| Agropyron is an important relative plant for genetic improvement of stress resistance in wheat. The development of molecular markers is of great significance to accelerate the efficiency of genetic breeding and innovative utilization of wheatgrass. In this study, based on transcriptome sequencing technology, the young roots, stems, leaves, flower ears and seeds of two wheatgrass have been bioinformatics analysis, and highly polymorphic EST-SSR markers were developed and verified. The results showed that a total of 33 993 Unigenes varieties were used as samples for bioinformatics analysis, and developed highly polymorphic were obtained, the total length was 29 309 958 bp, the GC content was 56.44%, and the Q30 was 93.04%. By comparing Unigenes with 9 functional databases, a total of 25 614 Unigenes were obtained functional annotations, among which 24 557 Unigenes were annotated in Nr database. 66 193 Unigenes were annotated in GO database, which were divided by function into 42 terms of 3 main categories. For KEGG analysis, a total of 14 412 Unigenes were assigned to 136 metabolic pathways. 11 321 Unigene were annotated in KOG database, which were grouped into 25 functional categories. 7 288 CDS of larger than 100bp, and 2 443 SSR loci were predicted, of which 1 439 were triple base repeats (58.90%). From this, 42 pairs of EST-SSR polymorphic primers were obtained, which showed high development efficiency and reliability. Ten pairs of EST-SSR primers were randomly selected to verify the effectiveness, and the average polymorphism rate was 48.33%. These results indicate that EST-SSR primers can be developed by RNA-seq technology, and can be effectively applied to the development of markers related to yield and quality traits, the fingerprint construction of excellent new germplasm fingerprinting and marker-assisted breeding with precision of tetraploid wheatgrass. |
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