HEMP PLANT NAMED 'CAF#13/2020'

Abstract

A vegetatively reproduced hemp (Cannabis sativa spp.) cultivar named ‘CAF#13/2020’ is distinguished from other commercially produced hemp varieties by its high cannabidiol (CBD) content, low Δ.sup.9-tetrahydrocannabinol (THC) content, and adaptation to growing conditions in Canada and the northern United States. It is also identifiable by its unique genotype, defined by 845,500 single nucleotide polymorphisms detected against the representative reference genome cs10 (GenBank: GCA_900626175.2; RefSeq: GCF_900626175.2) at the National Center for Biotechnology Information (NCBI).

Claims

1. A new and distinct hemp plant named ‘CAF#13/2020’ substantially as described herein.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0009] FIG. 1 is a photograph showing the claimed hemp plant; and,

[0010] FIG. 2 is a photograph showing the leaves and stalk of the claimed hemp plant.

DETAILED BOTANICAL DESCRIPTION OF THE VARIETY

[0011] The patent laws of the United States require that a description for a plant patent be complete and must distinguish the claimed variety. The inventors have sequenced the entirety of the nuclear genome of the ‘CAF#13/2020’ hemp plant, which provides a genotype of the variety at the nucleotide level.

[0012] The following methodology was used: [0013] 1. Genomic DNA (gDNA) was extracted from leaf material of ‘CAF#13/2020’ hemp plants. [0014] 2. A DNA-Seq library was prepared for 150 paired-end sequencing with an Illumina NovaSeq 6000 system. [0015] 3. Sequenced reads were checked for quality, and high quality reads were mapped to Cannabis sativa cs10, the representative genome for this species in the National Center for Biotechnology Information (NCBI). The RefSeq assembly accession is GCF_900626175.2 and the assembled genome was downloaded on Dec. 12, 2020. [0016] 4. The reads mapping at each nucleotide position were compiled and the corresponding nucleotides were annotated. If a nucleotide position in ‘CAF#13/2020’ had a different nucleotide than that of the reference genome cs10, a single nucleotide variant (SNP) was declared. [0017] 5. After applying quality filters to avoid false SNPs, the inventors compiled all SNPs located in genic regions and used to genotype ‘CAF#13/2020’. We found 845,500 high quality SNPs located in genic regions that can be used as a barcode or unique identifier for ‘CAF#13/2020’. The list of these SNPs is compiled in a .bed format file called “CAF#13/2020_genotype.bed”.

[0018] This application is accompanied by a compact disc containing the file “CAF#13/2020_genotype.bed” in a compressed format (bed.gz) as a unique identifier for the ‘CAF#13/2020’ hemp plant. Table 2 provides an example of the .bed format:

TABLE-US-00002 TABLE 2 Sequence Example Line Column 1 Column 2 Column 3 Column 4 Column 5 Column 6 Column 7 1 NC_044371.1 40280 40280 A G 91 0/1 2 NC_044371.1 40953 40953 G A 129 1/1 3 NC_044371.1 245388 245388 A T, C 24 1/2

[0019] Referring to Line 1 of Table 2 above, Column 1 (“NC_044371.1”) denotes the chromosome; Column 2 and Column 3 (“40280” and “40280”) identify the start and end chromosome coordinates of the SNP (which, in the case of SNPs, are equal); Column 4 (“A”) identifies the nucleotide in the cs10 reference genome; Column 5 (“G”) identifies the alternative nucleotide found in ‘CAF#13/2020’; Column 6 (“91”) identifies the number of reads mapping at that position; and, Column 7 (“0/1”) the called genotype. The allele values of the genotype are 0 for the cs10 reference allele (“A”) and 1 for the alternative allele (“G”). Thus, the genotype of ‘CAF#13/2020’ at position 40280 of chromosome NC_044371.1 is 0/1 or A/G (i.e., it is heterozygous). By comparison, it can be seen at Line 2 of Table 2 that the genotype of ‘CAF#13/2020’ at position 40953 of chromosome NC_044371.1 is 1/1 or A/A (i.e., it is homozygous for the alternative allele). Finally, there are some cases, such as the example shown in Line 3 of Table 2, where ‘CAF#13/2020’ has two alternative alleles (“T,C”) and the genotype is heterozygous. Homozygous sites in ‘CAF#13/2020’ for the cs10 reference allele are not included in the file. The submitted file includes 605,723 sites with genotype 0/1, 237,831 sites with genotype 1/1 and 1,946 sites with genotype 1/2.

[0020] The complete genotype description provided in the accompanying electronic file precisely defines the claimed variety and distinguishes it from other known hemp varieties.

[0021] Potency analysis of ‘CAF#13/2020’ was performed by CARO Analytical Services (Richmond, BC, Canada) using test method CR-TM-161 and instrument ID and AIN HPLC CR269-CR274. The analysis confirmed THC concentration of 0.277% by weight, and CBD concentration of 7.380% by weight.