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Proteome Changes Reveal the Protective Roles of Exogenous Citric Acid in Alleviating Cu Toxicity in Brassica napus L.

ABSTRACT: Citric acid (CA), as an organic chelator, plays a vital role in alleviating copper (Cu) stress-mediated oxidative damage, wherein a number of molecular mechanisms alter in plants. However, it remains largely unknown how CA regulates differentially abundant proteins (DAPs) in response to Cu stress in Brassica napus L. In the present study, we aimed to investigate the proteome changes in the leaves of B. L. seedlings in response to CA-mediated alleviation of Cu stress. Exposure of 21-day-old seedlings to Cu (25 and 50 μM) and CA (1.0 mM) for 7 days exhibited a dramatic inhibition of overall growth and considerable increase in the enzymatic activities (POD, SOD, CAT). Using a label-free proteome approach, a total of 6345 proteins were identified in differentially treated leaves, from which 426 proteins were differentially expressed among the treatment groups. Gene ontology (GO) and KEGG pathways analysis revealed that most of the differential abundance proteins were found to be involved in energy and carbohydrate metabolism, photosynthesis, protein metabolism, stress and defense, metal detoxification, and cell wall reorganization. Our results suggest that the downregulation of chlorophyll biosynthetic proteins involved in photosynthesis were consistent with reduced chlorophyll content. The increased abundance of proteins involved in stress and defense indicates that these DAPs might provide significant insights into the adaptation of Brassica seedlings to Cu stress. The abundances of key proteins were further verified by monitoring the mRNA expression level of the respective transcripts. Taken together, these findings provide a potential molecular mechanism towards Cu stress tolerance and open a new route in accelerating the phytoextraction of Cu through exogenous application of CA in B. napus.

SUBMITTER: Ju YH

PROVIDER: S-EPMC8198124 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Phytoremediation soil polluted by cadmium has drawn worldwide attention. However, how to improve the efficiency of plant remediation of cadmium contaminated soil remains unknown. Previous studies showed that nitrogen (N) significantly enhances cadmium uptake and accumulation in poplar plants. In order to explore the important role of nitrogen in plants' responses to cadmium stress, this study investigates the poplar proteome and phosphoproteome difference between Cd stress and Cd + N treatment. In total, 6573 proteins were identified, and 5838 of them were quantified. With a fold-change threshold of > 1.3, and a p-value < 0.05, 375 and 108 proteins were up- and down-regulated by Cd stress when compared to the control, respectively. Compared to the Cd stress group, 42 and 89 proteins were up- and down-regulated by Cd + N treatment, respectively. Moreover, 522 and 127 proteins were up- and down-regulated by Cd + N treatment compared to the CK group. In addition, 1471 phosphosites in 721 proteins were identified. Based on a fold-change threshold of > 1.2, and a p-value < 0.05, the Cd stress up-regulated eight proteins containing eight phosphosites, and down-regulated 58 proteins containing 69 phosphosites, whereas N + Cd treatment up-regulated 86 proteins containing 95 phosphosites, and down-regulated 17 proteins containing 17 phosphosites, when compared to Cd stress alone. N + Cd treatment up-regulated 60 proteins containing 74 phosphosites and down-regulated 37 proteins containing 42 phosphosites, when compared to the control. Several putative responses to stress proteins, as well as transcriptional and translational regulation factors, were up-regulated by the addition of exogenous nitrogen following Cd stress. Especially, heat shock protein 70 (HSP70), 14-3-3 protein, peroxidase (POD), zinc finger protein (ZFP), ABC transporter protein, eukaryotic translation initiation factor (elF) and splicing factor 3 B subunit 1-like (SF3BI) were up-regulated by Cd + N treatment at both the proteome and the phosphoproteome levels. Combing the proteomic data and phosphoproteomics data, the mechanism by which exogenous nitrogen can alleviate cadmium toxicity in poplar plants was explained at the molecular level. The results of this study will establish the solid molecular foundation of the phytoremediation method to improve cadmium-contaminated soil.

Project description:BACKGROUND:The ubiquitous signaling molecule melatonin (N-acetyl-5-methoxytryptamine) (MT) plays vital roles in plant development and stress tolerance. Selenium (Se) may be phytotoxic at high concentrations. Interactions between MT and Se (IV) stress in higher plants are poorly understood. The aim of this study was to evaluate the defensive roles of exogenous MT (0??M, 50??M, and 100??M) against Se (IV) (0??M, 50??M, 100??M, and 200??M) stress based on the physiological and biochemical properties, thiol biosynthesis, and antioxidant system of Brassica napus plants subjected to these treatments. RESULTS:Se (IV) stress inhibited B. napus growth and biomass accumulation, reduced pigment content, and lowered net photosynthetic rate (Pn) and PSII photochemical efficiency (Fv/Fm) in a dose-dependent manner. All of the aforementioned responses were effectively alleviated by exogenous MT treatment. Exogenous MT mitigated oxidative damage and lipid peroxidation and protected the plasma membranes from Se toxicity by reducing Se-induced reactive oxygen species (ROS) accumulation. MT also alleviated osmotic stress by restoring foliar water and sugar levels. Relative to standalone Se treatment, the combination of MT and Se upregulated the ROS-detoxifying enzymes SOD, APX, GR, and CAT, increased proline, free amino acids, and the thiol components GSH, GSSG, GSH/GSSG, NPTs, PCs, and cys and upregulated the metabolic enzymes ?-ECS, GST, and PCS. Therefore, MT application attenuates Se-induce oxidative damage in plants. MT promotes the accumulation of chelating agents in the roots, detoxifies Se there, and impedes its further translocation to the leaves. CONCLUSIONS:Exogenous MT improves the physiological traits, antioxidant system, and thiol ligand biosynthesis in B. napus subjected to Se stress primarily by enhancing Se detoxification and sequestration especially at the root level. Our results reveal better understanding of Se-phytotoxicity and Se-stress alleviation by the adequate supply of MT. The mechanisms of MT-induced plant tolerance to Se stress have potential implications in developing novel strategies for safe crop production in Se-rich soils.

Project description:Phytoremediation soil polluted by heavy metal has been drawn on a worldwide attention from human society. However, how to improve the efficiency of plant remediation of soil contaminated by cadmium remains unknown. Previous studies showed that nitrogen (N) significantly enhanced cadmium uptake in poplar plants. In order to further explore the key role of N in the detoxification against cadmium stress in plants, this study try to investigate the poplar proteome and phosphoproteome difference between Cd stress and Cd+N treatment. In total, 5838 of the 6573 identified were quantified. With a fold-change threshold >1.3 and p-value<0.05, 375 and 108 proteins were up- and down-regulated by Cd stress when compared to the control, 42 and 89 proteins were up- and down-regulated, respectively, in Cd+N / Cd group, 522 and 127 proteins were up- and down-regulated, respectively, in Cd+N / CK group. In addition, the phosphoproteome data was obtained after the proteomic difference was normalized, and 1471phosphosites in 721 proteins were quantified. Based on a fold-change threshold >1.2, P-value <0.05, the Cd stress up-regulated 8 phosphosites in 8 proteins and down-regulated 69 phosphosites in 58 proteins, whereas N+Cd treatment up-regulated 95 phosphosites in 86 proteins and down-regulated 17 phosphosites in 17 proteins when compared to sole Cd stress. In addition, N+Cd treatment up-regulated 74 phosphosites in 60 proteins and down-regulated 42 phosphosites in 37 proteins when compared to the control.Several putative responses to stress proteins, transcriptional and translational regulation factors were up-regulated by addition ofexogenous nitrogen followed Cd stress at the proteome and phosphoproteome levels. Especially, heat shock protein 70 (HSP70), peroxidase (POD), zinc finger protein (ZFP), ABC transporter protein (ABC), eukaryotic translation initiation factor (elF) and splicing factor 3B subunit 1-like (SF3BI) were up-regulated by Cd+N treatment whether at the proteome or at the phosphoproteome levels, which was need to further study. In a word, taken together of proteome and phosphoproteome data, nitrogen serves a protective role in plants treated with Cd by multiple ways.

Project description:BACKGROUND: Unlike maternal inheritance of mitochondria in sexual reproduction, somatic hybrids follow no obvious pattern. The introgressed segment orf138 from the mitochondrial genome of radish (Raphanus sativus) to its counterpart in rapeseed (Brassica napus) demonstrates that this inheritance mode derives from the cytoplasm of both parents. Sequencing of the complete mitochondrial genome of five species from Brassica family allowed the prediction of other extraneous sources of the cybrids from the radish parent, and the determination of their mitochondrial rearrangement. RESULTS: We obtained the complete mitochondrial genome of Ogura-cms-cybrid (oguC) rapeseed. To date, this is the first time that a heterogeneously composed mitochondrial genome was sequenced. The 258,473 bp master circle constituted of 33 protein-coding genes, 3 rRNA sequences, and 23 tRNA sequences. This mitotype noticeably holds two copies of atp9 and is devoid of cox2-2. Relative to nap mitochondrial genome, 40 point mutations were scattered in the 23 protein-coding genes. atp6 even has an abnormal start locus whereas tatC has an abnormal end locus. The rearrangement of the 22 syntenic regions that comprised 80.11% of the genome was influenced by short repeats. A pair of large repeats (9731 bp) was responsible for the multipartite structure. Nine unique regions were detected when compared with other published Brassica mitochondrial genome sequences. We also found six homologous chloroplast segments (Brassica napus). CONCLUSIONS: The mitochondrial genome of oguC is quite divergent from nap and pol, which are more similar with each other. We analyzed the unique regions of every genome of the Brassica family, and found that very few segments were specific for these six mitotypes, especially cam, jun, and ole, which have no specific segments at all. Therefore, we conclude that the most specific regions of oguC possibly came from radish. Compared with the chloroplast genome, six identical regions were found in the seven mitochondrial genomes, which show that the Brassica family has a stable chloroplast-derived source.

Project description:BackgroundCuticular waxes cover plant surface and play important roles in protecting plants from abiotic and biotic stresses. The variations of wax deposition and chemical compositions under changing environments have been shown to be related to plant adaptations. However, it is still not clear whether the wax depositions could be adjusted to increase plant adaptations to stressed conditions.MethodsIn this study, exogenous methyl jasmonate (MeJA), the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) and salicylic acid (SA) were applied to test their effects on cuticular wax deposition in two Brassica napus cultivars, Zhongshuang 9 (ZS9, low wax coverage ) and Yuyou 19 (YY19, high wax coverage). Next, we measured the water loss rate and the transcriptional expression of genes involved in wax biosynthesis as well as genes related to disease defense.ResultsSeven wax compound classes, including fatty acids, aldehydes, alkanes, secondary alcohols, ketones, and unbranched as well as branched primary alcohols, were identified in B. napus leaf wax mixtures. MeJA, SA and ACC treatments had no significant effect on total wax amounts in YY19, whereas ACC reduced total wax amounts in ZS9. Overall, hormone treatments led to an increase in the amounts of aldehydes and ketones, and a decrease of secondary alcohol in ZS9, whereas they led to a decrease of alkane amounts and an increase of secondary alcohol amounts in YY19. Concomitantly, both cultivars also exhibited different changes in cuticle permeability, with leaf water loss rate per 15 min increased from 1.57% (averaged across treatments) at 1.57% (averaged across treatments) at 15 min to 3.12% at 30 min for ZS9 (except for ACC treated plant) and decreased for YY19. MeJA-treated plants of both cultivars relatively had higher water loss rate per 15 min when compared to other treatments. Conclusion. Our findings that B. napus leaf wax composition and cuticle permeability are altered by exogenous SA, MeJA and ACC suggest that the hormone treatments affect wax composition, and that the changes in wax profiles would cause changes in cuticle permeability.

Project description:In this paper, the effect of isosteviol on the physiological metabolism of Brassica napus seedlings under salt stress is explored. Brassica napus seeds (Qinyou 2) were used as materials, and the seeds were soaked in different concentrations of isosteviol under salt stress. The fresh weight, dry weight, osmotic substance, absorption and distribution of Na+, K+, Cl-, and the content of reactive oxygen species (ROS) were measured, and these results were combined with the changes shown by Fourier transform infrared spectroscopy (FTIR). The results showed that isosteviol at an appropriate concentration could effectively increase the biomass and soluble protein content of Brassica napus seedlings and reduce the contents of proline, glycine betaine, and ROS in the seedlings. Isosteviol reduces the oxidative damage to Brassica napus seedlings caused by salt stress by regulating the production of osmotic substances and ROS. In addition, after seed soaking in isosteviol, the Na+ content in the shoots of the Brassica napus seedlings was always lower than that in the roots, while the opposite was true for the K+ content. This indicated that under salt stress the Na+ absorbed by the Brassica napus seedlings was mainly accumulated in the roots and that less Na+ was transported to the shoots, while more of the K+ absorbed by the Brassica napus seedlings was retained in the leaves. It is speculated that this may be an important mechanism for Brassica napus seedlings to relieve Na+ toxicity. The spectroscopy analysis showed that, compared with the control group (T1), salt stress increased the absorbance values of carbohydrates, proteins, lipids, nucleic acids, etc., indicating structural damage to the plasma membrane and cell wall. The spectra of the isosteviol seed soaking treatment group were nearly the same as those of the control group (T1). The correlation analysis shows that under salt stress the Brassica napus seedling tissues could absorb large amounts of Na+ and Cl- to induce oxidative stress and inhibit the growth of the plants. After the seed soaking treatment, isosteviol could significantly reduce the absorption of Na+ by the seedling tissues, increase the K+ content, and reduce the salt stress damage to the plant seedlings. Therefore, under salt stress, seed soaking with isosteviol at an appropriate concentration (10-9~10-8 M) can increase the salt resistance of Brassica napus seedlings by regulating their physiological and metabolic functions.

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Proteome Changes Reveal the Protective Roles of Exogenous Citric Acid in Alleviating Cu Toxicity in Brassica napus L.

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