PRODUCTION OF A PCR-BASED MARKER FOR DETECTING PSATHYROSTACHYS HUASHANICA KENG CHROMOSOMES IN A WHEAT BACKGROUND

Abstract

In this study, we developed a genome-specific DNA sequence for detecting the incorporation of Psathyrostachys huashanica Keng chromatin into wheat. Random amplified polymorphic DNA (RAPD) analysis was used to identify genome-specific DNA sequences of P. huashanica, which were selected using 21 different plant species. A 716-bp diagnostic band specific to P. huashanica (pHs24) was cloned, sequenced, and converted into a sequence-characterized amplified region (SCAR) marker, designated as RHS107. The sequence of pHs24 had no significant homology with any sequences deposited in NCBI databases, which showed that it was a novel repetitive P. huashanica sequence. A primer pair flanking this specific sequence was designed and a genome-specific SCAR marker for P. huashanica was developed and characterized. We validated its specificity using 21 different plant species and a complete set of wheat-P. huashanica disomic addition lines (1Ns–7Ns, 2n = 44 = 22 II) that carried different P. huashanica chromosomes. Our results indicated that the SCAR marker targeted only the Ns genome of P. huashanica and it was present in all seven of the P. huashanica chromosomes. The SCAR marker developed in this study was a reliable and rapid method for large-scale screening of the introgression of P. huashanica chromosomes in wheat-P. huashanica derivatives.