Sequence-tagged microsatellite site markers for chickpea (Cicer arietinum L.)

Two small-insert genomic libraries of chickpea (Cicer arietinum L. were scr eened with a set of microsatellite-specific oligonucleotide probes. A total of 121 positive clones were identified among 13 000 plated colonies. Thirt y-nine clones were recognized by (TAA)(5), 26 by (GA)(8), 18 by (GT)(8). 27 by a pool of AT-rich trinucleotide repeats [(CAA)(5), (CAT)(5), and (GAA)( 5)], and 11 by a pool of GC-rich trinucleotides [(TCC)(5), (CAC)(5), tCAG)( 5), and (CGA)(5)]. Of 53 clones selected for sequencing, 43 carried a micro satellite. Flanking primer pairs were designed for 28 loci, and used on a s mall test-set comprising one C. reticulatum and four C. arietinum accession s. Separation of the PCR products on agarose or polyacrylamide gels reveale d single bands of the expected size with 22 of the primer pairs. Sixteen of these "Cicer arietinum sequence-tagged microsatellite site" (CaSTMS) marke rs were polymorphic at an intraspecific level, detecting 2-4 alleles within the four accessions examined. Primer pairs CaSTMS10 and CaSTMS15 revealed 25 and 16 alleles among 63 C. arietinum accessions from different geographi c locations, reflecting gene diversity values of 0.937 and 0.922, respectiv ely. Mendelian inheritance of CaSTMS markers was demonstrated using a set o f recombinant inbred lines and their parents.