Efficient development of H aynaldia villosa chromosome 6 VS ‐specific DNA markers using a CISP ‐ IS strategy

Development of effective molecular markers linked to P m21 deriving from H aynaldia villosa is critical for wheat breeding of powdery mildew resistance. In this study, we designed 12 pairs of conserved‐intron scanning primers ( CISPs ), using intron‐containing conserved genes located on the short arm of B rachypodium distachyon chromosome 3 (3 B d S ) aligned with cDNA or expressed sequence tags ( ESTs ) of T riticeae crops. Of 12 CISP primer pairs, 11 amplified DNA both in H . villosa and in wheat, and four displayed H . villosa chromosome 6 VS ‐specific polymorphisms. Six non‐polymorphic DNA s were further sequenced for designing internal primers, and five additional 6 VS ‐specific markers were obtained. Of the total nine 6 VS ‐specific co‐dominant markers, six could effectively trace P m21 in F 2 population derived from the hybrid between the T 6 AL .6 VS line and ‘ Y angmai 158’. This study demonstrated that B rachypodium genomic information could be powerfully utilized to develop molecular markers in H . villosa or other Triticeae species.