Conversion of Existing AFLP Markers to SCAR Markers Linked to Globodera rostochiensis and Phytophthora infestans Resistance Could Be Performed Without Using Acrylamide Gel Electrophoresis

2021 ◽  
Author(s):  
Kengo Ohbayashi
Keyword(s):  
Phytophthora Infestans ◽  
Gel Electrophoresis ◽  
Globodera Rostochiensis ◽  
Aflp Markers ◽  
Scar Markers

scholarly journals Genetic variation among asexual progeny of Phytophthora infestans detected with RAPD and AFLP markers

2003 ◽  
Vol 52 (3) ◽  
pp. 314-325 ◽  
Author(s):  
F. M. Abu-El Samen ◽  
G. A. Secor ◽  
N. C. Gudmestad
Keyword(s):  
Genetic Variation ◽  
Phytophthora Infestans ◽  
Aflp Markers

scholarly journals Identification of Interspecific Potato Hybrids with Combined Resistance to Late Blight (Phytophthora Infestans) and Nematode (Globodera Rostochiensis)

2020 ◽  
Vol 74 (3) ◽  
pp. 188-195
Author(s):  
Nadezhda Zoteyeva ◽  
Guna Sprūde ◽  
Natalia Klimenko ◽  
Ieva Mežaka
Keyword(s):  
Molecular Markers ◽  
Late Blight ◽  
Phytophthora Infestans ◽  
Late Blight Resistance ◽  
Globodera Rostochiensis ◽  
Cyst Nematode ◽  
Potato Yields ◽  
Highly Correlated ◽  
Interspecific Potato Hybrids ◽  
Moderately Resistant

AbstractLate blight (agent Phytophthora infestans) and potato cyst nematode (PCN) caused by Globodera rostochiensis are economically important pathogens, which may significantly reduce potato yields. In this study interspecific potato hybrids were used as a source of resistance for combined resistance to economically important potato pathogens: late blight and cyst nematode. The aim of our study was to identify hybrid progenies with combined resistance to both pathogens and to verify the applicability of several molecular markers associated with resistance to G. rostochiensis pathotype Ro1 to identify resistant plants. Ninety-two clones of eleven original interspecific potato hybrids obtained in crosses with the cultivated S. tuberosum group tuberosum, S. tuberosum group Andigena, S. tuberosum group Phureja and wild S. guerreroense, S. microdontum, S. kurtzianum, S. neoantipoviczii and S. tarijense potato species were screened in bioassays and by molecular markers. PCN resistant or moderately resistant clones were found among the progenies of nine hybrids. Results were highly correlated with resistance status detected by molecular markers linked to the H1 (marker 57R) and Gro1-4 (marker Gro1) genes. Marker CP113 (linked to the H1 gene) was not polymorphic and failed to detect resistance status. Combination of foliar late blight resistance and resistance to PCN was identified in hybrids obtained in crosses with plants of species S. microdontum, S. tarijense and S. phureja and in the hybrid between S. guerreroense and Black’s P. infestans race differential carrying gene R-5.

scholarly journals Development of Pathotype-Specific SCAR Markers for Detection of Verticillium albo-atrum Isolates from Hop

Plant Disease ◽  
2004 ◽  
Vol 88 (10) ◽  
pp. 1115-1122 ◽  
Author(s):  
Sebastjan Radišek ◽  
Jernej Jakše ◽  
Branka Javornik
Keyword(s):  
Rapid Identification ◽  
Aflp Markers ◽  
Scar Markers ◽  
Specific Primer ◽  
Specific Sequence ◽  
Sequence Characterized Amplified Region ◽  
Wide Range ◽  
Rapid Polymerase Chain Reaction ◽  
Production Areas ◽  
Verticillium Albo Atrum

Rapid polymerase chain reaction (PCR) assays were developed for the identification and detection of Verticillium albo-atrum hop pathotypes PG1 and PG2 from Slovenia. Of 17 pathotype-linked amplified fragment length polymorphism (AFLP) markers, 11 were cloned successfully and sequenced. To convert polymorphic AFLP markers into pathotype-specific sequence-characterized amplified region (SCAR) markers, 22 PG2- and 10 PG1-specific primer pairs were designed from 16 sequences. When primer specificity was tested on a wide range of Verticillium isolates, 10 PG2- and 6 PG1-specific primer pairs retained amplification specificity for V. albo-atrum Slovene hop isolates, but also amplified sequences in V. albo-atrum and V. dahliae hop isolates from different hop production areas in Europe, as well as in some isolates from other hosts. Primer combinations obtained from the AFLP-9-1 marker were specific only for V. albo-atrum PG2 isolates. The highly specific primers were used in multiplex PCR and a nested PCR to detect the V. albo-atrum PG2 pathotype in xylem tissue of hop plants. These new SCAR markers provide a valuable tool for rapid identification of V. albo-atrum PG1 and PG2 hop pathotypes.

Autotetraploids and genetic mapping using common AFLP markers: the R2 allele conferring resistance to Phytophthora infestans mapped on potato chromosome 4

1998 ◽  
Vol 96 (8) ◽  
pp. 1121-1128 ◽  
Author(s):  
X. Li ◽  
H. J. van Eck ◽  
J. N. A. M. Rouppe van der Voort ◽  
D.-J. Huigen ◽  
P. Stam ◽  
...  
Keyword(s):  
Genetic Mapping ◽  
Phytophthora Infestans ◽  
Aflp Markers ◽  
Chromosome 4 ◽  
Potato Chromosome

Development of sequence-characterized amplified regions (SCARs) from amplified fragment length polymorphism (AFLP) markers tightly linked to the Vf gene in apple

Genome ◽  
2001 ◽  
Vol 44 (1) ◽  
pp. 63-70 ◽  
Author(s):  
Mingliang Xu ◽  
Ernesto Huaracha ◽  
Schuyler S Korban
Keyword(s):  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Apple Scab ◽  
Amplified Fragment Length Polymorphism ◽  
Genetic Distances ◽  
Aflp Markers ◽  
Scab Resistance ◽  
Scar Markers ◽  
Vf Gene

Amplified fragment length polymorphism (AFLP) markers have become widely used in saturating the region of a gene of interest for the ultimate goal of map-based cloning of the gene or for marker-assisted selection. However, conversion of AFLP markers into restriction fragment length polymorphism (RFLP) or polymerase chain reaction (PCR)-based markers will greatly expand their usefulness in genetic applications. Previously, we have identified 15 AFLP markers tightly linked to the Vf gene conferring scab resistance in apple. In this study, we have successfully converted 11 of these AFLPs into sequence-characterized amplified region (SCAR) markers. Of the remaining four nonconverted AFLP markers, one, ET2MC8-1, has been found to be very short (83 base pairs) and is an A/T rich (90%) marker; a second, EA2MG11-1, has shown identical sequences between Malus floribunda 821 (the original source of the Vf gene) and scab-susceptible apple cultivars; while the other two, EA12MG16-1 and ET8MG1-1, have not been cloned. Using the 11 converted SCAR markers along with 5 previously identified SCAR markers, a high-resolution linkage map around the Vf gene has been constructed, and found to be consistent with its corresponding AFLP map. Three converted SCAR markers (ACS-3, -7, and -9) are inseparable from the Vf gene; whereas one (ACS-6) is located left of, and the remaining seven (ACS-1, -2, -4, -5, -8, -10, and -11) are located right of the Vf gene at genetic distances of 0.4 and 0.2 cM, respectively. A reliable and robust procedure for development of SCAR markers from AFLP markers is presented.Key words: apple, AFLP, SCAR, apple scab disease, Vf gene.

Identification of two AFLP markers linked to bacterial wilt resistance in tomato and conversion to SCAR markers

2007 ◽  
Vol 36 (3) ◽  
pp. 479-486 ◽  
Author(s):  
Lixiang Miao ◽  
Senyan Shou ◽  
Jiayan Cai ◽  
Fang Jiang ◽  
Zhujun Zhu ◽  
...  
Keyword(s):  
Bacterial Wilt ◽  
Aflp Markers ◽  
Scar Markers ◽  
Wilt Resistance ◽  
Bacterial Wilt Resistance

Direct analysis of the secretions of the potato cyst nematode Globodera rostochiensis

Parasitology ◽  
1999 ◽  
Vol 119 (2) ◽  
pp. 167-176 ◽  
Author(s):  
L. ROBERTSON ◽  
W. M. ROBERTSON ◽  
J. T. JONES
Keyword(s):  
Superoxide Dismutase ◽  
Molecular Mass ◽  
Silver Staining ◽  
Gel Electrophoresis ◽  
Direct Analysis ◽  
Globodera Rostochiensis ◽  
Cyst Nematode ◽  
Potato Cyst Nematode ◽  
Western Blots ◽  
Gland Cells

Secretions were induced from second (invasive) stage juveniles (J2s) of the potato cyst nematode Globodera rostochiensis by exposing them to 5-methoxy-N,N-dimethyl tryptamine oxalate (DMT). Secretions were collected from J2s in sufficient quantity to allow direct analysis. Gel electrophoresis followed by monochromatic silver staining demonstrated the presence of at least 10 proteins. The presence of several enzymes, including superoxide dismutase and proteases, was demonstrated using Western blots and activity assays. Antisera raised against the secretions recognized bands on Western blots consistent in molecular mass with those identified on silver stained gels. The antisera recognized structures implicated in the production of secretions including the subventral gland cells and surface of J2s.

Protein variability among Portuguese and other populations of Globodera rostochiensis revealed by two-dimensional gel electrophoresis with computed image analysis

Nematology ◽  
2000 ◽  
Vol 2 (4) ◽  
pp. 461-471 ◽  
Author(s):  
Maria Susana Newton De Almeida Santos ◽  
Didier Mugniéry ◽  
Maria José Moreno Da Cunha ◽  
Isabel Maria De Oliveira Abrantes ◽  
Michel Bossis
Keyword(s):  
Image Analysis ◽  
Gel Electrophoresis ◽  
Gaussian Model ◽  
Globodera Rostochiensis ◽  
Computer Assisted ◽  
Two Dimensional ◽  
Protein Patterns ◽  
Two Dimensional Gel Electrophoresis ◽  
Bootstrap Analysis ◽  
Biological Studies

AbstractSilver-stained two dimensional gel electrophoresis (2-DGE) was used to characterize 46 populations of G. rostochiensis and, as an outgroup, one population of G. pallida. Protein patterns of white females were compared with the aid of computer assisted image analysis using Kepler software. A synthetic master pattern of G. rostochiensis, built with spots present in all populations and detected in at least two of three gels of each population, revealed the presence of 379 protein spots. The populations were compared taking into account the main spots present in each isolate (spots present in two of three gels, amplitude greater than 7, volume greater than 500 and with values fitting the Gaussian model). Comparison allowed the identification of 200 polymorphic spots. Similarity indices (F) and genetic distances (D = 1 ­ F) between populations were calculated on the basis of homologous polymorphic spots, and a dendrogram was constructed according to the UPGMA method. Bootstrap analysis was used to assess the significance of the results from the phenetic study. The presence or absence of specific spots in some G. rostochiensis populations is discussed. Further biochemical and biological studies are necessary to determine if specific proteins are linked to virulence. Variabilité protéique de populations portugaises et d'origines diverses de Globodera rostochiensis détectée par électrophorèse bidimensionnelle et analysée par informatique - L'électrophorèse bidimensionnelle (E2D) est utilisée pour caractériser 46 populations européennes de G. rostochiensis et une population de G. pallida, prise comme groupe extérieur. Les protéinogrammes obtenus à partir de femelles blanches sont comparés par analyse informatisée d'images à l'aide du logiciel Kepler. Une image de référence est constituée au vu des taches protéiques détectées dans l'ensemble des populations étudiées par transfert vers cette image synthétique de chaque protéine détectée au moins deux fois sur trois dans une population. Celle-ci permet de répertorier et de localiser 379 protéines. Les populations ont été comparées avec les taches pröteiques les plus importantes (taches présentes dans au moins deux gels sur trois, amplitude plus grande que 7, volume plus grand que 500 et valeurs dans le model Gaussien). La comparaison des populations a permis de détecter 200 protéines. Les indices de similarité (F) et les distances génétiques (D = 1 ­ F) ont été étudiés à partir des taches polymorphes homologues présentes dans chaque population. Le dendrogramme a été construit selon la méthode UPGMA. Une analyse des valeurs de bootstraps a été utilisée pour estimer la signification des résultats phénétiques. La présence ou l'absence de protéines spécifiques de certaines populations de G. rostochiensis est discutée. D'autres études biochimiques et biologiques sont en cours pour déterminer si les protéines spécifiques sont ou non liées à la virulence.

Ultrastructural localization of specific nucleoprotein fractions

1978 ◽  
Vol 36 (2) ◽  
pp. 204-205
Author(s):  
G. L. Brown
Keyword(s):  
Gel Electrophoresis ◽  
Mouse Liver ◽  
Polyacrylamide Gel Electrophoresis ◽  
Ultrastructural Localization ◽  
Polyacrylamide Gel ◽  
Acid Extraction ◽  
Acid Solutions ◽  
Low Salt ◽  
Liver Nuclei ◽  
Phosphate Groups

Bismuth (Bi) stains nucleoproteins (NPs) by interacting with available amino and primary phosphate groups. These two staining mechanisms are distinguishable by glutaraldehyde crosslinking (Fig. 1,2).Isolated mouse liver nuclei, extracted with salt and acid solutions, fixed in either formaldehyde (form.) or gl utaraldehyde (glut.) and stained with Bi, were viewed to determine the effect of the extractions on Bi stainina. Solubilized NPs were analyzed by SDS-polyacrylamide gel electrophoresis.Extraction with 0.14 M salt does not change the Bi staining characteristics (Fig. 3). 0.34 M salt reduces nucleolar (Nu) staining but has no effect on interchromatinic (IC) staining (Fig. 4). Proteins responsible for Nu and glut.- insensitive IC staining are removed when nuclei are extracted with 0.6 M salt (Fig. 5, 6). Low salt and acid extraction prevents Bi-Nu staining but has no effect on IC staining (Fig. 7). When nuclei are extracted with 0.6 M salt followed by low salt and acid, all Bi-staining components are removed (Fig. 8).

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