scholarly journals Target-Based Physiological Modulations and Chloroplast Proteome Reveals a Drought Resilient Rootstock in Okra (Abelmoschus esculentus) Genotypes

2021 ◽  
Vol 22 (23) ◽  
pp. 12996
Author(s):  
Kaukab Razi ◽  
Dong-Won Bae ◽  
Sowbiya Muneer
Keyword(s):  
Drought Stress ◽  
Large Scale ◽  
Polyacrylamide Gel Electrophoresis ◽  
Proteome Analysis ◽  
Specific Protein ◽  
Abelmoschus Esculentus ◽  
Carotenoid Content ◽  
Vegetable Crops ◽  
Total Carotenoid Content ◽  
Specific Proteins

As climate changes increase, drought stress is becoming a problem for all major horticultural crops; among them is okra (Abelmoschus esculentus). Despite its superior resilience to heat stress and high nutritional content, it is still underutilized in contrast to other vegetable crops. Moreover, the drought-resistant and drought-sensitive genotypes of okra are also not well known and require further exploration to improve their productivity. To investigate this in more detail, we performed comparative physiological and large-scale chloroplast proteomics on drought-stressed genotypes of okra. We evaluated four major genotypes of okra, viz., NS7774, NS7772, Green Gold, and OH3312 for drought resilient rootstock. The physiological modulations demonstrated a significant change by 50–76% in biomass, net-photosynthetic machinery, water transport, and absorption both in early and late stages of drought stress compared to well-watered crops in all genotypes. Maximum oxidative damage due to drought stress was observed for the genotypes NS7772, Green Gold and OH3312 as depicted by H2O2 and O2− determination. Greater oxidative stress was correlated to lesser antioxidant activity and expression of antioxidant enzymes, such as catalase and ascorbate peroxidase under stress in okra genotypes. The overall photosynthetic pigments, such as total chlorophyll, and total carotenoid content, were also decreased, and stomatal guard cells were disrupted and appeared closed compared to the control for the above three mentioned genotypes, except NS7774. A subsequent tissue-specific proteome analysis of chloroplasts and thylakoids analyzed by BN-PAGE (blue native polyacrylamide gel electrophoresis) revealed either over or under expression of specific proteins, such as ATPase, PSI, PSII core dimer, PSII monomer and ATP synthase. The expression of multiprotein complex proteins, including PSII-core dimer and PSII-core monomer, was slightly higher for the genotype NS7774 when compared to three other genotypes for both 5 and 10 days of drought stress. Further identification of specific proteins obtained in second dimension BN-PAGE provided descriptive detail of seven proteins involved in drought resistance across all genotypes. The identified proteins are majorly involved in photosynthesis under drought stress, suggesting NS7774 as a drought tolerant genotype. Further, the proteomic results were confirmed using Immunoblot by selecting specific protein such as PsaA. Overall, from our physiological modulations and chloroplast proteomics in all genotypes, we summarized NS7774 as a resilient rootstock and the other three genotypes (NS7772, OH3312, and Green Gold) as sensitive ones.

Proteome analysis of two soft-grained wheats of different processing quality: cultivar-specific proteins

2005 ◽  
Vol 56 (2) ◽  
pp. 145 ◽  
Author(s):  
D. J. Skylas ◽  
S. J. Cordwell ◽  
G. Craft ◽  
B. McInerney ◽  
M. J. Wu ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Starch Granule ◽  
Storage Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Proteome Analysis ◽  
Specific Protein ◽  
Processing Quality ◽  
Ph Gradients ◽  
Specific Proteins

Proteome analysis was conducted on the grain of 2 closely related soft biscuit-making wheats (Bowie and Rosella cultivars) differing in processing quality. Comparisons between these wheat cultivars were carried out on total wholemeal proteins, extracts with enriched starch granule proteins, and extracts enriched with gliadin storage proteins, with the intention of characterising, identifying, and cataloguing cultivar-specific proteins that could be used for segregation purposes. Initially, 2-dimensional gel electrophoresis was carried out on total wholemeal proteins using a broad range pH 3–10 immobilised pH gradient for the first dimension. Further screening was carried out using a combination of mid to narrow range immobilised pH gradients, including pH 4–7, 5.5–6.7, 5–8, 6–9, and 6–11. Best cultivar-specific protein fractionation was provided by the pH 5–8 range. Altogether, 4 unique cultivar-specific protein spots were excised from the pH 5–8 gels and identified by means of peptide mass fingerprinting, tandem mass spectrometry, or N-terminal sequencing. Starch granule protein extracts were prepared and fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A western blot was performed and probed with an anti-puroindoline-a antibody. Further to this, extracts enriched in gliadin storage proteins were isolated using 70% ethanol and analysed by 2-dimensional gel electrophoresis. The resulting gliadin protein maps showed 18 unique cultivar-specific gliadins. They were excised from the pH 6–9 gels and submitted for N-terminal amino acid sequencing. Overall, this study identified 23 proteins that could be used to distinguish between these closely related cultivars and may provide information on the molecular basis for the differences in processing exhibited by these wheats. The findings reported also contribute to a longer term objective of developing a broad and comprehensive knowledge base of commercial wheats, in regard to protein composition and their inherent processing qualities.

scholarly journals Optimization of Growth and Carotenoid Production by Haloferax mediterranei Using Response Surface Methodology

Marine Drugs ◽  
2018 ◽  
Vol 16 (10) ◽  
pp. 372 ◽  
Author(s):  
Zaida Montero-Lobato ◽  
Adrián Ramos-Merchante ◽  
Juan Fuentes ◽  
Ana Sayago ◽  
Ángeles Fernández-Recamales ◽  
...  
Keyword(s):  
Response Surface Methodology ◽  
Response Surface ◽  
Large Scale ◽  
Antioxidant Properties ◽  
Carotenoid Content ◽  
Scale Production ◽  
Optimal Conditions ◽  
Haloferax Mediterranei ◽  
Carotenoid Production ◽  
Total Carotenoid Content

Haloferax mediterranei produces C50 carotenoids that have strong antioxidant properties. The response surface methodology (RSM) tool helps to accurately analyze the most suitable conditions to maximize C50 carotenoids production by haloarchaea. The effects of temperature (15–50 °C), pH (4−10), and salinity (5–28% NaCl (w/v)) on the growth and carotenoid content of H. mediterranei were analyzed using the RSM approach. Growth was determined by measuring the turbidity at 600 nm. To determine the carotenoid content, harvested cells were lysed by freeze/thawing, then re-suspended in acetone and the total carotenoid content determined by measuring the absorbance at 494 nm. The analysis of carotenoids was performed by an HPLC system coupled with mass spectrometry. The results indicated the theoretical optimal conditions of 36.51 or 36.81 °C, pH of 8.20 or 8.96, and 15.01% or 12.03% (w/v) salinity for the growth of haloarchaea (OD600 = 12.5 ± 0.64) and production of total carotenoids (3.34 ± 0.29 mg/L), respectively. These conditions were validated experimentally for growth (OD600 = 13.72 ± 0.98) and carotenoid production (3.74 ± 0.20 mg/L). The carotenoid profile showed four isomers of bacterioruberin (89.13%). Our findings suggest that the RSM approach is highly useful for determining optimal conditions for large-scale production of bacterioruberin by haloarchaea.

scholarly journals Physiological and Proteomic Signatures Reveal Mechanisms of Superior Drought Resilience in Pearl Millet Compared to Wheat

2021 ◽  
Vol 11 ◽  
Author(s):  
Arindam Ghatak ◽  
Palak Chaturvedi ◽  
Gert Bachmann ◽  
Luis Valledor ◽  
Živa Ramšak ◽  
...  
Keyword(s):  
Drought Stress ◽  
Pearl Millet ◽  
Stress Responses ◽  
Root Morphology ◽  
Large Scale ◽  
Foxtail Millet ◽  
Proteome Analysis ◽  
Specific Marker ◽  
Stay Green ◽  
Drought Tolerant

Presently, pearl millet and wheat are belonging to highly important cereal crops. Pearl millet, however, is an under-utilized crop, despite its superior resilience to drought and heat stress in contrast to wheat. To investigate this in more detail, we performed comparative physiological screening and large scale proteomics of drought stress responses in drought-tolerant and susceptible genotypes of pearl millet and wheat. These chosen genotypes are widely used in breeding and farming practices. The physiological responses demonstrated large differences in the regulation of root morphology and photosynthetic machinery, revealing a stay-green phenotype in pearl millet. Subsequent tissue-specific proteome analysis of leaves, roots and seeds led to the identification of 12,558 proteins in pearl millet and wheat under well-watered and stress conditions. To allow for this comparative proteome analysis and to provide a platform for future functional proteomics studies we performed a systematic phylogenetic analysis of all orthologues in pearl millet, wheat, foxtail millet, sorghum, barley, brachypodium, rice, maize, Arabidopsis, and soybean. In summary, we define (i) a stay-green proteome signature in the drought-tolerant pearl millet phenotype and (ii) differential senescence proteome signatures in contrasting wheat phenotypes not capable of coping with similar drought stress. These different responses have a significant effect on yield and grain filling processes reflected by the harvest index. Proteome signatures related to root morphology and seed yield demonstrated the unexpected intra- and interspecies-specific biochemical plasticity for stress adaptation for both pearl millet and wheat genotypes. These quantitative reference data provide tissue- and phenotype-specific marker proteins of stress defense mechanisms which are not predictable from the genome sequence itself and have potential value for marker-assisted breeding beyond genome assisted breeding.

scholarly journals Sperm Proteomics: Road to Male Fertility and Contraception

2013 ◽  
Vol 2013 ◽  
pp. 1-11 ◽  
Author(s):  
Md Saidur Rahman ◽  
June-Sub Lee ◽  
Woo-Sung Kwon ◽  
Myung-Geol Pang
Keyword(s):  
Posttranslational Modifications ◽  
Gel Electrophoresis ◽  
Male Fertility ◽  
Large Scale ◽  
Polyacrylamide Gel Electrophoresis ◽  
Protein Biomarkers ◽  
Scientific Publications ◽  
Pubmed Database ◽  
Protein Functions ◽  
Specific Proteins

Spermatozoa are highly specialized cells that can be easily obtained and purified. Mature spermatozoa are transcriptionally and translationally inactive and incapable of protein synthesis. In addition, spermatozoa contain relatively higher amounts of membrane proteins compared to other cells; therefore, they are very suitable for proteomic studies. Recently, the application of proteomic approaches such as the two-dimensional polyacrylamide gel electrophoresis, mass spectrometry, and differential in-gel electrophoresis has identified several sperm-specific proteins. These findings have provided a further understanding of protein functions involved in different sperm processes as well as of the differentiation of normal state from an abnormal one. In addition, studies on the sperm proteome have demonstrated the importance of spermatozoal posttranslational modifications and their ability to induce physiological changes responsible for fertilization. Large-scale proteomic studies to identify hundreds to thousands of sperm proteins will ultimately result in the development of novel biomarkers that may help to detect fertility, the state of complete contraception, and beyond. Eventually, these protein biomarkers will allow for a better diagnosis of sperm dysfunctions and aid in drug development. This paper reviews the recent scientific publications available from the PubMed database to address sperm proteomics and its potential application to characterize male fertility and contraception.

Analysis of the cytosolic proteins of chick embryo gonads by two-dimensional gel electrophoresis

Development ◽  
1986 ◽  
Vol 94 (1) ◽  
pp. 221-230
Author(s):  
J. Samsel ◽  
B. Lorber ◽  
A. Petit ◽  
J. -P. Weniger
Keyword(s):  
Gel Electrophoresis ◽  
Developmental Stages ◽  
Polyacrylamide Gel Electrophoresis ◽  
Specific Protein ◽  
Chick Embryos ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Specific Proteins ◽  
Different Tissues

Gonads and mesonephros dissected from normal chick embryos, as well as a blood sample, were labelled in vitro with [35S]methionine and [14C]leucine. The patterns of cytosolic protein synthesis of the different tissues were analysed using two-dimensional (2-D) polyacrylamide gel electrophoresis. Three developmental stages, i.e. 8, 12 and 17 days of incubation, were investigated. Five sex-specific proteins were detected in the male and two in the female. In the testis, only one protein is already present at the 8-day stage. In the ovary, one protein exists since the 8-day stage, but it is also synthesized in the mesonephros. The second ovary-specific protein appeared only at the 12-day stage.

ВПЛИВ ДЖЕРЕЛ АЗОТНОГО ЖИВЛЕННЯ НА СИНТЕЗ КАРОТИНОЇДІВ ДЕЯКИМИ ШТАМАМИ БАЗИДІОМІЦЕТІВ

2014 ◽  
Vol 4 (01) ◽  
pp. 22
Author(s):  
A. K. Veligodska ◽  
O. V. Fedotov ◽  
A. S. Petreeva
Keyword(s):  
Culture Filtrate ◽  
Nutrient Medium ◽  
Culture Fluid ◽  
Carotenoid Content ◽  
Nitrogen Compounds ◽  
Total Carotenoid ◽  
Carotenoid Accumulation ◽  
Total Carotenoid Content ◽  
Peptone Medium ◽  
Acetone Extracts

<p>The influence of certain nitrogen compounds - components of glucose-peptone medium (GPM) on the accumulation of carotenoids by some strains was investigated by surface cultivating basidiomycetes. The total carotenoid content was set in acetone extracts of mycological material spectrophotometrically and calculated using the Vetshteyn formula.</p> <p>As the nitrogen-containing components used GPM with 9 compounds, such as peptone, DL-valine, L-asparagine, DL-serine, DL-tyrosine, L-proline, L-alanine, urea, NaNO<sub>3</sub>. The effect on the accumulation of specific compounds both in the mycelium and in the culture fluid of carotenoids by culturing certain strains of Basidiomycetes was identified.</p> <p>Adding to standard glucose-peptone medium peptone at 5 g/l causes an increase of carotenoid accumulation by strain <em>L. sulphureus</em> Ls-08, and in a concentration of 4 g/l by strains of <em>F. hepatica </em>Fh-18 and <em>F. fomentarius</em> Ff-1201.</p> <p>In order to increase the accumulation of carotenoids in the mycelium  we suggested to make a standard glucose-peptone medium with proline or valine for cultivating of <em>L. sulphureus</em> Ls- 08 strain; alanine for <em>F. fomentarius</em> Ff-1201 strain; proline, asparagine and serine - for strain Fh-18 of <em>F. hepatica</em>. The results can be implemented in further optimization of the composition of the nutrient medium for culturing strains of Basidiomycetes wich producing carotenoids.</p> <p><em>Keywords: nitrogen-containing substances, Basidiomycetes, mycelium</em><em>,</em><em> culture filtrate, carotenoids</em></p>

scholarly journals INTRACISTRONIC MAPPING OF ELECTROPHORETIC SITES IN DROSOPHILA MELANOGASTER: FIDELITY OF INFORMATION TRANSFER BY GENE CONVERSION

Genetics ◽  
1974 ◽  
Vol 76 (2) ◽  
pp. 289-299
Author(s):  
Margaret McCarron ◽  
William Gelbart ◽  
Arthur Chovnick
Keyword(s):  
Drosophila Melanogaster ◽  
Information Transfer ◽  
Large Scale ◽  
Genetic Basis ◽  
Xanthine Dehydrogenase ◽  
Specific Protein ◽  
Wild Type ◽  
Electrophoretic Variation ◽  
The Stability ◽  
Recombination Experiments

ABSTRACT A convenient method is described for the intracistronic mapping of genetic sites responsible for electrophoretic variation of a specific protein in Drosophila melanogaster. A number of wild-type isoalleles of the rosy locus have been isolated which are associated with the production of electrophoretically distinguishable xanthine dehydrogenases. Large-scale recombination experiments were carried out involving null enzyme mutants induced on electrophoretically distinct wild-type isoalleles, the genetic basis for which is followed as a nonselective marker in the cross. Additionally, a large-scale recombination experiment was carried out involving null enzyme rosy mutants induced on the same wild-type isoallele. Examination of the electrophoretic character of crossover and convertant products recovered from the latter experiment revealed that all exhibited the same parental electrophoretic character. In addition to documenting the stability of the xanthine dehydrogenase electrophoretic character, this observation argues against a special mutagenesis hypothesis to explain conversions resulting from allele recombination studies.

scholarly journals High-Throughput Phenotyping Methods for Breeding Drought-Tolerant Crops

2021 ◽  
Vol 22 (15) ◽  
pp. 8266
Author(s):  
Minsu Kim ◽  
Chaewon Lee ◽  
Subin Hong ◽  
Song Lim Kim ◽  
Jeong-Ho Baek ◽  
...  
Keyword(s):  
Drought Stress ◽  
High Throughput ◽  
Large Scale ◽  
Crop Yields ◽  
Plant Traits ◽  
Rapid Development ◽  
Plant Responses ◽  
Phenotypic Data ◽  
Agricultural Technologies ◽  
High Throughput Phenotyping

Drought is a main factor limiting crop yields. Modern agricultural technologies such as irrigation systems, ground mulching, and rainwater storage can prevent drought, but these are only temporary solutions. Understanding the physiological, biochemical, and molecular reactions of plants to drought stress is therefore urgent. The recent rapid development of genomics tools has led to an increasing interest in phenomics, i.e., the study of phenotypic plant traits. Among phenomic strategies, high-throughput phenotyping (HTP) is attracting increasing attention as a way to address the bottlenecks of genomic and phenomic studies. HTP provides researchers a non-destructive and non-invasive method yet accurate in analyzing large-scale phenotypic data. This review describes plant responses to drought stress and introduces HTP methods that can detect changes in plant phenotypes in response to drought.

scholarly journals Assessing the Potential of Algae Extracts for Extending the Shelf Life of Rainbow Trout (Oncorhynchus mykiss) Fillets

Foods ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 910
Author(s):  
María I. Sáez ◽  
María D. Suárez ◽  
Francisco J. Alarcón ◽  
Tomás F. Martínez
Keyword(s):  
Rainbow Trout ◽  
Cold Storage ◽  
Haematococcus Pluvialis ◽  
Radical Scavenging ◽  
Storage Period ◽  
Carotenoid Content ◽  
Total Phenolic ◽  
Antioxidant Power ◽  
Color Parameters ◽  
Total Carotenoid Content

This study evaluates the potential of different algae extracts (Crassiphycus corneus, Cc; Ulva ohnoi, Uo; Arthrospira platensis, Ap; Haematococcus pluvialis, Hp) as additives for the preservation of rainbow trout fillets. The extracts were prepared with different water to ethanol ratios from the four algae species. The highest ferric reducing antioxidant power (FRAP) was observed in Uo extracted in 80% ethanol. Ap aqueous extract also had considerable FRAP activity, in agreement with a high total phenolic content. Radical scavenging activity (DPPH) was higher in Cc 80% ethanol extract, in agreement with a high total carotenoid content. In fact, when the algae aqueous extracts were assayed on the fish fillets, their antioxidant activity exceeded that of ascorbic acid (ASC). All algae extracts delayed microbial growth and lipid oxidation processes in trout fillets throughout the cold storage period compared to controls, and also improved textural parameters, these effects being more evident for Ap and Hp. With respect to the color parameters, the Hp extract prevented the a* values (redness) from decreasing throughout cold storage, a key point when it comes to colored species, not least salmonids. On the other hand, the Ap extract was not as effective as the rest of treatments in avoiding a* and b* decrease throughout the storage period, and thereby the color parameters were impaired. The results obtained, together with the natural origin and the viability for large-scale cultivation, make algae extracts interesting fish preservative agents for the food industry.

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