RESUMO
PCR-based Landmark Unique Gene (PLUG) markers are EST-PCR markers developed based on the orthologous gene conservation between rice and wheat, and on the intron polymorphisms among the three orthologous genes derived from the A, B and D genomes of wheat. We designed a total of 960 primer sets from wheat ESTs that showed high similarity with 951 single-copy rice genes. When genomic DNA of Chinese Spring wheat was used as a template, 872 primer sets amplified one to five distinct products. Out of these 872 PLUG markers, 531 were assigned to one or more chromosomes by nullisomic-tetrasomic analysis. For each wheat chromosome, the number of loci detected ranged from 32 for chromosome 6A to 73 for chromosome 7D, with an average of 48 loci per chromosome. Several novel synteny perturbations were identified using deletion bin-mapping of markers. Furthermore, we demonstrated that PLUG markers can be used as probes to simultaneously identify BAC clones that contain homoeologous regions from all three genomes.
Assuntos
Cromossomos de Plantas , Oryza/genética , Triticum/genética , Cromossomos Artificiais Bacterianos/genética , Primers do DNA , DNA de Plantas , Etiquetas de Sequências Expressas , Marcadores Genéticos , Genoma de Planta , Reação em Cadeia da Polimerase , SinteniaRESUMO
We dissected barley chromosome 5H added to common wheat by a genetic method or the gametocidal system. Firstly, we induced chromosomal breaks in the offspring of a 5H addition line of common wheat carrying a gametocidal chromosome and cytologically screened for plants with structural chromosomal changes involving 5H, such as deletions and translocations. Secondly, we screened the progeny of such plants to establish common wheat lines carrying structurally changed chromosomes containing single segments of the dissected 5H. Using 23 representative 5H dissection lines, we physically mapped 97 barley EST markers assigned to 5H. The ESTs fell into 20 regions of 5H between the breakpoints of the 23 dissected segments, distributing rather evenly along the chromosome, with significantly higher frequency in the distal region of the long arm. The ESTs, in turn, allowed us to distinguish the breakpoints of dissected 5H segments. We demonstrated by PCR (polymerase chain reaction), as well as by in situ hybridization, that these dissected 5H segments were stably transmitted in the dissection lines. We discuss the usefulness of the 5H dissection lines for physical mapping of DNA markers. These 5H dissection lines are available from National BioResource Projects-Wheat, Japan.
Assuntos
Cromossomos de Plantas , Hordeum/genética , Mapeamento Físico do Cromossomo , Triticum/genética , Sequência de Bases , Bandeamento Cromossômico , Etiquetas de Sequências Expressas , Hibridização in Situ FluorescenteRESUMO
From about 10000 PCR-based EST markers of barley we chose 1421 EST markers that were demonstrated to be amplified differently by PCR between wheat (Triticum aestivum cv. Chinese Spring) and barley (Hordeum vulgare cv. Betzes). We assigned them to the seven barley chromosomes (1H to 7H) by PCR analysis using a set of wheat-barley chromosome addition lines. We successfully assigned 701 (49.3%) EST markers to the barley chromosomes: 75 to 1H, 127 to 2H, 119 to 3H, 94 to 4H, 108 to 5H, 81 to 6H and 97 to 7H. By using a set of Betzes barley telosomic addition lines of Chinese Spring, we could successfully determine the chromosome-arm (S or L) location of at least 90% of the EST markers assigned to each barley chromosome. We conducted a trial mapping using 90 EST markers assigned to 7HS (49) or 7HL (41) and 19 wheat lines carrying 7H structural changes. More EST markers were found in the distal region than in the proximal region.
