A novel alfalfa variety designated AFX145017 and seed, plants and plant parts thereof are provided. Methods for producing an alfalfa plant comprise crossing alfalfa variety AFX145017 with another alfalfa plant. Methods for producing an alfalfa plant containing in its genetic material one or more traits transgenes or locus conversions introgressed into AFX145017 through backcross conversion and/or transformation are provided and the alfalfa seed, plant and plant part produced thereby. Alfalfa seed, plants or plant parts produced by crossing alfalfa variety AFX145017 or a locus or trait conversion of AFX145017 with another alfalfa plant or population are disclosed. Alfalfa populations derived from alfalfa variety AFX145017, methods for producing other alfalfa populations derived from alfalfa variety AFX145017 and the alfalfa populations and their parts derived by the use of those methods.

A novel alfalfa variety designated AFX164041 and seed, plants and plant parts thereof are provided. Methods for producing an alfalfa plant comprise crossing alfalfa variety AFX164041 with another alfalfa plant. Methods for producing an alfalfa plant containing in its genetic material one or more traits transgenes or locus conversions introgressed into AFX164041 through backcross conversion and/or transformation are provided and the alfalfa seed, plant and plant part produced thereby. Alfalfa seed, plants or plant parts produced by crossing alfalfa variety AFX164041 or a locus or trait conversion of AFX164041 with another alfalfa plant or population are disclosed. Alfalfa populations derived from alfalfa variety AFX164041, methods for producing other alfalfa populations derived from alfalfa variety AFX164041 and the alfalfa populations and their parts derived by the use of those methods.

A novel alfalfa variety designated AFX164040 and seed, plants and plant parts thereof are provided. Methods for producing an alfalfa plant comprise crossing alfalfa variety AFX164040 with another alfalfa plant. Methods for producing an alfalfa plant containing in its genetic material one or more traits transgenes or locus conversions introgressed into AFX164040 through backcross conversion and/or transformation are provided and the alfalfa seed, plant and plant part produced thereby. Alfalfa seed, plants or plant parts produced by crossing alfalfa variety AFX164040 or a locus or trait conversion of AFX164040 with another alfalfa plant or population are disclosed. Alfalfa populations derived from alfalfa variety AFX164040, methods for producing other alfalfa populations derived from alfalfa variety AFX164040 and the alfalfa populations and their parts derived by the use of those methods.

A novel alfalfa variety designated AFX164041 and seed, plants and plant parts thereof are provided. Methods for producing an alfalfa plant comprise crossing alfalfa variety AFX164041 with another alfalfa plant. Methods for producing an alfalfa plant containing in its genetic material one or more traits transgenes or locus conversions introgressed into AFX164041 through backcross conversion and/or transformation are provided and the alfalfa seed, plant and plant part produced thereby. Alfalfa seed, plants or plant parts produced by crossing alfalfa variety AFX164041 or a locus or trait conversion of AFX164041 with another alfalfa plant or population are disclosed. Alfalfa populations derived from alfalfa variety AFX164041, methods for producing other alfalfa populations derived from alfalfa variety AFX164041 and the alfalfa populations and their parts derived by the use of those methods.

A novel alfalfa variety designated AFX164040 and seed, plants and plant parts thereof are provided. Methods for producing an alfalfa plant comprise crossing alfalfa variety AFX164040 with another alfalfa plant. Methods for producing an alfalfa plant containing in its genetic material one or more traits transgenes or locus conversions introgressed into AFX164040 through backcross conversion and/or transformation are provided and the alfalfa seed, plant and plant part produced thereby. Alfalfa seed, plants or plant parts produced by crossing alfalfa variety AFX164040 or a locus or trait conversion of AFX164040 with another alfalfa plant or population are disclosed. Alfalfa populations derived from alfalfa variety AFX164040, methods for producing other alfalfa populations derived from alfalfa variety AFX164040 and the alfalfa populations and their parts derived by the use of those methods.

Disclosed is a method for obtaining multileaflet Medicago sativa materials by means of MsPALM1 artificial site-directed mutants. The method comprises: selecting a target site from an exon region of a compound leaf developmental regulatory gene MsPALM1 of Medicago sativa and constructing a plant CRISPR/Cas9 editing recombinant vector MsCRISPR/Cas9::PALM1 and introducing the vector into Medicago sativa cells and regenerating into plants, cutting and repairing to cause a loss-of-function mutation in the MsPALM1 gene of Medicago sativa cells, and then screening the mutant plants by restriction endonuclease digestion and/or targeted deep sequencing of the target sites of the regenerated plants to obtain lines carrying four MsPALM1 allelic genes with simultaneous loss of function mutation. After phenotypic identification, it was confirmed that the compound leaves of the regenerated plants changed from three leaflets to five leaflets. The method can quickly obtain multileaflet Medicago sativa materials with a short breeding period and stable trait.

A method for site-specific mutagenesis of Medicago sativa genes by using a CRISPR/Cas9 system. The method comprises: first constructing a binary expression vector MsCRISPR/Cas9 that can be used for transforming Medicago sativa by Agrobacterium tumefaciens; then designing a target site for a target gene, and ligating the DNA fragment containing the guide sequence of the target site into MsCRISPR/Cas9 to construct a vector MsCRISPR/Cas9::target; and then transforming the Medicago sativa by Agrobacterium tumefaciens, and generating, by screening, a mutant transformed plant with the target gene mutated. According to the method, an MtU6 promoter is used for driving sgRNA transcription in the Medicago sativa.

The invention relates to the alfalfa variety designated R513W224S. Provided by the invention are the seeds, plants and derivatives of the alfalfa variety R513W224S. Also provided by the invention are tissue cultures of the alfalfa variety R513W224S and the plants regenerated therefrom. Still further provided by the invention are methods for producing alfalfa plants by crossing the alfalfa variety R513W224S with itself or another alfalfa variety and plants produced by such methods.

The invention relates to the alfalfa variety designated RRL43M118. Provided by the invention are the seeds, plants and derivatives of the alfalfa variety RRL43M118. Also provided by the invention are tissue cultures of the alfalfa variety RRL43M118 and the plants regenerated therefrom. Still further provided by the invention are methods for producing alfalfa plants by crossing the alfalfa variety RRL43M118 with itself or another alfalfa variety and plants produced by such methods.

The invention relates to the alfalfa variety designated RRL44M375. Provided by the invention are the seeds, plants and derivatives of the alfalfa variety RRL44M375. Also provided by the invention are tissue cultures of the alfalfa variety RRL44M375 and the plants regenerated therefrom. Still further provided by the invention are methods for producing alfalfa plants by crossing the alfalfa variety RRL44M375 with itself or another alfalfa variety and plants produced by such methods.