Construction Of EMS Mutant Library Of Medicago Truncatula And Development Of EST-SSR Primers Of Medicago Sativa

Alfalfa is the most widely cultivated forage legume and one of the most economically valuable crops inthe world. This research use the different concentrations of EMS (ethyl methyl sulfonic acid) solution treating alfalfa seeds A17, preliminary build alfalfa EMS mutant library, and developed107of EST-SSR primers using the Illumina GA IIx sequencing platform, Laid the foundation for alfalfa breeding research, the main results were as follows:1. Seeds of A17were treated with EMS of four concentrations and resulted in0.15%EMS treatment with induction effect better than others analyze the offspring mutant and mutation frequency.2085of M1generation and388of M2generation mutants were found. Leaf color mutation traits included speckle dark green leaves, leaf variegation, Red violet leaves, Dwarfed and yellow leaves, Albino, counting for5.37%in the population, leaf shape traits contained drape leaves, leaves with the mutation frequency of3.12%. Plant type mutation traits of9.11%, included long stem dwarf, moderate dwarf, Polytomy, subramose, etc. flowering stage type mutation traits of1.01%, included early flower, late flower, etc. The total mutant frequency was18.61%in the M2population.2. We performed de novo transcriptome sequencing of M. sativa L. subsp. Sativa using the Illumina GA IIx sequencing platform. A total of15tissue types wer included to identify the global and complete transcriptome of this species. The transcriptome coverage was5.64Gb of clean nucleotides, A total of40,433unigenes wer identified, and1,649SSRs were determined.400pairs of PCR primers was designed to developing molecular markers using domestic and overseas of10alfalfa germplasm primers screening, there are316pairs of primers can amplify the stripe,107of them can amplification high polymorphism bands.