scholarly journals RNAi-Mutants of Sorghum bicolor (L.) Moench with Improved Digestibility of Seed Storage Proteins

2021 ◽  
Author(s):  
Lev A. Elkonin ◽  
Valery M. Panin ◽  
Odissey A. Kenzhegulov ◽  
Saule Kh. Sarsenova
Keyword(s):  
Rna Interference ◽  
Storage Proteins ◽  
Marker Gene ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
Genetic Construct ◽  
Rnai Silencing ◽  
Sds Page ◽  
Developing Area

Modification of the composition of grain storage proteins is an intensively developing area of plant biotechnology, which is of particular importance for sorghum – high-yielding drought tolerant crop. Compared to other cereals, the majority of sorghum cultivars and hybrids are characterized by reduced nutritional value that is caused by a low content of essential amino acids in the seed storage proteins (kafirins), and resistance of kafirins to protease digestion. RNA interference (RNAi) by suppressing synthesis of individual kafirin subclasses may be an effective approach to solve this problem. In this chapter, we review published reports on RNAi silencing of the kafirin-encoding genes. In addition, we present new experimental data on phenotypic effects of RNAi-silencing of γ-KAFIRIN-1 gene in sorghum cv. Avans. To obtain RNAi mutants with γ-KAFIRIN-1 gene silencing we used Agrobacterium-mediated genetic transformation. Transgenic kernels had modified endosperm type with reduced vitreous layer and significantly improved in vitro protein digestibility (93% vs. 57%, according to the densitometry of SDS-PAGE patterns). SDS-PAGE of transgenic kernels showed lowered level of kafirins and appearance of globulin proteins, which were not observed in the original cultivar. For the first time, the cases of instability of inserted genetic construct were identified: elimination of ubi1-intron that is a constituent part of the genetic construct for RNAi silencing, or nos-promotor governing expression of the marker gene (bar) (in the RNAi mutants of cv. Zheltozernoe 10). The research findings presented in this chapter provide strong evidence that RNA interference can be used for improvement of the nutritional properties of sorghum grain.

scholarly journals Isolation and properties of a metalloproteinase from buckwheat (Fagopyrum esculentum) seeds

1990 ◽  
Vol 272 (3) ◽  
pp. 677-682 ◽  
Author(s):  
M A Belozersky ◽  
Y E Dunaevsky ◽  
N E Voskoboynikova
Keyword(s):  
Storage Proteins ◽  
Limited Proteolysis ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
Fagopyrum Esculentum ◽  
Gel Chromatography ◽  
Ph Optimum ◽  
Peptide Bonds ◽  
Sds Page ◽  
Homogeneous Preparation

A homogeneous preparation of metalloproteinase, purified 1000-fold, was obtained from buckwheat (Fagopyrum esculentum) seeds. The Mr of the enzyme, determined by SDS/PAGE, was 34,000 (it was 39,000 by gel chromatography). Its pH optimum was 8.0-8.2 with 13 S globulin, from buckwheat seeds, as substrate. Atomic-absorption spectroscopy revealed the presence of one Zn2+ ion per enzyme molecule. The enzyme was completely inhibited by EDTA (1 mM), zincone (1 mM) and 1, 10-phenanthroline (1 mM). The metalloproteinase performed limited proteolysis of the following seed storage proteins: 13 S globulin from buckwheat seeds and 11 S globulin from soybean (Glycine max) seeds. It hydrolysed three peptide bonds formed by the amino groups of Leu15, Tyr16 and Phe25 in the oxidized B-chain of insulin. In its main properties the enzyme is similar to metalloproteinases of animal and bacterial origin.

Revealing genetic diversity in land races and wild accessions of mungbean [Vigna radiata (L.) Wilczek] using SDS-PAGE of seed storage proteins

2015 ◽  
Author(s):  
Ananya Panda ◽  
Swapan K. Tripathy
Keyword(s):  
Storage Proteins ◽  
Seed Storage Protein ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
Land Races ◽  
Sds Page ◽  
Wild Accessions ◽  
Total Seed

Total seed storage protein profiles of 74 mungbean land races, three wild accessions and a popular variety ‘Jyoti’ of Odisha were analysed by Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). 32 genotypes could be clearly identified based on genotype-specific seed protein fingerprints while rest of the test genotypes were categorized into eight protein types. Genotypes included in each protein type had 100% homology and some of these could be duplicates. In this pursuit, a few specific polypeptide markers have been detected for identification of the land races/ genotypes. Dendrogram based on electrophoretic data clustered the genotypes into seven groups at 70% phenon level. Paralakhemundi local, Samarjhola local and Phulbani local-D; and three wild accessions (TCR 20, TCR 213 and TCR 243) were comparatively divergent from other genotypes. Besides, Jyoti, Kalahandi local 2A, Sikri local, kodala local A and TCR 20 were identified to be protein rich with high seed yield. TCR 20 being morphologically similar to mungbean, moderately high protein content and high yielding as well as resistant to drought and bruchids; it may serve as a valuable source genotype in recombination breeding

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

scholarly journals Subunit composition of seed storage proteins in high-protein soybean genotypes

2010 ◽  
Vol 45 (7) ◽  
pp. 721-729 ◽  
Author(s):  
Ksenija Taski-Ajdukovic ◽  
Vuk Djordjevic ◽  
Milos Vidic ◽  
Milka Vujakovic
Keyword(s):  
Protein Content ◽  
Storage Proteins ◽  
Seed Storage Protein ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
High Protein ◽  
Breeding Programs ◽  
Sds Page ◽  
Soybean Genotypes ◽  
Food Applications

The objective of this work was to quantify the accumulation of the major seed storage protein subunits, β-conglycinin and glycinin, and how they influence yield and protein and oil contents in high-protein soybean genotypes. The relative accumulation of subunits was calculated by scanning SDS-PAGE gels using densitometry. The protein content of the tested genotypes was higher than control cultivar in the same maturity group. Several genotypes with improved protein content and with unchanged yield or oil content were developed as a result of new breeding initiatives. This research confirmed that high-protein cultivars accumulate higher amounts of glycinin and β-conglycinin. Genotypes KO5427, KO5428, and KO5429, which accumulated lower quantities of all subunits of glycinin and β-conglycinin, were the only exceptions. Attention should be given to genotypes KO5314 and KO5317, which accumulated significantly higher amounts of both subunits of glycinin, and to genotypes KO5425, KO5319, KO539 and KO536, which accumulated significantly higher amounts of β-conglycinin subunits. These findings suggest that some of the tested genotypes could be beneficial in different breeding programs aimed at the production of agronomically viable plants, yielding high-protein seed with specific composition of storage proteins for specific food applications.

SDS–PAGE of seed proteins in Festuca (Poaceae): taxonomic implications

1991 ◽  
Vol 69 (7) ◽  
pp. 1425-1432 ◽  
Author(s):  
S. G. Aiken ◽  
S. E. Gardiner
Keyword(s):  
Native Species ◽  
Storage Proteins ◽  
Polyacrylamide Gel Electrophoresis ◽  
Seed Proteins ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
Banding Patterns ◽  
Ungerminated Seed ◽  
Sds Page ◽  
Species Specific

Taxonomically useful descriptors were provided by the banding patterns of seed storage proteins obtained when extracts of bulked, ungerminated seed samples from commercially available North American native species of Festuca were analyzed by sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS–PAGE). The banding patterns for three species of rough fescues (section Breviaristatae Krivot) indicate that although the taxa are closely related, F. campestris Rydb. (2n = 56) does not appear to be an autoploid of either F. altaica Trin. (2n = 28) or F. hallii (Vasey) Piper (2n = 28). A distinct band corresponding to a molecular weight of 57 000 occurred in the seed protein profiles of all native and commercial samples of Festuca L. analyzed. The profile for F. californica Vasey, questionably section Breviaristatae, also has a band at this position, and is very different from that of F. altaica, F. campestris, and F. hallii. Species-specific banding patterns were observed for F. brachyphylla Schultes, F. saximontana Rydb., F. idahoensis Elmer, and F. trachyphylla (Hackel) Krajina (F. ovina L. s.l., F. longifolia Thuill., F. ovina var. duriuscula auct. amer.). The results support the recognition of subgenus Schedonorus (Beauv.) Peter., and sections Breviaristatae Krivot and Festuca. Key words: Poaceae, Festuca, SDS–PAGE seed proteins.

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