Method of generating plants having white foliage
OwnerVicinius Vilperte, Thomas Debener +4inventors
InventorsVicinius Vilperte, Thomas Debener, Robert Boehm, Andrea Dohm, Guido Von Tubeuf, Ulrich Sander
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Abstract
The disclosure relates to a method for the generation of plants, such as Euphorbia pulcherrima, having a dysfunctional glutathione S-transferase (GST), and the seeds, plant parts or plant cells derived therefrom. The disclosure further relates to a molecular marker capable of identifying a dysfunctional GST gene, to isolated DNA encoding such a dysfunctional GST gene and to the use of such DNA for the preparation of a molecular marker and for use in methods of targeted mutagenesis to inactivate the GST gene to generate plants with a white foliage phenotype.
Claims30
A method for the generation of a Euphorbia pulcherrima (Poinsettia) plant having a white foliage phenotype comprising the steps of:
- a. providing a target E. pulcherrima plant without a white foliage phenotype comprising in its genome at least one functional allele of a glutathione S-transferase gene (EpGST) comprising a simple sequence repeat (SSR) in the region encoding amino acids at positions 40-50 of the protein of SEQ ID NO:
- 3, comprising a stretch of 12 nucleotides consisting of a threefold CTTC repeat;
- b. subjecting said E. pulcherrima plant to a mutagenesis treatment to produce a mutant E. pulcherrima plant;
- c. selecting a mutant E. pulcherrima plant, wherein at least one allele of the EpGST gene comprises a CTTC deletion in said SSR motif;
- d. repeating steps b. and c. until all alleles of the EpGST gene in the plant genome comprise said CTTC deletion in said SSR motif;
- and
- e. selecting a E. pulcherrima plant having a white foliage phenotype, wherein said plant is homozygous for said CTTC deletion in said SSR motif.
A white-foliaged E. pulcherrima plant derived from a non-white foliaged cultivated E. pulcherrima plant, wherein said non-white plant comprises in its genome a gene encoding a homolog or variant having at least 60% amino acid identity to SEQ ID NO:
- 3, said homolog or variant having a SSR comprising a stretch of 12 nucleotides consisting of a threefold CTTC repeat at positions 40-50 of the protein of SEQ ID NO:
- 3, wherein said derived white-foliaged E. pulcherrima plant comprises a CTTC deletion in said SSR, and wherein said white-foliaged E. pulcherrima plant is at least 99.9% genetically identical to said non-white foliaged E. pulcherrima plant.
An isolated nucleic acid of the EpGST gene described by SEQ ID NO:
- 61 or a variant thereof having at least 80% identity to the sequence described by SEQ ID NO:
- 1 and encoding a functional homolog or variant of the protein of SEQ ID NO:
- 3, wherein said homolog or variant has a first domain at positions 11-13 of SEQ ID NO:
- 3 being AAC, AGC or AAN, where N can be any amino acid (SEQ ID NO:
- 62), a second domain at positions 53-56 of SEQ ID NO:
- 3 being LVPA, QVPA (SEQ ID NO:
- 63) or QPVP (SEQ ID NO:
- 64), and a third amino acid domain at positions 65-68 of SEQ ID NO:
- 3 being FESR, and further comprising, in the region encoding amino acids at positions 40-50 of the protein of SEQ ID NO:
- 3, a stretch of 12 nucleotides consisting of a threefold CTTC repeat.
A method for producing a plant having reduced levels of anthocyanins comprising:
- a. providing a plant comprising in its genome at least one functional allele of a glutathione S-transferase gene;
- b. subjecting said plant to targeted mutagenesis treatment to produce a mutant GST gene therein, wherein said mutation is selected from the group consisting of loss-of-function, partial loss-of-function, a restored frameshift, an in-frame deletion, or a promoter deletion, and wherein said targeted mutagenesis uses at least one of the sequences of SEQ ID NO:
- 44-49, or variants thereof having at least 95% identity, to produce a guide RNA;
- and;
- c. selecting a plant having reduced levels of anthocyanins.
Dependencies are inferred from the claim text.