Enhanced transformation of recalcitrant monocots
AssigneeSyngenta Participations AG
InventorsQiudeng Que, David Nicholl
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Abstract
Provided herein are improved methods for transforming monocotyledonous plants, as well as an improved phosphomannose-isomerase (PMI) protein coding region and transformation vectors including the same.
Claims47
A method for increasing the transformation frequency of a monocotyledonous plant tissue, comprising:
- introducing a heterologous nucleic acid comprising a coding region having one or more maize-optimized codons into a cell of said plant tissue, wherein said coding region codes for a phosphomannose-isomerase (PMI) protein, thereby producing a transformed cell comprising the nucleic acid;
- whereby said transformation frequency of said monocotyledonous plant tissue is increased as compared to a transformation efficiency without maize-optimized codons in said coding region of said nucleic acid.
A method of transforming sugar cane using a phosphomannose-isomerase (PMI) protein as a selectable marker, said method comprising:
- (a) introducing a heterologous nucleic acid comprising a phosphomaimose-isomerase (PMI) protein coding region, said PMI coding region having one or more maize-optimized codons, into a cell of said plant tissue to thereby produce a transformed cell comprising the nucleic acid, wherein said introducing step is carried out by Agrobacterium-mediated transformation;
- and
- (b) regenerating a transformed plant from the transformed cell under conditions selective for PMI protein expression;
- to produce said transformed sugar cane.
A recombinant vector comprising a nucleic acid sequence, said nucleic acid sequence or the complement thereof comprising a coding region encoding:
- (a) the amino acid sequence of SEQ ID NO:3;
- or
- (b) an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO:3 and encoding a phosphomannose-isomerase (PMI) protein,
- wherein said coding region comprises one or more maize-optimized codons.
A nucleic acid sequence, said nucleic acid sequence or the complement thereof comprising a coding region encoding:
- (a) the amino acid sequence of SEQ ID NO:3;
- or
- (b) an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO:3 and encoding a phosphomannose-isomerase (PMI) protein,
- wherein said coding region comprises one or more maize-optimized codons.
Dependencies are inferred from the claim text.