Background: Sequencing of cDNA libraries for the development of expressed sequence tags (ESTs) as well as for the discovery of simple sequence repeats (SSRs) has been a common method of developing microsatellites or SSR-based markers. In this research, our objective was to further sequence and develop common bean microsatellites from leaf and root cDNA libraries derived from the Andean gene pool accession G19833 and the Mesoamerican gene pool accession DOR364, mapping parents of a commonly used reference map. The root libraries were made from high and low phosphorus treated plants.
View Article and Find Full Text PDFMicrosatellite markers are useful genetic tools for a wide array of genomic analyses although their development is time-consuming and requires the identification of simple sequence repeats (SSRs) from genomic sequences. Screening of non-enriched, small-insert libraries is an effective method of SSR isolation that can give an unbiased picture of motif frequency. Here we adapt high-throughput protocols for the screening of plasmid-based libraries using robotic colony picking and filter preparation.
View Article and Find Full Text PDFBackground: Gene-based (genic) microsatellites are a useful tool for plant genetics and simple sequence repeat loci can often be found in coding regions of the genome. While EST sequencing can be used to discover genic microsatellites, direct screening of cDNA libraries for repeat motifs can save on overall sequencing costs. The objective of this research was to screen a large cDNA library from and Andean common bean genotype for six di-nucleotide and tri-nucleotide repeat motifs through a filter hybridization approach and to develop microsatellite markers from positive clones.
View Article and Find Full Text PDFABSTRACT A polymerase chain reaction approach using degenerate primers that targeted the conserved domains of cloned plant disease resistance genes (R genes) was used to isolate a set of 15 resistance gene analogs (RGAs) from common bean (Phaseolus vulgaris). Eight different classes of RGAs were obtained from nucleotide binding site (NBS)-based primers and seven from not previously described Toll/Interleukin-1 receptor-like (TIR)-based primers. Putative amino acid sequences of RGAs were significantly similar to R genes and contained additional conserved motifs.
View Article and Find Full Text PDFBean golden yellow mosaic virus (BGYMV) is a whitefly-transmitted geminivirus of the Begomovirus family that causes important yield losses to common beans grown in tropical and sub-tropical countries of Latin America and the Caribbean. A major resistance gene that has been widely deployed in this region is the recessive locus bgm-1 that prevents the development of severe yellowing typical of the disease. In this study, we developed a co-dominant sequence-characterized amplified region (SCAR) marker, SR2, based on a previously identified random amplified polymorphic DNA (RAPD) marker that is tightly linked to the bgm-1 resistance gene and identified the position of the locus in the common bean genome through comparative mapping using two genetic maps for the species.
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