Development And Physical Mapping Of Specific Molecular Markers For Rye 4R Chromosome

Wheat(TriticumaestivumL.) is one of the most important crops worldwide. Rye (Secale cerealL.) is an important source of genes for wheat disease resistance, yield and quality traits,.So, intensive study on rye chromosomes is needed.In this study, wheat-rye 4R addition lines and 4R chromosome abberation lines were identified by using non-denaturing fluorescence in situ hybridization (ND-FISH) assays. And rye 4R-specific sequences have been developed by using specific length amplified fragment sequencing (SLAF-seq) technology. According to these 4R-specific sequences, PCR-based rye 4R chromosome-specific markers were developed and physically mapped by usingwheat-rye addition lines and 4R chromosome aberrationlines.Thedetail results are as follows:1? The wheat-rye 4RLditelosomic addition line DTA4RLKu and 4RS ditelosomic addition lineDTA4RSKufrom the progeny of wheat Mianyang 11(MY11) x Rye (Kustro) have× been identified. The 4R chromosome was divided into nine regions including S-1, S-2, S-3, S-4, L-1, L-2, L-3, L-4 and L-5by using the probes (AAC)6, Oligo-pSc119.2-1, Oligo-pSc200 and Oligo-pSc250..2?The progeny of 4RS-5DS.5DL,5DS-4RS.4RL translocation line 12FT2115 and some radiated progeny of DTA4RLKu and 12FT2115 have been analyzedusing ND-FISH. Homozygous 4RS-5DS.5DL translocation Hne14T190-1, homozygous 5DS-4RS.4RL translocation line 14T150-11 and homozygous 4RS/5DStranslocation line 14T150-27 have been identified from selfed progeny of 12FT2115. Homozygous 4BL.4BS-4RL, 4RL/5BLtranslocation line 14T201-11,4RL-5BL.5BStranslocation line 15T139-land 4RL/5BLtranslocation line 15T165-9 have been identified from the radiated progeny of DTA4RLKu. Homozygous 5DS-4RS.4RLtranslocation line 15T305-12 has been identified from the radiated progeny of 12FT2115. According to these wheat-rye 4R translocation aberration lines, seven regions of the 4R chromosome including S-1+S-2, S-3, S-4, L-1+L-2.1, L-2.2, L-2.3 and L-3+L-4+L-5 were divided.3?Rye 4R-specific sequence have been developed by using SLAF-seqtechnology, then 800 fragments were randomly selected to design primers.A total of 381 markers were located on 4R chromosomes of Kustro by using a set of wheat-rye monosomic addition lines MA1RKu-7RKu.4?314 of the 3814R chromosome-specific markers were physically mapped to7 specific regions (S-1+S-2, S-3, S-4, L-1+L-2.1, L-2.2, L-2.3 and L-3+L-4+L-5) on the 4R chromosome arm. The numbers of chromosome-specific markers on the seven regions were 109,9,19,27,18,6 and 126, respectively.5?Two sets of wheat-rye disomic addition lines DA1RKi-7RKi and DA1R1-7R1 were used to investigate the polymorphism of the 3814R-spesific markers.216 markers could amplify 4R-specific bands from 4R chromosomes of rye Kustro, King ? and Imperial indicating they are general usage.148 of the 3814R-spesific markers displayed polymorphic amplification in 4RKu?4RKi?and 4RI.