Plants with improved digestibility and marker haplotypes

A plant or plant part comprising

• a) a QTL allele associated with improved digestibility, said QTL allele comprising a nucleotide sequence of a gene encoding a cytochrome P450 flavonoid 3′,5′-hydroxylase (F35H), having a mutation, preferably a mutation leading to altered expression of the mRNA of the gene and/or the F35H protein, or a mutation leading to an F35H protein having altered enzymatic activity upon translation;
• or
• b) a nucleotide sequence of a gene encoding a cytochrome P450 flavonoid 3′,5′-hydroxylase (F35H), having a mutation, preferably a mutation leading to altered expression of the mRNA of the gene and/or the F35H protein, or a mutation leading to an F35H protein having altered enzymatic activity upon translation;
• or
• c) a nucleotide sequence of a gene encoding a cytochrome P450 flavonoid 3′,5′-hydroxylase (F35H) having altered expression of the mRNA of the gene and/or the F35H protein, or having altered enzymatic activity;
• or
• d) an RNAi molecule directed against, targeting, or hybridizing with a nucleotide sequence encoding an F35H protein, or comprising a polynucleotide sequence encoding an RNAi molecule directed against, targeting, or hybridizing with a nucleotide sequence encoding an F35H protein;
• or
• e) an RNA-specific CRISPR/Cas system directed against or targeting a nucleotide sequence encoding a cytochrome P450 flavonoid 3′,5′-hydroxylase (F35H) or one or more polynucleotide sequence(s) encoding said RNA-specific CRISPR/Cas system.

A method for improving digestibility of a plant or plant part, comprising introducing or introgressing into the genome of a plant or plant part

• (a) a nucleotide sequence of a gene encoding a cytochrome P450 flavonoid 3′,5′-hydroxylase (F35H), having a mutation, preferably a mutation leading to altered expression of the mRNA of the gene and/or the F35H protein, or a mutation leading to an F35H protein having altered enzymatic activity upon translation, or
• (b) a QTL allele associated with improved digestibility, and comprising a nucleotide sequence of a gene encoding a cytochrome P450 flavonoid 3′,5′-hydroxylase (F35H), having a mutation, preferably a mutation leading to altered expression of the mRNA of the gene and/or the F35H protein or a mutation leading to an F35H protein having altered enzymatic activity upon translation, or
• said method comprising
• (c) altering the expression of the mRNA of a gene encoding a cytochrome P450 flavonoid 3′,5′-hydroxylase (F35H) and/or the F35H protein, altering the enzymatic activity of a cytochrome P450 flavonoid 3′,5′-hydroxylase (F35H).

A method for producing a plant or plant part having improved digestibility, comprising

• (a) introducing or introgressing into the genome of a plant or plant part a nucleotide sequence of a gene encoding a cytochrome P450 flavonoid 3′,5′-hydroxylase (F35H), having a mutation, preferably a mutation leading to altered expression of the mRNA of the gene and/or the F35H protein, or a mutation leading to an F35H protein having altered enzymatic activity upon translation, or
• (b) introducing or introgressing into the genome of a plant or plant part a QTL allele, such as a QTL allele associated with improved digestibility, and comprising a nucleotide sequence of a gene encoding a cytochrome P450 flavonoid 3′,5′-hydroxylase (F35H), having a mutation, preferably a mutation leading to altered expression of the mRNA of the gene and/or the F35H protein, or a mutation leading to an F35H protein having altered enzymatic activity upon translation, or
• (c) introducing into a nucleotide sequence of an endogenous gene encoding the cytochrome P450 flavonoid 3′,5′-hydroxylase (F35H) a mutation, preferably a mutation leading to altered expression of the mRNA of the gene and/or the F35H protein, or a mutation leading to an F35H protein having altered enzymatic activity upon translation, or
• (d) introducing into the plant or the plant part an RNAi molecule directed against, targeting, or hybridizing with a nucleotide sequence encoding the F35H protein, or a polynucleotide sequence encoding an RNAi molecule directed against, targeting, or hybridizing with a nucleotide sequence encoding the F35H protein, or
• (e) introducing into the plant or the plant part an RNA-specific CRISPR/Cas system directed against or targeting a nucleotide sequence encoding the F35H protein, or one or more polynucleotide sequence(s) encoding said RNA-specific CRISPR/Cas system, or
• (f) introducing into the plant or the plant part a chemical compound or an antibody altering the enzymatic activity of the F35H protein upon interaction with said F35H;
• and
• (g) optionally, regenerating a plant from the plant part of any of (a) to (e).