scholarly journals Identification of Random Amplified Polymorphic DNA (RAPD) Markers for Self-incompatibility Alleles in Hazelnut

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 591e-591
Author(s):  
Kirk W. Pomper ◽  
Anita N. Azarenko ◽  
Joel W. Davis ◽  
Shawn A. Mehlenbacher
Keyword(s):  
Rapd Markers ◽  
Uv Light ◽  
Bulked Segregant Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Rapd Marker ◽  
Self Incompatibility ◽  
Ethidium Bromide Staining ◽  
Young Leaves ◽  
Incompatibility Alleles ◽  
Selection Of

Random amplified polymorphic DNA (RAPD) markers were identified for self-incompatibility (SI) alleles that will allow marker-assisted selection of desired S-alleles and assist in cloning the locus responsible for the sporophytic SI displayed in hazelnut (Corylus avellana L.). DNA was extracted from young leaves collected from field-planted parents and 27 progeny of the cross OSU 23.017 (S1 S12) × VR6-28 (S2 S26). Screening of 10-base oligonucleotide RAPD primers was performed using bulked segregant analysis. DNA samples from six trees each were pooled into four “bulks,” one for each of the following: S1 S2, S1 S26, S2 S12, and S12 S26. “Super bulks” of twelve trees each for S1, S2, S12, and S26 then were created for each allele by combining the appropriate bulks. The DNA from these four super bulks and also the parents was used as a template in the PCR assays. Amplification products were electrophoresed on 2% agarose gels and photographed under UV light after ethidium bromide staining. 200 primers were screened and one RAPD marker each was identified for alleles S2 (OPI-07700) and S1 (OPJ-141700).

scholarly journals Development of random amplified polymorphic DNA markers for authentication of Croton tiglium Linn.

2017 ◽  
Vol 6 (3) ◽  
pp. 164-166
Author(s):  
Shweta Vekariya ◽  
◽  
Krushnkumar Taviad ◽  
RN Acharya RN ◽  
CN Harisha ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Rapd Markers ◽  
Ricinus Communis ◽  
Random Amplified Polymorphic Dna ◽  
Rapd Marker ◽  
Natural Habitat ◽  
Drug Market ◽  
Croton Tiglium ◽  
Young Leaves

Background: Croton tiglium Linn., commonly known as Jayapala, in Ayurvedic pharmacopoeia, is wellknown for its purgative action. In the herbal raw drug market, seeds of Baliospermum montanum Blume., Ricinus communis Linn. and Croton roxburghii Wall. etc. are sold in the name of Jayapla seeds due to their morphological similarities. Hence their identification through molecular characters is need of the hour. Aim: Present study aims with the molecular characterization of young leaves of C. tiglium Linn. Materials & Methods: Fresh young leaves of C. tiglium Linn. were collected from its natural habitat Udupi, Karnnataka, during January 2017. It was used for molecular characterization and DNA fingerprints, by standard and most convenient Random Amplified Polymorphic DNA (RAPD) markers at Food testing laboratory, Junagadh Agriculture University, Gujarat, India. Results: All the primers gave good band patterns. Primer 5, 6, 8, 11 and 13 showed more number of light and bright bands matching characters with plant. Conclusion: Observed RAPD marker can be used to differentiate genuine as well as adulterated samples. The results may be used for the further research purposes and also required DNA Barcoding studies for further authentication.

Isolation of microsatellite and RAPD markers flanking the Yr15 gene of wheat using NILs and bulked segregant analysis

Genome ◽  
1999 ◽  
Vol 42 (6) ◽  
pp. 1050-1056 ◽  
Author(s):  
V Chagué ◽  
T Fahima ◽  
A Dahan ◽  
G L Sun ◽  
A B Korol ◽  
...  
Keyword(s):  
Resistance Gene ◽  
Stripe Rust ◽  
Rust Resistance ◽  
Rapd Markers ◽  
Bulked Segregant Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Rapd Marker ◽  
Rust Resistance Gene ◽  
Stripe Rust Resistance ◽  
Stripe Rust Resistance Gene

Microsatellite and random amplified polymorphic DNA (RAPD) primers were used to identify molecular markers linked to the Yr15 gene which confer resistance to stripe rust (Puccina striiformis Westend) in wheat. By using near isogenic lines (NILs) for the Yr15 gene and a F2 mapping population derived from crosses of these lines and phenotyped for resistance, we identified one microsatellite marker (GWM33) and one RAPD marker (OPA19800) linked to Yr15. Then, bulked segregant analysis was used in addition to the NILs to identify RAPD markers linked to the target gene. Using this approach, two RAPD markers linked to Yr15 were identified, one in coupling (UBC199700) and one in repulsion phase (UBC2121200). After Mapmaker linkage analysis on the F2 population, the two closest markers were shown to be linked to Yr15 within a distance of about 12 cM. The recombination rates were recalculated using the maximum likelihood technique to take into account putative escaped individuals from the stripe rust resistance test and obtain unbiased distance estimates. As a result of this study, the stripe rust resistance gene Yr15 is surrounded by two flanking PCR markers, UBC199700 and GWM33, at about 5 cM from each side.Key words: wheat, Triticum dicoccoides, Yr15 stripe rust resistance gene, genetic mapping, microsatellite markers, RAPD markers.

scholarly journals Segregation for Resistance to Eastern Filbert Blight in Progeny of `Zimmerman' Hazelnut

2006 ◽  
Vol 131 (6) ◽  
pp. 731-737 ◽  
Author(s):  
China F. Lunde ◽  
Shawn A. Mehlenbacher ◽  
David C. Smith
Keyword(s):  
Rapd Markers ◽  
Chromosome Region ◽  
Random Amplified Polymorphic Dna ◽  
Rapd Marker ◽  
Segregation Ratio ◽  
Enzyme Linked Immunosorbent Assay ◽  
Resistance Locus ◽  
Resistance Allele ◽  
Reciprocal Translocations ◽  
Eastern Filbert Blight

Eastern filbert blight (EFB), caused by the fungus Anisogramma anomala (Peck) E. Müller, is an important disease of european hazelnut (Corylus avellana L.) in the Pacific northwestern United States. In 1989, a chance seedling free of EFB was discovered adjacent to a severely diseased orchard near Troutdale, Ore. This selection, subsequently named `Zimmerman', was crossed with three susceptible selections. Based on morphological characters and incompatibility alleles, we speculated that `Zimmerman' (S1 S3) was a hybrid between `Barcelona' (S1 S2) and `Gasaway' (S3 S26). The three seedling populations were inoculated with spores of the pathogen in a greenhouse test and assayed by indirect enzyme-linked immunosorbent assay (ELISA) and by observation of canker incidence. The observed segregation fit a 3 resistant : 1 susceptible ratio in all three progenies, in contrast to the 1 : 1 ratio found when the resistant pollinizer `Gasaway' was crossed to susceptible genotypes. Random amplified polymorphic DNA (RAPD) marker UBC 152800 linked to the resistance gene in `Gasaway' co-segregated with the resistant phenotype in all three populations with 2%, 4%, and 6% recombination, respectively. Seed germination and transplanting records did not provide evidence of selection in favor of resistant seedlings. Pollen germination was 71% in `Gasaway', 29% in `Zimmerman', and 18% in `Barcelona', indicating possible selection at the gametophytic level. Subsequently 16 resistant seedlings of `Zimmerman' were crossed with the highly susceptible selection OSU 313.078. Segregation fit a 3 : 1 ratio in 14 of the 16 progenies, and showed a surplus of resistant seedlings in the other two. None showed a 1 : 1 segregation. Resistance co-segregated with two RAPD markers that flank the `Gasaway' resistance allele. To test allelism of resistance from `Gasaway' and `Zimmerman', VR 6-28 with resistance from `Gasaway' was crossed with `Zimmerman'. Eight resistant selections from this progeny were crossed with OSU 313.078. Five of the eight progenies segregated 3 : 1, two progenies segregated 1 : 1, and OSU 313.078 × OSU 720.056 gave only resistant offspring. The ratios indicate that OSU 720.056 is homozygous resistant and that `Zimmerman' and `Gasaway' share a common resistance allele. Reciprocal translocations have been reported in hazelnut cultivars, including `Barcelona', the leading cultivar in Oregon. `Zimmerman' appears to be a hybrid of `Barcelona' and `Gasaway', but because of cytogenetic abnormalities, `Zimmerman' may have inherited two copies of the chromosome region that contain the resistance locus and flanking RAPD markers. If the region containing the resistance were attached to two independent centromeres, a 3 : 1 segregation ratio for disease response and flanking markers would be expected, and we propose this as the most likely explanation. Resistance from `Gasaway' and `Zimmerman' has been called “immunity” or “complete resistance.” However, we noted a few seedlings with small cankers, nearly all of which lacked sporulating stromata. Flanking RAPD markers indicate that the resistance allele is present in these seedlings. Although not “immune” or “completely resistant,” `Gasaway' and `Zimmerman' transmit a very high level of resistance.

scholarly journals Molecular Markers for the Scab Resistance (Vf) Region in Apple

1998 ◽  
Vol 123 (6) ◽  
pp. 992-996 ◽  
Author(s):  
Minou Hemmat ◽  
Norman F. Weeden ◽  
Herb S. Aldwinckle ◽  
Susan K. Brown
Keyword(s):  
Rapd Markers ◽  
Bulked Segregant Analysis ◽  
Venturia Inaequalis ◽  
Apple Scab ◽  
Scab Resistance ◽  
Resistant Plant ◽  
Tight Linkage ◽  
Test Population ◽  
Rate Selection ◽  
Selection Of

Bulked segregant analysis was used to identify RAPD markers that display tight linkage to the Vf gene in apple (Malus sp.) that confers resistance to five races of apple scab [Venturia inaequalis (Cke.) Wint.]. We identified several new RAPD markers linked to Vf. The most tightly linked marker in the test population, S52500, was cloned and sequenced. A linkage map of the Vf region was developed using these markers, RAPD markers previously described by other laboratories, and the isozyme locus Pgm-1. An assay was developed for Vf by multiplexing the two markers closely flanking the Vf locus. This assay has a theoretical `escape' value (discarding a resistant plant) of 3% and an error rate (selection of a susceptible plant) of 0.02%.

Segregation of random amplified polymorphic DNA markers and strategies for molecular mapping in tetraploid alfalfa

Genome ◽  
1993 ◽  
Vol 36 (5) ◽  
pp. 844-851 ◽  
Author(s):  
K. F. Yu ◽  
K. P. Pauls
Keyword(s):  
Chromosome Segregation ◽  
Dna Markers ◽  
Rapd Markers ◽  
Molecular Mapping ◽  
Random Amplified Polymorphic Dna ◽  
Rapd Marker ◽  
Female Parent ◽  
Linkage Groups ◽  
Double Reduction ◽  
Chromatid Segregation

An F1 population was used to analyze the inheritance of random amplified polymorphic DNA (RAPD) markers in tetraploid alfalfa. Of the 32 RAPD markers that were used for a segregation analysis in this study, 27 gave ratios that are consistent with random chromosome and random chromatid segregation at meiosis. However, among all of the RAPD markers (121) that were screened in this study, only one example of a double reduction, that is typical of chromatid segregation, was observed. These results indicate that random chromosome segregation is likely the predominant but not the exclusive mode of inheritance for tetraploid alfalfa. χ2 analyses of cosegregation for RAPD marker pairs derived from the female parent revealed nine linkages that fell into four linkage groups. The recombination fractions among linked marker pairs ranged from 1 to 37%. These are the first molecular linkage groups reported in tetraploid alfalfa. In addition, various strategies for molecular mapping in the tetraploid alfalfa genome are proposed that should be of interest to plant breeders who are planning to use molecular markers for alfalfa or other tetraploid species.Key words: RAPD markers, tetraploid alfalfa, segregation, linkage groups.

scholarly journals Phenotype Instability in Botrytis cinerea in the Absence of Benzimidazole and Dicarboximide Fungicides

2001 ◽  
Vol 91 (3) ◽  
pp. 307-315 ◽  
Author(s):  
L. F. Yourman ◽  
S. N. Jeffers ◽  
R. A. Dean
Keyword(s):  
Botrytis Cinerea ◽  
High Frequency ◽  
Rapd Markers ◽  
Random Amplified Polymorphic Dna ◽  
Rapd Marker ◽  
Potato Dextrose Agar ◽  
Multiple Generations ◽  
Rapd Fingerprints ◽  
Growth Room ◽  
Conidium Germination

Stability of phenotypes of isolates of Botrytis cinerea that were sensitive or resistant to benzimidazole and dicarboximide fungicides was examined in the absence of fungicides in laboratory and growth room experiments. Twelve greenhouse isolates of B. cinerea were subcultured on potato dextrose agar (PDA) for 20 generations and on geranium seedlings for 15 generations. Three isolates of each of the following four phenotypes were used: sensitive to the fungicides thiophanate-methy1 (a benzimidazole) and vinclozolin (a dicarboximide) (STSV), resistant to both fungicides (RTRV), resistant to thiophanate-methy1 and sensitive to vinclozolin (RTSV), and sensitive to thiophanate-methy1 and resistant to vinclozolin (STRV). In three trials on PDA, 36 populations were subcultured; 8 populations changed phenotypes by the end of 20 generations, as determined by conidium germination on fungicide-amended medium. Five of the eight initially were STRV; the resulting phenotypes were STSV, RTSV, and RTRV. Populations from eight other isolates exhibited temporary changes in phenotype during intermediate generations on PDA but reverted to initial phenotypes by the twentieth generation; five of these populations changed to phenotype RTRV. In two geranium seedling trials, each of the 12 greenhouse isolates was inoculated onto a set of three seedlings for each generation, and diseased tissue that developed was used to initiate the next generation. Therefore, a total of 72 populations of B. cinerea were subcultured in the two trials; 5 of these populations changed phenotype at the end of 15 generations. Three of the five initially were STRV; these changed to phenotypes STSV or RTRV. In each of the two trials on geranium seedlings, a population subcultured from one STSV isolate changed phenotype one to phenotype RTRV and one to phenotype RTSV. In all trials, no population resistant to thiophanate-methy1 changed to a thiophanate-methy1-sensitive phenotype, and no population changed to phenotype STRV. Random amplified polymorphic DNA (RAPD) fingerprints were generated with the 12 initial isolates and 49 isolates subcultured on PDA or geranium seedlings. Cluster analyses of RAPD markers showed that subcultured isolates exhibiting the same phenotype clustered together and that subcultured isolates derived from a common greenhouse isolate but with different phenotypes were in different clusters. Some populations that did not change phenotype exhibited considerable differences in RAPD marker patterns. The results of this study indicate that, in the absence of fungicides, sensitive populations of B. cinerea can develop resistance to thiophanate-methy1 and vinclozolin, and this resistance can be maintained in populations through multiple generations. Populations resistant only to vinclozolin (STRV) exhibited a high frequency of phenotype change, and populations resistant to both fungicides (RTRV) were stable.

Development of DNA fingerprinting keys for the identification of radish cultivars

2004 ◽  
Vol 44 (1) ◽  
pp. 95 ◽  
Author(s):  
A. Pradhan ◽  
G. Yan ◽  
J. A. Plummer
Keyword(s):  
Dna Fingerprinting ◽  
Random Amplified Polymorphic Dna ◽  
Rapd Marker ◽  
Morphological Characteristics ◽  
Group Method ◽  
Base Pairs ◽  
Difference Matrix ◽  
Pair Group ◽  
Young Leaves ◽  
Polymerase Chain

Identification of cultivars is extremely important both for cultivation and breeding of crop plants. Cultivar identification based on morphological characteristics can be difficult and complicated. Polymerase chain reaction technologies, such as random amplified polymorphic DNA (RAPD) analysis, can readily and quickly identify cultivars using seeds and young leaves. Sixty individuals representing 7 radish cultivars were examined for RAPD marker polymorphism. Based on the polymorphism generated, 5 primers were selected, out of the 14��examined, to fingerprint the cultivars. The 5 primers produced a total of 52 fragments, 6 monomorphic and 46�polymorphic fragments, ranging in size from 206 to 2258 base pairs. A total and mean character difference matrix was calculated based on the RAPD data and a dendrogram was constructed using the unweighted pair-group method with arithmetic averages (UPGMA). Three DNA fingerprinting keys were developed for the 7 cultivars and 5 markers derived from 3 primers was the minimum required to distinguish cultivars. Results demonstrated that RAPD markers could be effectively used for the identification of radish cultivars.

Identification of coupling and repulsion phase RAPD markers for powdery mildew resistance gene er-1 in pea

Genome ◽  
1998 ◽  
Vol 41 (3) ◽  
pp. 440-444 ◽  
Author(s):  
K R Tiwari ◽  
G A Penner ◽  
T D Warkentin
Keyword(s):  
Powdery Mildew ◽  
Resistance Gene ◽  
Rapd Markers ◽  
Powdery Mildew Resistance ◽  
Bulked Segregant Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Powdery Mildew Resistance Gene ◽  
Erysiphe Pisi ◽  
Mildew Resistance ◽  
Repulsion Phase

Powdery mildew is a serious disease of pea caused by the obligate parasite Erysiphe pisi Syd. Random amplified polymorphic DNA (RAPD) analysis has emerged as a cost-effective and efficient marker system. The objective of this study was to identify RAPD markers for powdery mildew resistance gene er-1. The resistant cultivar Highlight (carrying er-1) and the susceptible cultivar Radley were crossed, and F3 plants were screened with Operon (OP) and University of British Columbia (UBC) primers, using bulked segregant analysis. A total of 416 primers were screened, of which amplicons of three Operon primers, OPO-18, OPE-16, and OPL-6, were found to be linked to er-1. OPO-181200 was linked in coupling (trans to er-1) and no recombinants were found. OPE-161600 (4 ± 2 cM) and OPL-61900 (2 ± 2 cM) were linked in repulsion (cis to er-1). The fragments OPO-181200 and OPE-161600 were sequenced and specific primers designed. The specific primer pair Sc-OPO-181200 will be useful in identifying homozygous resistant individuals in F2 and subsequent segregating generations. Sc-OPE-161600 will have greatest utility in selecting heterozygous BC\dn6 nF1 individuals in backcross breeding programs.Key words: bulked segregant analysis,Erysiphe pisi, pea, RAPD.

scholarly journals 666 PB 110 RANDOM AMPLIFIED POLYMORPHIC DNA MARKERS ARE INADEQUATE FOR FINGERPRINTING GRAPE ROOTSTOCKS

HortScience ◽  
1994 ◽  
Vol 29 (5) ◽  
pp. 528c-528
Author(s):  
Alan T. Bakalinsky ◽  
Hong Xu ◽  
Diane J. Wilson ◽  
S. Arulsekar
Keyword(s):  
Dna Markers ◽  
Rapd Markers ◽  
Restriction Site ◽  
Random Amplified Polymorphic Dna ◽  
Rapd Marker ◽  
Base Length ◽  
Specific Primer ◽  
Annealing Temperatures ◽  
Individual Reaction ◽  
Length Variant

A total of eight random amplified polymorphic DNA (RAPD) markers were generated in a screen of 77 primers of 10-base length and were detected reproducibly among nine different grape (Vitis) rootstocks. Occasional failed amplifications could not be explained rationally nor easily corrected by systematic replacement of individual reaction components. In an effort to improve their reliability, the RAPD markers were cloned, their termini sequenced, and new sequence-specific primer pairs were synthesized based on addition of 10 to 14 bases to the 3' termini of the original 10-mers. Six pairs of the new primers were evaluated at their optimal and higher-than optimal annealing temperatures. One primer pair amplified a product the same size as the original RAPD marker in all rootstocks, resulting in loss of polymorphism. Post-amplification digestion with 7 different restriction endonucleases failed to reveal restriction site differences. Three primer pairs amplified an unexpected length variant in some accessions. Two other pairs of primers amplified a number of unexpected bands. Better approaches for exploiting the sequence differences that account for the RAPD phenomenon will be discussed.

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