Dataset Information

Na2CO3-responsive proteins in Puccinellia tenuiflora leaves

ABSTRACT: Puccinellia tenuiflora is a monocotyledonous halophyte species belonging to the Gramineae. It has strong ability for surviving in the extreme saline-alkali soil (pH range of 9-10). In the present study, proteins from leaves under various Na2CO3 treatments were separated and visualized on Coomassie Brilliant Blue-stained two-dimensional gel electrophoresis (2-DE) gels. After gel image analysis on the base of the calculated average vol% values of each protein spot, 174 protein spots were detected as abundance changed protein spots (1.5-fold changes, p < 0.05). Among them, 104 protein spots were identified using MALDI-TOF MS/MS and Mascot database searching. Thus the 104 protein identities represented 80 unique proteins were taken as Na2CO3-responsive proteins in leaves. After integrative analysis by BLAST alignment, Kyoto Encyclopedia of Genes and Genomes, and referring information from related literature, they were classified into 10 functional categories, including photosynthesis, carbohydrate and energy metabolism, other metabolisms, stress and defense, membrane and transport, signaling, protein synthesis, folding, and turnover, cell wall mechanism, cell cycle, and miscellaneous or unknown.

INSTRUMENT(S): TripleTOF 5600

ORGANISM(S): Oryza Sativa (rice)

TISSUE(S): Plant Cell

SUBMITTER: Shaojun Dai

LAB HEAD: Shaojun Dai

PROVIDER: PXD005455 | Pride | 2021-08-30

REPOSITORIES: Pride

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Publications

Na2CO3-responsive Photosynthetic and ROS Scavenging Mechanisms in Chloroplasts of Alkaligrass Revealed by Phosphoproteomics.

Suo Jinwei J Zhang Heng H Zhao Qi Q Zhang Nan N Zhang Yongxue Y Li Ying Y Song Baohua B Yu Juanjuan J Cao Jianguo J Wang Tai T Luo Ji J Guo Lihai L Ma Jun J Zhang Xumin X She Yimin Y Peng Lianwei L Ma Weimin W Guo Siyi S Miao Yuchen Y Chen Sixue S Qin Zhi Z Dai Shaojun S

Genomics, proteomics & bioinformatics 20200601 3

Alkali-salinity exerts severe osmotic, ionic, and high-pH stresses to plants. To understand the alkali-salinity responsive mechanisms underlying photosynthetic modulation and reactive oxygen species (ROS) homeostasis, physiological and diverse quantitative proteomics analyses of alkaligrass (Puccinellia tenuiflora) under Na2CO3 stress were conducted. In addition, Western blot, real-time PCR, and transgenic techniques were applied to validate the proteomic results and test t ...[more]

PMID: 32683046

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Project description:Data analysis. Spot detection and matching were performed with a comparative cross analysis of all the gels using DeCyder software v.6.5 (GE Healthcare). 178 spots were selected based on 1.15-fold for protein ratio cut-off, allowing for the appearance of the spots in 23 out of 28 gels (69 out of 84 total images). Data from 95 spots were submitted. 93 spots were identified with high confidence. Spot picking and Trypsin digestion. The spots of interest were picked up by Ettan Spot Picker (GE Healthcare) based on the in-gel analysis and spot picking design by DeCyder software. The gel spots were washed a few times then digested in-gel with modified porcine trypsin protease (Promega, Fitchburg, WI). The digested tryptic peptides were desalted using a Zip-tip C18 (Millipore, Billerica, MA). Peptides were eluted from the Zip-tip with 0.5 uL of matrix solution (alpha-cyano-4-hydroxycinnamic acid, 5 mg/mL in 50% acetonitrile, 0.1% trifluoroacetic acid, 25mM ammonium bicarbonate) and spotted on a MALDI plate. Mass Spectrometry. MALDI-TOF MS and TOF/TOF tandem MS/MS were performed on AB SCIEX TOF/TOF 5800 System (AB SCIEX). MALDI-TOF mass spectra were acquired in reflectron positive ion mode, averaging 4000 laser shots per spectrum. TOF/TOF tandem MS fragmentation spectra were acquired for each sample, averaging 4000 laser shots per fragmentation spectrum on each of the 7-10 most abundant ions present in each sample (excluding trypsin autolytic peptides and other known background ions). Database search. Both the resulting peptide mass and the associated fragmentation spectra were submitted to GPS Explorer workstation equipped with MASCOT search engine (Matrix Science, Boston, MA) to search the Swiss-Prot database. Searches were performed without constraining protein molecular weight or isoelectric point, with variable carbamidomethylation of cysteine and oxidation of methionine residues, and with one missed cleavage also allowed in the search parameters. Candidates with either protein score C.I.% or Ion C.I.% greater than 95 were considered significant. When multiple IDs were significant for a given spot, the selection was made by evaluating apparent molecular weight, isoelectric point, the location of the spot in the gel, and the presence of strips of multiple protein isoforms in the adjacent spots.

Project description:Spermatophyte pollen tubes and root hairs have been used as single-cell-type model systems toward understanding the molecular processes underlying polar growth of plant cells. Horsetail (Equisetum arvense L.) is a perennial herb species in Equisetopsida, which creates separately growing spring and summer stems in its life cycle. The mature chlorophyllous spores produced from spring stems can germinate without dormancy. Here we report the cellular features and protein expression patterns in five stages of horsetail spore germination (mature spores, rehydrated spores, double-celled spores, germinated spores, and spores with protonemal cells). Using 2-DE combined with mass spectrometry, 80 proteins were found to be differentially expressed upon spore germination. Among them, proteins involved in photosynthesis, protein turnover, and energy supply were over-represented. Thirteen proteins appeared as isoforms on the gels, indicating the potential importance of post-translational modification. In addition, the dynamic changes of ascorbate peroxidase, peroxiredoxin, and dehydroascorbate reductase implied that reactive oxygen species homeostasis is critical in regulating cell division and tip-growth. The time course of germination and diverse expression patterns of proteins in photosynthesis, energy supply, lipid and amino acid metabolism indicated that heterotrophic and autotrophic metabolism were necessary in light-dependent germination of the spores. Twenty-six proteins were involved in protein synthesis, folding, and degradation, indicating that protein turnover is vital to spore germination and rhizoid tip-growth. Furthermore, the altered abundance of small G protein Ran1, 14-3-3 protein, actin, and Caffeoyl-CoA O-methyltransferase revealed that signaling transduction, vesicle trafficking, cytoskeleton dynamics, and cell wall modulation were critical to cell division and polar growth. These findings provide up-to-date evidences for understanding fern spore asymmetric division and rhizoid polar growth.

Dataset Information

Na2CO3-responsive proteins in Puccinellia tenuiflora leaves

Publications

Na<sub>2</sub>CO<sub>3</sub>-responsive Photosynthetic and ROS Scavenging Mechanisms in Chloroplasts of Alkaligrass Revealed by Phosphoproteomics.

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