Development of flow cytogenetics and physical genome mapping in chickpea ( Cicer arietinum L.)

摘要: Procedures for flow cytometric analysis and sorting of mitotic chromosomes (flow cytogenetics) have been developed chickpea (Cicer arietinum). Suspensions intact were prepared from root tips treated to achieve a high degree metaphase synchrony. The optimal protocol consisted treatment roots with 2 mmol/L hydroxyurea 18 h, 4.5-h recovery in hydroxyurea-free medium, h incubation 10 µmol/L oryzalin, ice-water overnight. This procedure resulted an average index 47%. Synchronized fixed 2% formaldehyde 20 min, chromosome suspensions by mechanical homogenization tips. More than 4×105 morphologically could be isolated 15 Flow DAPI-stained histograms relative fluorescence intensity karyotypes) containing eight peaks, representing individual and/or groups similar DNA content. Five peaks assigned (A, B, C, G, H). purity sorted fractions was high, B H 100% purity. PCR on flow-sorted primers sequence-tagged microsatellite site (STMS) markers permitted assignment the genetic linkage group LG8 smallest H. study extends number legume species which cytogenetics is available, demonstrates potential genome mapping chickpea.