Dataset Information

Salinity-induced Palmella Formation Mechanism in Dunaliella salina Revealed by Quantitative Proteomics

ABSTRACT: The halotolerant algae Dunaliella salina is a good single-cell model for studying plant adaptation to high salinity. We found 151 salinity-responsive proteins were involved in multiple signaling and metabolic pathways upon palmella formation. The patterns of protein accumulation exhibited changes, including dynamics of cytoskeleton and cell membrane curvature, accumulation and transport of exopolycsccharides, photosynthesis and energy supplying (i.e. photosystem II stability and activity, cyclic electron transport, photorespiration, and C4 pathway), nuclear/chloroplastic gene expression regulation and protein processing, reactive oxygen species homeostasis, and salt signaling transduction.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Dunaliella Salina

TISSUE(S): Whole Body

SUBMITTER: Shaojun Dai

LAB HEAD: Shaojun Dai

PROVIDER: PXD005443 | Pride | 2017-06-12

REPOSITORIES: Pride

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Publications

Salinity-Induced Palmella Formation Mechanism in Halotolerant Algae Dunaliella salina Revealed by Quantitative Proteomics and Phosphoproteomics.

Wei Sijia S Bian Yangyang Y Zhao Qi Q Chen Sixue S Mao Jiawei J Song Chunxia C Cheng Kai K Xiao Zhen Z Zhang Chuanfang C Ma Weimin W Zou Hanfa H Ye Mingliang M Dai Shaojun S

Frontiers in plant science 20170523

Palmella stage is critical for some unicellular algae to survive in extreme environments. The halotolerant algae Dunaliella salina is a good single-cell model for studying plant adaptation to high salinity. To investigate the molecular adaptation mechanism in salinity shock-induced palmella formation, we performed a comprehensive physiological, proteomics and phosphoproteomics study upon palmella formation of D. salina using dimethyl labeling and Ti4+-immobilized metal i ...[more]

PMID: 28588593

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Project description:Transcriptional variation, also called expression level polymorphism (ELP), contributes to intra-specific phenotypic variation in many organisms. Differentially expressed transcripts are typically enriched for stress-related genes, suggesting that differences in response to the environment are a particularly common point of divergence among gentoypes. Analysis of ELPs also has been suggested as a way to assess unintended consequences of transgene introduction; however, it is important that interpretation of transcriptional changes be performed within the context of potential fitness effects. In these studies we sought to examine differential gene expression in response to salinity for two widely used Arabidopsis thaliana ecotypes, Wassilewskija (Ws) and Columbia (Col), and a single gene mutation (glabrous, gl1-1) in the Col background (Col(gl)), in relation to genetic, phenotypic, and fitness differences. Growth analyses were performed with seedlings germinated on culture media and growth chamber-grown plants carried through the full life cycle. Transcriptome analyses were performed with salt treated and control growth-chamber grown plants six days post initiation of salt stress. Ws plants had the least salt injury and highest dry matter accumulation and seed production in salt stressed conditions. ELPs among genoytypes and in response to 100 mM NaCl were enriched for genes associated with response to stress, including stress-associated transcription factors, heat shock and redox metabolism genes, and R genes. Application of salt resulted in many more transcripts up- or down-regulated in Col and Ws than in Col(gl). Many of the transcripts influenced by salt in Col were already altered in gl1-1 plants in the absence of salt, although Col(gl) plants did not show any detectable signs of stress, or effects on fecundity in the absence of salt treatment. The majority of salt-induced transcriptional changes that occurred in Ws also occurred in Col, suggesting common salt stress responses in these two ecotypes. Many more genes were affected by salt in Col than Ws, however, possibly reflecting the greater salt injury observed for Col. There was minimal overlap between the transcripts that differed for Ws and Col prior to salt treatment and those that were subsequently affected by salt stress. Thus, many genes conferring comparative salt stress tolerance in Ws likely differ from those whose expression levels are modified in response to salt stress. These studies demonstrate transcriptional variation among Arabidopsis genotypes in response to salt stress. Greater transcriptome differences did not necessarily correspond with greater genetic difference or phenotypic differences in morphology, fecundity, and resistance to salt stress. These results suggest that depending on circumstance, transcriptional changes can reflect response to injury, facilitate adaptive expression of fitness-associated traits, or allow for phenotypic buffering to minimize the impact of genetic changes. Three Arabidopsis genotypes were grown in the growth chamber in the absence and presence of salt stress. Plants from 20 days after sowing (6 days after salt treatment) were used for RNA extraction and hybridization on Affymetrix microarrays. There were two biological replicates for each genotype and salt treatment combination.

Project description:Mannitol is a sugar alcohol that serves as a compatible solute contributing to exceptional salt tolerance in several plant species. Arabidopsis plants transformed with the mannose-6-phosphate reductase (M6PR) gene from celery were dramatically more salt tolerant. Following treatment with 100mM NaCl, transgenic plants, relative to wild type (WT), were more successful in bolting, flowering, and production of viable seed, were less chlorotic/necrotic, and with less inhibition of growth (leaf number, rosette diameter, plant height, stalk number, and dry weight). When irrigated with 200 mM NaCl, both transformants and WT plants died without producing seeds, but M6PR transformants bolted and showed significantly less chlorosis and necrosis and had higher survival rates and dry weights than those of WT. Under normal growth conditions there were no negative effects of the M6PR transgene on overall growth (leaf number, rosette diameter, plant height, stalk number, and dry weight), or time to bolting/flowering or seed production compared to wild type (WT). Despite the lack of effects on phenotype in the absence of salt stress, genome wide expression analyses indicated that expression levels of more than 2000 genes were altered by presence of the M6PR transgene: however, there were many fewer differences observed between the M6PR and non-transgenic plants in the presence of salt stress (459), suggesting that M6PR pre-conditioned the plants for exposure to salinity. Gene categories most affected in the M6PR transgenic plants were involved in DNA binding, signal transduction, metabolism/energy, cell structure, membrane transport, defense response, and transcription. Notably, a large number of known stress genes were induced, including those related to cell walls, biotic stress, and ABA- and ethylene-responses. Our work and that in other labs has suggested that mannitol acts as an osmoprotectant, but mannitol levels are invariably quite low, and perhaps inadequate to explain its effects as an osmoprotectant. The gene expression data here indicate that stress tolerance of mannitol-producing Arabidopsis is also due, at least in part, to enhanced expression of a number of stress inducible genes related to both biotic and abiotic stress tolerance. Both M6PR transgenic and Col WT plants were grown in the growth chamber in the absence and presence of salt stress. Plants from 20 days after sowing (6 days after salt treatment) were used for RNA extraction and hybridization on Affymetrix microarrays. There were two biological replicates for each genotype and salt treatment combination.

Project description:Arabidopsis ecotypes of Sha and Ler showed differences in tolerance to salinity stress. A previous study indicated that a premature stop codon resulting in a truncated Response to ABA and Salt 1 (RAS1) protein in Sha contributes to the increased salt tolerance relative to Ler ecotype. Sha exhibited higher germination rates and longer roots on MS plate, presumably due to the decreased ABA sensitivity in Sha. More Sha plants also survived in soil after salt treatment with relatively lower electrolyte leakage when compared to Ler. Transcriptome analysis revealed that expression levels of many genes were changed between Sha and Ler ecotypes and by salt treatments. About 500 transcripts were commonly changed by at least one salinity effect and one ecotype effect, and 171 of them were co-regulated by all four comparisons. Transcripts involved in redox, secondary metabolism, auxin metabolism, photosynthesis, cell wall, and protein synthesis were mainly down-regulated by salinity effects, while transposable element genes, microRNA and antisense sequences, histone superfamily genes, and biotic stress related genes were significantly changed by Sha ecotype effects and only slightly by salinity. Several metabolic pathways such as stress, TCA, hormone/lipid/secondary metabolism, redox, development, and GO terms involved in stress, oxidation, and defense response were enriched by both salinity and ecotype effects. Ninety-five highly inducible genes were identified as candidates of RAS1 target genes and the functions involved hormone metabolism, biotic stress, RNA, DNA synthesis, protein metabolism, cell, and microRNA metabolism. All these results indicated that the Sha ecotype was possibly preconditioned to abiotic stress relative to Ler through regulation of signaling pathways and stress responsive gene expression. These comparative transcriptomic and analytical results also confirm the complexity of ABA responses and salt stress tolerance mechanisms, and they suggest additional targets for improving tolerance. Ten days old seedlings of two Arabidopsis ecotypes, Sha and Ler, were treated with 100 mM NaCl on MS plate. Plant materials were collected for RNA extraction at 4th days after treatments.

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Dataset Information

Salinity-induced Palmella Formation Mechanism in Dunaliella salina Revealed by Quantitative Proteomics

Dataset's files

Publications

Salinity-Induced Palmella Formation Mechanism in Halotolerant Algae <i>Dunaliella salina</i> Revealed by Quantitative Proteomics and Phosphoproteomics.

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