Transfer of sequence tagged site PCR markers between wheat and barley

Genome ◽  
1996 ◽  
Vol 39 (4) ◽  
pp. 802-810 ◽  
Author(s):  
J. E. Erpelding ◽  
N. K. Blake ◽  
T. K. Blake ◽  
L. E. Talbert
Keyword(s):  
Fragment Length Polymorphism ◽  
Pcr Markers ◽  
Chain Reaction ◽  
Chromosome Group ◽  
Sequence Tagged Site ◽  
Sequence Tagged Sites ◽  
Probability Of Success ◽  
Polymerase Chain ◽  
Primer Sets ◽  
Mapping Information

Transfer of mapping information between related species has facilitated the development of restriction fragment length polymorphism (RFLP) maps in the cereals. Sequence tagged site (STS) primer sets for use in the polymerase chain reaction may be developed from mapped RFLP clones. For this study, we mapped 97 STS primer sets to chromosomes in wheat and barley to determine the potential transferability of the primer sets and the degree of correspondence between RFLP and STS locations. STS products mapped to the same chromosome group in wheat and barley 75% of the time. RFLP location predicted STS location 69% of the time in wheat and 56% of the time in barley. Southern hybridizations showed that most primer sets amplified sequences homologous to the RFLP clone, although additional sequences were often amplified that did not hybridize to the RFLP clone. Nontarget sequences were often amplified when primer sets were transferred across species. In general, results suggest a good probability of success in transferring STSs between wheat and barley, and that RFLP location can be used to predict STS location. However, transferability of STSs cannot be assumed, suggesting a need for recombinational mapping of STS markers in each species as new primer sets are developed. Key words : sequence tagged sites, PCR, wheat, barley.

Identification of barley genome segments introgressed into wheat using PCR markers

Genome ◽  
2001 ◽  
Vol 44 (1) ◽  
pp. 38-44 ◽  
Author(s):  
J D Sherman ◽  
L Y Smith ◽  
T K Blake ◽  
L E Talbert
Keyword(s):  
Seed Set ◽  
Barley Chromosome ◽  
Progeny Testing ◽  
Pcr Markers ◽  
Chain Reaction ◽  
Sequence Tagged Site ◽  
Chinese Spring Wheat ◽  
Polymerase Chain ◽  
Primer Sets ◽  
Disomic Addition Lines

Barley has several important traits that might be used in the genetic improvement of wheat. For this report, we have produced wheat-barley recombinants involving barley chromosomes 4 (4H) and 7 (5H). Wheat-barley disomic addition lines were crossed with 'Chinese Spring' wheat carrying the ph1b mutation to promote homoeologous pairing. Selection was performed using polymerase chain reaction (PCR) markers to identify lines with the barley chromosome in the ph1b background. These lines were self pollinated, and recombinants were identified using sequence-tagged-site (STS) primer sets that allowed differentiation between barley and wheat chromosomes. Several recombinant lines were isolated that involved different STS-PCR markers. Recombination was confirmed by allowing the lines to self pollinate and rescreening the progeny via STS-PCR. Progeny testing confirmed 9 recombinants involving barley chromosome 4 (4H) and 11 recombinants involving barley chromosome 7 (5H). Some recombinants were observed cytologically to eliminate the possibility of broken chromosomes. Since transmission of the recombinant chromosomes was lower than expected and since seed set was reduced in recombinant lines, the utility of producing recombinants with this method is uncertain.Key words: introgression, sequence-tagged-site, recombination.

scholarly journals Rare Pyrenophora teres Hybridization Events Revealed by Development of Sequence-Specific PCR Markers

2017 ◽  
Vol 107 (7) ◽  
pp. 878-884 ◽  
Author(s):  
Barsha Poudel ◽  
Simon R. Ellwood ◽  
Alison C. Testa ◽  
Mark McLean ◽  
Mark W. Sutherland ◽  
...  
Keyword(s):  
Host Plants ◽  
Fragment Length Polymorphism ◽  
Putative Hybrid ◽  
Pyrenophora Teres ◽  
Net Blotch ◽  
Pcr Markers ◽  
Chain Reaction ◽  
Specific Pcr ◽  
Polymerase Chain ◽  
Pathogenic Variability

Pyrenophora teres f. teres and P. teres f. maculata cause net form and spot form, respectively, of net blotch on barley (Hordeum vulgare). The two forms reproduce sexually, producing hybrids with genetic and pathogenic variability. Phenotypic identification of hybrids is challenging because lesions induced by hybrids on host plants resemble lesions induced by either P. teres f. teres or P. teres f. maculata. In this study, 12 sequence-specific polymerase chain reaction markers were developed based on expressed regions spread across the genome. The primers were validated using 210 P. teres isolates, 2 putative field hybrids (WAC10721 and SNB172), 50 laboratory-produced hybrids, and 7 isolates collected from barley grass (H. leporinum). The sequence-specific markers confirmed isolate WAC10721 as a hybrid. Only four P. teres f. teres markers amplified on DNA of barley grass isolates. Amplified fragment length polymorphism markers suggested that P. teres barley grass isolates are genetically different from P. teres barley isolates and that the second putative hybrid (SNB172) is a barley grass isolate. We developed a suite of markers which clearly distinguish the two forms of P. teres and enable unambiguous identification of hybrids.

scholarly journals Detection of New Ethylene-Producing Bacteria, Pseudomonas syringae pvs. cannabina and sesami, by PCR Amplification of Genes for the Ethylene-Forming Enzyme

1997 ◽  
Vol 87 (12) ◽  
pp. 1192-1196 ◽  
Author(s):  
M. Sato ◽  
K. Watanabe ◽  
M. Yazawa ◽  
Y. Takikawa ◽  
K. Nishiyama
Keyword(s):  
Pseudomonas Syringae ◽  
Fragment Length Polymorphism ◽  
Pcr Amplification ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Enzyme Gene ◽  
Indigenous Plasmid ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Primer Sets

Strains of Pseudomonas syringae (78 strains and 43 pathovars) and other strains (79) of plant and insect origin were examined for the presence of the ethylene-forming enzyme gene (efe) by polymerase chain reaction (PCR) assay. The sequence of the efe gene of P. syringae pv. phaseolicola PK2 was used to design two primer sets for amplification of the gene. In addition to P.syringae pv. phaseolicola (the “kudzu strain”) and P.syringae pv. glycinea, which were efficient ethylene producers, several strains of P.syringae pvs. sesami and cannabina generated PCR products of the predicted size. A DNA probe of the efe gene, isolated from strain PK2, hybridized to these PCR products, indicating homology to the P.syringae pv. phaseolicola efe gene. PCR restriction fragment length polymorphism analyses suggested that these four pathovars harbor a similar efe gene. Furthermore, the probe hybridized to an indigenous plasmid of P.syringae pv. cannabina, suggesting that the efe gene could be located on a plasmid in this pathovar, but did not hybridize to plas-mids of P.syringae pv. sesami strains. P.syringae pvs. sesami and cannabina strains produced ethylene in King's medium B at levels similar to those of P.syringae pvs. phaseolicola and glycinea. Thus, two new ethylene-producing bacteria were detected by the PCR assay.

Properties of sequence-tagged-site primer sets influencing repeatability

Genome ◽  
2000 ◽  
Vol 43 (1) ◽  
pp. 47-52 ◽  
Author(s):  
A Vanichanon ◽  
N K Blake ◽  
J M Martin ◽  
L E Talbert
Keyword(s):  
Genomic Dna ◽  
Standard Procedure ◽  
Gc Content ◽  
Pcr Primers ◽  
Gene Identification ◽  
Sequence Information ◽  
Chain Reaction ◽  
Sequence Tagged Site ◽  
Polymerase Chain ◽  
Primer Sets

The polymerase chain reaction (PCR) has become a standard procedure in plant genetics, and is the basis for many emerging genomics approaches to mapping and gene identification. One advantage of PCR is that sequence information for primer sets can be exchanged between laboratories, obviating the need for exchange and maintenance of biological materials. Repeatability of primer sets, whereby the same products are amplified in different laboratories using the same primer set, is important to successful exchange and utilization. We have developed several hundred sequence-tagged site (STS) primer sets for wheat and barley. The ability of the primer sets to generate reproducible amplifications in other laboratories has been variable. We wished to empirically determine the properties of the primer sets that most influenced repeatability. A total of 96 primer sets were tested with four genomic DNA samples on each of four thermocyclers. All major bands were repeatable across all four thermocyclers for approximately 50% of the primer sets. Characteristics most often associated with differences in repeatability included primer GC content and 3'-end stability of the primers. The propensity for primer-dimer formation was not a factor in repeatability. Our results provide empirical direction for the development of repeatable primer sets. Key words: STS-PCR primers, wheat, barley.

scholarly journals Phytoplasma occurrence in apple trees in the Czech Republic

2011 ◽  
Vol 39 (No. 1) ◽  
pp. 7-12 ◽  
Author(s):  
R. Fialová ◽  
M. Navrátil ◽  
P. Válová
Keyword(s):  
Fragment Length Polymorphism ◽  
Rrna Gene ◽  
Polymorphism Analysis ◽  
Apple Proliferation ◽  
Chain Reaction ◽  
Apple Trees ◽  
The Czech Republic ◽  
High Incidence ◽  
Phytoplasma Infection ◽  
Polymerase Chain

The presence of phytoplasmas in apple trees with proliferation symptoms, rubbery wood symptoms and no symp­toms was determined by using polymerase chain reaction assays with primers amplifying phytoplasma 16S rRNA gene. Phytoplasmas were detected in all trees with proliferation symptoms. Positive tests for phytoplasma in the group of trees with rubbery wood symptoms and of those without symptoms revealed a relatively high incidence of latent phytoplasma infection. Using restriction fragment length polymorphism analysis, phytoplasma of the same identity – apple proliferation phytoplasma (subgroup 16SrX-A) – was recorded in all positively tested trees.  

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