MetaboLightsapplication/xmlftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14777/m_MTBLS14777_LC-MS_negative_hilic_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14777/m_MTBLS14777_LC-MS_positive_hilic_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14777/a_MTBLS14777_LC-MS_negative_hilic.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14777/s_MTBLS14777.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14777/a_MTBLS14777_LC-MS_positive_hilic.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14777/i_Investigation.txtprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14777<p>After that, the table will be further analyzed and processed. Software infor: Compound Discoverer Version:v.3.3 Paramenter:Parent ion mass deviation: &lt; 5ppm Mass deviation of fragment ions: &lt; 10ppm Retention time deviation: &lt; 0.2min Offical Website:https://mycompounddiscoverer.com/</p>MetaboLightsPublicLiquid Chromatography MS - positive - hilicLiquid Chromatography MS - negative - hilic<p>In this experiment, waters 2777c UPLC (waters, USA) in series with Q exactive HF high resolution mass spectrometer (Thermo Fisher Scientific, USA) was used for the separation and detection of metabolites.</p><p>Chromatographic conditions:Chromatographic separation was performed on Hypersil GOLD aQ Dim column (1.9 μm 2.1*100 mm, Thermo Fisher Scientific, USA), with mobile phase A consisting 0.1% formic acid in water and mobile phase B consisting 0.1 formic acid in acetonitrile. The column temperature was maintained at 40 °C. The gradient conditions were as follows: 5% B over 0.0-2.0 min, 5-95% B over 2.0-22.0min, held constant at 95% B over 22.0-27.0 min and washed with 95% B over 27.1-30 min. The flow rate was 0.3 mL/min and the injection volume was 5 μL.</p>Elevated temperature accelerates leaf senescence and promotes leaf nitrogen transport to improve rice grain protein synthesis. 10.1093/plphys/kiag310. PMID:42230303Yufei ZhaoNanjing Agriculture UniversityLeaf and grain mixturemass spectrometry assay<p>50 μg of samples were weighed into 1.5 mL Eppendorf tubes respectively and soaked with 800 μL of precooling extraction solution (methanol : H2O = 7:3, v/v) and 20 μL Internal Standard 1 (IS1). Homogenization was conducted with weaving grinder at 50 Hz for 10 minutes and then water bath ultrasonication at 4 °C for 30 minutes. Following the standing still at -20°C for 1 hour, the extracts were centrifuged at the speed of 14000 rpm at 4 °C for 15 minutes. 600 μL of the supernatant was filtered with 0.22 μm membrane and 20 μL of filtered solution from each sample composited the mixed QC sample to evaluate the repeatability and stability of LC/MS analysis. Filtered samples and mixed QC samples were transferred to the 1.5 mL sample vials for instrument running.</p>Oryza sativa L.https://www.ebi.ac.uk/metabolights/MTBLS14777Yufei Zhao. Nanjing Agriculture University. 2022201093@stu.njau.edu.cn.<p>After importing the off-line data of mass spectrometry into compound discoverer 3.3 (Thermo Fisher Scientific, USA) software and analyzing the mass spectrometry data in combination with bmdb (BGI metabolome database), mzcloud database and chemspider online database, a data matrix containing information such as metabolite peak area and identification results will be obtained.&nbsp;</p>Treatment2022201093@stu.njau.edu.cn<p>At 15 and 30 days after flowering, 30 whole rice plants were collected from each plot under ambient and warming treatments. The plants were brought to the laboratory, separated by tissue type, bagged, and immediately immersed in liquid nitrogen. For strong and weak grains, glumes were removed on dry ice. Leaf and grain samples were ground using a ball mill and then stored at -80 °C for subsequent metabolomic analysis.</p>MetabolomicsOryza sativa L.MetabolomicsOryza sativaSenescenceLeaf and grain mixtureuntargeted analysisWaters ACQUITY UPLC systemexperimental blankThermo Scientific Q Exactive HFuntargeted metabolite profilingOryza sativa L.MetabolomicsOryza sativaSenescenceLeaf and grain mixtureuntargeted analysisWaters ACQUITY UPLC systemexperimental blankThermo Scientific Q Exactive HFuntargeted metabolite profiling<p>Mass spectrometry conditions:Using Q Exactive HF (Thermo Fisher Scientific, USA) perform primary and secondary mass spectrometry data acquisition. The scan range was125~1500m/z for positive ion and 100-1500 m/z for negative ion with a resolution of 120,000, and the automatic gain control (AGC) target for MS acquisitions was set to 1e6 with a maximum ion injection time of 100 ms. Top 3 precursors were selected for subsequent MSMS fragmentation with a maximum ion injection time of 50 ms and resolution of 30000, the AGC was 2e5. The stepped normalized collision energy was set to 20, 40 and 60 eV. ESI parameters were setting as : Sheath gas flow rate was 40, Aux gas flow rate was 10, positive-ion mode Spray voltage(|KV|) was 3.80, negative-ion mode Spray voltage(|KV|) was 3.20, Capillary temperature was 320°C, Aux gas heater temperature was 350°C.</p>Benzoic acidCorchorifatty acid FGlycerol 3-phosphateD-alpha-Hydroxyglutaric acidGuanosineNicotinic acid6-Aminocaproic acidMethylsuccinic acidalpha-LactoseGlycylglycine(2R)-2,3-Dihydroxypropanoic acidD-(+)-MannoseDiosmetinItaconic acid5-Aminolevulinic acid16-Hydroxyhexadecanoic acidalpha,alpha-TrehaloseVanillic acid4-Coumaric acidL-HistidineL-(-)-Malic acidReserpineJasmonic acidD-Glucose 6-phosphateGluconic acidN-Acetyl-DL-glutamic acidEsculinThreonineD-(-)-Quinic acidGallic acid2-Hydroxycaproic acid3-Coumaric acid5-Hydroxyindole-3-acetic acid2-Ketobutyric acid2-Furoic acidD-(+)-Galactosebeta-D-Fructose 6-phosphatedelta-Ribono-1,4-lactoneD-RaffinoseIbuprofen metabolite ASyringic acidUridineSuccinic acidL-Aspartic acid2-Oxoglutaric acid3-Hydroxyanthranilic acidPhenylacetaldehyde4-Dodecylbenzenesulfonic acidD-(-)-FructoseFlavin mononucleotide (FMN)AICA ribonucleotide3-Methylglutaric acidSuberic acidtrans-Aconitic acidL-Threonic acidDantroleneLauryl gallate4-NitrophenolIsocitric acidMevalonic acid2-MethoxyestroneKaempferitrincis-Aconitic acidSalicylic acidLevulinic acidDodecanedioic acid(+/-)12(13)-DiHOMEPimelic acid2,5-Furandicarboxylic acidOleoyl-L-alpha-lysophosphatidic acid2-Methylglutaric acidCitric acid3-Hydroxy-3-methylglutaric acidD-Sedoheptulose 7-phosphateL-TyrosineCynarosideFumaric acidCitraconic acid2,4-Dihydroxybenzoic acidL-SerinePantothenic acidElevated temperatures during grain filling severely constrain rice yield and quality. Although additional nitrogen can mitigate the adverse effects of elevated temperature, both factors increase grain protein content, and the underlying physiological mechanisms remain poorly understood. Here, we conducted actual field warming (2.28°C day/4.33°C night) during the grain-filling period and applied an additional 60 kg N ha-1. The grain weight of superior spikelets (SS) was 2.1% lower under elevated temperature (ET), while ET increased the grain weight by 4.23% and protein content by 2.95% in inferior spikelets (IS). ET promoted the level of free amino acids, improved the activities of glutamine synthetase and glutamate synthase, and up-regulated the expression of an amino acid transporter gene (OsLHT1), mainly 9-15 days after flowering (DAF), in the leaf, ultimately accelerating leaf senescence post 20 DAF. Regardless of the temperature, the effect of nitrogen on leaves was similar to that of ET, while delaying leaf senescence and further increasing protein content in SS and IS. Metabolomic analysis further confirmed that ET accelerated leaf senescence and amino acid depletion in the leaves. Furthermore, the higher levels of L-histidine, along with increased levels of stress-responsive metabolites (D-raffinose and gentisic acid), collectively contributed to the improved protein content in IS under ET. Overall, the study provides insight into grain protein accumulation under warming.Elevated temperature accelerates leaf senescence and promotes leaf nitrogen transport to improve rice grain protein synthesis.Zhao Yufei Y, Zhang Chen C, Shi Wentao W, Liu Ke K, Wu Wei W, Wang Yifan Y, Li Ruiqi R, Peng Yuxuan Y, Shen Yingying Y, Liu Wenzhe W, Ding Yanfeng Y, Xi Min M, Tang She SfalseElevated temperature accelerates leaf senescence and promotes leaf nitrogen transport to improve rice grain protein synthesisElevated temperatures during grain filling severely constrain rice yield and quality. Although additional nitrogen can mitigate the adverse effects of elevated temperature, both factors increase grain protein content, and the underlying physiological mechanisms remain poorly understood. Here, we conducted actual field warming (2.28°C day/4.33°C night) during the grain-filling period and applied an additional 60 kg N ha-1. The grain weight of superior spikelets (SS) was 2.1% lower under elevated temperature (ET), while ET increased the grain weight by 4.23% and protein content by 2.95% in inferior spikelets (IS). ET promoted the level of free amino acids, improved the activities of glutamine synthetase and glutamate synthase, and up-regulated the expression of an amino acid transporter gene (OsLHT1), mainly 9-15 days after flowering (DAF), in the leaf, ultimately accelerating leaf senescence post 20 DAF. Regardless of the temperature, the effect of nitrogen on leaves was similar to that of ET, while delaying leaf senescence and further increasing protein content in SS and IS. Metabolomic analysis further confirmed that ET accelerated leaf senescence and amino acid depletion in the leaves. Furthermore, the higher levels of L-histidine, along with increased levels of stress-responsive metabolites (D-raffinose and gentisic acid), collectively contributed to the improved protein content in IS under ET. Overall, the study provides insight into grain protein accumulation under warming.2026-06-172026-06-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