US 12,336,470 B2
Method for selecting seed size in watermelon and generating modifications in the tomato-seed size gene
Richard Bernard Berentsen, Paterna (ES); Courtney Hu, Sacramento, CA (US); and Elena Chiapparino, Sant Agata Bolognese Emilia-Romagna (IT)
Assigned to NUNHEMS B.V., Nunhem (NL)
Appl. No. 17/800,388
Filed by NUNHEMS B.V., Nunhem (NL)
PCT Filed Feb. 9, 2021, PCT No. PCT/EP2021/053032
§ 371(c)(1), (2) Date Aug. 17, 2022,
PCT Pub. No. WO2021/165091, PCT Pub. Date Aug. 26, 2021.
Claims priority of provisional application 62/977,834, filed on Feb. 18, 2020.
Prior Publication US 2023/0092854 A1, Mar. 23, 2023
Int. Cl. A01H 1/04 (2006.01); A01H 5/10 (2018.01); A01H 6/34 (2018.01)
CPC A01H 1/045 (2021.01) [A01H 5/10 (2013.01); A01H 6/342 (2018.05)] 9 Claims
 
1. A method of producing F1 hybrid seed, comprising
crossing a first parent line being homozygous for a haplotype for a gene named Ts-gene selected from:
a) a deletion of all or part of the Ts-gene, or a mutant allele of the Ts-gene, said mutant allele results in no expression of the Ts-gene or the mutant allele encodes a Ts-protein having loss-of-function compared to the wild type Ts-protein (chromosome 2 haplotype ‘NO’);
b) a mutant allele of the Ts-gene, which mutant allele encodes a mutant Ts-protein comprising one or more amino acids inserted, deleted or replaced compared to the wild type Ts-protein, said mutant Ts-protein having reduced function compared to the wild type Ts-protein (chromosome 2 haplotype ‘RE’); or
c) a wild type allele of the Ts-gene, encoding a functional Ts-protein (chromosome 2 haplotype ‘WT’);
and
being homozygous for a haplotype for SNP_01 and INDEL_02 selected from i) the combination of SEQ ID NO: 4 and SEQ ID NO: 6, or ii) the combination of SEQ ID NO: 4 and SEQ ID NO: 7 or the combination of SEQ ID NO: 5 and SEQ ID NO: 6 or iii) the combination of SEQ ID NO: 5 and SEQ ID NO: 7;
with a second parent line being homozygous for a haplotype for the Ts-gene selected from:
a) a deletion of all or part of the Ts-gene, or a mutant allele of the Ts-gene, said mutant allele results in no expression of the Ts-gene or the mutant allele encodes a Ts-protein having loss-of-function compared to the wild type Ts-protein (chromosome 2 haplotype ‘NO’),
b) a mutant allele of the Ts-gene, which mutant allele encodes a mutant Ts-protein comprising one or more amino acids inserted, deleted or replaced compared to the wild type Ts-protein, said mutant Ts-protein having reduced function compared to the wild type Ts-protein (chromosome 2 haplotype ‘RE’); or
c) a wild type allele of the Ts-gene, encoding a functional Ts-protein (chromosome 2 haplotype ‘WT’);
and
being homozygous for a haplotype for SNP_01 and INDEL_02 selected from i) the combination of SEQ ID NO: 4 and SEQ ID NO: 6, or ii) the combination of SEQ ID NO: 4 and SEQ ID NO: 7 or the combination of SEQ ID NO: 5 and SEQ ID NO: 6 or iii) the combination of SEQ ID NO: 5 and SEQ ID NO: 7;
and obtaining the F1 hybrid seeds, optionally cleaning, drying and packaging the F1 hybrid seeds,
wherein said Ts-gene is the gene which encodes a wild type, functional Ts-protein of SEQ ID NO: 1 or a protein comprising at least 95% sequence identity to SEQ ID NO: 1.