Proteomic investigations to assess the impact of salinity on Vigna radiata L. genotypes

2020 ◽  
Vol 17 ◽  
Author(s):  
Hesham F. Alharby ◽  
Khalid Rehman Hakeem
Keyword(s):  
Vigna Radiata ◽  
Elongation Factor ◽  
Rubisco Activase ◽  
Nacl Stress ◽  
Expression Change ◽  
Two Dimensional Gel Electrophoresis ◽  
Sds Page ◽  
Scatter Plots ◽  
Oxygen Evolving ◽  
The Impact

Background: In our previous study, six cultivars of Mungo (Vigna radiata) were exposed to 100-250 mM NaCl and studied for changes in growth and biomass. Among them AEM-96 cultivar of the Mung bean [Vigna radiata (L.) Wilczek cv.] was found to tolerate the NaCl stress at 250 mM. Objective: The soluble proteome of salt-tolerant mungo cultivar (AEM-96) was compared to the proteome of control mungo to work out the possible mechanism of salinity tolerance. Methods: Gel-based two-dimensional gel electrophoresis was employed for comparative proteomics. PDQuest-based image analysis of 2D SDS-PAGE was performed. Scatter plots were prepared and total spots were analysed for 2-fold changes in abundance of protein spot intensities in control and treated gels. Results: In total 517 protein spots were detected; 36 with high significance. Among these 36 spots, 2-fold expression change was analysed in 27 protein spots. Seven spots were upregulated, eight spots were down-regulated, 3 were newly induced and 9 were silenced, while as 9 did not change their abundance 2-fold under salinity. Protein spots (9 in total) which were 2-fold upregulated and newly induced were excised from the respective gels. The spots were tryptically digested and run on LC-MS/MS for generating peptides and performing a comparative fingerprinting of the proteins. The peptide signal data was loaded on the Mascot (Swissprot) database to retrieve protein IDs. Proteins with the best score were selected, namely isomers of oxygen-evolving enhancer protein 1 (S1-S3), RuBisCO (S4), oxygen-evolving enhancer protein 2 (S5), Heat shock protein 70 isomers (S6-S7), RuBisCO activase (S8), rubber elongation factor (S9) and pathogen-related protein 10 (S10). Conclusion: The identified proteins were found to play important roles in photosynthesis, stress response and plant growth.

scholarly journals Identification of Components of the Sigma B Regulon in Listeria monocytogenes That Contribute to Acid and Salt Tolerance

2008 ◽  
Vol 74 (22) ◽  
pp. 6848-6858 ◽  
Author(s):  
F. Abram ◽  
E. Starr ◽  
K. A. G. Karatzas ◽  
K. Matlawska-Wasowska ◽  
A. Boyd ◽  
...  
Keyword(s):  
Cell Wall ◽  
Listeria Monocytogenes ◽  
Stress Tolerance ◽  
Intact Cell ◽  
Microscopic Analysis ◽  
Phenotypic Characterization ◽  
Carbon Utilization ◽  
Two Dimensional Gel Electrophoresis ◽  
Sigma B ◽  
The Impact

ABSTRACT Sigma B (σB) is an alternative sigma factor that controls the transcriptional response to stress in Listeria monocytogenes and is also known to play a role in the virulence of this human pathogen. In the present study we investigated the impact of a sigB deletion on the proteome of L. monocytogenes grown in a chemically defined medium both in the presence and in the absence of osmotic stress (0.5 M NaCl). Two new phenotypes associated with the sigB deletion were identified using this medium. (i) Unexpectedly, the strain with the ΔsigB deletion was found to grow faster than the parent strain in the growth medium, but only when 0.5 M NaCl was present. This phenomenon was independent of the carbon source provided in the medium. (ii) The ΔsigB mutant was found to have unusual Gram staining properties compared to the parent, suggesting that σB contributes to the maintenance of an intact cell wall. A proteomic analysis was performed by two-dimensional gel electrophoresis, using cells growing in the exponential and stationary phases. Overall, 11 proteins were found to be differentially expressed in the wild type and the ΔsigB mutant; 10 of these proteins were expressed at lower levels in the mutant, and 1 was overexpressed in the mutant. All 11 proteins were identified by tandem mass spectrometry, and putative functions were assigned based on homology to proteins from other bacteria. Five proteins had putative functions related to carbon utilization (Lmo0539, Lmo0783, Lmo0913, Lmo1830, and Lmo2696), while three proteins were similar to proteins whose functions are unknown but that are known to be stress inducible (Lmo0796, Lmo2391, and Lmo2748). To gain further insight into the role of σB in L. monocytogenes, we deleted the genes encoding four of the proteins, lmo0796, lmo0913, lmo2391, and lmo2748. Phenotypic characterization of the mutants revealed that Lmo2748 plays a role in osmotolerance, while Lmo0796, Lmo0913, and Lmo2391 were all implicated in acid stress tolerance to various degrees. Invasion assays performed with Caco-2 cells indicated that none of the four genes was required for mammalian cell invasion. Microscopic analysis suggested that loss of Lmo2748 might contribute to the cell wall defect observed in the ΔsigB mutant. Overall, this study highlighted two new phenotypes associated with the loss of σB. It also demonstrated clear roles for σB in both osmotic and low-pH stress tolerance and identified specific components of the σB regulon that contribute to the responses observed.

PROTEIN PROFILING OF DIFFERENT PLANT TISSUES FROM HERB PHYLLANTHUS NIRURI

2015 ◽  
Vol 77 (24) ◽  
Author(s):  
Ainul Mardhiah Mohd Nail ◽  
Noor Hasniza Md Zin
Keyword(s):  
Molecular Weight ◽  
Protein Band ◽  
Protein Profiling ◽  
Protein Extract ◽  
Phyllanthus Niruri ◽  
Two Dimensional Gel Electrophoresis ◽  
Plant Parts ◽  
Sds Page ◽  
Isoelectric Points

Herb Phyllanthus niruri (P. niruri) is known to have various pharmacological functions including anticancer, antibacterial, antioxidant, anti-hypertensive and also anti-diabetic properties.  In this research, the proteomic part of P. niruri was studied to determine the bioactive peptides that responsible for specific characteristics. Total soluble proteins from different plant parts of freshly collected P. niruri were extracted using TCA/acetone method and then quantified using Bradford assay. Fruits part was found to have a significantly higher amount of proteins (4.91µg/µl + 0.21) compared to leaves (4.18µg/µl + 0.15). To determine the quality of proteins in the crude extract, SDS-Page was carried out which separates proteins in the basis of molecular weight. Proteins extracted from leaves were widely distributed between the range of 3.5 kDa to 160 kDA. Meanwhile, proteins in fruits mainly distributed within the range of 15 kDa to 80 kDa. The most highly expressed protein band was found in fruit, located in between 30 to 40 kDa. The protein extracts were then further analyzed based on the molecular weight and isoelectric points using two-dimensional gel electrophoresis (2D-GE) approach. Based on the profile pattern obtained from 2D-GE analysis, protein extract from fruits seems to express more protein spots compared to protein extract from leaves. Protein spots from fruit are seen to be intensely resolved within pH 4 to 10 at molecular weight between 10 kDa to 80 kDa. On the other hand, protein spots from leaves were moderately resolved at pH 4 to 10 at molecular weight within 10 kDa to 50 kDa.

scholarly journals Se Nanoparticles Induce Changes in the Growth, Antioxidant Responses, and Fruit Quality of Tomato Developed under NaCl Stress

Molecules ◽  
2019 ◽  
Vol 24 (17) ◽  
pp. 3030 ◽  
Author(s):  
Mónica Carolina Morales-Espinoza ◽  
Gregorio Cadenas-Pliego ◽  
Marissa Pérez-Alvarez ◽  
Alma Delia Hernández-Fuentes ◽  
Marcelino Cabrera de la Fuente ◽  
...  
Keyword(s):  
Fruit Quality ◽  
Positive Impact ◽  
Nacl Stress ◽  
Selenium Nanoparticles ◽  
Human Diet ◽  
Tomato Crop ◽  
Antioxidant Responses ◽  
The Impact ◽  
Positive Effect

Nanotechnology represents an opportunity to improve the use of elements in agriculture. Selenium is an element that is beneficial to plants and essential to the human diet. The size of nanoparticles gives them characteristics that can enhance the benefits that selenium provides to plants. The objective of the present study was to determine the effects of selenium nanoparticles on the growth, antioxidant responses, and fruit quality of tomato developed under NaCl stress. Four doses of selenium nanoparticles (1, 5, 10, and 20 mg L−1) under NaCl stress, only NaCl, and a control were evaluated. The results showed that the impact of salinity on the growth of the tomato crop can be reduced with the application of selenium nanoparticles. However, the amount of both enzymatic and non-enzymatic compounds significantly increased in the leaves and fruits of tomato. The results suggest that the application of selenium nanoparticles generated a positive effect against salinity in the tomato crop; moreover, it had a positive impact on the content of beneficial biocompounds for human health in tomato fruits.

scholarly journals Revealing the Influence of the Degumming Process in the Properties of Silk Fibroin Nanoparticles

Polymers ◽  
2019 ◽  
Vol 11 (12) ◽  
pp. 2045 ◽  
Author(s):  
Guzmán Carissimi ◽  
A. Abel Lozano-Pérez ◽  
Mercedes G. Montalbán ◽  
Salvador D. Aznar-Cervantes ◽  
José Luis Cenis ◽  
...  
Keyword(s):  
Silk Fibroin ◽  
Nanoparticle Synthesis ◽  
Biochemical Properties ◽  
Strong Impact ◽  
Lack Of Information ◽  
Sds Page ◽  
Silk Fibroin Nanoparticles ◽  
Mean Size ◽  
Degumming Process ◽  
The Impact

Several studies have stated that the process used for sericin removal, or degumming, from silk cocoons has a strong impact in the silk fibroin integrity and consequently in their mechanical or biochemical properties after processing it into several biomaterials (e.g. fibers, films or scaffolds) but still, there is a lack of information of the impact on the features of silk nanoparticles. In this work, silk cocoons were degummed following four standard methods: autoclaving, short alkaline (Na2CO3) boiling, long alkaline (Na2CO3) boiling and ultrasounds. The resultant silk fibroin fibers were dissolved in the ionic liquid 1-ethyl-3-methylimidazolium acetate and used for nanoparticle synthesis by rapid desolvation in polar organic solvents. The relative efficiencies of the degumming processes and the integrity of the resulting fibroin fibers obtained were analyzed by mass loss, optical microscopy, thermogravimetric analysis, infrared spectroscopy and SDS-PAGE. Particle sizes and morphology were analyzed by Dynamic Light Scattering and Field Emission Scanning Electronic Microscopy. The results showed that the different treatments had a remarkable impact on the integrity of the silk fibroin chains, as confirmed by gel electrophoresis, which can be correlated with particle mean size and size distribution changes. The smallest nanoparticles (156 ± 3 nm) and the most negative Z potential (−30.2 ± 1.8 mV) were obtained with the combination of long treatment (2 h) of boiling in alkaline solution (Na2CO3 0.02 eq/L). The study confirms that parameters of the process, such as composition of the solution and time of the degumming step, must be controlled in order to reach an optimum reproducibility of the nanoparticle production.

scholarly journals Composition of Pseudorabies Virus Particles Lacking Tegument Protein US3, UL47, or UL49 or Envelope Glycoprotein E

2006 ◽  
Vol 80 (3) ◽  
pp. 1332-1339 ◽  
Author(s):  
Kathrin Michael ◽  
Barbara G. Klupp ◽  
Thomas C. Mettenleiter ◽  
Axel Karger
Keyword(s):  
Protein Interactions ◽  
Pseudorabies Virus ◽  
Cellular Protein ◽  
Tegument Protein ◽  
Virion Protein ◽  
Two Dimensional Gel Electrophoresis ◽  
Glycoprotein E ◽  
Tegument Proteins ◽  
Glycoprotein H ◽  
The Impact

ABSTRACT Proteins located in the tegument layer of herpesvirus particles play important roles in the replicative cycle at both early and late times after infection. As major constituents of the virion, they execute important functions in particular during formation of progeny virions. These functions have mostly been elucidated by construction and analysis of mutant viruses deleted in single or multiple tegument protein-encoding genes (reviewed in the work of T. C. Mettenleiter, Virus Res. 106:167-180, 2004). However, since tegument proteins have been shown to be involved in numerous protein-protein interactions, the impact of single protein deletions on the composition of the virus particle is unknown, but they could impair correct interpretation of the results. To analyze how the absence of single virion constituents influences virion composition, we established a procedure to assay relative amounts of virion structural proteins in deletion mutants of the alphaherpesvirus Pseudorabies virus (PrV) in comparison to wild-type particles. The assay is based on the mass spectrometric quantitation of virion protein-derived peptides carrying stable isotope mass tags. After deletion of the US3, UL47, UL49, or glycoprotein E gene, relative amounts of a capsid protein (UL38), a capsid-associated protein (UL25), several tegument proteins (UL36 and UL47, if present), and glycoprotein H were unaffected, whereas the content of other tegument proteins (UL46, UL48, and UL49, if present) varied significantly. In the case of the UL48 gene product, a specific increase in incorporation of a smaller isoform was observed after deletion of the UL47 or UL49 gene, whereas a larger isoform remained unaffected. The cellular protein actin was enriched in virions of mutants deficient in any of the tegument proteins UL47, UL49, or US3. By two-dimensional gel electrophoresis multiple isoforms of host cell-derived heat shock protein 70 and annexins A1 and A2 were also identified as structural components of PrV virions.

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