scholarly journals The GAMYB gene in rye: sequence, polymorphisms, map location, allele-specific markers, and relationship with α-amylase activity

BMC Genomics ◽  
2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Anna Bienias ◽  
Magdalena Góralska ◽  
Piotr Masojć ◽  
Paweł Milczarski ◽  
Beata Myśków
2019 ◽  
Author(s):  
Anna Bienias ◽  
Magdalena Góralska ◽  
Piotr Masojć ◽  
Paweł Milczarski ◽  
Beata Myśków

Abstract Background. A master GA-induced regulatory protein, crucial for development and germination of cereal grain and involved in anther formation is MYB transcription factor GAMYB, activating a vast number of genes including high-molecular-weight glutenin and α-amylase gene families. This paper presents the first attempt to characterize rye gene encoding GAMYB in relation to its sequence, polymorphisms and phenotypic effects. Results. ScGAMYB gene was identified and mapped on rye chromosome 3R using high-density DArT/DArTseq-based maps developed in two mapping populations. Comparative analysis of the gene sequence revealed its high level of homology to wheat and barley orthologues. Single nucleotide polymorphisms detected among rye inbred lines allowed developing AS-PCR markers for ScGAMYB (ten pairs of primers) which might be used to detect this gene in wide genetic stocks of rye and triticale. Segregation of ScGAMYB alleles showed significant relationship with quantitative traits including plant height, thousand grain weight, α-amylase activity, earliness per se and leaf rolling. Conclusions. The research showed the strong similarity of rye GAMYB sequence to its orthologues in other Graminae and confirmed the position in the genome consistent with the collinearity rule of cereal genomes. The statistically significant, however moderate association of ScGAMYB with many agronomic features has been pointed out, which proved that this gene is a QTL of pleiotropic character. The effect of ScGAMYB on flowering time was statistically the most significant. Developed sequence-based, allele-specific PCR markers could be useful in research and application purposes.


1994 ◽  
Vol 71 (05) ◽  
pp. 651-654 ◽  
Author(s):  
Rainer Kalb ◽  
Sentot Santoso ◽  
Katja Unkelbach ◽  
Volker Kiefel ◽  
Christian Mueller-Eckhardt

SummaryAlloimmunization against the human platelet alloantigen system Br (HPA-5) is the second most common cause of neonatal alloimmune thrombocytopenia (NAIT) in Caucasian populations. We have recently shown that a single base polymorphism at position 1648 on platelet mRNA coding for GPIa results in an aminoacid substitution at position 505 on the mature GPIa which is associated with the two serological defined Br phenotypes.Since DNA-typing of platelet alloantigens offers possibilities for useful clinical applications, we designed genomic DNA-based restriction fragment length polymorphism (RFLP) typing for Br alloantigens. To establish this technique we analyzed the genomic organization of GPIa adjacent to the polymorphic base. Using the polymerase chain reaction (PCR) of blood cell DNA we have identified two introns (approximately 1.7 and 1.9 kb) flanking a 144 bp coding sequence of the GPIa gene encompassing the polymorphic base 1648. Based on the in- tron sequence, a PCR primer was constructed to amplify a 274 bp fragment which was used for allele-specific RFLP to determine the Br genotypes. The results of RFLP analysis using Mnll endonuclease obtained from 15 donors (2 Br37*, 2 Br^ and 11 Brb/b) correlate perfectly with serological typing by monoclonal antibody-specific immobilization of platelet antigens (MAIPA) assay.


1996 ◽  
Vol 75 (05) ◽  
pp. 757-759 ◽  
Author(s):  
Rainer Blasczyk ◽  
Markus Ritter ◽  
Christian Thiede ◽  
Jenny Wehling ◽  
Günter Hintz ◽  
...  

SummaryResistance to activated protein C is the most common hereditary cause for thrombosis and significantly linked to factor V Leiden. In this study, primers were designed to identify the factor V mutation by allele-specific PCR amplification. 126 patients with thromboembolic events were analysed using this technique, PCR-RFLP and direct sequencing. The concordance between these techniques was 100%. In 27 patients a heterozygous factor VGln506 mutation was detected, whereas one patient with recurrent thromboembolism was homozygous for the point mutation. Due to its time- and cost-saving features allele-specific amplification should be considered for screening of factor VGln506.


1997 ◽  
Vol 77 (06) ◽  
pp. 1154-1155 ◽  
Author(s):  
Gary D Sinclair ◽  
Sandra Low ◽  
Man-Chiu Poon

SummaryWe describe a novel hemi-nested, allele specific whole blood PCR assay for detection of the factor V Leiden mutation associated with the plasma defect, activated protein C resistance. This assay utilizes 5 μl of whole blood without prior DNA extraction. The hemi-nested design, employing an outer primer pair in combination with nested, allele specific primers obviates the need for restriction enzyme digestion. PCR reactions are analysed directly on agarose or polyacrylamide minigels. The assay confirmed the genotypes of 50 individuals previously categorized by PCR and Mnll digestion, and has been subsequently utilized in the genotyping of 445 individuals referred for thrombosis studies.


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