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.

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.

Protein disulphide isomerase promoter sequence analysis of Triticum urartu, Aegilops speltoides and Aegilops tauschii

2011 ◽  
Vol 9 (2) ◽  
pp. 338-341 ◽  
Author(s):  
Arun Prabhu Dhanapal ◽  
Mario Ciaffi ◽  
Enrico Porceddu ◽  
Elisa d'Aloisio
Keyword(s):  
Sequence Analysis ◽  
Storage Proteins ◽  
Aegilops Tauschii ◽  
Seed Storage ◽  
Seed Storage Proteins ◽  
Aegilops Speltoides ◽  
Secretory Proteins ◽  
Triticum Urartu ◽  
Specific Expression ◽  
Protein Disulphide Isomerase

Protein disulphide isomerase (PDI) catalyses the formation, reduction and isomerization of disulphide bonds in the newly synthesized secretory proteins. Plant PDIs have been shown to be involved in the folding and deposition of seed storage proteins, which makes this enzyme particularly interesting in wheat, as flour quality is strongly affected by composition and structure of seed storage proteins. In hexaploid wheat cultivar (AABBDD) Chinese Spring (CS), the genomic, complementary DNA and promoter sequences of the three homoeologous gene encoding PDI had been isolated and characterized in a previous study revealing high levels of sequence conservation. In this study, we report the isolation and sequencing of a ~700 bp region, comprising ~600 bp of the putative promoter region and 88 bp of the first exon of the typical PDI gene, in five accessions each from Triticum urartu (AA), Aegilops speltoides (BB) and Aegilops tauschii (DD). Sequence analysis indicated large variation among sequences belonging to the different genomes, while close similarity was found within each species and with the corresponding homoeologous PDI sequences of Triticum aestivum cv. CS (AABBDD) resulting in an overall high conservation of the regulatory motifs conferring endosperm-specific expression.

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.

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.

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
Sign in / Sign up

Export Citation Format

Share Document