Characterisation of a methionine-rich storage protein cDNA from perilla (Perilla frutescens) seeds

2000 ◽  
Vol 27 (7) ◽  
pp. 701
Author(s):  
Un-Ho Jin ◽  
Byung-Rae Jin ◽  
Jin-Woo Lee ◽  
Young-Su Cho ◽  
O-Chang Kwon ◽  
...  
Keyword(s):  
Storage Protein ◽  
Storage Proteins ◽  
Expression Patterns ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
Perilla Frutescens ◽  
Amino Acid Sequences ◽  
Significant Feature ◽  
Transcriptional Level ◽  
Specific Expression

We have cloned and characterised a cDNA (PrLeg) coding for a methionine-rich storage protein, which is reported for the first time in perilla (Perilla frutescens (L.) Britt. var. Japonica) seeds, homologous to the 11S legumin-like storage proteins. The most significant feature of the PrLeg precursor protein is that it has the highest content of methionine residues among the 11S legumin-like storage proteins examined so far. Another feature is that the deduced amino acid sequences of the PrLeg protein are phylogenetically close to the sequence groups derived from evolutionally ancient states of the 11S legumin-like storage proteins, or from gymnospermous seed storage proteins, such as Magnolia, Asarum, Dioscorea, Cryptomeria, Metasequoia and Ginkgo. In contrast, with the exception of sesame, relatively low phylogenetic relationships are determined between the PrLeg sequence group and those derived from crop plants, such as soybean, pea, broad bean, rape, pumpkin, rice, coffee and citrus. Southern blot analysis suggests that there may be several copy numbers of thePrLeg genes and their seed-specific expression patterns at the transcriptional level were confirmed by northern hybridisation analysis.

scholarly journals Functional characterization of an endosperm specific promoter p1062 from common buckwheat (Fagopyrum esculentum Moench) for driving tissue specific gene expression / Funkcijske lastnosti endospermskega promotorja p1062 navadne ajde ...

2020 ◽  
Vol 61 (1) ◽  
pp. 45-54
Author(s):  
Lashaihun Dohtdong ◽  
Nikhil Kumar Chrungoo
Keyword(s):  
Storage Proteins ◽  
Functional Characterization ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
35S Promoter ◽  
Specific Gene ◽  
World Population ◽  
Nucleotide Position ◽  
Specific Expression ◽  
Camv 35S

Seed storage proteins of grain crops meet the major dietary protein requirement of over half of the world population. PCR based genome walking the 5’UTR of the gene coding for a lysine rich legumin type protein amplified a 1.1kb DNA fragment representing the promoter region of the gene. Clustal alignment of this sequence with other sequences in the Genbank database clearly showed 100 percent complementary base match of 282 bases at the 3’ end of the sequence, corresponding to nucleotide position 780-1062 with correspondingly similar number of bases on the 5’ end of the 1.7kb Bwleg gene.We detected one prolamin box and three RY-repeat motifs in the sequence. Seven deletion fragments of the putative promoter were generated by 5’ nested PCR and cloned in pCAMBIA1304 upstream of GUS gene after excising the CaMV 35S promoter from the vector. Arabidopsis plants plants harbouring the deletion construct pBwlDF1 to pBwlDF6 clearly showed seed specific expression of the reporter gene. Seeds harbouring the constructs pBwlDF3, pBwlDF4 and pBwlDF5 showed a nearly threefold decrease in GUS activity than those harbouring the construct with full length promoter. Key words: buckwheat, DNA, promoter, constructs   Izvleček Založne beljakovine semen zrnastih poljščin ustrezajo glavnim potrebam po beljakovinah za več kot polovico svetovnega prebivalstva. S PCR in 5’UTR so za kodiranje kakovostnih beljakovin leguminskega tipa pomnožili odlomek 1,1 kb DNK, ki je promotorsko gensko območje. Vzporejanje te sekvence z drugimi sekvencami podatkovne baze genske banke jasno pokaže popolno komplementarnost 282 baz na 3’ koncu sekvence, kar ustreza pozicijam 780-1062 z ustreznim številom baz na 5’ koncu gena 1,7 kb Bwleg. V sekvenci smo odkrili eno prolaminsko škatljo in tri RY-ponovljene motive. Sedem delecijskih fragmentov putativnega promotorja smo generirali z 5’ PCR kloniranjem pCAMBIA1304 navzgor od GUS gena po izločitvi promotorja CaMV 35S iz vektorja. Semena s konstrukti pBwlDF3, pBwlDF4 in pBwlDF5 so izražali skoraj trikratno zmanjšanje GUS aktivnosti v primerjavi s konstrukti, ki so vsebovali polne dolžine promotorjev.  Ključne besede: ajda, DNK, promotor, konstrukti

scholarly journals Engineering sulfur storage in maize seed proteins without apparent yield loss

2017 ◽  
Vol 114 (43) ◽  
pp. 11386-11391 ◽  
Author(s):  
Jose Planta ◽  
Xiaoli Xiang ◽  
Thomas Leustek ◽  
Joachim Messing
Keyword(s):  
Yield Loss ◽  
Storage Proteins ◽  
Seed Proteins ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
Sulfur Assimilation ◽  
Specific Expression ◽  
Feeding Trials ◽  
The Cost ◽  
Reduction Capacity

Sulfur assimilation may limit the pool of methionine and cysteine available for incorporation into zeins, the major seed storage proteins in maize. This hypothesis was tested by producing transgenic maize with deregulated sulfate reduction capacity achieved through leaf-specific expression of the Escherichia coli enzyme 3′-phosphoadenosine-5′-phosphosulfate reductase (EcPAPR) that resulted in higher methionine accumulation in seeds. The transgenic kernels have higher expression of the methionine-rich 10-kDa δ-zein and total protein sulfur without reduction of other zeins. This overall increase in the expression of the S-rich zeins describes a facet of regulation of these proteins under enhanced sulfur assimilation. Transgenic line PE5 accumulates 57.6% more kernel methionine than the high-methionine inbred line B101. In feeding trials with chicks, PE5 maize promotes significant weight gain compared with nontransgenic kernels. Therefore, increased source strength can improve the nutritional value of maize without apparent yield loss and may significantly reduce the cost of feed supplementation.

Profiling the Seed Storage Protein Among Different Genotypes of Trigonella foenum-graecum L. (Fenugreek)

2019 ◽  
Author(s):  
Nisha . ◽  
Priyanka Khati ◽  
P B Rao
Keyword(s):  
Storage Protein ◽  
Gel Electrophoresis ◽  
Storage Proteins ◽  
Seed Storage Protein ◽  
Sodium Dodecyl ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
Protein Band ◽  
Trigonella Foenum Graecum ◽  
Trigonella Foenum Graecum L

A qualitative as well as quantitative categorization of seed storage proteins profiles of 23 genotypes of Trigonella foenum- graecum L. were performed by using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) for exploring the level of genetic discrepancy at seed storage protein level. Total soluble proteins were resolved on 10% resolving gel. A dendrogram was constructed on the basis of weight of seed storage proteins, which divide total genotypes into two groups further classified into different sub groups containing different genotypes in them. The bands obtained from gel electrophoresis can serve as a potent tool in discrimination of different genotypes on the basis of their protein content. Proteins with molecular weight 66, 43 and 35 kDa were found in all the genotypes except Fgk-76, PR, Rmt-303, PEB and Rmt-361, The 43 kDa protein band was found missing in Fgk-67, AFg-2, AM-2, AFg-4, Fgk-73, although the protein with 35 kDa weight was present in all the genotypes but not in Rmt-303 same as 63 kDa which is not present in Fgk-70 and 55 kDa protein band was found missing in Fgk-67, Afg-4 and Rmt-361.

scholarly journals Characterization of Seed Storage Proteins from Chickpea Using 2D Electrophoresis Coupled with Mass Spectrometry

2016 ◽  
Vol 2016 ◽  
pp. 1-6 ◽  
Author(s):  
Pramod Kumar Singh ◽  
Nidhi Shrivastava ◽  
Krishna Chaturvedi ◽  
Bechan Sharma ◽  
Sameer S. Bhagyawant
Keyword(s):  
Mass Spectrometry ◽  
Storage Proteins ◽  
Seed Storage Protein ◽  
Expression Patterns ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
Differentially Expressed ◽  
2D Electrophoresis ◽  
Differentially Expressed Proteins ◽  
Amino Acid Homology

Proteomic analysis was employed to map the seed storage protein network in landrace and cultivated chickpea accessions. Protein extracts were separated by two-dimensional gel electrophoresis (2D-GE) across a broad range 3.0–10.0 immobilized pH gradient (IPG) strips. Comparative elucidation of differentially expressed proteins between two diverse geographically originated chickpea accessions was carried out using 2D-GE coupled with mass spectrometry. A total of 600 protein spots were detected in these accessions. In-gel protein expression patterns revealed three protein spots as upregulated and three other as downregulated. Using trypsin in-gel digestion, these differentially expressed proteins were identified by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) which showed 45% amino acid homology of chickpea seed storage proteins withArabidopsis thaliana.

scholarly journals Genetic variation in common bean (Phaseolus vulgaris L.) using seed protein markers

2020 ◽  
Vol 12 (1) ◽  
pp. 58-69
Author(s):  
Henok Ayelign ◽  
Eleni Shiferaw ◽  
Faris Hailu
Keyword(s):  
Common Bean ◽  
Storage Protein ◽  
Storage Proteins ◽  
Seed Storage Protein ◽  
Gene Diversity ◽  
Seed Proteins ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
Protein Markers ◽  
Mesoamerican Gene Pool

AbstractThe genetic diversity of common bean accessions were assessed using seed storage protein markers. At regional level, accessions from the two major growing regions showed the highest level of gene diversity (H = 0.322, I = 0.485, and H = 0.312, I = 0.473), which can be exploited for the future improvement of the crop. Based on phaseolin, the major storage protein in common bean, the majority of the accessions (86%) were grouped under Mesoamerican gene pool. Seed proteins were also used to differentiate various Phaseolus species, indicating the usefulness of seed storage proteins in species identification in this genus.

scholarly journals Identification and Characterization of the Seed Storage Proteins and Related Genes of Cannabis sativa L.

2021 ◽  
Vol 8 ◽  
Author(s):  
Xin Sun ◽  
Yao Sun ◽  
Yao Li ◽  
Qiong Wu ◽  
Lei Wang
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Storage Protein ◽  
Cannabis Sativa ◽  
Storage Proteins ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
Hemp Seed ◽  
Abundant Protein ◽  
Molecular Weights

Hemp (Cannabis sativa L.) seed is emerging as a novel source of plant protein owing to its rich protein content and reasonable nutritional structure. In the current study, the storage proteins of hemp seed were extracted using different methods. The modified Osborne method yielded maximum extraction of the hemp seed storage proteins, while degreasing had little effect on the hemp seed protein (HSP) extraction. Protein identification results revealed that 11S globulin (edestin) was the most abundant protein in hemp seed, and the molecular weights of the two subunits of this protein were ~35 and 20 kDa, respectively. The second most abundant protein was 2S albumin (Cs2S), with a molecular weight of ~14–15 kDa. The least abundant protein was 7S vicilin-like protein (Cs7S), with a molecular weight of ~47 kDa. Subsequently, gene families encoding these three storage protein classes, including three genes for edestin, two for Cs2S, and one for Cs7S, were cloned and then analyzed for amino acid composition and structure. The three edestins were different in their amino acid sequences and calculated molecular weights. The analysis of coding sequences revealed a higher percentage of similarity (62.7%) between Edestin1 and Edestin3, while the similarity decreased significantly to ~57% between Edestin1 and Edestin2, and 58% between Edestin2 and Edestin3. The calculated protein molecular weight was the highest for the protein encoded by Edestin1 and the smallest for the protein encoded by Edestin2. All three edestins were rich in arginine, while Edestin3 had a higher methionine content relative to that in the other two, which proved that Edestin3 had a better nutritional value. Cs2S and Cs7S were different from those reported in previous studies. Therefore, it could be inferred that amino acid composition varies with different hemp cultivars. The current research brought significant theoretical advance in illuminating the understanding of hemp seed storage protein and would have significance for future research on improving the nutritional quality of hemp seed and developing bioactive peptides.

scholarly journals Assessment of Genetic Diversity in Bambara Groundnut (Vigna subterranea) Landraces Based on Seed Storage Proteins Electrophoretic Pattern

2021 ◽  
Vol 38 (1) ◽  
pp. 40-47
Author(s):  
N.M. Saminu ◽  
B.G. Kurfi ◽  
Y.Y. Muhammad
Keyword(s):  
Storage Protein ◽  
Storage Proteins ◽  
Seed Storage Protein ◽  
Protein Profile ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
Bambara Groundnut ◽  
Vigna Subterranea ◽  
Research Attention ◽  
Major Cluster

Bambara groundnut (Vigna subterranea) is a leguminous crop that is considered underutilized and has previously received little research attention. Variability in a number of physiological factors, including germination rate, widely affects its production. Seed storage protein, its fractions and protein profile of six Bambara groundnut local landraces were studied to assess their genetic relatedness. Total seed storage protein and its fractions were estimated by Bradford’s method. SDS-PAGE analysis was used to evaluate storage protein profile. The results showed significant differences (p<0.05) in protein contents among the landraces. The major seed storage proteins were found to be globulins (0.048 to 0.088mg/mL ), albumins (0.023 to 0.038mg/mL ), glutelins (0.007 to 0.013mg/mL ) and prolamins (0.002 to 0.004mg/mL ). Five peptide bands were detected with molecular weights corresponding to 97.4 kDa, 45 kDa, 29 kDa, 20.1 kDa and 18 kDa, respectively. Three peptide bands corresponding to 97.4 kDa, 45 kDa and 18 kDa were detected in all the landraces and two peptide bands between 29 kDa and 20.1 kDa were detected in five landraces. Dendrogram generated by UPGMA grouped the six landraces into one major cluster with two sub-clusters. The observed diversity in storage protein pattern of the landraces indicated their potential as materials for crop improvement.

Seed storage proteins in cultivars and subspecies of alfalfa (Medicago sativa L.)

2000 ◽  
Vol 10 (4) ◽  
pp. 423-434 ◽  
Author(s):  
Joan E. Krochko ◽  
J. Derek Bewley
Keyword(s):  
Medicago Sativa ◽  
Storage Protein ◽  
Species Complex ◽  
Storage Proteins ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
Protein Patterns ◽  
2S Albumin ◽  
Two Dimensional Gel Electrophoresis ◽  
11S Globulin

AbstractSeed storage proteins were analysed in 27 varieties of alfalfa (Medicago sativaL.); these included five subspecies (glomerata, caerulea, falcata, hemicycla, praefalcata), seven of the nine sources of Medicago germplasm introduced into North America and a sample of additional cultivars. The protein patterns were remarkably consistent for all of these taxa. One-dimensional and two-dimensional gel electrophoresis revealed only minor differences in polypeptide composition within each of the three major classes of storage protein (7S globulin, 11S globulin, 2S albumin). The slight variations that were found provided no information on either parentage or evolutionary relationships amongst these particular taxa. Nonetheless, persistent and reproducible heterogeneity of some minor polypeptides of 11S globulin (medicagin) may be useful under other circumstances for cultivar identification in alfalfa. Both subfamilies (I and II) of the 11S globulin were strongly expressed in all of the cultivars and subspecies examined. It was concluded that this structural divergence within the 11S storage protein family predated the evolution of the M. sativa L. species complex. Most of the variability in storage proteins was quantitative. However, even this variability was reduced when data were standardized with respect to seed dry weights. The consistent similarities in qualitative and quantitative expression of seed storage proteins amongst all of these taxa suggest a high degree of uniformity in both seed physiology and genetics within the alfalfa species complex.

The role of mRNA and protein sorting in seed storage protein synthesis, transport, and deposition

2005 ◽  
Vol 83 (6) ◽  
pp. 728-737 ◽  
Author(s):  
Andrew J Crofts ◽  
Haruhiko Washida ◽  
Thomas W Okita ◽  
Mio Satoh ◽  
Masahiro Ogawa ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Storage Protein ◽  
Storage Proteins ◽  
Seed Storage Protein ◽  
Protein Localization ◽  
Protein Sorting ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
Rna Localization ◽  
Endomembrane System

Rice synthesizes and accumulates high levels of 2 distinct classes of seed storage proteins and sorts them to separate intracellular compartments, making it an ideal model system for studying the mechanisms of storage protein synthesis, transport, and deposition. In rice, RNA localization dictates the initial site of storage protein synthesis on specific subdomains of the cortical endoplasmic reticulum (ER), and there is a direct relation between the RNA localization site and the final destination of the encoded protein within the endomembrane system. Current data support the existence of 3 parallel RNA localization pathways leading from the nucleus to the actively synthesizing cortical ER. Additional pathways may exist for the synthesis of cytoplasmic and nuclear-encoded proteins targeted to organelles, the latter located in a stratified arrangement in developing endosperm cells. The study of rice mutants, which accumulate unprocessed glutelin precursors, indicates that these multiple pathways prevent nonproductive interactions between different classes of storage proteins that would otherwise disrupt protein sorting. Indeed, it appears that the prevention of disruptive interactions between different classes of storage proteins plays a key role in their biosynthesis in rice. In addition to highlighting the unique features of the plant endomembrane system and describing the relation between RNA and protein localization, this minireview will attempt to address a number of questions raised by recent studies on these processes.Key words: mRNA localization, protein localization, endomembrane system, seed storage proteins, rice.

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