Efficient CRISPR/Cas9 Genome Editing in Alfalfa Using a Public Germplasm

Because its ability to acquire large amounts of nitrogen by symbiosis, tetraploid alfalfa is the main source vegetable proteins in meat and milk production systems temperate regions. Alfalfa cultivation also adds fixed soil, improving non-legumes crop rotation reducing use fertilizers derived from fossil fuel. Despite economic ecological relevance, genetics remains poorly understood, limiting development public elite germplasm. In this brief article, we reported high-efficiency mutagenesis using clone C23 CRISPR/Cas9 system. Around half GUS overexpressing plants (35S- under genomic background) transformed with an editing plasmid containing two sgRNAs against gene Cas9 nuclease exhibited absence activity. Nucleotide analysis showed that inactivation CRISPR/Cas9-editing events were produced via different modifications gene, including frameshift non-sense mutations. Using system sgRNAs, have edited NOD26 , generating doses alleles at locus, complete knockout high efficiency (11%). Finally, discuss potential applications genome-editing technologies polyploid research improvement programs.