Development Of Molecular Markers For Lophopyrum Elongatum And Establishment Of Triticum Durum-Lophopyrum Elongatum Alien Disomic Addition Lines

Wheat(Triticum aestivum) is an important crop food in the world, its acreage is biggest in the world and the total output only less than corn. In recent years, the wheat variety improvement work has achieved rapid development, its production has been significantly improved, but its quality and resistance trait improvement is not obviously. Species of wild relatives of wheat have many good genes that wheat do not have, import of these wild relatives excellent exogenous genes to be used in wheat on the rich wheat breeding and genetic improvement of great is significance. Agropyron elongatum(Lophopyrum elongatum) is one of the important wheat wild relatives, has many good traits, one of the most valuable use of wild species for genetic improvement of common wheat. In this experiment, the material is Wheat-Lophopyrum elongatum disomic addition and substitution lines, use iPBS technology to get the Lophopyrum elongatum E group chromosome specific DNA fragments, and tanslate to the corresponding molecular markers. And use durum wheat Langdon(Triticum durum) and heterologous hexaploid wheat8801(AABBEE)hybrid offspring for materials, comprehensive utilization of cytology and molecular markers tracking exogenous chromosomes, breeding Triticum durum-Lophopyrum elongatum alien disomic addition lines. The results were as follows:1Development of E-chromosome specific SCAR markers for Lophopyrum elongatumUse iPBS technology synthetic25primers were used to screen Triticum aestivum cv. Chinese Spring, Lophopyrum elongatum and Chinese Spring-Lophopyrum elongatum disomic additional lines for PCR amplification, it was showed that31fragments which were specific for1E to7E were obtained. These specific fragments were cloned and sequenced, the sequencing results comparison homology with wheat in the NCBI GenBank,16fragments have no homology or lower homology with wheat. Design PCR primers for these16specific fragments, respectively, using Chinese Spring-Lophopyrum elongatum disomic additional and substitutional lines, Chinese spring and Lophopyrum elongatum for PCR reaction. Primer iPBS8amplified fragment only appear in the Chinese spring-Lophopyrum elongatum3E additional line and substitution line and Lophopyrum elongatum, size is consistent with the theoretical value, indicating that is located on chromosome3E, primer iPBS15amplified fragments appear in the China spring-Lophopyrum elongatum all disomic addition line and substitution line, Lophopyrum elongatum, size is consistent with the theoretical value, indicating that is located on Lophopyrum elongatum1E to7E chromosome. Primer iPBS10and iPBS5amplified fragment appear in the Chinese spring-Lophopyrum elongatum2and4disomic additions and substitution lines, Lophopyrum elongatum, size is consistent with the theoretical value, that is located in2and4chromosomes. These molecular markers have good repeatability, amplification stability, easy and simple to handle, and cost effective, provides an excellent marker for molecular marker-assisted breeding.2Identification of Triticum durum-Lophopyrum elongatum alien disomic addition lines by cytological and molecular markersMeiosis inspection wheat durum and allohexaploid8801hybrid F1, found that10of the15offsprings were true hybrids, indicating successful hybridization.Extraction DNA for17plants F2generation that preliminary examination of the chromosome number was2n=29, use of Lophopyrum elongatum1E-7E chromosome-specific molecular markers on these DNA for PCR amplification, the results show that the2plants containing Lophopyrum elongatum1E chromosome,3plants containing Lophopyrum elongatum5E chromosome,2plants containing Lophopyrum elongatum chromosome7E.The preliminary chromosome number examination of the F3generation hybrid as2n=29of the40plants for DNA extraction, tmolecular markers verification results show that which contains3E,4E,5E,7E chromosome number of plants were2,6,8,17plants, no plants containing1E,2E,6E chromosome.Identification of F4generation by PCR, in which F1-10-25-3contain Lophopyrum elongatum3E chromosome, F1-1-28-2and F1-1-28-3contain Lophopyrum elongatum4E chromosome, F1-3-9-4and F1-3-9-5contain Lophopyrum elongatum5E chromosome, F1-3-12-6-1and F-1-23-2contain Lophopyrum elongatum7E chromosome, these plants have Lophopyrum elongatum chromosome, initially identified as Triticum durum-Lophopyrum elongatumt alien monomer or disomic addition lines. F-1-23-2chromosome examination showed that the chromosomes number of30, initially identified as Triticum durum-Lophopyrum elongatum7E alien disomic addition lines.