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PCR Markers for Triticum speltoides Leaf Rust Resistance Gene Lr51 and Their Use to Develop Isogenic Hard Red Spring Wheat Lines

^a INTA EEA Marcos Juárez, CC 21, 2580-Marcos Juárez, Argentina
^b Dep. of Agronomy and Range Science, One Shields Ave., Univ. California, Davis, CA 95616, USA
^c USDA-ARS, Cereal Disease Lab., 1551 Lindig Ave, Univ. of Minnesota, St. Paul, MN 55108

View larger version (28K): [in this window] [in a new window]   Fig. 1. Inferred RFLP map of the translocated chromosomes in Neepawa F-7-3 and F-7-12. The T. speltoides segment is indicated in black and regions involved in additional translocations among chromosomes from homeologous group 1 are indicated in gray. The + and – signs in the right panels indicate the presence or absence of RFLP fragments from the A, B, D, and S genomes. Underlined ± signs indicate a relatively higher hybridization intensity. The order and distances among markers were inferred from a T. monococcum map including the same markers (Dubcovsky et al., 1996).

View larger version (96K): [in this window] [in a new window]   Fig. 2. Best-fit alignment of partial nucleotide sequences from wheat clones from the A (pNF7-17), D (pNB-25), S (pNF7-16), and B (pNee-2) genomes. Gaps were introduced to maximize nucleotide alignment and are indicated with dashes. Locations of PCR primers AGA7-342F and S30-13L are indicated with arrows and sequences from the primers are italicized. BamHI and PstI restriction sites used to develop the CAPS marker are underlined with letters in gray color. Exons are numbered from the first transcribed exon and indicated above the sequences.

View larger version (54K): [in this window] [in a new window]   Fig. 3. Undigested and PstI digested PCR amplification products with CAPS primers S30-13L and AGA7-759R. 1-3) Chinese Spring nullisomic-tetrasomic lines. 1) N1AT1B, 2) N1BT1D, 3) N1DT1A, 4) Neepawa, 5) Neepawa*6/Triticum speltoides F-7-3. The gray arrowhead indicates the 780-bp product band from the B genome. The black arrowhead indicates the PstI digested product from the S genome. "M" indicates the molecular markers in bp (100-bp ladder, Biodynamics Corp., Seattle, WA).

View larger version (56K): [in this window] [in a new window]   Fig. 4. BamHI digestion of PCR products obtained from genomic DNAs amplified using CAPS primers S30-13L and AGA7-759R. Genomic DNAs were extracted from Neepawa (Nee) and Neepawa*6/Triticum speltoides F-7-3 (NeeF7), and from five BC6F2 plants. Letters "BS" and "SS" indicates plants heterozygous and homozygous for the presence of Lr51, respectively. Parental line Neepawa (Lane 7) is the only BB plant in this figure. The black arrowhead indicates the 819-bp PCR amplification product from the S genome allele and the gray arrowhead indicates the larger of the two BamHI digested fragments from the B genome allele of XAga7. "M" indicates the size molecular marker (100-bp ladder, Biodynamics).

View larger version (43K): [in this window] [in a new window]   Fig. 5. PstI and BamHI digestion of PCR products obtained with XAga7 CAPS primers S30-13L and AGA7-759R. Genomic DNAs were extracted from 1) Neepawa*6/Triticum speltoides F-7-3, 2) Neepawa, 3) Madsen, 4) Hyak, 5) Penawawa, 6) Buck Manantial, 7) Prointa Milenium, 8) Prointa Granar, and 9) Klein Pegaso. Gray and black arrowheads indicate amplification products from the B and S genome alleles, respectively. "M" indicates the size molecular marker (100-bp ladder, Biodynamics).