scholarly journals A Single Seed Protein Extraction Protocol for Characterizing Brassica Seed Storage Proteins

Agronomy ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 107
Author(s):  
Mahmudur Rahman ◽  
Lei Liu ◽  
Bronwyn J. Barkla
Keyword(s):  
Seed Protein ◽  
Storage Proteins ◽  
Protein Extraction ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
Protein Yield ◽  
Tissue Type ◽  
Single Seed ◽  
Material Optimization ◽  
Hazardous Chemical

Rapeseed oil-extracted expeller cake mostly contains protein. Various approaches have been used to isolate, detect and measure proteins in rapeseeds, with a particular focus on seed storage proteins (SSPs). To maximize the protein yield and minimize hazardous chemical use, isolation costs and the loss of seed material, optimization of the extraction method is pivotal. For some studies, it is also necessary to minimize or avoid seed-to-seed cross-contamination for phenotyping and single-tissue type analysis to know the exact amount of any bioactive component in a single seed, rather than a mixture of multiple seeds. However, a simple and robust method for single rapeseed seed protein extraction (SRPE) is unavailable. To establish a strategy for optimizing SRPE for downstream gel-based protein analysis, yielding the highest amount of SSPs in the most economical and rapid way, a variety of different approaches were tested, including variations to the seed pulverization steps, changes to the compositions of solvents and reagents and adjustments to the protein recovery steps. Following SRPE, 1D-SDS-PAGE was used to assess the quality and amount of proteins extracted. A standardized SRPE procedure was developed and then tested for yield and reproducibility. The highest protein yield and quality were obtained using a ball grinder with stainless steel beads in Safe-Lock microcentrifuge tubes with methanol as the solvent, providing a highly efficient, economic and effective method. The usefulness of this SRPE was validated by applying the procedure to extract protein from different Brassica oilseeds and for screening an ethyl methane sulfonate (EMS) mutant population of Brassica rapa R-0-18. The outcomes provide useful methodology for identifying and characterizing the SSPs in the SRPE.

scholarly journals Study on Seed Protein and Protein Profile Pattern of Chickpea (Cice arietinum L.) by SDS-PAGE under Drought Stress and Fertilization

2015 ◽  
Vol 9 (5) ◽  
pp. 87-90 ◽  
Author(s):  
Vahid Ashrafi ◽  
Hoorieh Pourbozorg ◽  
Nasroallah Moradi Kor ◽  
Abasalt Rostami Ajirloo ◽  
Morteza Shamsizadeh ◽  
...  
Keyword(s):  
Drought Stress ◽  
Seed Protein ◽  
Storage Proteins ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
Protein Yield ◽  
Severe Drought ◽  
Stress Treatment ◽  
Sds Page ◽  
Effects Of Drought

A field experiment was performed in order to evaluate the effects of drought stress and application of starter nitrogen fertilizer on seed storage proteins and protein pattern in chickpea cultivars. Experiment was performed in split-factorial using randomized complete block design with three replications. The experiment was laid out in a split-factorial design with drought stress in main plots and cultivar with nitrogen fertilizer in subplots with three replications. The experimental treatments consisted of three levels of drought stress [severe drought stress (S2), moderate drought stress (S1) and no drought stress(S0)] and two cultivars of chickpea consist of Azad and Bivanij and 2 N levels. Plants were either not given any N fertilizer (N0), or fertilized by N fertilizer at the rate of 25 kg ha-1 (N25). The results of this study showed that the effects of drought stress on seed storage proteins and protein yield, effect of cultivars on protein yield were significant. With increase of drought stress seed storage proteins was increased and protein yield decreased. Severe drought stress treatment has the highest seed protein and the control treatment has the lowest seed protein. Non stress treatment has the highest protein yield and the severe  drought stress treatment has the lowest protein yield. The SDS- PAGE results revealed that no effects treatments on the protein banding patterns but the related severe drought stress bands were chromatic, because they have highest protein concentration in some protein bands.DOI: http://dx.doi.org/10.3126/ijls.v9i5.12704

Karyotype and seed protein analyses of Lycium(Solanaceae) in Iran

1999 ◽  
Vol 56 (2) ◽  
pp. 253-264 ◽  
Author(s):  
M. Sheidai ◽  
Z. Narengi ◽  
M. Khatamsaz
Keyword(s):  
Cluster Analysis ◽  
Statistical Methods ◽  
Chromosome Numbers ◽  
Close Relationships ◽  
Seed Protein ◽  
Storage Proteins ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
Multivariate Statistical Methods ◽  
Multivariate Statistical

Twelve populations of six Lycium species were analysed for karyotypie characters and seed storage proteins using multivariate statistical methods. All reported chromosome numbers (mostly new) are 2n = 24 (x = 12). Karyotypes were symmetrical and placed in 1A and 2A classes of Stebbins karyotype classification. Cluster analysis of karyological and protein data revealed variations among the populations of L. depressum and L. ruthenicum, and supports close relationships of L. kopetdaghi and L. depressum with L. makranicum, and L. shawii with L. edgeworthii.

Genetic and environment interactions of seed storage proteins in narrow-leafed lupin (Lupinus angustifolius)

2012 ◽  
Vol 63 (12) ◽  
pp. 1066 ◽  
Author(s):  
Shahidul Islam ◽  
Wujun Ma ◽  
Bevan J. Buirchell ◽  
Rudi Appels ◽  
Guijun Yan
Keyword(s):  
Seed Protein ◽  
Storage Proteins ◽  
Animal Feed ◽  
Environmental Influences ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
Variable Number ◽  
Mass Peak ◽  
Protein Mass ◽  
Specific Proteins

This study investigated the genetic and environmental influences and their interactions on seed protein profiles of five narrow-leafed lupin cultivars grown under three different environmental conditions. High throughput MALDI-TOF mass spectrometry revealed 133 reproducible seed protein mass peaks. Thirty-one seed protein mass peaks were detected in all 15 combinations of cultivar × environment. Twenty mass peaks were influenced by cultivars irrespective of environment. Only six protein mass peaks were influenced by environments. Seventy-six mass peaks were highly variable. Number of mass peaks of lupin seed protein is mostly genetically controlled (P = 0.008) with no significant influence of the environment (P = 0.131). Environment and cultivar interactions were not significant (P = 0.889). Multivariate analyses of mass peak profiles supported the above analysis showing that protein mass peak profile was significantly (P = 0.001) influenced by cultivar but not by environment (P = 0.053). This result indicates the possibility of breeding new lupin cultivars targeting specific proteins for human food and animal feed without being too concerned about environmental influences.

SEED PROTEIN AMINO ACID COMPOSITIONS RESULTING FROM CROSSES BETWEEN TWO BRASSICA CAMPESTRIS CULTIVARS

1972 ◽  
Vol 52 (1) ◽  
pp. 63-71 ◽  
Author(s):  
L. A. GODING ◽  
R. K. DOWNEY ◽  
A. J. FINLAYSON
Keyword(s):  
Amino Acid ◽  
Amino Acid Composition ◽  
Seed Protein ◽  
Brassica Campestris ◽  
Storage Proteins ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
Protein Amino Acid ◽  
Amino Acid Compositions ◽  
The Individual

Amino acid compositions were determined on the seed meal and two storage proteins (the 12S globulin and 1.7S protein) from each of the parents of two cultivars of the Brassica campestris L. species (Arlo and Yellow sarson), from the progeny of the cultivar cross, and seven individual F1 plants. All plants examined contained the 12S globulin and the 1.7S protein. The proteins from the parents are indistinguishable chromatographically and electrophoretically but have slight differences in amino acid compositions, whereas those from the individual F1 plants are different from their parental counterparts. The results show that the amino acid composition of seed storage proteins may be changed by intercultivar crossing and the cross in this case has produced a wide array of phenotypes.

scholarly journals Wheat (Triticum aestivum L.) TaHMW1D Transcript Variants Are Highly Expressed in Response to Heat Stress and in Grains Located in Distal Part of the Spike

Plants ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 687
Author(s):  
Chan Seop Ko ◽  
Jin-Baek Kim ◽  
Min Jeong Hong ◽  
Yong Weon Seo
Keyword(s):  
Heat Stress ◽  
High Temperature ◽  
Temperature Stress ◽  
Storage Proteins ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
Grain Filling ◽  
High Temperature Stress ◽  
Two Dimensional Gel Electrophoresis ◽  
Transcript Variants

High-temperature stress during the grain filling stage has a deleterious effect on grain yield and end-use quality. Plants undergo various transcriptional events of protein complexity as defensive responses to various stressors. The “Keumgang” wheat cultivar was subjected to high-temperature stress for 6 and 10 days beginning 9 days after anthesis, then two-dimensional gel electrophoresis (2DE) and peptide analyses were performed. Spots showing decreased contents in stressed plants were shown to have strong similarities with a high-molecular glutenin gene, TraesCS1D02G317301 (TaHMW1D). QRT-PCR results confirmed that TaHMW1D was expressed in its full form and in the form of four different transcript variants. These events always occurred between repetitive regions at specific deletion sites (5′-CAA (Glutamine) GG/TG (Glycine) or (Valine)-3′, 5′-GGG (Glycine) CAA (Glutamine) -3′) in an exonic region. Heat stress led to a significant increase in the expression of the transcript variants. This was most evident in the distal parts of the spike. Considering the importance of high-molecular weight glutenin subunits of seed storage proteins, stressed plants might choose shorter polypeptides while retaining glutenin function, thus maintaining the expression of glutenin motifs and conserved sites.

Seed storage proteins in developing somatic embryos of alfalfa: defects in accumulation compared to zygotic embryos

1994 ◽  
Vol 45 (6) ◽  
pp. 699-708 ◽  
Author(s):  
Joan E. Krochko ◽  
David J. Bantroch ◽  
John S. Greenwood ◽  
J. Derek Bewley
Keyword(s):  
Somatic Embryos ◽  
Storage Proteins ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
Zygotic Embryos
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