Manganese (Mn), chemical element, one of the silvery white, hard, brittle metals of Group 7 (VIIb) of the periodic table. It was recognized as an element in 1774 by the Swedish chemist Carl Wilhelm Scheele while working with the mineral pyrolusite and was isolated the same year by his associate, Johan Gottlieb Gahn. Although it is rarely used in pure form, manganese is essential to steelmaking.
Manganese combined with other elements is widely distributed in Earths crust. Manganese is second only to iron among the transition elements in its abundance in Earths crust; it is roughly similar to iron in its physical and chemical properties but is harder and more brittle. It occurs in a number of substantial deposits, of which the most important ores (which are mainly oxides) consist primarily of manganese dioxide (MnO2) in the form of pyrolusite, romanechite, and wad. Manganese is essential to plant growth and is involved in the assimilation of nitrates in green plants and algae. It is an essential trace element in higher animals, in which it participates in the action of many enzymes. Lack of manganese causes testicular atrophy. An excess of this element in plants and animals is toxic.
Manganese ores are primarily produced by Australia, South Africa, China, Gabon, and Brazil. Large areas of the ocean floor are covered with manganese nodules, also called polymetallic nodules, concretions of manganese with some iron, silicon, and aluminum. The amount of manganese in the nodules is estimated to be much more than that in land reserves.
Most of the manganese produced is used in the form of ferromanganese and silicomanganese alloys for iron and steel manufacture. Manganese ores containing iron oxides are first reduced in blast furnaces or electric furnaces with carbon to yield ferromanganese, which in turn is used in steelmaking. Adding manganese, which has a greater affinity for sulfur than does iron, converts the low-melting iron sulfide in steel to high-melting manganese sulfide. Produced without manganese, steel breaks up when hot-rolled or forged. Steels generally contain less than 1 percent manganese. Manganese steel is used for very rugged service; containing 1114 percent manganese, it provides a hard, wear-resistant, and self-renewing surface over a tough unbreakable core. Pure manganese produced electrolytically is used mostly in the preparation of nonferrous alloys of copper, aluminum, magnesium, and nickel and in the production of high-purity chemicals. Practically all commercial alloys of aluminum and magnesium contain manganese to improve corrosion resistance and mechanical properties. Aluminum cans contain about 1.5 percent manganese. (For detailed information on the extraction, refining, and applications of manganese, see manganese processing.)
All natural manganese is the stable isotope manganese-55. It exists in four allotropic modifications; the complex cubic structure of the so-called alpha phase is the form stable at ordinary temperatures. Manganese somewhat resembles iron in general chemical activity. The metal oxidizes superficially in air and rusts in moist air. It burns in air or oxygen at elevated temperatures, as does iron; decomposes water slowly when cold and rapidly on heating; and dissolves readily in dilute mineral acids with hydrogen evolution and the formation of the corresponding salts in the +2 oxidation state.
Manganese is quite electropositive, dissolving very readily in dilute nonoxidizing acids. Although relatively unreactive toward nonmetals at room temperature, it reacts with many at elevated temperatures. Thus, manganese burns in chlorine to give manganese(II) chloride (MnCl2), reacts with fluorine to give manganese(II) fluoride (MnF2) and manganese(III) fluoride (MnF3), burns in nitrogen at about 1,200 C (2,200 F) to give manganese(II) nitride (Mn3N2), and burns in oxygen to give manganese(II,III) oxide (Mn3O4). Manganese also combines directly with boron, carbon, sulfur, silicon, or phosphorus but not with hydrogen.
Of the wide variety of compounds formed by manganese, the most stable occur in oxidation states +2, +6, and +7. These are exemplified, respectively, by the manganous salts (with manganese as the Mn2+ ion), the manganates (MnO42), and the permanganates (MnO4). As in the case of titanium, vanadium, and chromium, the highest oxidation state (+7) of manganese corresponds to the total number of 3d and 4s electrons. That state occurs only in the oxo species permanganate (MnO4), manganese heptoxide (Mn2O7), and manganese trioxide fluoride (MnO3F), which show some similarity to corresponding compounds of the halogensfor example, in the instability of the oxide. Manganese in oxidation state +7 is powerfully oxidizing, usually being reduced to manganese in the +2 state. The intermediate oxidation states are known, but, except for some compounds in the +3 and +4 states, they are not particularly important.
The principal industrial compounds of manganese include several oxides. Manganese(II) oxide, or manganese monoxide (MnO), is used as a starting material for the production of manganous salts, as an additive in fertilizers, and as a reagent in textile printing. It occurs in nature as the green mineral manganosite. It also can be prepared commercially by heating manganese carbonate in the absence of air or by passing hydrogen or carbon monoxide over manganese dioxide.
The most important manganese compound is manganese dioxide, in which manganese is in the +4 oxidation state, and the black mineral pyrolusite is the chief source of manganese and all of its compounds. It is also widely used as a chemical oxidant in organic synthesis. Manganese dioxide is used as the cathode material in dry-cell batteries. It is produced directly from the ore, although substantial amounts are also prepared synthetically. The synthetic oxide is prepared by decomposition of manganous nitrate; by reaction of manganous sulfate, oxygen, and sodium hydroxide; or by electrolysis of an aqueous solution of manganese sulfate.
Various manganese salts also have commercial importance. Manganese sulfate (MnSO4) is added to soils to promote plant growth, especially of citrus crops. In addition, it is a good reducing agent, particularly useful in the manufacture of paint and varnish dryers. The deep-purple compound potassium permanganate (KMnO4) has many uses, most notably as a disinfectant, water purifier, and antiseptic.
Asurvey byindependent pollster Levada Center in2013 found that 52 percent ofRussians believed inomens, prophetic dreams andastrology. Admittedly, this was adecline from57 percent in2000 but still represented more than half ofthe population atthe time.
Forforeigners visiting Russia, it is useful tobe aware ofthese "do's" and"don't's" oflocal culture. TheMoscow Times highlights 10 ofthe most common superstitions travelers toRussia are likely toencounter.
Russians believe that shaking hands or kissing aguest across adoorway is abig no-no. InRussian folklore, thethreshold is where the"house spirit" is believed toreside, andbridging this gap with ahandshake is therefore extremely bad luck.
Instead, you should wait until completely entering aRussian home before shaking hands, or have theperson inside thehome come completely out before you greet them. Fora more definitive guide onshaking hands inRussia, click here.
It's never amistake totake abouquet offlowers when invited tosomeone's home or fora birthday or other celebration. However, make sure that bouquets forsuch festive occasions are filled with anodd number offlowers. Bouquets with aneven numbers offlowers are reserved forfunerals.
Before embarking onany journey, superstition dictates that all members ofthe group should sit down insilence even if not everyone is traveling. This doesn't have tobe fora long time, but will ensure that thetrip is asafe one. It's also agood opportunity tomake sure you have everything you need forthe journey.
Legend has it that thepractice started when Cossack soldiers drove Napoleon back toFrance in1814. Thesoldiers worked out that Parisian restaurateurs charged customers per empty bottle left onthe table rather than per bottle ordered, andso theCossacks cunningly hid them under thetable. When thesoldiers returned toRussia, they brought thecustom with them.
Inorder toavoid putting thecurse onsomething, Russians will knock onwood, spit three times over their left shoulder, or do both. If you don't want tospit, you can always mimic thesound bysaying "fu-fu-fu."
Unmarried people should avoid sitting atthe corner ofa table otherwise they will never get married, according toRussian superstition. Others believe that this will only hold true forseven years, making it possible foryounger children tosit atthe table corner.
Andanother thing. Make sure that you never sit directly onthe cold ground, or frankly any cold surface otherwise you will become infertile (or so Russians believe). This is particularly true if you are ayoung woman.
It often seems that there are only about 10 names inRussian. Nearly every woman you meet is Natasha or Masha or Ira andevery man, Alexander or Dmitry or Alexei. There is asuperstition related tomeeting people who have thesame name, but fortunately as it happens so often this superstition brings good, rather than bad, luck.
The Moscow Times team of journalists has been first with the big stories on the coronavirus crisis in Russia since day one. Our exclusives and on-the-ground reporting are being read and shared by many high-profile journalists.
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There is no one single or best structure for the periodic table but by whatever consensus there is, the form used here is very useful and the most common. The periodic table is a masterpiece of organised chemical information and the evolution of chemistry's periodic table into the current form is an astonishing achievement.
On 1 May 2014 a paper published in Phys. Rev. Lett by J. Khuyagbaatar and others states the superheavy element with atomic number Z = 117 (ununseptium) was produced as an evaporation residue in the 48Ca and 249Bk fusion reaction at the gas-filled recoil separator TASCA at GSI Darmstadt, Germany. The radioactive decay of evaporation residues and their -decay products was studied using a detection setup that allows measurement of decays of single atomic nuclei with very short half-lives. Two decay chains comprising seven -decays and a spontaneous fission each were identified and assigned to the isotope 294Uus (element 117) and its decay products.
Manganese is one of the most abundant elements on Earth. The oxidation of manganese has long been theorized1yet has not been demonstrated2,3,4to fuel the growth of chemolithoautotrophic microorganisms. Here we refine an enrichment culture that exhibits exponential growth dependent on Mn(II) oxidation to a co-culture of two microbial species. Oxidation required viable bacteria at permissive temperatures, which resulted in the generation of small nodules of manganese oxide with which the cells associated. The majority member of the culturewhich we designate Candidatus Manganitrophus noduliformansis affiliated to the phylum Nitrospirae (also known as Nitrospirota), but is distantly related to known species of Nitrospira and Leptospirillum. We isolated the minority member, a betaproteobacterium that does not oxidize Mn(II) alone, and designate it Ramlibacter lithotrophicus. Stable-isotope probing revealed 13CO2 fixation into cellular biomass that was dependent upon Mn(II) oxidation. Transcriptomic analysis revealed candidate pathways for coupling extracellular manganese oxidation to aerobic energy conservation and autotrophic CO2 fixation. These findings expand the known diversity of inorganic metabolisms that support life, and complete a biogeochemical energy cycle for manganese5,6 that may interface with other major global elemental cycles.
All sequencing data has been deposited at the NCBI under BioProject PRJNA562312. The cloned 16S rRNA gene sequences of Candidatus Manganitrophus noduliformans (species A) and R.lithotrophicus (species B) from the co-culture have been deposited at GenBank under accession numbers MN381734 and MN381735, respectively. The iTAG sequences from the different enrichments have been deposited at the Sequence Read Archive (SRA) under accession numbers SRR10031198, SRR10031199 and SRR10031200. Genome sequences of the co-culture, from which the genome of Candidatus Manganitrophus noduliformans was reconstructed, have been deposited under BioSample SAMN12638105 with raw sequences deposited at SRA under accession number SRR10032644; the reconstructed genome of Candidatus Manganitrophus noduliformans has been deposited at DDBJ/ENA/GenBank under accession number VTOW00000000. Genome sequences of R.lithotrophicus strain RBP-1 have been deposited under BioSample SAMN12638106, with raw sequences deposited at SRA under accession number SRR10031379; the reconstructed genome of R.lithotrophicus strain RBP-1 has been deposited at DDBJ/ENA/GenBank under accession number VTOX00000000. Additionally, reconstructed genomes have been deposited in Joint Genome Institute (JGI) Genomes Online Database Study ID Gs0134339, with Integrated Microbial Genome ID 2784132095 for Candidatus Manganitrophus noduliformans and ID 2778260901 for R. lithotrophicus strain RBP-1. Transcriptome sequence data for the seven biological replicates have been deposited at SRA under accession numbers SRR10060009, SRR10060010, SRR10060011, SRR10060012, SRR10060013, SRR10060017 and SRR10060018. Unique biological materials are available from the corresponding author upon reasonable request. Source data are provided with this paper.
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This work was supported by NASA Astrobiology Institute Exobiology grant #80NSSC19K0480; and by Caltechs Center for Environmental Microbial Interactions and Division of Geological and Planetary Sciences. We thank S. Connon for assistance with iTag sequencing preparations; G. Rossman and U. Lingappa for spectroscopic analyses and minerology insights; G. Chadwick for discussions on physiology and bioenergetics; I. Antoshechkin and V. Kumar for assistance with nucleic acid library preparation and sequencing at the Millard and Muriel Jacobs Genetics and Genomics Laboratory; N. Dalleska for assistance with ICPMS analyses at the Environmental Analysis Center; F. Gao for inputs on RNA data analysis using kallisto software at the Bioinformatics Resource Center in the Beckman Institute; C. Ma for assistance with SEM analyses at the GPS Analytical Facility; Y. Guan for assistance with nanoSIMS analyses at the GPS Microanalysis Center; and multiple colleagues for feedback before publication.
a, Incubation temperature influences oxidation. An optimum between 34C and 40C was observed, but above these temperatures oxidation was inhibited. By contrast, non-biological reactions would generally be predicted to continue to increase in rate with increasing temperature. b, Sensitivity of Mn(II) oxidation to the presence of either of two antibiotics, or to prior pasteurization before extended incubation at 32C. c, When amended to active co-cultures at concentrations >2.0 mM, MnCl2 appeared to inhibit MnCO3 oxidation when an active culture containing about2.2 mM unreacted MnCO3 was used as the inoculum. The number of points for each experimental condition represents independent cultivation experiments.
a, DNA yield of the two-species co-culture incubated in MOPS-buffered basal medium in the absence of Mn(II) substrate. No statistically significant changes in the mean DNA yields (P=0.06, day 0 vs 10; P=0.70, day 10 vs 21; P=0.20, day 0 vs 21; two-tailed t-test with equal variance) are observed over the incubation period. b, c, Exponential increase in bacteria and biomass yields in a Mn(II)-oxidizing culture, which is coupled to exponential increases Mn(II) oxidation (same culture analysed in Fig. 2). Bacteria were measured via 16S rRNA gene copies using a general bacteria probe in quantitative PCR; points represent 3 technical replicates. Biomass was measured via DNA yield from same culture volumes. d, Exponential increases in Mn(II) oxidation (Fig. 2a) and DNA yields by this same culture (1 mM nitrate replicate 1, c) correlate. Similar relationships were observed in samples from independent cultivation experiments (n=2). el, Kinetics of Mn(II) oxidation by the co-culture in basal medium; two phases of exponential Mn(II) oxidation were observed. eg, Basal medium with 1 mM nitrate (n=4; for replicate 1, see bd and Fig. 2). hl, Basal medium with 1 mM ammonia (n=5). m, Exponential growth of species A and species B paralleled Mn(II) oxidation in basal medium with 1 mM ammonia as the nitrogen source (1 mM ammonia replicate 5, l), rather than 1 mM nitrate. n, Linear relationship between cell growth and the amount of Mn(II) oxidized (1 mM ammonia replicate 5, l and m). Values in n were normalized by subtracting the initial cell number and Mn oxide concentrations at the onset of the experiment, and negative values after normalization were excluded from the analysis. All data points included in the line fits are used to calculate the doubling times (Td), unless otherwise noted by x symbols.
a, Estimations of the relative ratio between species A and species B. Slow-growing microorganisms, in particular species A (which also has a smaller cell volume than species B or Escherichia coli) could have a lower number of ribosomes, resulting in lower signal intensity from rRNA-targeted fluorescent probes, relative to the fluorescent signal from DNA stain DAPI. The two species together account for 99.7% of assigned sequence reads (Supplementary Table 1). The two species together account for 97.54% of the sequence reads in the metagenome (f). The two species together account for 99.576% (s.d.=0.005%, n=7) of the rRNA sequence reads and 100.1700% (s.d.=0.0005%, n=7) of the non-rRNA sequence reads in the co-culture metatranscriptomes (h). be, Possible metabolic interactions that may be occurring between species A (orange) and species B (blue). f, Genome statistics for species A and species B. g, Observed rates and yields of Mn(II) oxidation by the co-culture, in comparison to the literature values5,23,26,39,95,96,97,98,99,100 reported for other physiologically or phylogenetically related lithotrophs or metal-active heterotrophs. ||Conversion estimate based on Escherichia coli biomass of 2.81013 g dry cell weight per cell, of which 55% is protein27. Co-culture values correspond to results from the single independent culture with nitrate as the nitrogen source for which extensive data on both oxidation kinetics and growth (genome copies) were collected. h, Transcriptome statistics for 7 co-cultures sampled at different degrees of Mn(II) oxidation.
ae, Epifluorescence microscopy reveals distribution of cells of species A and species B associated with dissolved Mn oxide nodules. DAPI (blue) was used to stain DNA, in addition to applying species-specific FISH probes targeting the 16S rRNA of species A (magenta) and species B (green). Probe fluorescence for species A was dim and faded rapidly, but was associated with the cells that otherwise appear in photomicrographs to only be DAPI-stained. No third species is present, as observed in independent cultivation experiments (n=2), and confirmed via independent methods (Extended Data Fig. 3a). fp, Scanning electron micrographs of Mn(II)CO3 substrate (f, g) and Mn oxide nodules collected from liquid cultures (hp). Representative nodules are from independent cultivation experiments (n=4).
a, 16S rRNA gene phylogram, based on a Bayesian analysis of 1,532 aligned nucleotide positions. NCBI82 taxonomic classifications are used, and sequences shown are all from the phylum Nitrospirae. The names and known physiologies for the previously described genera in this phylum are shown on the right. NCBI accession numbers for 16S rRNA sequences are included in the node names. Source environment for the sequences are shown in brackets. b, Multilocus phylogram, based on a Bayesian analysis of 5,036 aligned amino acid positions concatenated from 120 bacterial protein markers62. GTDB62 taxonomic classifications are used, and sequences shown are from the phylum under the headings Nitrospirota and Nitrospirota_A. The names and known physiologies for the previously described classes in this phylum are shown on the right. NCBI accession numbers for genome assemblies are included in the node names. For a, b, the dots on the branches indicate posterior probabilities greater than 0.80. c, Phylogenetic analyses of the phylum Nitrospirae (Nitrospirota) limited to only those species with reconstructed genomes yield a topology different from that observed in a and Fig. 3a. Bayesian phylogram based on 1,532 aligned 16S rRNA nucleotide positions (left); multilocus Bayesian phylogram, based on 5,036 aligned amino acid positions of 120 concatenated bacterial protein markers (right). Sequences clustering within the three previously described classes within this phylum are collapsed into separate nodes. d, Protein sequence phylogeny of dihydroxy-acid and 6-phosphogluconate dehydratases. Sequences were selected based on a previous study101, with the addition of homologues found in Nitrospira inopinata, Leptospirillum ferriphilum and species A (red). All 770 aligned amino acid positions were used in the maximum likelihood analysis. Protein accession numbers from the NCBI database or gene identifiers from the IMG database of the 3 new sequences are shown in parentheses. Black dots on the branches represent bootstrap values equal to 100%. Although dihydroxy-acid dehydratase and 6-phosphogluconate dehydratase are homologous, they form separate clusters phylogenetically as reported101. The homologues in Nitrospirae all belong to the dihydroxy-acid dehydratase clade, therefore are unlikely candidates for 6-phosphogluconate dehydratase activity and function in the ED pathway. All scale bars show evolutionary distance (0.1 substitutions-per-site average).
a, 16S rRNA gene phylogram, based on a Bayesian analysis of 1,532 aligned nucleotide positions. NCBI82 taxonomic classifications are used, with sequences selected from the class Betaproteobacteria. The genus Ramlibacter, consistently identified in two phylogenetic approaches, is shaded in grey, with species B in bold. Source environments for the species in Ramlibacter are shown in brackets. The order and family classifications are included to the right separated by a semicolon. The black dots on the branches indicate posterior probabilities greater than 0.90. b, Multilocus phylogram, based on a maximum-likelihood analysis of 5,035 aligned amino acid positions concatenated from 120 bacterial protein markers62. GTDB62 taxonomic classifications are used, and sequences shown are from the order Betaproteobacteriales. The GTDB family classifications are included to the right of species names. NCBI accession numbers for 16S rRNA sequences or the genome assemblies are included after the species names. The black dots on the branches indicate bootstrap values greater than 90%. Scale bars shown evolutionary distance (0.1 substitutions-per-site average). c, d, Kinetics of species B growth basal media with either 5 g/l of tryptone (n=3 biological replicates) (c) or 10 mM acetate (n=2 biological replicates) (d).
Only cytochrome bd-like oxidases were identified in species A, in contrast to other classes in the phylum Nitrospirae (Nitrospirota). a, Unrooted maximum-likelihood tree, constructed using 242 amino acid positions shared between cytochrome bd and bd-like oxidases, using RAxML89 (model LGF). Deduced proteins from the genome of species A are in red, with their IMG gene identifiers and clade numbering (as shown in Fig. 3b) included in brackets. Other proteins from the phylum Nitrospirae (Nitrospirota) are coloured blue, orange or brown for classes Nitrospiria, Leptospirillia or Thermodesulfovibrionia, respectively. Cytochrome bd oxidase of species B, with its IMG identifier, is in green; it belongs to the cyanide insensitive oxidase clade in purple. b, Phylogenetic analysis of cytochrome bd-like oxidases from species A. Unrooted maximum-likelihood tree was constructed using 242 amino acid positions shared between different clades of cytochrome bd-like oxidases. Cytochrome bd-like oxidases are assigned to different clades, based on the phylogeny and their gene cluster structures. Species A encodes 8 cytochrome bd-like oxidases (bold), representing clades I, II, IIIb, Va and Vb; clade numbering as shown in Fig. 3b are included in brackets after the IMG identifiers. Black dots on branches represent bootstrap values greater than 90%. Scale bars show evolutionary distance (substitutions-per-site average).
Cytochrome bd-like oxidase in species A (sequence names starting with A, followed by their IMG gene identifier and clade numbering as shown in brackets in Fig. 3b) and cytochrome bd oxidase subunit I in species B (sequence name starting with B, followed by its IMG gene identifier) are aligned to characterized cytochrome bd oxidases in Escherichia coli (sequence name starting with Eco, followed by its NCBI identifier) and Geobacillus thermodenitrificans (sequence name starting with Geo, followed by its NCBI identifier). Key features as revealed by structure102 are indicated at the top of the alignment, using E. coli protein residue numbering. The cytochrome bd oxidase subunit I sequence from species B shows conservation of all key residues. By contrast, cytochrome bd-like oxidases in species A do not show conservation of many key residues; instead, they are predicted to have up to 14 transmembrane helixes (compared to 9 in E. coli). One cytochrome bd-like oxidase in species A has a C-terminus extension with a haem c binding motif (CXXCH).
a, Summary of stable isotope probing analysis of cells dissolved from Mn oxide nodules, either with paraformaldehyde fixation and FISH, or without (to avoid dilution with natural abundance isotopes). Cells of species A and species B were either identified by FISH or by elemental composition (species B cells were observed to have higher 14N/15N ratios), and their isotopic compositions were obtained via nanoSIMS (n=the total number of cell regions of interest analysed in the nanoSIMS images). For FISHnanoSIMS analyses, a total of 2 and 5 nanoSIMS images from single cultures incubated with either MnCO3 or Mn13CO3, respectively, was examined. For nanoSIMS analyses without paraformaldehyde fixation and FISH, a total of 3 and 17 nanoSIMS images from single cultures incubated with either MnCO3 or Mn13CO3, respectively, was examined. bu, Individual secondary ion images from nanoSIMS showing incorporation of inorganic 13C and 15N into the cells of both species (dissolved from Mn oxide nodules grown in the presence of MnCO3 and 15NO3 (bk) or Mn13CO3 and 15NO3 (lu)), and species B cells could have higher 14N content than species A. Secondary ions 12C2 (mass 24 for 12C), 13C12C (mass 25 for 13C), 14N12C (mass 26 for 14N), 15N12C (mass 27 for 15N), 32S (mass 32 for 32S) were simultaneously measured. The counts of the secondary ions are shown in brackets (minimummaximum) and displayed using the colour scale shown to the right of the images. bf and lp correspond to the top and bottom panels in Fig. 4, respectively. White arrows indicate species B cells identified in FISH showing high 14N in nanoSIMS. v, NanoSIMS measurement of residual Mn associated with cells grown with Mn13CO3 and 15NO3, after dissolving from Mn oxide nodules. The same nanoSIMS image area was analysed as in lp, except 55Mn16O (mass 71 for 55Mn) was measured (n=1 nanoSIMS image) in addition to other secondary ions. Negligible amount of Mn was found in the biomass, indicating that any remaining Mn13CO3 substrate had been completely dissolved away during sample preparation, and thus did not interfere with the 13C analyses.
ad, Evaluation of FISH oligonucleotide probes. Three probes (NLT499, black circles; BET359, white circles; BET867, white squares) were tested in different probe combinations and formamide concentrations, using 16S rRNA gene clones of species A (a, b) or species B (c, d). Each point in the dissociation profile represents the mean of fluorescence intensities of at least 100 different single cells in 5 distinct microscopic fields of 1 biological replicate. Lines connect the 95% confidence intervals of the points. No interference was found when targeting either species A or species B with different probe combinations and formamide concentrations. RU, relative units of fluorescence intensity. e, Evaluation of ICPMS method to measure Mn compounds with different oxidation states. Mn(II) in its various forms can be almost entirely measured in the acid-soluble fraction with little in the acid-insoluble fraction, and any increase in the acid-insoluble fraction is an indication of oxidized Mn(II). We refer to the acid-soluble fraction as Mn(II), and the acid-insoluble fraction as Mn(II) oxidized representing Mn(III/IV). Supplementary Note4 provides more details. MnO2 was synthesized according to two previously published methods103,104. f, Evaluation of transcriptome analysis software kallisto93. Average fragment length for RNA libraries was measured to be 230 bp. However, using 230 bp as the input parameter for fragment length caused a kallisto93 expression evaluation issue for genes <230 bp in length; thus, the fragment length was adjusted downward to 100 bp to evaluate the expression of genes <230 bp. This parameter change does not affect the overall transcript expression for genes >230 bp as seen in the correlation analysis, performed using transcriptome sample Mn03. g, h, Evaluation of quantification range and efficiency of quantitative PCR oligonucleotide probes. Three quantitative PCR oligonucleotide probes (bacteria (g) or species A- or species B-specific (h)) were tested using cloned 16S rRNA gene of either species A (open squares, solid lines) or species B (open triangles, dashed lines) as DNA templates. Threshold cycle (CT) versus gene copies show that all three probes had amplification efficiencies between 90105% in the quantification ranges plotted. Points represent 3 technical replicates. i, j, Evaluation of specificity of quantitative PCR oligonucleotide probes. The percentage of species A (i) and species B (j) was estimated in reactions containing a mixture of cloned 16S rRNA genes from both species A and species B as DNA templates. Dashed lines represent theoretical 100% match in the expected versus measured values. The results indicate that the species-specific probes quantified their targeted species with minimal interference. Points represent 4 technical replicates.
This file contains the Supplementary Taxonomic Proposal; Supplementary Notes; and Supplementary Information References. Together, these provide additional support and information underlying the main text and methods.
The genus Denhamia(Celastraceae) includes fifteen Australian species, many of which have a propensity for manganese (Mn) (hyper)accumulation. Among the key aims of this study were to: i) elucidate Mn accumulation in D. bilocularis, D. celastroides, D. pittosporoides and D. silvestris under controlled conditions; ii) examine the in situ distributions of Mn and other elements in tissues of i) above, and also in two other species growing in their natural habitat, i.e. D. silvestris and D. cunninghamii; iii) test numerous Denhamia herbarium specimens for Mn accumulation using portable X-ray fluorescence spectroscopy (XRF).
Portable XRF and laboratory micro-XRF were used to examine Mn accumulation and foliar distribution patterns in several Australian Denhamia species. These techniques were variously applied to fresh field material and experimentally-raised plants treated with Mn (250gg1, 500gg1 and 1000gg1) and dry herbarium material.
The findings revealed D. bilocularis as a new Mn hyperaccumulator, with foliar Mn concentrations of up to 15,300gg1 in herbarium samples, and 13,700gg1 in experimentally grown plants. Laboratory XRF maps consistently showed that foliar Mn accumulation was localized at the extremities of wild D. cunninghamii and D. silvestris leaves; whereas it was found in the vasculature of experimentally grown D. bilocularis and D. pittosporoides leaves.
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Farida Abubakari is the recipient of a UQ Graduate School Scholarship (UQGSS) from The University of Queensland. We thank Lachlan Casey (Centre for Microscopy and Microanalysis at the University of Queensland) for technical support with the XRF analysis. We acknowledge the support of the AMMRF at the Center for Microscopy and Microanalysis at the University of Queensland.
Abubakari, F., Nkrumah, P.N., Erskine, P.D. et al. Manganese (hyper)accumulation within Australian Denhamia (Celastraceae): an assessment of the trait and manganese accumulation under controlled conditions. Plant Soil 463, 205223 (2021). https://doi.org/10.1007/s11104-021-04833-z