Citrus sinensis manganese tolerance: Insight from manganese-stimulated secretion of root exudates and rhizosphere alkalization.

We used manganese (Mn)-tolerant 'Xuegan' (Citrus sinensis) seedlings as materials and examined the characterization of Mn uptake and Mn-activated-release of root exudates under hydroponic conditions. We observed that root and shoot Mn bioaccumulation factor (BCF) reduced with the increase of Mn supply, and that Mn transfer factor (Tf) reduced greatly as Mn supply increased from 0 to 500 µM, beyond which Tf slightly increased with increasing Mn supply, suggesting that Mn supply reduced the ability to absorb and accumulate Mn in roots and shoots, as well as root-to-shoot Mn translocation. Without Mn, roots alkalized the solution pH from 5.0 to above 6.2, while Mn supply reduced root-induced alkalization. As Mn supply increased from 0 to 2000 µM, the secretion of root total phenolics (TPs) increased, while the solution pH decreased. Mn supply did not alter the secretion of root total free amino acids, total soluble sugars, malate, and citrate. Mn-activated-release of TPs was inhibited by low temperature and anion channel inhibitors, but not by protein biosynthesis inhibitor. Using widely targeted metabolome, we detected 48 upregulated [35 upregulated phenolic compounds + 13 other secondary metabolites (SMs)] and three downregulated SMs, and 39 upregulated and eight downregulated primary metabolites (PMs). These findings suggested that reduced ability to absorb and accumulate Mn in roots and shoots and less root-to-shoot Mn translocation in Mn-toxic seedlings, rhizosphere alkalization, and Mn-activated-release of root exudates (especially phenolic compounds) contributed to the high Mn tolerance of C. sinensis seedlings.

Variation of Cicer Germplasm to Manganese Toxicity Tolerance.

After aluminum, manganese toxicity is the most limiting factor for crops grown in acidic soils worldwide. But overall, research on Mn toxicity is still limited. The poor acid tolerance of chickpea may be related to Mn toxicity, but there has been no previous screening of chickpea germplasm (nor in its wild Cicer relatives, Cicer reticulatum and Cicer echinospermum) for tolerance to Mn toxicity. A screening technique was developed for tolerance to Mn toxicity using three released cultivars of chickpea (Cicer arietinum L), Ambar, PBA HatTrick, and PBA Striker; one accession each of C. reticulatum and C. echinospermum; and lupin (Lupinus angustifolius) as a Mn-tolerant check, with eight Mn concentrations of 2, 25, 50, 100, 150, 200, 250, and 500 µM Mn as MnSO4 in a low-ionic-strength nutrient solution. The plants were harvested at 14 and 28 days after Mn treatments. The nutrient uptake in shoots (young, old leaves, and the rest of the shoot) and roots was investigated. The best discrimination between tolerant and intolerant Cicer genotypes based on relative shoot dry weight, root dry weight, total root length, and scoring of toxicity symptoms was achieved at 150 µM Mn after 14 days of growth in Mn solution. Among the chickpea cultivars, the greater relative plant growth (both shoot and root) of Ambar and PBA Striker at 100-200 µM Mn contrasted with that of PBA HatTrick, while the C. echinospermum accession was more tolerant to Mn toxicity than C. reticulatum. Manganese tolerance in both domestic cultivars and wild accessions was associated with internal tolerance to excess Mn following greater uptake of Mn and translocation of Mn from roots to shoots.

Identification of MTP gene family in tea plant (Camellia sinensis L.) and characterization of CsMTP8.2 in manganese toxicity.

Cation diffusion facilitators (CDFs) play central roles in metal homeostasis and tolerance in plants, but the specific functions of Camellia sinensis CDF-encoding genes and the underlying mechanisms remain unknown. Previously, transcriptome sequencing results in our lab indicated that the expression of CsMTP8.2 in tea plant shoots was down-regulated exposed to excessive amount of Mn2+ conditions. To elucidate the possible mechanisms involved, we systematically identified 13 C. sinensis CsMTP genes from three subfamilies and characterized their phylogeny, structures, and the features of the encoded proteins. The transcription of CsMTP genes was differentially regulated in C. sinensis shoots and roots in responses to high concentrations of Mn, Zn, Fe, and Al. Differences in the cis-acting regulatory elements in the CsMTP8.1 and CsMTP8.2 promoters suggested the expression of these two genes may be differentially regulated. Transient expression analysis indicated that CsMTP8.2 was localized to the plasma membrane in tobacco and onion epidermal cells. Moreover, when heterologously expressed in yeast, CsMTP8.2 conferred tolerance to Ni and Mn but not to Zn. Additionally, heterologous expression of CsMTP8.2 in Arabidopsis thaliana revealed that CsMTP8.2 positively regulated the response to manganese toxicity by decreasing the accumulation of Mn in plants. However, there was no difference in the accumulation of other metals, including Cu, Fe, and Zn. These results suggest that CsMTP8.2 is a Mn-specific transporter that contributes to the efflux of excess Mn2+ from plant cells.

Complete genome sequence of the highly Mn(II) tolerant Staphylococcus sp. AntiMn-1 isolated from deep-sea sediment in the Clarion-Clipperton Zone.

Staphylococcus sp. AntiMn-1 is a deep-sea bacterium inhabiting seafloor sediment in the Clarion-Clipperton Zone (CCZ) that is highly tolerant to Mn(II) and displays efficient Mn(II) oxidation. Herein, we present the assembly and annotation of its genome.

Physiological response of Polygonum perfoliatum L. following exposure to elevated manganese concentrations.

Polygonum perfoliatum L. is a Mn-tolerant plant as considered having the potential to revegetate in manganese mine wasteland. The glasshouse experiments were carried out to evaluate its tolerance and physiological response in different Mn concentrations (5, 500, 1000, 2000, 5000, and 10,000 µmol L-1). Absorption bands of P. perfoliatum differed greatly in lipids, proteins, and carbohydrates. With elevated levels of Mn (5-2000 µmol L-1), absorbance changed little, which demonstrated that lower Mn concentrations had negligible influence on transport functions. As Mn concentrations in excess of 2000 µmol L-1, absorbance increased slightly but eventually decreased. Furthermore, a hydroponic culture was carried out in order to study its changes of ultrastructure with the increasing Mn concentrations (5, 1000, and 10,000 µmol L-1). Lower Mn levels with 5 and 1000 µmol L-1 had no breakage function to the ultrastructure of P. perfoliatum. However, as Mn concentration was up to 10,000 µmol L-1, visible damages began to appear, the quantity of mitochondria in root cells increased, and the granum lamellae of leaf cell chloroplasts presented a disordered state. In comparison with the controls, black agglomerations were found in the cells of P. perfoliatum under the controlling concentration of Mn with 1000 and 10,000 µmol L-1 for 30 days, which became obvious at higher Mn concentrations. As Mn concentration was 10,000 µmol L-1, a kind of new acicular substance was developed in leaf cells and intercellular spaces, possibly indicating a resistance mechanism in P. perfoliatum. These results confirm that P. perfoliatum shows potential for the revegetation of abandoned manganese tailings.

Early induction of Fe-SOD gene expression is involved in tolerance to Mn toxicity in perennial ryegrass.

Manganese (Mn) toxicity limits plant growth in acid soils. Although Mn toxicity induces oxidative stress, the role of superoxide dismutase (SOD, EC.1.15.1.1) isoforms in conferring Mn tolerance remains unclear. Seedlings of ryegrass cultivars Nui (Mn-sensitive) and Kingston (Mn-tolerant) were hydroponically grown at 2.4 (optimal) or 750 µM Mn (toxic) concentration, and harvested from 2 to 48 h. Kingston showed higher shoot Mn than Nui at 2.4 µM Mn. At toxic supply, shoot Mn concentration steadily increased in both cultivars, with Kingston having the highest accumulation at 48 h. An early (2 h) increase in lipid peroxidation under Mn excess occurred, but it returned (after 6 h) to the basal level in Kingston only. Kingston exhibited higher SOD activity than Nui, and that difference increased due to toxic Mn. In general, Mn-induced gene expression of Mn- and Cu/Zn-SOD isoforms was higher in Nui than Kingston. Nevertheless, under Mn excess, we found a greater Fe-SOD up-regulation (up to 5-fold) in Kingston compared to Nui. Thus, Fe-SOD induction in Kingston might explain, at least partly, its high tolerance to Mn toxicity. This is the first evidence that Mn toxicity causes differential gene expression of SOD isoforms in ryegrass cultivars in the short-term.

Mn tolerance in rice is mediated by MTP8.1, a member of the cation diffusion facilitator family.

Manganese (Mn) is an essential micronutrient for plants, but is toxic when present in excess. The rice plant (Oryza sativa L.) accumulates high concentrations of Mn in the aerial parts; however, the molecular basis for Mn tolerance is poorly understood. In the present study, genes encoding Mn tolerance were screened for by expressing cDNAs of genes from rice shoots in Saccharomyces cerevisiae. A gene encoding a cation diffusion facilitator (CDF) family member, OsMTP8.1, was isolated, and its expression was found to enhance Mn accumulation and tolerance in S. cerevisiae. In plants, OsMTP8.1 and its transcript were mainly detected in shoots. High or low supply of Mn moderately induced an increase or decrease in the accumulation of OsMTP8.1, respectively. OsMTP8.1 was detected in all cells of leaf blades through immunohistochemistry. OsMTP8.1 fused to green fluorescent protein was localized to the tonoplast. Disruption of OsMTP8.1 resulted in decreased chlorophyll levels, growth inhibition in the presence of high concentrations of Mn, and decreased accumulation of Mn in shoots and roots. However, there was no difference in the accumulation of other metals, including Zn, Cu, Fe, Mg, Ca, and K. These results suggest that OsMTP8.1 is an Mn-specific transporter that sequesters Mn into vacuoles in rice and is required for Mn tolerance in shoots.