Evaluation of automated analysis of 15N and total N in plant material and soil

Simultaneous determination of ¹⁵N and total N using an automated nitrogen analyser interfaced to a continuous-flow isotope ratio mass spectrometer (ANA-MS method) was evaluated. The coefficient of variation (CV) of repeated analyses of homogeneous standards and samples at natural abundance was lower than 0.1%. The CV of repeated analyses of ¹⁵N-labelled plant material and soil samples varied between 0.3% and 1.1%. The reproductibility of repeated total N analyses using the automated method was comparable to results obtained with a semi-micro Kjeldahl procedure. However, the automated method gave results which were 3% to 5% higher than those obtained with the Kjeldahl procedure. Since only small samples can be analysed, careful sample homogenization and fine grinding are very important. Evaluation of a diffusion method for preparing nitrate and ammonium in solution for automated ¹⁵N analysis showed that the recovery of inorganic N in the NH₃ trap was lower when the N was diffused from water than from 2 M KCl. The results also indicated that different proportions of the NO₃ ^- and the NH₄ ⁺ in aqueous solution were recovered in the trap after combined diffusion. The method is most suited for diffusing either NO₃ ^- or NH₄ ⁺ alone, but can be used for combined diffusion of the two ions.     

Tissue-Characteristic Expression of Mouse Proteome

The mouse is a valuable model organism for biomedical research. Here, we established a comprehensive spectral library and the data-independent acquisition–based quantitative proteome maps for 41 mouse organs, including some rarely reported organs such as the cornea, retina, and nine paired organs. The mouse spectral library contained 178,304 peptides from 12,320 proteins, including 1678 proteins not reported in previous mouse spectral libraries. Our data suggested that organs from the nervous system and immune system expressed the most distinct proteome compared with other organs. We also found characteristic protein expression of immune-privileged organs, which may help understanding possible immune rejection after organ transplantation. Each tissue type expressed characteristic high-abundance proteins related to its physiological functions. We also uncovered some tissue-specific proteins which have not been reported previously. The testis expressed highest number of tissue-specific proteins. By comparison of nine paired organs including kidneys, testes, and adrenal glands, we found left organs exhibited higher levels of antioxidant enzymes. We also observed expression asymmetry for proteins related to the apoptotic process, tumor suppression, and organ functions between the left and right sides. This study provides a comprehensive spectral library and a quantitative proteome resource for mouse studies.     

A caterpillar (Callopistria floridensis G. (Lepidoptera: Noctuidae)) accumulates arsenic from an arsenic‐hyperaccumulating fern (Pteris vittata L.).

1. The consumption of arsenic is toxic to most biota. However, a noctuid caterpillar was recently reported feeding on a plant known to hyperaccumulate arsenic. 2. The aim of this study was to investigate the effects of arsenic‐rich Pteris vittata L. consumption by Callopistria floridensis G., and measure differences in arsenic concentrations at various stages of development (larval and adult), and associated with exuviae and frass. 3. Callopistria floridensis accumulated extraordinary concentrations of arsenic. The relative accumulation of arsenic was highest in exuviae and larvae. Larvae invariably preferred P. vittata grown on low arsenic soil to P. vittata grown on higher soil arsenic concentrations, and appeared able to selectively forage on lower arsenic concentrations within each treatment. 4. These findings show that C. floridensis is tolerant of arsenic, and successfully develops to adulthood containing elevated concentrations of arsenic. Callopistria floridensis represents the only known terrestrial animal capable of accumulating arsenic, and may have developed novel physiological and behavioural adaptations to regulate the negative effects of arsenic.     

Nitrogen and phosphorus content of some temperate and tropical freshwater fishes

The nitrogen and phosphorus content of two temperate fishes, Rutilus rutilus and Perca fluviatilis , and six tropical fishes, Oreochromis niloticus , Cichla monoculus , Serrassalmus rhombeus , Plagioscion squamosissimus , Prochilodus brevis and Hoplias malabaricus , were investigated to test the hypothesis that variation in body P content and N:P ratio is related to body size. Regressions of %P and N:P ratios against fish size (length and mass) confirmed the hypothesis for P. fluviatilis and P. squamosissimus , suggesting that body size is an important factor driving body P content and N:P ratios in some fishes. Moreover, significant increases in %N and N:P ratio with body size was found for H. malabaricus , a common piscivorous fish of the Neotropics. Interspecific variation in %P and N:P ranged two-fold and significant differences ( P < 0·05) were found among the tested species. The mean ± s . d . elemental content across all fishes ( n = 170) was 10·35 ± 1·29% for N and 3·05 ± 0·82% for P, while the N:P ratio was 8·00 ± 2·14. Data on fish body nutrient content and ratio will improve parameterization of bioenergetics and mass balance models and help clarify the role of fishes in nutrient cycles in both temperate and tropical freshwaters.     

The fate of nitrogen (15N) released from different plant materials during decomposition under field conditions

Release of N, retention in soil, availability to a subsequent crop and total recovery of N derived from different¹⁵N-labelled plant materials decomposing in soil was investigated in two field experiments. In the first experiment five different plant species (white clover, red clover, subterranean clover, field bean and timothy) and in the second subterranean clover of different maturity (2,3 and 4 months old) were buried in mesh bags in the soil and allowed to decompose for 10 and 4 months, respectively. Most of the N released from the decaying plant materials was retained in the soil (27–46% of input). The subsequent crop (barley) took up 6–25% of input. The uptake correlated with the amount of N released from the decomposing material (r=0.936^*, I experiment). Similar amounts of subterranean clover N were taken up by barley regardless to whether the material was buried in soil in the previous autumn or just before sowing of the crop. At the end of the experiments, the total recovery of the introduced plant-derived N varied between 89 and 102%. The results present evidence that the ability of the soil to retain plant-derived N is strong in comparison with the ability of the subsequent crop and different loss mechanisms to remove it.     

Distribution and redistribution of extension growth along vertical and horizontal gravireacting maize roots

Horizontal primary roots of Zea mays L. were photographed during the course of their gravireaction and during a preceding growth period in the vertical orientation. The displacement, by root elongation, of marker particles on the root surface was recorded. The particle-displacement rates were used to estimate the distribution of elemental elongation rates along opposite sides of the growing root apex. In the temperature range 21–25°C there was a stimulation of local elongation rates along the upper side of a gravireacting root and a reduction (and sometimes a cessation) of elongation along the lower side. Elemental elongation rates have been related to the development of root curvature, and the magnitude of the differential growth between upper and lower sides required for a particular rate of bending has also been estimated. The results complement, and are compatible with, findings relating to the distribution of certain endogenous growth regulators believed to participate in the gravireaction.Abbreviation RELEL relative elemental rate of elongation     

DIA-Based Proteomics Identifies IDH2 as a Targetable Regulator of Acquired Drug Resistance in Chronic Myeloid Leukemia

Drug resistance is a critical obstacle to effective treatment in patients with chronic myeloid leukemia. To understand the underlying resistance mechanisms in response to imatinib mesylate (IMA) and adriamycin (ADR), the parental K562 cells were treated with low doses of IMA or ADR for 2 months to generate derivative cells with mild, intermediate, and severe resistance to the drugs as defined by their increasing resistance index. PulseDIA-based (DIA [data-independent acquisition]) quantitative proteomics was then employed to reveal the proteome changes in these resistant cells. In total, 7082 proteins from 98,232 peptides were identified and quantified from the dataset using four DIA software tools including OpenSWATH, Spectronaut, DIA-NN, and EncyclopeDIA. Sirtuin signaling pathway was found to be significantly enriched in both ADR-resistant and IMA-resistant K562 cells. In particular, isocitrate dehydrogenase (NADP(+)) 2 was identified as a potential drug target correlated with the drug resistance phenotype, and its inhibition by the antagonist AGI-6780 reversed the acquired resistance in K562 cells to either ADR or IMA. Together, our study has implicated isocitrate dehydrogenase (NADP(+)) 2 as a potential target that can be therapeutically leveraged to alleviate the drug resistance in K562 cells when treated with IMA and ADR.     

Alteration of plankton communities and biogeochemical cycles by harmful Cochlodinium polykrikoides (Dinophyceae) blooms

Cochlodinium polykrikoides is a globally distributed, ichthyotoxic, bloom-forming dinoflagellate. Blooms of C. polykrikoides manifest themselves as large (many km²) and distinct patches with cell densities exceeding 10³ ml⁻¹ while water adjacent to these patches can have low cell densities (<100 cells ml⁻¹). While the effect of these blooms on fish and shellfish is well-known, their impacts on microbial communities and biogeochemical cycles are poorly understood. Here, we investigated plankton communities and the cycling of carbon, nitrogen, and B-vitamins within blooms of C. polykrikoides and compared them to areas in close proximity (<100 m) with low C. polykrikoides densities. Within blooms, C. polykrikoides represented more than 90% of microplankton (>20 μm) cells, and there were significantly more heterotrophic bacteria and picoeukaryotic phytoplankton but fewer Synechococcus. Terminal restriction fragment length polymorphism analysis of 16S and 18S rRNA genes revealed significant differences in community composition between bloom and non-bloom samples. Inside the bloom patches, concentrations of vitamin B₁₂ were significantly lower while concentrations of dissolved oxygen were significantly higher. Carbon fixation and nitrogen uptake rates were up to ten times higher within C. polykrikoides bloom patches. Ammonium was a more important source of nitrogen, relative to nitrate and urea, for microplankton within bloom patches compared to non-bloom communities. While uptake rates of vitamin B₁ were similar in bloom and non-bloom samples, vitamin B₁₂ was taken up at rates five-fold higher (>100 pmol⁻¹ L⁻¹ d⁻¹) in bloom samples, resulting in turn-over times of hours during blooms. This high vitamin demand likely led to the vitamin B₁₂ limitation of C. polykrikoides observed during nutrient amendment experiments conducted with bloom water. Collectively, this study revealed that C. polykrikoides blooms fundamentally change microbial communities and accelerate the cycling of carbon, some nutrients, and vitamin B₁₂.     

The chemical composition of Astragalus: a comparison of seleniferous and non-seleniferous plants growing side by side

COWGILL, U. M. & LANDENBERGER, B.D., 1992. The chemical composition of Astragalus: a comparison of seleniferous and non-seleniferous plants growing side by side. This paper describes how non-seleniferous plant species coexist with seleniferous ones and what chemical changes occur in non-seleniferous species that allow the toleration of large quantities of volatilized Se-bearing compounds. These compounds are known for their phytotoxicity as well as for their toxicity to mammals and insects. Twenty-three sites (Colorado, Utah, New Mexico, U.S.A.) were examined over a 6-year period. Plants collected from these sites were divided into four groups: seleniferous astragali, seleniferous Brassicaceae, non-seleniferous astragali and non-seleniferous associated genera. Furthermore, it was possible to categorize these sites: Type 1 sites supported all four groups of plants, Type 2 sites contained only seleniferous astragali, whereas Type 3 sites sustained only non-seleniferous astragali. When concentrations of Cu, Zn (P <0.0001), Si (P<0.02), As, Pb (P<0.03) and Cs (P<0.05) were measured, the values for those seleniferous astragali that coexisted with other species were significantly different from the values found for those seleniferous astragali that grew alone. There were four areas that in some years supported all four groups of plants (Type 1 site) and in other years contained only seleniferous astragali. In this way, the latter may be used as a chemical control for the former. The seleniferous astragali collected at Type 1 sites always contained more Cu, As, Si and Pb than the solitary astragali of Type 2 sites. When seleniferous astragali of Type 1 sites that support all four groups of plants develop in a year where the usually associated species of past years are absent, they contain in their tissues concentrations of Cu, As, Si, Pb, Zn and Cs typical of Type 2 sites where the seleniferous astragali grew alone. Non-seleniferous astragali of Type 1 sites that support all four groups of plants have a chemical composition that differs significantly in the quantity of 15 elements from non-seleniferous astragali of Type 3 sites where the latter grew alone. Since allelochemicals such as phenolic acids and flavonoids are well known to be able to alter mineral absorption by plants and since the astragali are known to produce such substances, it is suggested that the possible production of phenolic acids and flavonoids may permit coexistence of seleniferous plants with non-seleniferous ones and thus explain the chemical differences among the plants of the four types of sites.