Proteomics
Samira Mardani Zonouz; Mahmoud Toorchi; Shaghayegh aslzad
Abstract
Drought is one of the most important factors causing abiotic stress in plants. Wheat, as a vital crop, is extensively cultivated in regions that face water scarcity at least during one period of the year. Proteomic approach is one of the ways to identify proteins involved in plant tolerance to water ...
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Drought is one of the most important factors causing abiotic stress in plants. Wheat, as a vital crop, is extensively cultivated in regions that face water scarcity at least during one period of the year. Proteomic approach is one of the ways to identify proteins involved in plant tolerance to water stress. In order to investigate the effect of water deficit stress on the root proteome pattern of desert-tolerant wheat, an experiment was conducted in the form of a completely randomized design with seven replications. Root proteins were extracted by TCA/acetone method and the protein expression pattern was analyzed using two-dimensional electrophoresis. Potential proteins involved in the response to water deficiency were identified by comparing the protein expression patterns under water deficit stress with the expression pattern in control conditions. The results revealed significant differences in root weight and root length at a 5% probability level, indicating the detrimental effects of water stress on plant roots. The proteomic analysis identified 98 reproducible protein spots, of which 10 exhibited statistically significant changes, with eight spots showing increased expression and one showing decreased expression. These protein spots were identified based on their molecular weight (MW) and isoelectric point (pI) through database searches. The identified proteins were classified into various functional categories related to stress response, including protein synthesis and accumulation, oxidative stress, response and defense against stress and metabolic pathways.
Proteomics
Mohammad Reza Naghavi; Marouf Khalili
Abstract
In order to investigate the mechanism of tolerance to salt stress in wheat, a factorial experiment basis of completely randomized design with four replications in greenhouse media were conducted. The factors of this experiment were included salinity of sodium chloride in four levels of zero (control), ...
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In order to investigate the mechanism of tolerance to salt stress in wheat, a factorial experiment basis of completely randomized design with four replications in greenhouse media were conducted. The factors of this experiment were included salinity of sodium chloride in four levels of zero (control), 100, 200 and 300 mM and two wheat cultivars namely Niknejhad and Pishtaz as a representative of tolerant and susceptible cultivars, respectively. Leaf samples were prepared two weeks after the starting of salt stress. Then, extraction of protein from leaf tissue was done and two-dimensional electrophoresis in control plants and under salt stress plants were carried out. The results of proteome analysis revealed that 15 replicated protein spots with different expression variations were common between two tolerant and susceptible cultivars, and five protein spots of unique to each tolerant and susceptible cultivar, had significantly expression variations under stress. Protein spots were detected by mass spectrometry and the results showed that more number from commonly detected proteins were classified in functional groups including antioxidant defense and Calvin cycle, while other proteins in each cultivar had more roles in antioxidant activity. Overall, the results showed that there was a significant difference between two cultivars in terms of morpho-physiological response to salt stress, and the Niknejhad cultivar showed a better protein response under stress.
Proteomics
Mehdi Ghaffari; Saadat Sharifi; Gholamreza Bakhshi Khaniki
Volume 8, Issue 22 , September 2018, , Pages 15-26
Abstract
In order to understand of the molecular mechanisms of drought tolerance in sunflower, proteomic pattern of roots in two drought sensitive and drought-tolerant lines were evaluated under limited and favorable water conditions. After 2DE and comparison of relative abundance of protein spots using t test, ...
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In order to understand of the molecular mechanisms of drought tolerance in sunflower, proteomic pattern of roots in two drought sensitive and drought-tolerant lines were evaluated under limited and favorable water conditions. After 2DE and comparison of relative abundance of protein spots using t test, 12 of 417 protein spots in sensitive and 17 of 467 in tolerant line were affected by drought stress significantly. Following nano-LC MS/MS the protein spots were identified using Mascot search engine in NCBI protein database considering more than 10 % sequence coverage and score of above 80. Cytoplasmic and nuclear proteins were the most proteins which were affected by water deficiency. Three protein spots i.e. Enolase, Glyceraldehyde 3-phosphate dehydrogenase and Chalcone synthase were expressed differentially in these lines. Reduction of Enolase as a sign of metabolic impairment could be resulted in downstream process under drought stress. Increased expression of Glyceraldehyde 3-phosphate dehydrogenase and Chalcone synthase could have a role in detoxification/removal of oxidative destruction and antioxidant capability of the tolerant line. Increased level of heat shock protein, dihydroflavonol reductase, Seed linoleate 9S-lipoxygenase, Ubiquitin carboxyl-terminal hydrolase and G protein indicated crucial role of defensive, protective and transductive process in reduction of drought injuries.
Proteomics
Maryam Jamshidnia; Sayed Kamal Kazemitabar; Christian Lindermayr; Hamid Najafi Zarini
Volume 6, Issue 16 , March 2017, , Pages 1-12
Abstract
Recently, transient gene expression has been developed to provide a more rapid means of assessing plant tissues as a protein production platform without the labor-intensive and time-consuming process of generating stably transformed transgenic plants. This study reports the expression of HDA19 gene in ...
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Recently, transient gene expression has been developed to provide a more rapid means of assessing plant tissues as a protein production platform without the labor-intensive and time-consuming process of generating stably transformed transgenic plants. This study reports the expression of HDA19 gene in two species of tobacco plants (Nicotiana tabacum and Nicotiana bentamiana) by means of transient transformation. Specific primers were designed and used for PCR amplification and cloning of HDA19 gene in the plant expression vector pB2GW7. The recombinant construct was transferred into Agrobacterium tumefaciens strain GV3101, and was used for Agrobacterium mediated transformation of tobacco plants. The presence of the desired gene in transgenic lines was confirmed through colony PCR. The expression of the protein in transgenic lines was confirmed by immune-dot blot assay and ELISA. Although the transformation of the two species was confirmed by immune-dot blot assay and SDS-PAGE, recombinant protein production in Nicotiana tabacum plants was confirmed by ELISA and it was estimated 400 µg per gram wet weight of tobacco leaves. According to the results, this species is the appropriate host for the production of recombinant HDA19, one of the histone deacetylases, rather than Nicotiana bentamiana.
Proteomics
Marouf Khalili; Mohammad Reza Naghavi
Volume 6, Issue 16 , March 2017, , Pages 31-44
Abstract
In this research, proteome analysis was done by two-dimensional electrophoresis and stainig by Commassie brilliant blue method for two cultivars of Kavir as tolerant and Bahar as susceptible was done. After of be quantitative spots with Same spot progenesis software, 10 common protein spots with significant ...
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In this research, proteome analysis was done by two-dimensional electrophoresis and stainig by Commassie brilliant blue method for two cultivars of Kavir as tolerant and Bahar as susceptible was done. After of be quantitative spots with Same spot progenesis software, 10 common protein spots with significant difference between control and drought stress conditions in the Kavir and Bahar was diagnosed. Using MALDI-TOF/TOF mass spectrometry of common proteins, nine common protein spots were identified and the type of activity and mode of action of these proteins in the cells was determined. Based on the results, proteins involved in light reactions of photosynthesis (three protein spots Chlorophyll ab binding protein 8, chloroplastic, Cytochrome b6-f complex iron-sulfur subunit, chloroplasti and Peptidyl-prolyl cis-trans isomerase CYP38, chloroplastic) and Calvin cycle (two protein spots include Fructose-1,6-bisphosphatase, chloroplastic and ribulose-bisphosphate carboxylase small chain precursor) in both cultivars were the greatest functional groups and in other words, the most important common proteins for maintain of efficiency under drought stress were. Since these proteins in both tolerant and susceptible cultivars showed changes in expression, seems to be the most sensitive proteins in response to drought stress in wheat. In other words, these results show that it is important to maintain the efficiency of photosynthesis under drought stress.
Proteomics
Mohammad Reza Ghaffari; Nicolaus von Wirén; Klaus Humbeck; Phillip Franken
Volume 6, Issue 13 , May 2016, , Pages 27-39
Abstract
Deciphering of network correlation operating in the plant biomass accumulation and yield production is a pre-requisite for understanding the relationships between omics data and growth rate in plants. To investigate the relationship among transcripts and their regulation for lignocellulose biomass formation ...
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Deciphering of network correlation operating in the plant biomass accumulation and yield production is a pre-requisite for understanding the relationships between omics data and growth rate in plants. To investigate the relationship among transcripts and their regulation for lignocellulose biomass formation at the generative stage of barley, transcript profiling was applied on three contrasting spring barley lines two weeks after flowering. A custom barley cDNA Microarray (Agilent Technologies, Germany) containing 56000 barley oligonucleotides was used for transcriptome analysis on flag leaves of spring barley. The network correlation of transcripts involved in secondary and RNA metabolism revealed a higher number of positive than negative correlations of which a signal molecule, ABH1-Cap binding protein showed the highest node degree centrality. Statistical test showed a strong positive interrelation between ABH1-Cap binding protein and a key gene of phenylpropanoid pathway, Cinnamoyl-CoA reductase. The integrated data suggested Cinnamoyl-CoA reductase (CCR) might be used as putative biomarker for engineering of lignocellulose biomass improvement at the generative stage in barley.
Proteomics
Mohammad Reza Azimi; Ghasem Hosseini Salekdeh
Volume 4, Issue 7 , December 2015, , Pages 1-13
Abstract
Water shortage is one of the most important environmental factors in limiting plant production worldwide. Molecular breeding may help to develop drought tolerant plants. Proteomics approach can help in comprehensive analysis of stress responsive genes and identification of drought signaling pathways. ...
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Water shortage is one of the most important environmental factors in limiting plant production worldwide. Molecular breeding may help to develop drought tolerant plants. Proteomics approach can help in comprehensive analysis of stress responsive genes and identification of drought signaling pathways. These drought tolerance candidate genes or their regulatory genes may be further analyzed for their possible implication in increase plant tolerance to drought stress. In this studied we analyzed the changes in proteome of wheat flag leaf in response to drought. We compared a drought tolerant with drought susceptible genotypes under normal and stress well-watered and stress conditions. Stressed plants were exposed to 50% field capacity before anthesis. Out of 900 proteins analyzed across two dimensional gels, 57 protein spots showed significant differences in response to stress. Of these, 42 protein spots could be identified using mass spectrometry analysis. Differentially expressed proteins and enzymes could be grouped in different functional groups. Our results showed that tolerant genotype may use various mechanisms particularly the up-regulation of genes involved in oxidative stress defense in flag leaf. This may help the tolerant genotypes to better remove reactive oxygen species generate by stress in flag leaf and maintain its physiological and photosynthetic activities.
Proteomics
Ghasem Hosseini Salkade; Davood Nasr Abadi
Volume 1, Issue 1 , March 2012, , Pages 1-11
Abstract
Rice is an excellent model cereal for molecular biology and genetics research. Salinity is a major factor limiting rice production world wide. The analysis of stress-responsiveness in plants is an important route to the discovery of genes conferring stress tolerance and their use in breeding programs. ...
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Rice is an excellent model cereal for molecular biology and genetics research. Salinity is a major factor limiting rice production world wide. The analysis of stress-responsiveness in plants is an important route to the discovery of genes conferring stress tolerance and their use in breeding programs. To further understand the mechanism of plant response to salinity we employed a proteomic approach to profile the protein changes of rice 3th leaf and root under salt stress. Plants were grown in Yoshida nutrient solution and salt stress imposed after 25 days. Plants were treated by 100¬mM NaCl for 10. After that 3th leaves and total root were collected from control and salt stressed plants. The Na+ and K+ content of leaves/roots and several yield components changed significantly in response to short-term salt stress and their proteome patterns were analyzed using 2-DE in triplicates. The expression pattern of proteins significantly changed in all leaves/roots in response to stress. More than 488 and 345 protein were detected repeatedly in root and leaf 2Dgels respectively by software package. 107 proteins in root and 86 proteins in leaf of two genotypes showed significant response to stress. 3 protein in leaf gels and 2 protein in root gels were selected and identified by ESI-Q-TOF. The most important were Ferritin, Rubisco activase and ascorbat¬peroxidase in leaf and Peroxidase and Ascorbat¬peroxidase in root. All of them were enzyme and involved in detoxification and removal of reactive oxygen species (peroxidase, ascorbat¬peroxidase) Iron homeostasis (ferritin) or activation of other enzymes (rubisco¬activase).
