Cloning Of P-Genome Specific Repetitive DNA Sequences From Agropyron Gaertn And Its Application

In this study, Agropyron Gaertn(P genome) and other genome of Triticum were used to isolate P-genome specific repetitive DNA sequences. SCAR markers were transformed from RAPD and ISSR markers through P genome-specific repetitive sequences. These markers were used to detect alien chromatin. The main results were described as follows: 1. The genomes of part plants of Triticum were screened by RAPD markers. P-genome specific fragments were reclaimed. A specific DNA repetitive sequence of P genome in Agropyron Gaertn, OPC08578, was isolated and cloned. Its length was 578bp. The G+C content was composed with 44%. Homologous comparison with the 2,587,412 DNA sequences registered in gene bank (GenBank + EMBL + PDB) indicated that it was a new P-genome specific DNA repetitive sequence. Southern hybridization showed that OPC08578 had the same distribution in the genome W,H and F. But it has no hybridization signals detected in ABD,R,V,E,St,Ns,I,D and A genomes. The results suggested that OPC08578 had strong specificity of P-genome 2. A pair of specific primers SC-OPC08 were designed according to the sequence of OPC08578. This primer could not only distinguish P genome from genomes ABD,R,V,E,St ,W ,Ns,D,A,C,S,U,I,M and AG, but also could be used in the detection of chromatin of P genome from Wheat-Agropyron additions. The results of PCR amplification were consistent with the results of Southern hybridization. Then RAPD marker was transformed into SCAR marker. 3. The genomes of part plants of Triticum were screened by ISSR markers. Specific fragments were reclaimed. A P-genome specific DNA repetitive sequence in Agropyron Gaertn, UBC811535, was isolated and cloned. Its length was 535bp. The G+C content was composed with 44%. Homologous comparition with the 2,587,412 DNA sequences registered in gene bank...