Cloning of the Telomere Associated Sequences of Rye B-chromosomes

The short arm terminal segments of 10 rye ( Secale cereal L. ) B-chromosomes were isolated by means of needle microdissection technology. A two-step single primer PCR method was used to amplify the telomere associated sequences of rye B-chromosomes. The PCR products were located in the terminal region of the short ann of rye B-chromosomes by chromosome in situ hybridization, and most A-chromosomes also showed clear signal dots. Some PCR products were cloned in pUC19 vector and one clone pp3 was sequenced. Sequence analysis demonstrated that it has high similarity with the maize subtelomeric clone pBF266. Further utility of this mierodissection-PCR system in construction of high density RFLP map was discussed.