PCR analysis of the wheat varieties and near-isogenic wheat lines with the use of allele-specific primers for the Gli-1 and Glu-3 loci

2010 ◽  
Vol 44 (6) ◽  
pp. 345-353
Author(s):  
A. M. Polishchuk ◽  
S. V. Chebotar ◽  
E. M. Blagodarova ◽  
N. A. Kozub ◽  
I. A. Sozinov ◽  
...  
Keyword(s):  
Pcr Analysis ◽  
Specific Primers ◽  
Wheat Varieties ◽  
Allele Specific ◽  
Gli 1 ◽  
Wheat Lines

scholarly journals Short Communication: Genotyping of kdr allele in insecticide resistant-Aedes aegypti populations from West Sumatra, Indonesia

2018 ◽  
Vol 19 (2) ◽  
pp. 502-508
Author(s):  
HASMIWATI HASMIWATI ◽  
SUPARGIYONO SUPARGIYONO
Keyword(s):  
Aedes Aegypti ◽  
Dengue Fever ◽  
Short Communication ◽  
Point Mutations ◽  
Pcr Analysis ◽  
Chemical Insecticide ◽  
Paper Test ◽  
Specific Primers ◽  
West Sumatra ◽  
Allele Specific

Hasmiwati, Supargiyono. 2018. Short Communication: Genotyping of kdr allele in insecticide resistant-Aedes aegypti populations from West Sumatra, Indonesia. Biodiversitas 19: 502-508. Aedes aegypti, the primary vector of Dengue fever, is widely distributed in Indonesia including in West Sumatra. Chemical insecticide is an effective way of shedding the chain of Dengue fever transmission. Long-term applications of insecticides have resulted in the development of resistance in Ae. aegypti populations. Knockdown resistance (kdr) allele as a result of point mutations has been found in Voltage-Gated Sodium Channel (VGSC) gene. This study aimed to design allele-specific primers to detect the kdr allele mutations. Ae aegypti samples were collected from several dengue endemic areas in West Sumatra. The susceptibility of adult mosquitos to insecticides was determined by bioassays using impregnation paper test with 0.18% malathion and 0.75% permethrin. In this study, we successfully confirmed that the A. populations have point mutations in the VGSC gene corresponding to S989P and V1016G amino acid substitutions. To genotype S989P and V1016G alleles, we designed kdr allele-specific primers based on the sequence of VGSC gene (NCBI Accession number No. EU399179.1. PCR analysis using the kdr allele-specific primers successfully identified the genotype of Ae. aegypti populations resistant to malathion and permethrin in West Sumatra.

scholarly journals Identification of allelic combinations of the Ppd-D1, Vrn-A1, Vrn-B1 and Vrn-D1 genes in common wheat lines obtained in the National Center of Grain named after P. P. Lukyanenko

2020 ◽  
Author(s):  
Yu.S. Zubanova ◽  
◽  
V.A. Filobok ◽  
E.A. Guenkova ◽  
E.R. Davoyan ◽  
...  
Keyword(s):  
Common Wheat ◽  
Recessive Allele ◽  
Dominant Allele ◽  
Specific Primers ◽  
Photoperiodic Sensitivity ◽  
Allelic Composition ◽  
Vrn1 Gene ◽  
Allele Specific ◽  
Wheat Lines

An analysis of the allelic composition of the genes determining photoperiodic sensitivity (Ppd-D1) and the need for vernalization (Vrn-A1, Vrn-B1, Vrn-D1) was carried out in 286 common wheat lines obtained in the National Center of Grain named after P. P. Lukyanenko with the use of allele-specific primers. The analyzed samples were distributed over 21 haplotypes; the dominant allele of the Ppd-D1a gene prevailed in the studied material. 123 lines of common wheat carry a combination of D-RRD alleles. The lines that can be attributed to the group of alternate wheat (R-RDR, R-RRD) were identified. All studied samples carry the recessive allele of at least one VRN1 gene.

scholarly journals Molecular labeling of Vrn, Ppd genes and vernalization response of the ultra-early lines of spring bread wheat Triticum aestivum L.

2021 ◽  
Vol 4 (3) ◽  
pp. 26-36
Author(s):  
B. V. Rigin ◽  
E. V. Zuev ◽  
I. I Matvienko ◽  
A. S. Andreeva
Keyword(s):  
Bread Wheat ◽  
Dominant Gene ◽  
Wheat Breeding ◽  
High Rate ◽  
Pcr Analysis ◽  
Vernalization Response ◽  
Spring Bread Wheat ◽  
Allele Specific ◽  
Wheat Lines ◽  
Northwestern Russia

Background. The knowledge of genetic control of vernalization response in the ultra-early accessions can facilitate bread wheat breeding for a high adaptive capacity. Materials and methods. The study involved the ultra-early lines Rico (k-65588) and Rimax (k-67257) as the earliest maturing lines in the VIR bread wheat collection, as well as 10 Rifor lines (k-67120, k-67121, k-67250-67256) with a high rate of development before heading. A late ripening accession ‘Forlani Roberto’ (k-42641) and ‘Leningradskaya 6’ variety (k-64900), regionally adapted to Northwestern Russia, were also studied. The alleles of the Vrn and Ppd genes were identified by the PCR analysis using the allele-specific primers published in literature sources. The response to vernalization (30 days at 3°C) and a short 12-hour day were determined using a methodology accepted at VIR. Results. The ultra-early lines respond to a short 12-hour day and 30-day vernalization very poorly. The genotype of ultra-early wheat lines is mainly represented by three genes, Vrn-A1, Vrn-B1a, and Vrn-D1, which ensure insensitivity to vernalization alongside with the expression of Ppd-D1a, which controls the response to photoperiod. The ultra-early lines Rifor 4 and Rifor 5 have a recessive allele vrn-A1a, like the original ‘Forlani Roberto’ accession. The lines Rifor 4 and Rifor 5 are vernalization-insensitive under the long day and have a very weak response under the short day (3.5±0.42 days and 4.0±0.61 days, respectively). However, ‘Forlani Roberto’ with the vrn-A1a gene responds to vernalization in the same way under any photoperiod (12.3±1.58 days and 12.2±0.74 days). Conclusion The ultra-early lines of bread wheat Rifor 4 and Rifor 5 with the vrn-A1a gene can have no response to vernalization or have a low level response. This effect can be a reason for the formation of a complex of modifier genes along with the dominant gene Vrn-D1, which forms during the hybridization of F7-8 Rico × Forlani Roberto. The ultra-early lines of bread wheat Rico, Rimax and Rifor (k-67120, k-67121, k-67250-67256) can serve as effective sources of genes for earliness in common wheat breeding.

scholarly journals PCR-analysis of photoperiod sensitivity genes in bread wheat varieties from Bilatserkovska experimental breeding station

2019 ◽  
Vol 16 (2) ◽  
pp. 217-226
Author(s):  
V. M. Filimonov ◽  
A. A. Bakuma ◽  
G. A. Chebotar ◽  
L. A. Burdenyuk-Tarasevich ◽  
S. V. Chebotar
Keyword(s):  
Winter Wheat ◽  
Dna Isolation ◽  
Photoperiod Sensitivity ◽  
Pcr Analysis ◽  
Classification Methods ◽  
Wheat Varieties ◽  
Breeding Station ◽  
Allele Specific ◽  
Photoperiod Sensitivity Genes

The aim. Determination of alleles of the photoperiod sensitivity genes Ppd-1 in 16 winter wheat varieties of the Bilatserkovska Experimental Breeding Station and identification of the haplotypes of Ppd-D1 gene according to the accepted classification. Methods. DNA isolation, allele-specific and nested PCR, electrophoresis in agarose and polyacrylamide gels, determination of the significant differences in the time of heading. Results. Plants of the varieties Vodohrai bilotserkivs’kiy; Bilotserkivs’ka napivkarlykova, Olesia, Perlyna lisostepu, Elehiia, Yasochka, Lybid’, Tsarivna, Lisova pisnia, Romantyka, Vidrada, Schedra nyva, Charodiika bilotsekivs’ka, Russa, Driada 1 have the genotype PpdA1b Ppd-B1b Ppd-D1a and belong to the VII haplotype of the Ppd-D1 gene. The genotype of the variety Legenda bilotsekivs’ka is characterized by the presence of the alleles Ppd-A1b Ppd-B1b Ppd-D1b and belongs to the IV haplotype of the Ppd-D1 gene. The significant differences (P = 0.01) in the time of heading was detected only between the varieties Russa and Legenda bilotsekivs’ka and was 12.4 days. Conclusions. Most BEBS varieties are characterized by genotype — PpdA1b Ppd-B1b Ppd-D1a, in which the allele Ppd-D1a determines insensitivity to the photoperiod and leads to early heading. Only variety Legenda bilotsekivs’ka is the carrier of the recessive allele of Ppd-D1 gene, belongs to the IV haplotype, has the latest time of heading. Keywords: Ppd-A1, Ppd-B1, Ppd-D1 genes, winter wheat, PCR analysis, photoperiod sensitivity.

A Hemi-nested, Allele Specific, Whole Blood PCR Assay for the Detection of the Factor V Leiden Mutation

1997 ◽  
Vol 77 (06) ◽  
pp. 1154-1155 ◽  
Author(s):  
Gary D Sinclair ◽  
Sandra Low ◽  
Man-Chiu Poon
Keyword(s):  
Whole Blood ◽  
Protein C ◽  
Activated Protein C ◽  
Factor V Leiden ◽  
Restriction Enzyme Digestion ◽  
Factor V ◽  
Pcr Assay ◽  
Specific Primers ◽  
Factor V Leiden Mutation ◽  
Allele Specific

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.

VIRULENCE OF THE NORTH CAUCASIAN POPULATION OF PUCCINIA GRAMINIS PERS. F. SP. TRITICI

1970 ◽  
pp. 40-45
Author(s):  
G. V. Volkova ◽  
Е. V. Gladkova ◽  
O. O. Miroshnichenko
Keyword(s):  
High Efficiency ◽  
Caucasian Population ◽  
Puccinia Graminis ◽  
North Caucasus ◽  
Wheat Varieties ◽  
Caucasus Region ◽  
Pathogen Virulence ◽  
The North ◽  
New Varieties ◽  
Wheat Lines

The aim of the study was to monitor the virulence of Puccinia graminis Pers. f. sp. tritici in the North Caucasus.  The objectives of the study were to collect P. graminis infectious material from sown winter wheat varieties and evaluate the long-term dynamics of the pathogen virulence in the North Caucasus region in 2014-2019. As a result, an analysis of the virulence of the stem rust pathogen population of wheat collected in Krasnodar, Stavropol Territories, and Rostov Region was carried out. 81 mono-empty mushroom isolates were isolated and differentiated.  The genes Sr5, Sr31, Sr38 were characterized by high efficiency.  On the lines with the genes Sr7b, Sr8b, Sr9f, Sr9g, Sr10, Sr11, Sr12, Sr13, Sr14, Sr21, Sr22, Sr23, Sr26, Sr29, Sr32, Sr33, S35, Sr37, SrDp2, SrWLD, a variation in the virulence frequencies of P. graminis was observed.  Significant changes (in the direction of increasing occurrence) in the North Caucasian population 2014-2019  the pathogen was noted in the frequency of clones virulent to wheat lines with resistance genes Sr11, Sr21, Sr22, Sr26, Sr32, Sr33. A decrease in the frequency of clones virulent to Sr8b, Sr9g, Sr10, Sr12, Sr14, Sr35. At approximately the same level, the occurrence of clones virulent to the genes Sr6, Sr7a, Sr8а, Sr9a, Sr9b, Sr9d, Sr9e, Sr13, Sr15, Sr16, Sr17, Sr19, Sr20, Sr24, Sr25, Sr27, Sr30, Sr36, Sr39, Sr40, Sr44, SrGt, SrTmp. Effective genes that have shown their resistance to P. graminis in the seedling phase are proposed for use in breeding in southern Russia to create new varieties of wheat.

Design of allele-specific primers and detection of the human ABO genotyping to avoid the pseudopositive problem

2008 ◽  
Vol 29 (20) ◽  
pp. 4130-4140 ◽  
Author(s):  
Hidenobu Yaku ◽  
Tetsuo Yukimasa ◽  
Shu-ichi Nakano ◽  
Naoki Sugimoto ◽  
Hiroaki Oka
Keyword(s):  
Specific Primers ◽  
Allele Specific

Triticum polonicum L. as potential source material for the biofortification of wheat with essential micronutrients

2018 ◽  
Vol 17 (03) ◽  
pp. 213-220 ◽  
Author(s):  
Teresa Bieńkowska ◽  
Elżbieta Suchowilska ◽  
Wolfgang Kandler ◽  
Rudolf Krska ◽  
Marian Wiwart
Keyword(s):  
Durum Wheat ◽  
Common Wheat ◽  
Principal Component ◽  
Small Scale ◽  
Wheat Varieties ◽  
Triticum Polonicum ◽  
Iron And Zinc ◽  
Modern Wheat ◽  
Wheat Lines ◽  
Source Materials

AbstractThe grain of modern wheat cultivars has a significantly lower mineral content, including the content of copper, iron, magnesium, manganese, phosphorous, selenium and zinc. For this reason cereal breeders, are constantly searching for new genetic sources of minerals that are essential in human nutrition. Triticum polonicum, which is grown on a small scale in Spain, southern Italy, Algeria, Ethiopia and warm regions of Asia, deserves special attention in this context. The micronutrient and macronutrient content of T. polonicum versus T. durum and T. aestivum was compared in this study. Polish wheat grain was characterized by the significantly highest content of phosphorus (4.55 g/kg), sulphur (1.82 g/kg), magnesium (1.42 g/kg), zinc (49.5 mg/kg), iron (39.1 mg/kg) and boron (0.56 mg/kg) as well as a low content of aluminium (only 1.04 mg/kg). The macronutrient profile of most T. polonicum lines differed completely from that of common wheat and durum wheat. The principal component analysis supported discrimination of seven Polish wheat lines with a particularly beneficial micronutrient profile (P2, P3, P5, P7, P9, P22 and P25). These lines were characterized by the highest content of copper, iron and zinc, as well as the lowest concentrations of strontium, aluminium and barium which are undesirable in food products. The above lines can be potentially applied as source materials for breeding new wheat varieties. The results of this study indicate that Polish wheat could be used in genetic biofortification of durum wheat and common wheat.

scholarly journals ASSESSMENT OF THE INTEGRATED CHITINASE GENE STABILITY IN WHEAT LINES AFTER BIOBALLISTIC TRANSFORMATION

REPORTS ◽  
2020 ◽  
Vol 6 (334) ◽  
pp. 5-13
Author(s):  
N.P. Malakhova ◽  
◽  
Y.A. Skiba ◽  
E.R. Maltseva ◽  
G.A. Iskakova ◽  
...  
Keyword(s):  
Plant Immunity ◽  
Dna Amplification ◽  
Chitinase Gene ◽  
Fungal Diseases ◽  
Pr Proteins ◽  
Pcr Analysis ◽  
Soil Conditions ◽  
Wheat Lines ◽  
Resistance To Fungal Diseases ◽  
Regenerant Plants

The synthesis of PR-proteins (pathogenesis related proteins), the most studied of which are chitinases and β-1,3-glucanases, occurs in response to infection with pathogens in plants. Information about the exact role of individual PR proteins within plant immunity makes it possible to use certain specific antifungal proteins for the development of transgenic plants with increased resistance to fungal diseases. At the same time, it is important not only to obtain a plant with the desired trait, but also to fix in it the stable expression of the transferred gene and the inheritance of the acquired trait in generations. Herein we have studied the stability of the chitinase gene insertion in T1, T2 and T3 generations of transformed wheat lines obtained by the method of cis-gene transfer. Primary transformed regenerant plants were obtained as a result of bioballistic transformation of the chitinase gene into immature wheat germ of Saratovskaya 29 and Kazakhstanskaya 19 varieties. Following screening of regenerant plants by PCR for the presence of the target gene made it possible to select 6 lines presumably carrying the insert based on variety Saratovskaya 29 and 2 lines based on variety Kazakhstanskaya 19. The seed material of the selected regenerant plants was cultivated in soil conditions and the seeds of the T1 generation were obtained. DNA amplification of 8 selected lines St-29№25, St -29№43, St-29№44, St-29№33, St-29№26, St-29№35, Кz-19№1, Кz-19№2 with specific primers revealed insert-carrying lines, partially cleaved lines and lines with a high degree of insert instability. According to the results of the T2 and T3 generations PCR analysis, a complete absence of insertion in the St-29№35 and St-29№33 lines was revealed, a partial cleavage of the trait in the St-29№43 and Kz-19№2 lines was revealed, and the stable inheritance of the chitinase gene in four lines St-29№25, St-29№44, St-29№26 and Kz-19№1.was confirmed. These lines were selected as promising for in-depth study of their resistance to fungal diseases and further replication.

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