Pre-Harvest MeJA Application Counteracts the Deleterious Impact of Al and Mn Toxicity in Highbush Blueberry Grown in Acid Soils

Volcanic ash-derived soils are characterized by low pH (pH ≤ 5.5) with increased concentrations of aluminum (Al3+) and manganese (Mn2+), which decreases plant growth, fruit quality, and yield. Methyl jasmonate (MeJA) improves abiotic stress tolerance. Our work aimed to evaluate the application of MeJA’s impact on the growth, antioxidant defense, and fruit quality of highbush blueberry grown under Al and Mn toxicity. A field assay was conducted with four-year-old bushes of highbush blueberry cultivar Legacy under eight treatments (Control, Al (87% of Al saturation), Mn (240 mg kg−1), and Al–Mn with and without MeJA application). Physiological, biochemical, and fruit quality parameters were measured. Growth rate significantly decreased with Al (20%), Mn (45%), and Al–Mn (40%). MeJA application recovered the growth rate. Photosynthetic parameters were not affected. Antioxidant activity increased under all treatments compared with controls, being higher with MeJA application. Total phenols (TP) were decreased in plants under Al (43%) and Mn (20%) compared with controls. MeJA application increased TP in all treatments. Fruits of bushes under Al and Mn toxicity with MeJA applications exhibited an increase in fruit firmness and weight, maintaining suitable contents of soluble solids. Our results provide insights about the beneficial effect of MeJA application on growth, antioxidant properties, and fruit quality of highbush blueberry plants grown in acid soils under Al and Mn toxicity.

Characterization of Metal Tolerance Proteins and Functional Analysis of GmMTP8.1 Involved in Manganese Tolerance in Soybean

Manganese is an essential micronutrient for plant growth but can be toxic to plants when it reaches excessive levels. Although metal tolerance proteins (MTPs), which belong to the cation diffusion facilitator (CDF) family, have been demonstrated to play critical roles in manganese (Mn) tolerance in plants, the characteristics and functions of GmMTP members in the response of soybean (Glycine max) to Mn toxicity have not been documented. In this study, growth inhibition was observed in soybean plants that were exposed to a toxic level of Mn in hydroponics, as reflected by the generation of brown spots, and decreased leaf chlorophyll concentration and plant fresh weight. Subsequent genome-wide analysis resulted in the identification of a total of 14 GmMTP genes in the soybean genome. Among these GmMTPs, 9 and 12 were found to be regulated by excess Mn in leaves and roots, respectively. Furthermore, the function of GmMTP8.1, a Mn-CDF homologue of ShMTP8 identified in the legume Stylosanthes hamata that is involved in Mn detoxification, was characterized. Subcellular localization analysis showed that GmMTP8.1 was localized to the endoplasmic reticulum (ER). Heterologous expression of GmMTP8.1 led to the restoration of growth of the Mn-hypersensitive yeast (Saccharomyces cerevisiae) mutant Δpmr1, which is made defective in Mn transport into the Golgi apparatus by P-type Ca/Mn-ATPase. Furthermore, GmMTP8.1 overexpression conferred tolerance to the toxic level of Mn in Arabidopsis (Arabidopsis thaliana). Under excess Mn conditions, concentrations of Mn in shoots but not roots were decreased in transgenic Arabidopsis, overexpressing GmMTP8.1 compared to the wild type. The overexpression of GmMTP8.1 also led to the upregulation of several transporter genes responsible for Mn efflux and sequestration in Arabidopsis, such as AtMTP8/11. Taken together, these results suggest that GmMTP8.1 is an ER-localized Mn transporter contributing to confer Mn tolerance by stimulating the export of Mn out of leaf cells and increasing the sequestration of Mn into intracellular compartments.

Effects of manganese toxicity on the growth of soybean (Glycine max L.) at the seedling stage

Effects of manganese (Mn) toxicity stress on the growth of soybean, the number of Mn spots on leaves and the absorption of iron and magnesium were studied by nutrient solution hydroponics. The results showed that the presence of Mn spots on leaves was the main symptom of Mn toxicity in soybean. When the concentration of exogenous Mn was 25 μmol/l, the leaf generated obvious Mn oxidation spots; when the concentration of exogenous Mn exceeded 50 μmol/l, the growth of soybean was inhibited, and the number of Mn spots increased significantly. With the increase in exogenous Mn concentration, the Mn concentration in the roots, young leaves and old leaves of soybean increased significantly. When the concentration of exogenous Mn reached 200 μmol/l, the number of Mn spots on primary leaves, old leaves and young leaves increased significantly. Although the iron concentration in the roots remained the same, the iron content in the old and young leaves decreased significantly. On the other hand, although Mn toxicity significantly reduced the concentration of magnesium in soybean roots, it increased the concentration of magnesium in old and young leaves. Bangladesh J. Bot. 50(3): 803-811, 2021 (September) Special

Manganese toxicity disrupts indole acetic acid homeostasis and suppresses the CO2 assimilation reaction in rice leaves

Despite the essentiality of Mn in terrestrial plants, its excessive accumulation in plant tissues can cause growth defects, known as Mn toxicity. Mn toxicity can be classified into apoplastic and symplastic types depending on its onset. Symplastic Mn toxicity is hypothesised to be more critical for growth defects. However, details of the relationship between growth defects and symplastic Mn toxicity remain elusive. In this study, we aimed to elucidate the molecular mechanisms underlying symplastic Mn toxicity in rice plants. We found that under excess Mn conditions, CO2 assimilation was inhibited by stomatal closure, and both carbon anabolic and catabolic activities were decreased. In addition to stomatal dysfunction, stomatal and leaf anatomical development were also altered by excess Mn accumulation. Furthermore, indole acetic acid (IAA) concentration was decreased, and auxin-responsive gene expression analyses showed IAA-deficient symptoms in leaves due to excess Mn accumulation. These results suggest that excessive Mn accumulation causes IAA deficiency, and low IAA concentrations suppress plant growth by suppressing stomatal opening and leaf anatomical development for efficient CO2 assimilation in leaves.

Management of Iron and Manganese Toxicities of Lentil Crops Grown in Central Chile

Iron (Fe) and manganese (Mn) toxicity is a widespread problem in lentil production in the coastal dryland of Chile. Increasing the soil pH by liming with CaCO3 or incrementing grain yields through nitrogen fertilization can help the plants to reduce metal concentration. Thus, the main objective of this work was to evaluate two different fertilization strategies (lime (CaCO3) and nitrogen (N) additions) to reduce Fe and Mn toxicities in lentils. Lentils grown under field conditions with the highest Fe and Mn concentrations showed toxicity symptoms, but without grain yield reductions. In a pot experiment using the same soil as in the field with toxicity symptoms, the dry matter (DM) produced at the end of the trial was higher in the plants that received N while the lowest DM production was recorded in those plants treated with lime. In particular, higher root DM sustained the growth of the N-fertilized shoots, which also positively affected the grain yields being 33% higher than the control treatment (no fertilization addition). In the plants fertilized with N, the Fe and Mn levels in the shoots were lower than the control plants and those grown in soils treated with lime, but showed higher concentrations of Fe and Mn in roots. In parallel, roots exhibited high concentrations of Fe and Mn that were 13- and 9-fold higher than in the shoots. Additionally, a significant decrease of 29% in Mn concentration in the grains of plants treated with N was reported. Overall, our results suggest that an increase in DM of lentils by the addition of N can reduce the Mn concentration on leaves to a level that is likely under the threshold that causes toxicity in plant tissues. Finally, we conclude that the increase of Fe and Mn in the roots may be connected to the reduction of these metals on leaves.

Effects of Manganese on Genomic Integrity in the Multicellular Model Organism Caenorhabditis elegans

Although manganese (Mn) is an essential trace element, overexposure is associated with Mn-induced toxicity and neurological dysfunction. Even though Mn-induced oxidative stress is discussed extensively, neither the underlying mechanisms of the potential consequences of Mn-induced oxidative stress on DNA damage and DNA repair, nor the possibly resulting toxicity are characterized yet. In this study, we use the model organism Caenorhabditis elegans to investigate the mode of action of Mn toxicity, focusing on genomic integrity by means of DNA damage and DNA damage response. Experiments were conducted to analyze Mn bioavailability, lethality, and induction of DNA damage. Different deletion mutant strains were then used to investigate the role of base excision repair (BER) and dePARylation (DNA damage response) proteins in Mn-induced toxicity. The results indicate a dose- and time-dependent uptake of Mn, resulting in increased lethality. Excessive exposure to Mn decreases genomic integrity and activates BER. Altogether, this study characterizes the consequences of Mn exposure on genomic integrity and therefore broadens the molecular understanding of pathways underlying Mn-induced toxicity. Additionally, studying the basal poly(ADP-ribosylation) (PARylation) of worms lacking poly(ADP-ribose) glycohydrolase (PARG) parg-1 or parg-2 (two orthologue of PARG), indicates that parg-1 accounts for most of the glycohydrolase activity in worms.

Gut Microbiota as a Potential Player in Mn-Induced Neurotoxicity

Manganese (Mn) is an essential metal, which at high exposures causes neurotoxic effects and neurodegeneration. The neurotoxic effects of Mn are mediated by neuroinflammation, oxidative and endoplasmic reticulum stress, mitochondrial dysfunction, and other mechanisms. Recent findings have demonstrated the potential impact of Mn overexposure on gut microbiota dysbiosis, which is known to contribute to neurodegeneration via secretion of neuroactive and proinflammatory metabolites. Therefore, in this review, we discuss the existing data on the impact of Mn exposure on gut microbiota biodiversity, bacterial metabolite production, and gut wall permeability regulating systemic levels. Recent data have demonstrated that Mn exposure may affect gut microbiota biodiversity by altering the abundance of Shiegella, Ruminococcus, Dorea, Fusicatenibacter, Roseburia, Parabacteroides, Bacteroidetes, Firmicutes, Ruminococcaceae, Streptococcaceae, and other bacterial phyla. A Mn-induced increase in Bacteroidetes abundance and a reduced Firmicutes/Bacteroidetes ratio may increase lipopolysaccharide levels. Moreover, in addition to increased systemic lipopolysaccharide (LPS) levels, Mn is capable of potentiating LPS neurotoxicity. Due to the high metabolic activity of intestinal microflora, Mn-induced perturbations in gut microbiota result in a significant alteration in the gut metabolome that has the potential to at least partially mediate the biological effects of Mn overexposure. At the same time, a recent study demonstrated that healthy microbiome transplantation alleviates Mn-induced neurotoxicity, which is indicative of the significant role of gut microflora in the cascade of Mn-mediated neurotoxicity. High doses of Mn may cause enterocyte toxicity and affect gut wall integrity through disruption of tight junctions. The resulting increase in gut wall permeability further promotes increased translocation of LPS and neuroactive bacterial metabolites to the systemic blood flow, ultimately gaining access to the brain and leading to neuroinflammation and neurotransmitter imbalance. Therefore, the existing data lead us to hypothesize that gut microbiota should be considered as a potential target of Mn toxicity, although more detailed studies are required to characterize the interplay between Mn exposure and the gut, as well as its role in the pathogenesis of neurodegeneration and other diseases.

Up-regulation of the manganese transporter SLC30A10 by hypoxia-inducible factors defines a homeostatic response to manganese toxicity

Manganese (Mn) is an essential metal that induces incurable parkinsonism at elevated levels. However, unlike other essential metals, mechanisms that regulate mammalian Mn homeostasis are poorly understood, which has limited therapeutic development. Here, we discovered that the exposure of mice to a translationally relevant oral Mn regimen up-regulated expression of SLC30A10, a critical Mn efflux transporter, in the liver and intestines. Mechanistic studies in cell culture, including primary human hepatocytes, revealed that 1) elevated Mn transcriptionally up-regulated SLC30A10, 2) a hypoxia response element in the SLC30A10 promoter was necessary, 3) the transcriptional activities of hypoxia-inducible factor (HIF) 1 or HIF2 were required and sufficient for the SLC30A10 response, 4) elevated Mn activated HIF1/HIF2 by blocking the prolyl hydroxylation of HIF proteins necessary for their degradation, and 5) blocking the Mn-induced up-regulation of SLC30A10 increased intracellular Mn levels and enhanced Mn toxicity. Finally, prolyl hydroxylase inhibitors that stabilize HIF proteins and are in advanced clinical trials for other diseases reduced intracellular Mn levels and afforded cellular protection against Mn toxicity and also ameliorated the in vivo Mn-induced neuromotor deficits in mice. These findings define a fundamental homeostatic protective response to Mn toxicity—elevated Mn levels activate HIF1 and HIF2 to up-regulate SLC30A10, which in turn reduces cellular and organismal Mn levels, and further indicate that it may be possible to repurpose prolyl hydroxylase inhibitors for the management of Mn neurotoxicity.

Short Report: Using Targeted Urine Metabolomics to Distinguish Between Manganese Exposed and Unexposed Workers in a Small Occupational Cohort

Objectives: Despite the widespread use of manganese (Mn) in industrial settings and its association with adverse neurological outcomes, a validated and reliable biomarker for Mn exposure is still elusive. Here, we utilize targeted metabolomics to investigate metabolic differences between Mn-exposed and -unexposed workers, which could inform a putative biomarker for Mn and lead to increased understanding of Mn toxicity.Methods: End of shift spot urine samples collected from Mn exposed (n = 17) and unexposed (n = 15) workers underwent a targeted assay of 362 metabolites using LC-MS/MS; 224 were quantified and retained for analysis. Differences in metabolite abundances between exposed and unexposed workers were tested with a Benjamini-Hochberg adjusted Wilcoxon Rank-Sum test. We explored perturbed pathways related to exposure using a pathway analysis.Results: Seven metabolites were significantly differentially abundant between exposed and unexposed workers (FDR ≤ 0.1), including n-isobutyrylglycine, cholic acid, anserine, beta-alanine, methionine, n-isovalerylglycine, and threonine. Three pathways were significantly perturbed in exposed workers and had an impact score >0.5: beta-alanine metabolism, histidine metabolism, and glycine, serine, and threonine metabolism.Conclusion: This is one of few studies utilizing targeted metabolomics to explore differences between Mn-exposed and -unexposed workers. Metabolite and pathway analysis showed amino acid metabolism was perturbed in these Mn-exposed workers. Amino acids have also been shown to be perturbed in other occupational cohorts exposed to Mn. Additional research is needed to characterize the biological importance of amino acids in the Mn exposure-disease continuum, and to determine how to appropriately utilize and interpret metabolomics data collected from occupational cohorts.