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Homeostasis of the essential micronutrient manganese (Mn) is crucially determined through availability and uptake efficiency in all organisms. Mn deficiency of plants especially occurs in alkaline and calcareous soils, seriously restricting crop yield. However, the mechanisms underlying the sensing and signaling of Mn availability and conferring regulation of Mn uptake await elucidation. Here, we uncover that Mn depletion triggers spatiotemporally defined long-lasting Ca

Free fatty acids are involved in many metabolic regulations in the human body. In this work, an ultra-fast screening method was developed for the analysis of free fatty acids using trapped ion mobility spectrometry coupled with mass spectrometry. Thirty-three free fatty acids possessing different unsaturation degrees and different carbon chain lengths were baseline separated and characterized within milliseconds. Saturated, monounsaturated, and polyunsaturated free fatty acids showed different linearities between collision cross-section values and m/z. The establishment of correlations between structures and collision cross-section values provided additional qualitative information and made it possible to determine free fatty acids which were out of the standards pool but possessed the confirmed linearity. The gas-phase separation made the quantitative analysis reliable and repeatable at a much lower time cost than chromatographic methods. The sensitivity was comparable to and even better than the reported results. The method was validated and applied to profiling free fatty acids in human plasma. Saturated free fatty acids abundance in the fasting state was found to be lower than that in the postprandial state, while unsaturated species abundance was found higher. The method was fast and robust with minimum sample pretreatment, so it was promising in the high-throughput screening of free fatty acids.

Manganese (Mn) is an essential micronutrient for all living organisms. However, excess supply of Mn as occurring in acid or waterlogged soils has toxic effects on plant physiology and development. Although a variety of Mn transporter families has been characterized, our understanding of how these transporters are regulated to uphold and adjust Mn homeostasis in plants remains rudimentary. Here, we identify two calcineurin-B-like proteins, CBL2/3, and their interacting kinases, CIPK3/9/26, as key regulators for plant Mn homeostasis. We found that Arabidopsis mutants lacking CBL2 and 3 or their interacting protein kinases CIPK3/9/26 exhibit remarkable high-Mn tolerance. Intriguingly, CIPK3/9/26 interact with and phosphorylate the tonoplast-localized Mn and iron (Fe) transporter MTP8 primarily at Ser35, which is conserved among MTP8 proteins from various species. Mn transport complementation assays in yeast combined with multiple physiological assays indicate that CBL-CIPK-mediated phosphorylation of MTP8 negatively regulates its transport activity from the cytoplasm to the vacuole. Moreover, we report that sequential phosphorylation of MTP8 initially at Ser31/32 by the calcium-dependent protein kinases CPK5 and subsequently by CIPK26 at Ser35 provides an activation/deactivation fine-tuning mechanism for differential Mn transport regulation. Collectively, our studies define a two-tiered calcium-controlled mechanism for dynamically orchestrating Mn homeostasis under conditions of fluctuating Mn supply.

Homeostasis of the essential micronutrient manganese (Mn) is crucially determined through availability and uptake efficiency in all organisms. Mn deficiency of plants especially occurs in alkaline and calcareous soils, seriously restricting crop yield. However, the mechanisms underlying the sensing and signaling of Mn availability and conferring regulation of Mn uptake await elucidation. Here, we uncover that Mn depletion triggers spatiotemporally defined long-lasting Ca 2+ oscillations in Arabidopsis roots. These Ca 2+ signals initiate in individual cells, expand, and intensify intercellularly to transform into higher-order multicellular oscillations. Furthermore, through an interaction screen we identified the Ca 2+ -dependent protein kinases CPK21 and CPK23 as Ca 2+ signal-decoding components that bring about translation of these signals into regulation of uptake activity of the high-affinity Mn transporter natural resistance associated macrophage proteins 1 (NRAMP1). Accordingly, a cpk21 / 23 double mutant displays impaired growth and root development under Mn-limiting conditions, while kinase overexpression confers enhanced tolerance to low Mn supply to plants. In addition, we define Thr498 phosphorylation within NRAMP1 as a pivot mechanistically determining NRAMP1 activity, as revealed by biochemical assays and complementation of yeast Mn uptake and Arabidopsis nramp1 mutants. Collectively, these findings delineate the Ca 2+ -CPK21/23-NRAMP1 axis as key for mounting plant Mn homeostasis.

Manganese (Mn) is an essential micronutrient in plants. However, excessive Mn absorption in acidic and waterlogged soils can lead to Mn toxicity. Despite their essential roles in Mn homeostasis, transcriptional and post-transcriptional modifications of Mn transporters remain poorly understood. Here, we demonstrated that high-Mn stress induces an obvious Ca2+ signature in Arabidopsis. We identified four calcium-dependent protein kinases, CPK4/5/6/11, that interact with the tonoplast-localized Mn and iron (Fe) transporter MTP8 in vitro and in vivo. The cpk4/5/6/11 quadruple mutant displayed a dramatic high-Mn-sensitive phenotype similar to that of the mtp8 mutant. CPKs phosphorylated the N-terminal domain of MTP8 primarily at the Ser31 and Ser32 residues. Transport assays combined with multiple physiological experiments on phospho-dead variant MTP8S31/32A and phospho-mimetic variant MTP8S31/32D plants under different Mn and Fe conditions suggested that Ser31 and Ser32 are crucial for MTP8 function. In addition, genetic analysis showed that CPKs functioned upstream of MTP8. In summary, we identified a tonoplast-associated calcium signaling cascade that orchestrates Mn homeostasis and links Mn toxicity, Ca2+ signaling, and Mn transporters. These findings provide new insight into Mn homeostasis mechanisms and Ca2+ signaling pathways in plants, providing potential targets for engineering heavy metal toxicity-tolerant plants.

Bio-oils, produced by biomass pyrolysis, have become promising candidates for feedstocks of high value-added chemicals and alternative sources for transportation fuels. Bio-oil is such a complicated mixture that contains nonpolar hydrocarbons and polar components which cover almost all kinds of organic oxygenated compounds such as carboxylic acids, alcohols, aldehydes, ketones, esters, furfurals, phenolic compounds, sugar-like material, and lignin-derived compounds. Comprehensive characterization of bio-oil and its subfractions could provide insight into the conversion process of biomass processing, as well as its further utilization as transportation fuels or chemical raw materials. This review focuses on advanced analytical strategies on in-depth characterization of bio-oil, which is concerned with gas chromatography, high-resolution mass spectrometry, FTIR spectroscopy and NMR spectroscopy, offering complementary information for previous reviews.

Phosphorus acts as an essential macroelement in plant growth and development. A lack of phosphate (Pi) in arable soil and phosphate fertilizer resources is a vital limiting factor in crop yields. Calcineurin B-like proteins (CBLs) act as one of the most important calcium sensors in plants; however, whether CBLs are involved in Pi deficiency signaling pathway remains largely elusive. In this study, we utilized a reverse genetic strategy to screen Arabidopsis thaliana T-DNA insertion mutants belonging to the CBL family under Pi deficiency conditions. The cbl1 mutant exhibited a relatively tolerant phenotype, with longer roots, lower anthocyanin content, and elevated Pi content under Pi deficiency, and a more sensitive phenotype to arsenate treatment compared with wild-type plants. Moreover, CBL1 was upregulated, and the mutation of CBL1 caused phosphate starvation-induced (PSIs) genes to be significantly induced under Pi deficiency. Histochemical staining demonstrated that the cbl1 mutant has decreased acid phosphatase activity and hydrogen peroxide concentrations under Pi deficiency. Collectively, our results have revealed a novel role of CBL1 in maintaining Pi homeostasis.

Measurements of photosynthetically active radiation (PAR), leaf photosynthesis, canopy leaf area index (LAI) and crop yield were carried from flowering to maturity to study the effects of tree shading on crop yield in a Paulownia-wheat intercropping system in China. We found that the tree shading reduced the amount of incoming PAR within the intercropping system by 22%, 44% and 56% during flowering, grain-filling and maturing, respectively. The amount of PAR intercepted by the wheat crop inside the intercropping system was 34%, 55% and 68% less than the crop outside the intercropping system during flowering, grain-filling and maturing. Estimates of PAR-saturated leaf photosynthetic rate were not affected by tree shading, and the differences between the wheat yield inside the intercropping system and outside the intercropping system can be explained by the difference in the amount of PAR intercepted. Total grain numbers and grain dry weight per 1000 grains were linearly correlated with the amount of PAR intercepted during 7 days prior to anthesis and during anthesis and grain-filling, respectively. As compared with the wheat crop outside the intercropping system, grain numbers and grain dry weight were reduced by 36% and 25%, respectively. As a result wheat yield inside the intercropping system was 51% lower than that outside the intercropping system. Our results suggest a much higher reduction in crop yield and much smaller economic gain of the Paulownia-wheat intercropping system than previous studies on the similar Paulownia-wheat intercropping systems in China.