Screening And Iedntification Of Wheat-Dasypyrum Breviaristatum Alien Chromosome Lines

Dasypyrum breviaristatum includes two species, tetraploid D. breviaristatum(VbVbVbVb)and diploid D. breviaristatum (VbVb). These speices mainly distributed innorthwest Africa, Greece and Morocco. As the rare and important wild related spices ofwheat. They offer potential to increase the genetic variability and desirable charactersfor wheat improvement, such as, resistant to stripe rust, powdery mildew,eyespot,andthey have strong tillering ability, produce more spicules, and high content of seedsprotein.In order to further transfer the good agronomic traits from D. breviaristatum towheat, wheat-D. breviaristatum amphiploid was crossed to wheat and then backcrossedby recurrent wheat, as a result, a series of wheat-D. breviaristatum cross progenies areobtained and need to be screened. In the present research, Dasypyrum specificmolecular markers OPH12, A-PAGE and SDS-PAGE are used to screen these offsprings,molecular marker screening displayed that60progenies could be amplified by thismarker, indicating that these60progenies contain Dasypyrum chromatin which needsfurther identification. A-PAGE and SDS-PAGE result showed that，the majority of theselines are genetically stable, however, compared to Chinese Spring, new high molecularweight and low molecular weight protein subunits arose due to the introgression of D.breviaristatum chromatin into wheat. Combined with wheat chromosome1DSmolecular marker data, we found that the1DS chromosomes of10offerings are missing,while1BL chromosomes of7offerings are missing.In order to establish Dasypyrum chromosome arm specific marker, PCR using145pairs of PLUG primers were performed on wheat-D. breviaristatum amphiploid, a set ofwheat-D. villosum addition lines, a set of wheat-D. villosum translocation lines andwheat controls, as a result,13chromosome (arm) specific molecular markers aredeveloped. Among these13markers,1,2,2,1,1,2,3and1markers are specific forDasypyrum chromosome arm1VS,1VL,2VS,3VL,3VS,4VL,6VL and7VL,respectively.Chromosome2VS marker TNAC1142and7VL marker TNAC1811are used to screen wheat-D. breviaristatum cross progenies, the result indicated that25progeniescould amplified by TNAC1142and1plant could amplified by TNAC1811. Genomic insitu hybridization suggested that, a pair D. breviaristatum chromosomes in progeneyV252(2n＝42) showed hybridization signal. Molecular markers data confirmed that theintroduced D. breviaristatum chromosomes in V252are2Vb, therefore, this line is awheat-D. breviaristatum substitution line, the substituted wheat chromosome are underidentification. Stripe rust resistance test showed that V252is highly resistant to wheatstripe rust, therefore, it can be used as stripe rust resistance source for wheat breeding.In summary, by protein electrophoresis, in situ hybridization, molecular markeranalyze material, find that the materials have a high breeding value for chromosomeengineering breeding and good application of D. breviaristatum gene pool.