Molecular And Cytogenetic Identification And Transcriptome Analysis Of Wheat-barley Intermedium Alien Lines

Barley?Hordeum vulgare L.?consists of wild barley?Hordeum vulgare ssp.spontaneum?and cultivated barley?H.vulgare ssp.vulgare?.It possesses novel agronomically important traits as early-maturing,multiple disease resistance,and wide adaption,thus it has a large range of cultivation.As the genetic background of wheat becomes narrower and the diversity decreases,new genes transfering strategies from wheat relatives has been used as one of the main breeding methods.These novel genes of barley have become an important source for broadening the genetic basis of wheat.However,it is difficult to obtain successful hybridization betwwen barley and wheat,,and the precise identification of stable progenies from wheat-barley hybrids and the varieties with good agronomic traits have received continuous attention.In this research,common wheat Chinese spring?CS?hybridizes with cultivated barley Morex to obtain a series of wheat-barley addition lines after several crosses,backcrosses and self-crossing.By using fluorescence in situ hybridization?FISH?,the karyotype of individual barley chromosomes was established.In combined with molecular marker technique,a series of wheat-barley addition lines were preisely identified.At the same time,the agronomic traits were statistically analyzed for the wheat-barley stable lines.The transcriptomic analysis of the materials with good agronomic traits were further carried out.The main results of this research are as follows:?1?Using fluorescence in situ hybridization to identify the parental barley material,it was found that the probe Oligo-?GAA?₇ can distinguish the barley chromosome,but the hybridization signals are not scattered enough.Further experiments have obtained two relatively widespread distribution on the barley chromosome.The probes combination are Oligo-?GAA?₇?red?and Oligo-?TAG?₆?green?,Oligo-HVT01?red?and Oligo-?ACT?₈?green?,respectively.The accuracy of the two probe combinations was verified by two sets of Israeli barley,and the results showed that the two sets of probes can accurately identify the barley chromosomes with different genotypes.?2?Using the probe Oligo-pSc119.2 and Oligo-pTa535,coupled with the probe Oligo-?GAA?₇,the FISH was carried out to identify the progenies of the wheat-barley hybrids,and a series of disomic addition lines,which containing only the single-form chromosome of barley?1HS,2H,2HS,2HL,3H,3H,3HL,4H,4HL,5H,5HS,5HL,6H,6HS,6HL,7H,7H??.The chromosomes 2H,2HS,2HL,6H addition lines can be stably transmitted in wheat background.Karyotype analysis of the above stable materials in the addition lines revealed the frequent changes in the wheat chromosomes.The wheat-2H addition lines were screened using CINAU markers,the 43 markers of which were localized to barley 2HL,while other 17 markers were localized to barley 2HS.?3?The agronomic traits of the addition materials were measured,and it was found that the 2H and 2HL addition lines had good stripe rust resistance,but the 2HS addition line was highly susceptible to stripe rust.It is likely that the stripe rust resistance gene was derived from 2HL at adults plant stages.?4?The introgression of barley chromosome 2H into wheat background has an effect on the genes expression in wheat background.We found that the most significant is the transcriptional differences of a large number of genes occurred on chromosome 2A.The function of gene down-regulated in 2A chromosome and gene up-regulated in 2H is enriched.It is found that functional terms mainly focuses on protein metabolism,intracellular macromolecular metabolism.Therefore,the introgression of 2H chromosome has some compensation for the loss of wheat 2A chromosome.In summary,this research identified a number of addition lines containing only single-type chromosomes of barley,as well as molecular markers localized on the chromosome of barley,and comparative analysis of the deferentially expressed genes in some materials,which can be applied to later researches and breeding of introgression of barley novel genes into wheat.