Table_1_Seasonal changes in foliar calcium oxalate concentrations in conifer and hardwood trees: a potentially bioavailable source of cellular calcium and/or oxalate under stress.DOCX

The present study compared seasonal changes in the concentrations of calcium oxalate (CaOx) crystals and total calcium (Ca) in the foliage of red spruce (Picea rubens Sarg.), white pine (Pinus strobus L.), black oak (Quercus velutina L.), and sugar maple (Acer saccharum Marshall) trees. Samples were collected from the same four replicate trees of each species starting in June 2014 through September 2015 for a total of six times for conifers and four times for hardwoods. Calcium oxalate was extracted from tissues using a method developed in our laboratory in 2015. The purity of the extracted CaOx was indicated by an r2 of 0.98 between Ca and oxalate (Ox) for the data pooled across all species and all sampling times. As expected, the concentrations of CaOx varied between species. We hypothesized that the only role of CaOx crystals is to bind excess Ca, so based on this hypothesis the concentrations of CaOx would increase over the growing season both in conifer and hardwood trees, and in conifers, its quantities would be higher in the older relative to the younger needles. However, we found, that for most species, CaOx concentrations were not significantly different from each other for all collection times. In addition, relative to total Ca, the percent of Ca that existed in the form of CaOx varied widely with species, time of collection within a species, and needle age. Thus, no specific trend was observed for CaOx accumulations with changes in seasons. Concentrations of CaOx were indeed higher in older spruce and pine needles. Based on the available literature on this topic and our data, this could mean that CaOx amounts are dynamic and are continuously being adjusted according to the metabolic needs of cells for either Ca or Ox while still performing the function of shedding off excess Ca.</p

DataSheet_1_Is foliar tissue drying and grinding required for reliable and reproducible extraction of total inorganic nutrients? A comparative study of three tissue preparation methods.pdf

In response to abiotic and biotic stress or experimental treatment(s), foliar concentrations of inorganic nutrients and metabolites often change in concert to maintain a homeostatic balance within the cell’s environment thus allowing normal functions to carry on. Therefore, whenever possible, changes in cellular chemistry, metabolism, and gene expressions should be simultaneously evaluated using a common pool of tissue. This will help advance the knowledge needed to fill the gaps in our understanding of how these variables function together to maintain cellular homeostasis. Currently, foliar samples of trees for total inorganic nutrients and metabolic analyses are often collected at different times and are stored and processed in different ways before analyses. The objective of the present study was to evaluate whether a pool of wet (previously frozen) intact tissue that is used for metabolic and molecular work would also be suitable for analyses of foliar total inorganic nutrients. We compared quantities of nutrients extracted from wet-intact, dried-intact, and dried-ground tissues taken from a common pool of previously frozen foliage of black oak (Quercus velutina L.), sugar maple (Acer saccharum Marshall), red spruce (Picea rubens Sarg.), and white pine (Pinus strobus L.). With a few exceptions in the case of hardwoods where concentrations of total Ca, Mg, K, and P extracted from wet-intact tissue were significantly higher than dry tissue, data pooled across all collection times suggest that the extracted nutrient concentrations were comparable among the three tissue preparation methods and all for species. Based on the data presented here, it may be concluded that drying and grinding of foliage may not be necessary for nutrient analyses thus making it possible to use the same pool of tissue for total inorganic nutrients and metabolic and/or genomic analyses. To our knowledge, this is the first report on such a comparison.</p

DataSheet_2_Is foliar tissue drying and grinding required for reliable and reproducible extraction of total inorganic nutrients? A comparative study of three tissue preparation methods.pdf

In response to abiotic and biotic stress or experimental treatment(s), foliar concentrations of inorganic nutrients and metabolites often change in concert to maintain a homeostatic balance within the cell’s environment thus allowing normal functions to carry on. Therefore, whenever possible, changes in cellular chemistry, metabolism, and gene expressions should be simultaneously evaluated using a common pool of tissue. This will help advance the knowledge needed to fill the gaps in our understanding of how these variables function together to maintain cellular homeostasis. Currently, foliar samples of trees for total inorganic nutrients and metabolic analyses are often collected at different times and are stored and processed in different ways before analyses. The objective of the present study was to evaluate whether a pool of wet (previously frozen) intact tissue that is used for metabolic and molecular work would also be suitable for analyses of foliar total inorganic nutrients. We compared quantities of nutrients extracted from wet-intact, dried-intact, and dried-ground tissues taken from a common pool of previously frozen foliage of black oak (Quercus velutina L.), sugar maple (Acer saccharum Marshall), red spruce (Picea rubens Sarg.), and white pine (Pinus strobus L.). With a few exceptions in the case of hardwoods where concentrations of total Ca, Mg, K, and P extracted from wet-intact tissue were significantly higher than dry tissue, data pooled across all collection times suggest that the extracted nutrient concentrations were comparable among the three tissue preparation methods and all for species. Based on the data presented here, it may be concluded that drying and grinding of foliage may not be necessary for nutrient analyses thus making it possible to use the same pool of tissue for total inorganic nutrients and metabolic and/or genomic analyses. To our knowledge, this is the first report on such a comparison.</p

Data_Sheet_1_Contribution of Maize Polyamine and Amino Acid Metabolism Toward Resistance Against Aspergillus flavus Infection and Aflatoxin Production.PDF

Polyamines (PAs) are ubiquitous polycations found in plants and other organisms that are essential for growth, development, and resistance against abiotic and biotic stresses. The role of PAs in plant disease resistance depends on the relative abundance of higher PAs [spermidine (Spd), spermine (Spm)] vs. the diamine putrescine (Put) and PA catabolism. With respect to the pathogen, PAs are required to achieve successful pathogenesis of the host. Maize is an important food and feed crop, which is highly susceptible to Aspergillus flavus infection. Upon infection, the fungus produces carcinogenic aflatoxins and numerous other toxic secondary metabolites that adversely affect human health and crop value worldwide. To evaluate the role of PAs in aflatoxin resistance in maize, in vitro kernel infection assays were performed using maize lines that are susceptible (SC212) or resistant (TZAR102, MI82) to aflatoxin production. Results indicated significant induction of both PA biosynthetic and catabolic genes upon A. flavus infection. As compared to the susceptible line, the resistant maize lines showed higher basal expression of PA metabolism genes in mock-inoculated kernels that increased upon fungal infection. In general, increased biosynthesis and conversion of Put to Spd and Spm along with their increased catabolism was evident in the resistant lines vs. the susceptible line SC212. There were higher concentrations of amino acids such as glutamate (Glu), glutamine (Gln) and γ-aminobutyric acid (GABA) in SC212. The resistant lines were significantly lower in fungal load and aflatoxin production as compared to the susceptible line. The data presented here demonstrate an important role of PA metabolism in the resistance of maize to A. flavus colonization and aflatoxin contamination. These results provide future direction for the manipulation of PA metabolism in susceptible maize genotypes to improve aflatoxin resistance and overall stress tolerance.</p

Reduced Silver Nanoparticle Phytotoxicity in Crambe abyssinica with Enhanced Glutathione Production by Overexpressing Bacterial γ‑Glutamylcysteine Synthase

Silver nanoparticles (Ag NPs) are widely used in consumer products, and their release has raised serious concerns about the risk of their exposure to the environment and to human health. However, biochemical mechanisms by which plants counteract NP toxicity are largely unknown. We have previously engineered Crambe abyssinica plants expressing the bacterial γ-glutamylecysteine synthase (γ-ECS) for enhancing glutathione (GSH) levels. In this study, we investigated if enhanced levels of GSH and its derivatives can protect plants from Ag NPs and AgNO₃ (Ag⁺ ions). Our results showed that transgenic lines, when exposed to Ag NPs and Ag⁺ ions, were significantly more tolerant, attaining a 28%–46% higher biomass and 34–49% more chlorophyll content, as well as maintaining 35–46% higher transpiration rates as compared to those of wild type (WT) plants. Transgenic γ-ECS lines showed 2–6-fold Ag accumulation in shoot tissue and slightly lower or no difference in root tissue relative to levels in WT plants. The levels of malondialdehyde (MDA) in γ-ECS lines were also 27.3–32.5% lower than those in WT Crambe. These results indicate that GSH and related peptides protect plants from Ag nanotoxicity. To our knowledge, this is the first direct report of Ag NP detoxification by GSH in transgenic plants, and these results will be highly useful in developing strategies to counteract the phytotoxicty of metal-based nanoparticles in crop plants

Additional file 2 of INFERR-Iron infusion in haemodialysis study: INtravenous iron polymaltose for First Nations Australian patients with high FERRitin levels on haemodialysis—a protocol for a prospective open-label blinded endpoint randomised controlled trial

Additional file 2

Additional file 3 of INFERR-Iron infusion in haemodialysis study: INtravenous iron polymaltose for First Nations Australian patients with high FERRitin levels on haemodialysis—a protocol for a prospective open-label blinded endpoint randomised controlled trial

Additional file 3

Additional file 5 of INFERR-Iron infusion in haemodialysis study: INtravenous iron polymaltose for First Nations Australian patients with high FERRitin levels on haemodialysis—a protocol for a prospective open-label blinded endpoint randomised controlled trial

Additional file 5

Additional file 1 of INFERR-Iron infusion in haemodialysis study: INtravenous iron polymaltose for First Nations Australian patients with high FERRitin levels on haemodialysis—a protocol for a prospective open-label blinded endpoint randomised controlled trial

Additional file 1

Additional file 4 of INFERR-Iron infusion in haemodialysis study: INtravenous iron polymaltose for First Nations Australian patients with high FERRitin levels on haemodialysis—a protocol for a prospective open-label blinded endpoint randomised controlled trial

Additional file 4