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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, spikes at grain-filling stage, and seeds of two wheatgrass accessions have been bioinformatics analyzed, and highly polymorphic EST-SSR markers were developed and verified. The results showed that a total of 23 491 793 high-quality sequences were obtained, with a base length of 7.03 Gb, the GC content of 56.44%, and the Q30 content of 93.04%. 33 993 Unigenes were obtained, with a total length of 29 309 958 bp. A total of 25 614 Unigene annotations were obtained by comparing Unigene with nine functional databases, including 24 557 annotations in the Nr database and 66 193 annotations in the GO database, which were divided into 3 categories and 42 subcategories according to their functions. A total of 14 412 Unigene pathways were annotated into 136 metabolic pathways in the KEGG database. A total of 11 321 Unigenes were annotated in the KOG database, categorized into 25 functional categories. A total of 7 288 CDSs with 2 443 SSR sites greater than 100 bp were predicted, of which 1 439 were tribase repeats, accounting for 58.90% of the total. A total of 42 pairs of primers with EST-SSR polymorphisms were obtained, and 10 pairs were randomly selected for validation using 30 F2 individuals. The average polymorphism ratio was 48.33%. The results indicated that the EST-SSR primers, which were developed in large quantities by RNA-seq technology, can be efficiently applied to the development of markers related to yield and quality traits of tetraploid wheatgrass, the construction of excellent new germplasm fingerprints, and precise molecular marker-assisted breeding.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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