Precise Identification And Genetic Diversity Study Of Chromosome 1R With Different Rye Origins In Wheat Background

Rye(Secale cereale L,2n = 2x = 14,RR)is a natural cross-pollination related species of common wheat.It has significant genetic diversity and abundant valuable genes.It can play an important role in wheat improvement.Accurate and convenient identification of rye chromatin will facilitate the transfer and utilization of novel genes from rye in wheat breeding.In order to discover cytological and molecular markers for the identification of rye genetic material from different sources,this study mainly includes the following aspects:(1)Discovery and application of repeat sequences specific fluorescence in situ hybridization(FISH)probes for chromosomes identification of different triticale and rye,(2)Development of rye specific molecular markers for genetic diversity observation of chromosome 1R of rye with different origins,(3)Transmission of chromosome 1R and agronomic traits analysis in new wheat-rye progenies.The results are as follows:1.Discovery of rye chromosome-specific probes: using non-deformation fluorescence in situ hybridization(ND-FISH)method,11 oligonucleotide probes synthesized in the laboratory were used to screen 9 triticale and 3 rye accessions.Four probes displayed reproducible signals for identifying chromosomes 1R-7R.Oligo-3A1 has specific signals for 5RL,and Oligo-1RS-6 and Oligo-K566 have strong hybridization sites of 1R,4R and 7R.The Oligo-1RS-4 probe produced specific signals at the terminal regions of seven rye chromosomes.The probes give rise to different signal distribution of 1R-7R,which allows to fast and efficient identification of rye chromosomes in triticale.At the same time,through the comparison of different triticale and rye chromosome hybridization signals,it was found that the signal distribution of Oligo-pSc119.2 is similar,but Oligo-1RS-4 displayed large difference in signal distribution variation among these 12 triticale and rye materials,which lays the foundation for the subsequent genetic diversity analysis.2.Development and application of rye chromosome-specific molecular markers:(1)Screening of rye chromosome-specific molecular markers using 164 linkage group 1 markers,including 73 pairs of CINAU primers,rye first homology 72 ssrR primers and 19 glk primers.Finally,51 PLUG primers,64 ssrR primers and 17 pairs of glk primers were obtained.These primers were used to perform PCR verification in different rye 1R.100 pairs of primers were polymorphic and 89 pairs were rye 1R specific primers.The genetic diversity of 13 test materials was analyzed and clustered according to polymorphic markers.The results showed that the genetic difference of rye in the 1R additional line was large,and the genetic difference of the 1RS.1BL translocation line material was small,indicating that the cultivation material was single Genetic basis,so introducing the diversity of different rye into wheat is an effective way to further expand rye germplasm resources.3.Using chromosome engineering technology and cytological identification methods,six types of wheat-rye new chromosome introduction line materials were obtained and identified in this study.The transmission of the 1R chromosome introduction line was analyzed and found to have a high transmission rate.In this study,additional lines and substitution lines of 1R chromatin,additional and translocation lines involving 1RS and 1RL were obtained.Their offspring are expected to carry higher frequency small fragment translocation lines.The agronomic traits of materials from different sources of 1R hetero-dyeing system were found,and it was found that this batch of materials showed excellent agronomic traits and disease resistance,and had potential application value in wheat breeding.The rye chromosome-specific cell and molecular markers developed in this study can be used to study the diversity of wheat varieties and identify rye chromatin in the wheat background.Chromosome segment-specific markers can assist the introduction of excellent genes on the rye chromosome into the wheat background breeding process.In addition,the obtained new wheat-rye chromosome addition lines,substitution lines and fragment introduction lines have rich genetic variation and can be used as excellent germplasm for subsequent breeding.