Chromosome 1V~b Of Dasypyrum Breviaristatum Were Introduced Into Wheat And Molecular And Cytogenetic Identification

Dasypyrum is one of the wild relatives of wheat.It possesses many important agronomic traits,such as resistance to powdery mildew,leaf and stem rusts,as well as vigorous tillering ability,more spikelets,high grain proteins content,and drought tolerance.Dasypyrum includes two species of D.villosum and D.breviaristatum.There have been many studies on D.villosum,and some excellent genes have been introduced into wheat and applied to breeding.However,there are few studies on D.breviaristatum.The main purpose of this study are to identify and analyze the genetic material and investigate agronomic traits of the genetic material of the offspring of the common wheat-D.breviaristatum partial amphiploid and the common wheat after backcross and it is also a reference for wheat molecular chromosome engineering breeding.It is a prerequisite for the study and utilization of the excellent gene resources of D.breviaristatum for the accurate identification of its chromosomes introduced into the common wheat background.This study found that the oligonucleotide probes OligopSc119.2 and Oligo-pTa535 Oligo-(GAA)7 developed using two repeating sequences and Oligo-(GAA)7 from a microsatellite sequence can distinguish differnt chromosomes between common wheat-D.breviaristatum partial amphiploid and dur wheat-D.villosum amphiploid.Using these three probes and Oligo-pDb12 H which is a D.breviaristatum specific probe,the chromosome karyotype of the common wheat-D.breviaristatum partial amphiploid has been constructed.Using Oligo-pScA9.2,OligopTa535 and Oligo-(GAA)7 to identify the progeny of the common wheat-D.breviariestatum partial amphiploid and Mianyang 11,some materials only having 1V~b,2V~b,3V~b,5V~b,6V~bS or 7V~b were obtained by FISH.A total of 17 wheat cultivars with 1V~b chromosome were identified.Among the 17 strains,the offspring materials after 2 hybrids crossing with Mianyang 11 contained 1V~b chromosomes,but the chromosomal variation of wheat background was large;all the offspring materials fron 1 hybrids crossing with Mianyang 11 have one pair of 1V~bL.5V~bL with additional wheat chromosome,and the wheat background chromosome was stable.In order to further confirm the identified 1V~b chromosome in the 1V~b material,it was confirmed by Oligo-pDb12 H and the specific molecular markers to 5V~bL.The offspring materials from the crossing between the other 14 strains and Mianyang11 were not stable.To track the identification of 1V~b chromosomes and help the genetic-assisted breeding,wheat materials with only 1V~bwere screened by group 1 primers of PCR-based Landmark Unique Gene(PLUG),and 12 specific molecular markers were obtained.Wheat seed protein is a major factor affecting the quality of wheat,in which glutenin,especially high molecular weight glutenin subunits,has a great influence on wheat quality.the high molecular weight glutenin gene is mainly located on the first homologous group of the wheat.The analysis of seed protein subunits was carried out on the identified wheat with only 1V~b,and a specific high molecular weight glutenin subunit and gliadin subunit were obtained from 1V~b chromosome expression of D.breviaristatum.The degenerate primers which were previously used to isolate the homologous HMW-GS genes from wheat and its relative species were used to isolate the homologous genes from1 V~b chromosome of D.breviaristatum.Ten sequences were obtained in the clone.An protein that include 577 amino acid sequences that predicted by an contact 1737 bp ORF that is highly similar to high molecular weight glutenin subunit gene coding sequences have similar weight with specific high molecular weight glutenin subunit from 1V~b chromosome expression of D.breviaristatum.In summary,The wheat materials in this study that chromosomes of D.breviaristatum were introduced into the common wheat and the chromosomal specific molecular markers of D.breviaristatum can be used in the breeding practice of D.breviaristatum excellent gene into wheat.