Lanthanum interferes with the fundamental rhythms of stomatal opening, expression of related genes, and evapotranspiration in Arabidopsis thaliana.

The accumulation of rare earth elements (REEs) in the global environment poses a threat to plant health and ecosystem stability. Stomata located on leaves serve as the primary site for plant responses to REE-related threats. This study focused on lanthanum [La(III)], a prevalent REE in the atmospheric environment. Using interdisciplinary techniques, it was found that La(III) (≤80 µM) interfered with the fundamental rhythms of stomatal opening, related gene expression, and evapotranspiration in plants. Specifically, when exposed to low concentrations of La(III) (15 and 30 µM), the expression levels of six genes were increased, stomatal opening was enhanced, and the evapotranspiration rate was accelerated. The interference on stomatal rhythms was enhanced with higher concentrations of La(III) (60 and 80 µM), increasing the expression levels of six genes, stomatal opening, and evapotranspiration rate. To counter the interference of low concentrations of La(III) (15 and 30 µM), plants accelerated nutrient replenishment through La(III)-induced endocytosis, which the redundant nutrients enhanced photosynthesis. However, replenished nutrients failed to counter the disruption of plant biological rhythms at higher concentrations of La(III) (60 and 80 µM), thus inhibiting photosynthesis due to nutrient deficit. The interference of La(III) on these biological rhythms negatively affected plant health and ecosystem stability.

Early-life noise exposure causes cognitive impairment in a sex-dependent manner by disrupting homeostasis of the microbiota-gut-brain axis.

Epidemiological investigations show that noise exposure in early life is associated with health and cognitive impairment. The gut microbiome established in early life plays a crucial role in modulating developmental processes that subsequently affect brain function and behavior. Here, we examined the impact of early-life exposure to noise on cognitive function in adolescent rats by analyzing the gut microbiome and metabolome to elucidate the underlying mechanisms. Chronic noise exposure during early life led to cognitive deficits, hippocampal injury, and neuroinflammation. Early-life noise exposure showed significant difference on the composition and function of the gut microbiome throughout adolescence, subsequently causing axis-series changes in fecal short-chain fatty acid (SCFA) metabolism and serum metabolome profiles, as well as dysregulation of endothelial tight junction proteins, in both intestine and brain. We also observed sex-dependent effects of microbiota depletion on SCFA-related beneficial bacteria in adolescence. Experiments on microbiota transplantation and SCFA supplementation further confirmed the role of intestinal bacteria and related SCFAs in early-life noise-exposure-induced impairments in cognition, epithelial integrity, and neuroinflammation. Overall, these results highlight the homeostatic imbalance of microbiota-gut-brain axis as an important physiological response toward environmental noise during early life and reveals subtle differences in molecular signaling processes between male and female rats.

Arabinogalactan protein-rare earth element complexes activate plant endocytosis.

Endocytosis is essential to all eukaryotes, but how cargoes are selected for internalization remains poorly characterized. Extracellular cargoes are thought to be selected by transmembrane receptors that bind intracellular adaptors proteins to initiate endocytosis. Here, we report a mechanism for clathrin-mediated endocytosis (CME) of extracellular lanthanum [La(III)] cargoes, which requires extracellular arabinogalactan proteins (AGPs) that are anchored on the outer face of the plasma membrane. AGPs were colocalized with La(III) on the cell surface and in La(III)-induced endocytic vesicles in Arabidopsis leaf cells. Superresolution imaging showed that La(III) triggered AGP movement across the plasma membrane. AGPs were then colocalized and physically associated with the µ subunit of the intracellular adaptor protein 2 (AP2) complexes. The AGP-AP2 interaction was independent of CME, whereas AGP's internalization required CME and AP2. Moreover, we show that AGP-dependent endocytosis in the presence of La(III) also occurred in human cells. These findings indicate that extracellular AGPs act as conserved CME cargo receptors, thus challenging the current paradigm about endocytosis of extracellular cargoes.

Low-dose lanthanum activates endocytosis, aggravating accumulation of lanthanum or/and lead and disrupting homeostasis of essential elements in the leaf cells of four edible plants.

Rare earth elements (REEs) are emerging as a serious threat to ecological safety due to their increasing accumulation in environments. The accumulation of REEs in environments has significantly increased its accumulation in the leaves of edible plants. However, the accumulation pathway of REEs in the leaves of edible plants are still unknown. In this study, lanthanum [La(III), a widely used and accumulated REE] and four edible plants (soybean, lettuce, pakchoi, and celery) with short growth cycles were selected as research objects. By using interdisciplinary research techniques, we found that low-dose La(III) activated endocytosis (mainly the clathrin-mediated endocytosis) in the leaf cells of four edible plants, which provided an accumulation pathway for low-dose La in the leaf cells of these edible plants. The accumulation of La in the leaf cells was positively correlated with the intensity of endocytosis, while the intensity of endocytosis was negatively correlated with the density of leaf trichomes. In addition to the accumulation of La, low-dose La(III) also brought other risks. For example, the harmful element (Pb) can also be accumulated in the leaf cells via La(III)-activated endocytosis; the homeostasis of the essential elements (K, Ca, Fe, Mg) was disrupted, although the chlorophyll synthesis and the growth of these leaf cells were accelerated; and the expression of stress response genes (GmNAC20, GmNAC11) in soybean leaves was increased. These results provided an insight to further analyze the toxicity and mechanism of REEs in plants, and sounded the alarm for the application of REEs in agriculture.

A detection method in living plant cells for rapidly monitoring the response of plants to exogenous lanthanum.

The pollution of rare earth elements (REEs) in ecosystem is becoming more and more serious, so it is urgent to establish methods for monitoring the pollution of REEs. Monitoring environmental pollution via the response of plants to pollutants has become the most stable and accurate method compared with traditional methods, but scientists still need to find the primary response of plants to pollutants to improve the sensitivity and speed of this method. Based on the facts that the initiation of endocytosis is the primary cellular response of the plant leaf cells to REEs and the detection of endocytosis is complex and expensive, we constructed a detection method in living plant cells for rapidly monitoring the response of plants to exogenous lanthanum [La(III), a representative of REEs] by designing a new immuno-electrochemical method for detecting the content change in extracellular vitronectin-like protein (VN) that are closely related to endocytosis. Results showed that when 30 µM La(III) initiated a small amount of endocytosis, the content of extracellular VN increased by 5.46 times, but the structure and function of plasma membrane were not interfered by La(III); when 80 µM La(III) strongly initiated a large amount of endocytosis, the content of extracellular VN increased by 119 times, meanwhile, the structure and function of plasma membrane were damaged. In summary, the detection method can reflect the response of plants to La(III) via detecting the content change in extracellular VN, which provides an effective and convenient way to monitor the response of plants to exogenous REEs.

The stimulatory effect and mechanism of low-dose lanthanum on soybean leaf cells.

Light rare earth elements (LREEs) have been long used in agriculture (i.e., mainly via aerially applied LREE fertilizers) based on the fact that low-dose LREEs promote plant growth. Meanwhile, the toxic effects of low-dose LREEs on organisms have also been found. However, the cellular and molecular mechanism of low-dose LREEs acting on organisms remain unclear. Plants are at the beginning of food chains, so it is critical to uncover the cellular and molecular mechanism of low-dose LREEs on plants. Here, lanthanum (La) and soybean were the representatives of LREEs and plants, respectively. The effects of low-dose La on soybean leaves were investigated, and the stimulatory effect and mechanism of low-dose LREEs on leaf cells were revealed. Specifically, clathrin-mediated endocytosis (CME) activated by low-dose La is an influx channel for La in soybean leaf cells. The intracellular La and La-activated CME jointly disturbed multiple forms of intracellular homeostasis, including metallic element homeostasis, redox homeostasis, gene expression homeostasis. The disturbed homeostasis either stimulated cell growth or caused damage to the plasma membrane of soybean leaf cells. These results provide new insights for clarifying the cellular and molecular mechanisms of low-dose LREEs as a class of stimulators instead of nutrients to stimulate plants.

Sharply and simultaneously increasing pollutant accumulations in cells of organisms induced by rare earth elements in the environment of Nanjing.

Exploring the factors that simultaneously increase the accumulation of various pollutants in cells of organisms to restrict the toxic effects of pollutants on organisms has become a focus of research aimed at protecting ecosystems. Here, we found that the accumulation of organic [e.g., benzo(a)pyrene (BaP)], inorganic [e.g., cadmium (Cd)] and emerging [e.g., rare earth elements (REEs)] pollutants in leaf cells of different plants grown in Nanjing was 567-1022%, 547-922% and 972-1392% of those grown in Haikou, respectively, when the concentration of REEs in rainwater of Nanjing and Haikou was 4.31 × 10-3 µg/L and 3.04 × 10-6 µg/L. Unprecedentedly, endocytosis in leaf cells of different plants grown in Nanjing was activated by REEs, and then extracellular BaP, Cd and REEs (e.g. terbium) were transported into these leaf cells together via endocytic vesicles. Particularly, the co-accumulation of those pollutants in these leaf cells was sharply increased, thus magnifying their toxic effects on these plants. Furthermore, the co-accumulation of those pollutants in human cells was also significantly increased by REEs, in a similar way to these leaf cells. Therefore, REEs in environments are key factors that greatly increase the co-accumulation of various pollutants in cells of organisms. These results provide new insights into how pollutants are accumulated in cells of organisms in ecosystems, informing a reference for making policy to ensure the safety of ecosystems.

Cerium exposure in Lake Taihu water aggravates microcystin pollution via enhancing endocytosis of Microcystis aeruginosa.

Aggravating the pollution of microcystins (MCs) in freshwater environments is detrimental to aquatic living organisms and humans, and thus threatens the stability of ecosystems. Some environmental factors have been verified to promote the production of MCs in Microcystis aeruginosa, thereby aggravating the pollution of MCs. However, the effects of cerium (Ce), the most abundant rare earth element in global water environments, on the production of MCs in M. aeruginosa are unknown. Here, Lake Taihu water was selected as a representative of freshwater environments. By using interdisciplinary methods, it was found that: (1) the exposure level of Ce [Ce(III) and Ce(IV)] in Lake Taihu water is in the range of 0.271-0.282 µg/L; (2) Ce exposure in Lake Taihu water promoted the contents of three main MCs (MC-LR, MC-LW and MC-YR) in M. aeruginosa and water; (3) a cellular mechanism of Ce promoting the production of MCs in M. aeruginosa in Lake Taihu water was suggested: Ce enhanced endocytosis in cells of M. aeruginosa to promote the essential element uptake by M. aeruginosa for MC synthesis. Thus, Ce exposure in Lake Taihu water aggravates the pollution of MCs via enhancing endocytosis in cells of M. aeruginosa. The results provide reference for assessing the environmental risk of Ce in water environments, investigating the mechanism of the pollution of MCs induced by environmental factors, and developing strategies aimed at preventing and controlling the pollution of MCs.

A new mechanism by which environmental hazardous substances enhance their toxicities to plants.

The coexistence of hazardous substances enhances their toxicities to plants, but its mechanism is still unclear due to the unknown cytochemical behavior of hazardous substance in plants. In this study, by using interdisciplinary methods, we observed the cytochemical behavior of coexisting hazardous substances {terbium [Tb(III)], benzo(a)pyrene (BaP) and cadmium [Cd(II)] in environments} in plants and thus identified a new mechanism by which coexisting hazardous substances in environments enhance their toxicities to plants. First, Tb(III) at environmental exposure level (1.70 × 10-10 g/L) breaks the inert rule of clathrin-mediated endocytosis (CME) in leaf cells. Specifically, Tb(III) binds to its receptor [FASCICLIN-like arabinogalactan protein 17 (FLA17)] on the plasma membrane of leaf cells and then docks to an intracellular adaptor protein [adaptor protein 2 (AP2)] to form ternary complex [Tb(III)-FLA17-AP2], which finally initiates CME pathway in leaf cells. Second, coexisting Tb(III), BaP and Cd(II) in environments are simultaneously transported into leaf cells via Tb(III)-initiated CME pathway, leading to the accumulation of them in leaf cells. Finally, these accumulated hazardous substances simultaneously poison plant leaf cells. These results provide theoretical and experimental bases for elucidating the mechanisms of hazardous substances in environments poisoning plants, evaluating their risks, and protecting ecosystems.

Lanthanum(III) triggers AtrbohD- and jasmonic acid-dependent systemic endocytosis in plants.

Trivalent rare earth elements (REEs) are widely used in agriculture. Aerially applied REEs enter leaf epidermal cells by endocytosis and act systemically to improve the growth of the whole plant. The mechanistic basis of their systemic activity is unclear. Here, we show that treatment of Arabidopsis leaves with trivalent lanthanum [La(III)], a representative of REEs, triggers systemic endocytosis from leaves to roots. La(III)-induced systemic endocytosis requires AtrbohD-mediated reactive oxygen species production and jasmonic acid. Systemic endocytosis impacts the accumulation of mineral elements and the development of roots consistent with the growth promoting effects induced by aerially applied REEs. These findings provide insights into the mechanistic basis of REE activity in plants.

A living plant cell-based biosensor for real-time monitoring invisible damage of plant cells under heavy metal stress.

Heavy metals inevitably cause invisible or visible damage to plants, leading to significant economic losses. Therefore, it is necessary to develop a method for timely monitoring the damage of plants under the stress of heavy metals. Here, vitronectin-like proteins (VN) on the surface of plant cells is as an important biomarker for monitoring damage of plants under the stress of heavy metals. A living plant cell-based biosensor is constructed to monitor invisible damage of plant cells induced by cadmium [Cd(II)] or lead [Pb(II)]. To fabricate this sensor, l-cysteine was first modified on the glassy carbon electrode followed by the modification of anti-IgG-Au antibody. Then, the living plant cells, incubated with the anti-VN, were modified onto the electrode. The sensor worked by determining the change in electrochemical impedance. Cd(II) and Pb(II) was detected in the linear dynamic range of 45-210 and 120-360 µmol·L-1, respectively. And the detection limit of Cd(II) and Pb(II) of this biosensor was 18.5 nmol·L-1 [with confidence interval (95%) 18.4-18.6 nmol·L-1] and 25.6 nmol·L-1 [with confidence interval (95%) 25.4-25.8 nmol·L-1], respectively. In both Arabidopsis and soybean, when the content of VN increased by about 20 times under the stress of Cd(II) or Pb(II), which means when the electron-transfer resistance increased by 35%, chlorophyll content showed significant decrease about 17%. Therefore, by establishing a quantitative relationship among the content of biomarker, the electron-transfer resistance and chlorophyll content in plant cells, the invisible damage of plants under the stress of heavy metals was detected. These results can provide a reference method for early-onset warning systems for heavy metal pollution in the environment.

Combined acid rain and lanthanum pollution and its potential ecological risk for nitrogen assimilation in soybean seedling roots.

Rare earth elements (REEs) are used in various fields, resulting in their accumulation in the environment. This accumulation has affected the survival and distribution of crops in various ways. Acid rain is a serious global environmental problem. The combined effects on crops from these two types of pollution have been reported, but the effects on crop root nitrogen assimilation are rarely known. To explore the impact of combined contamination from these two pollutants on crop nitrogen assimilation, the soybean seedlings were treated with simulated environmental pollution from acid rain and a representative rare earth ion, lanthanum ion (La3+), then the indexes related to plant nitrogen assimilation process in roots were determined. The results showed that combined treatment with pH 4.5 acid rain and 0.08 mM La3+ promoted nitrogen assimilation synergistically, while the other combined treatments all showed inhibitory effects. Moreover, acid rain aggravated the inhibitory effect of 1.20 or 0.40 mM La3+ on nitrogen assimilation in soybean seedling roots. Thus, the effects of acid rain and La3+ on crops depended on the combination levels of acid rain intensity and La3+ concentration. Acid rain increases the bioavailability of La3+, and the combined effects of these two pollutants were more serious than that of either pollutant alone. These results provide new evidence in favor of limiting overuse of REEs in agriculture. This work also provides a new framework for ecological risk assessment of combined acid rain and REEs pollution on soybean crops.

Toxic effects of environmental rare earth elements on delayed outward potassium channels and their mechanisms from a microscopic perspective.

The wide applications cause a large amount of rare earth elements (REEs) to be released into the environment, and ultimately into the human body through food chain. Toxic effects of REEs on humans have been extensively studied, but their toxic effects and binding targets in cells are not understood. Delayed outward potassium channels (K+ channels) are good targets for exogenous substances or clinical drugs. To evaluate cellular toxicities of REEs and clarify toxic mechanisms, the toxicities of REEs on the K+ channel and their structural basis were investigated. The results showed that delayed outward potassium channels on the plasma membrane are the targets of REEs acting on living organisms, and the changes in the thermodynamic and kinetic characteristics of the K+ channel are the reasons of diseases induced by REEs. Two types of REEs, a light REE La3+ and a heavy REE Tb3+, displayed different intensity of toxicities on the K+ channel, in which the toxicity of Tb3+ was stronger than that of La3+. More interestingly, in comparison with that of heavy metal Cd2+, the cytotoxicities of the light and heavy REEs showed discriminative differences, and the cytotoxicity of Tb3+ was higher than that of Cd2+, while the cytotoxicity of La3+ was lower than that of Cd2+. These different cytotoxicities of La3+, Tb3+ and Cd2+ on human resulted from the varying binding abilities of the metals to this channel protein.

Responses of plant calmodulin to endocytosis induced by rare earth elements.

The wide application of rare earth elements (REEs) have led to their diffusion and accumulation in the environment. The activation of endocytosis is the primary response of plant cells to REEs. Calmodulin (CaM), as an important substance in calcium (Ca) signaling systems, regulating almost all of the physiological activities in plants, such as cellular metabolism, cell growth and division. However, the response of CaM to endocytosis activated by REEs remains unknown. By using immunofluorescence labeling and a confocal laser scanning microscope, we found that trivalent lanthanum [La(III)], an REE ion, affected the expression of CaM in endocytosis. Using circular dichroism, X-ray photoelectron spectroscopy and computer simulations, we demonstrated that a low concentration of La(III) could interact with extracellular CaM by electrostatic attraction and was then bound to two Ca-binding sites of CaM, making the molecular structure more compact and orderly, whereas a high concentration of La(III) could be coordinated with cytoplasmic CaM or bound to other Ca-binding sites, making the molecular structure more loose and disorderly. Our results provide a reference for revealing the action mechanisms of REEs in plant cells.