants. When grown in Pb-contaminated paddy soil, OsNRAMP5 knockout mutants accumulated approximately 50 and 70% lower Pb concentrations in the grain and straw, respectively, than the wild type. OsNRAMP5 expression in rice roots was not affected by Pb exposure. These results indicate that OsNRAMP5 is a major transporter for Pb uptake in rice, in addition to its role in the uptake of manganese and cadmium. This study provides a mechanistic understanding of Pb uptake in rice plants and a potential strategy to limit Pb accumulation in rice grains.

OsNRAMP1 transporter contributes to cadmium and manganese uptake in rice.

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.

Overexpression of the manganese/cadmium transporter OsNRAMP5 reduces cadmium accumulation in rice grain.

Rice is a major dietary source of the toxic metal cadmium (Cd), and reducing its accumulation in the grain is therefore important for food safety. We selected two cultivars with contrasting Cd accumulation and generated transgenic lines overexpressing OsNRAMP5, which encodes a major influx transporter for manganese (Mn) and Cd. We used two different promoters to control the expression, namely OsActin1 and maize Ubiquitin. Overexpression of OsNRAMP5 increased Cd and Mn uptake into the roots, but markedly decreased Cd accumulation in the shoots, whilst having a relatively small effect on Mn accumulation in the shoots. The overexpressed OsNRAMP5 protein was localized to the plasma membrane of all cell types in the root tips and lateral root primordia without polarity. Synchrotron X-ray fluorescence mapping showed that the overexpression lines accumulated more Cd in the root tips and lateral root primordia compared with the wild-type. When grown in three Cd-contaminated paddy soils, overexpression of OsNRAMP5 decreased concentration of Cd in the grain by 49-94% compared with the wild type. OsNRAMP5-overexpression plants had decreased Cd translocation from roots to shoots as a result of disruption of its radial transport into the stele for xylem loading, demonstrating the effect of transporter localization and polarity on ion homeostasis.

Producing Cd-safe rice grains in moderately and seriously Cd-contaminated paddy soils.

Rice grains produced on cadmium (Cd) contaminated paddy soils often exceed the maximum permissible limit. A number of mitigation methods have been proposed to decrease Cd accumulation in rice grain in contaminated acidic soils, including altering water management regimes, liming, and genetic engineering. In the present study, we conducted a pot experiment to compare these methods for their effectiveness at decreasing grain Cd concentrations in both acidic (pH 5.1-5.2) and alkaline (pH 7.5-7.9) paddy soils that varied in the degree of Cd contamination. In moderately Cd-contaminated acidic soils (with Cd concentrations lower than the intervention value of Chinese soil standard, GB15618-2018), any of the three methods was effective, reducing grain Cd concentration by 80-90% to levels below the Chinese maximum permissible limit (0.2 mg/kg). However, in the highly Cd-contaminated soils (with soil Cd concentrations exceeding the intervention value) with elevated concentrations of extractable Cd, although both liming and alternation of the water management regime (continuous flooding) was effective at decreasing grain Cd accumulation, grain Cd concentrations still exceeded the Chinese limit. Genetic engineering of rice, such as knockout of OsNramp5 (encoding the plasma membrane transporter responsible for Cd uptake into root cells) or overexpression of OsHMA3 (encoding a tonoplast Cd transporter sequestering Cd into the vacuoles), produced dramatic decreases (≥90%) in grain Cd concentration. Even in seriously contaminated soils, overexpression of OsHMA3 alone produced grain with Cd concentrations below the Chinese limit, offering a highly effective approach to produce Cd-safe rice especially in seriously Cd-contaminated paddy soils without affecting grain biomass or the concentrations of essential micronutrients.

Effects of elevated CO2 and temperature on Gynostemma pentaphyllum physiology and bioactive compounds.

Recently, an important topic of research has been how climate change is seriously threatening the sustainability of agricultural production. However, there is surprisingly little experimental data regarding how elevated temperature and CO2 will affect the growth of medicinal plants and production of bioactive compounds. Here, we comprehensively analyzed the effects of elevated CO2 and temperature on the photosynthetic process, biomass, total sugars, antioxidant compounds, antioxidant capacity, and bioactive compounds of Gynostemma pentaphyllum. Two different CO2 concentrations [360 and 720µmolmol(-1)] were imposed on plants grown at two different temperature regimes of 23/18 and 28/23°C (day/night) for 60days. Results show that elevated CO2 and temperature significantly increase the biomass, particularly in proportion to inflorescence total dry weight. The chlorophyll content in leaves increased under the elevated temperature and CO2. Further, electron transport rate (ETR), photochemical quenching (qP), actual photochemical quantum yield (Yield), instantaneous photosynthetic rate (Photo), transpiration rate (Trmmol) and stomatal conductance (Cond) also increased to different degrees under elevated CO2 and temperature. Moreover, elevated CO2 increased the level of total sugars and gypenoside A, but decreased the total antioxidant capacity and main antioxidant compounds in different organs of G. pentaphyllum. Accumulation of total phenolics and flavonoids also decreased in leaves, stems, and inflorescences under elevated CO2 and temperature. Overall, our data indicate that the predicted increase in atmospheric temperature and CO2 could improve the biomass of G. pentaphyllum, but they would reduce its health-promoting properties.