Using Rye 6R Deletion Lines To Physically Locate New PCR-Based Specific Markers Of 6RL

Distant hybridization is a widely used method in wheat breeding program for transferring valuable genes of rye into wheat, among which a variety of elite genes are located on 6RL arm of rye. For effective utilization of elite genes of rye on 6RL arm, precisely identification of the 6RL chromosome or chromosome segments from wheat background is important. In this study, based on Specific Length Amplified Fragment Sequencing (SLAF-seq) technology, PCR-based 6RL-specific markers were developed and located on four regions of 6RL by using wheat-rye 6R and 6RL deletion lines. Meanwhile, three sets of wheat-rye monosomic or disomic addition lines were used to detect polymorphism of these markers. The results are as follows.1. Seven wheat-rye monosomic addition lines (MA1RKu-7RKu lines), A 6RS monotelosomic addition line (MTA6RSKu), a 6RL monotelosomic addition line (MTA6RLKu), a 6R deletion line (DEL6RKu) and a 6RL deletion line (DEL6RLKu) were detected.2. A total of 300 new PCR-based 6RL-specific markers have been developed by using Specific Length Amplified Fragment Sequencing (SLAF-seq) technology.3. These 300 new PCR-based 6RL-specific markers were physically located in four regions on 6RL arm by using MTA6RSKu, MTA6RLKu, DEL6RKu and DEL6RLKu.4. The powdery mildew resistant gene on 6RL was located on the region from the breaking point between 2.3 and 2.5 to the telomere and 127 PCR-based 6RL-specific markers were physical mapping in this region.5. A total of 95 markers from these 300 new developed 6RL-spesific markers displayed polymorphic amplification in different rye species, which could be used to identify the origin of rye 6RL arm.The developed markers in this study could be used to identify a given segment of 6RL in wheat background that have important significance for transferring good genes of 6RL into wheat, and it would promote the utilization of elite genes on 6RL for wheat breeding.