RESUMO
Direct estimates of sequence diversity provides an abundant source of DNA polymorphisms based on single nucleotide polymorphisms (SNPs). The frequency and distribution of nucleotide diversity within 23 genes associated with grain germination in barley were determined in a sample of accessions representing European cultivars, landraces, and wild barley accessions from throughout the fertile crescent. The overall nucleotide diversity ranged from 0.0021 to 0.0189 with a single nucleotide change being detected every 78 bp and insertion-deletion events being observed every 680 bp. Within the cultivated (H. vulgare) genepool, a small number of haplotypes were detected, the total number of haplotypes observed in H. spontaneum was almost double that detected in H. vulgare (46 and 26, respectively). Distinct haplotypes were observed in the H. spontaneum and landrace genepools, which are highly divergent from H. vulgare. A comparison of SNP-based haplotype data with EST-derived SSRs and genomic SSRs revealed a similar trend of decreasing variability in the cultivated genepool. However, the number of unique alleles identified in the cultivated sample was much greater with genomic SSRs (18%) compared with only 2.1% for SNPs and 3.8% for EST-derived SSRs. The potential utility of SNPs and EST-derived SSRs for association mapping in barley is discussed.
Assuntos
Genoma de Planta , Hordeum/genética , Polimorfismo de Nucleotídeo Único , Sequência de Bases , Etiquetas de Sequências Expressas , Biblioteca Gênica , Genes de Plantas , Marcadores Genéticos , Germinação/genética , Haplótipos/genética , Repetições de Microssatélites , Dados de Sequência Molecular , Alinhamento de Sequência , Transcrição GênicaRESUMO
Microspores at late uninucleate/early binucleate stages were isolated from flower buds of tobacco (Nicotiana tabacum L.) and in vitro culture methods optimised for their maturation to fully functional viable pollen which, after application to the stigma of emasculated plants in situ, led to the generation of large numbers of seed. Efficient protocols were established for the biolistic introduction of a construct containing a reporter gene and selectable marker into these microspores and hence, after in vitro maturation and in situ fertilisation, for the generation of transgenic plants. Stable transformants of low copy number were generated by this procedure. The efficiency of transformation achieved allows the production of large numbers of transgenic plants without selection, dispensing with the requirement for a selectable marker in plant transgenesis.
Assuntos
Nicotiana/genética , Pólen/genética , Marcadores Genéticos/genética , Glucuronidase/genética , Glucuronidase/metabolismo , Plantas Geneticamente Modificadas , Pólen/citologia , Pólen/crescimento & desenvolvimento , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Nicotiana/fisiologia , Transformação GenéticaRESUMO
Radiation hybrid (RH) mapping, a somatic cell genetic technique, has been developed in animal systems as a general approach for the construction of long-range physical maps of chromosomes. This statistical method relies on X-ray induced breakage of chromosomes to determine the physical distance between markers, as well as their order on the chromosome. The method can be applied to single chromosomes or across the whole genome. The generation of plant (barley) radiation hybrids and their culture in vitro is described here. PCR-based marker systems are used to verify hybrid status and to demonstrate genome coverage. RH panels of the type generated can be used for physical mapping, map-based cloning, or sequence contig assembly. RH resources will greatly aid the physical characterisation of crop plants with large genomes.
