Increasing gene editing and site-directed integration events utilizing developmental promoters

A plant comprising:

• (a) (i) a first nucleic acid sequence encoding a guided nuclease capable of generating a staggered cut in a double-stranded DNA molecule operably linked to a heterologous floral tissue-preferred promoter or floral cell-preferred promoter;
• and
• (ii) a second nucleic acid sequence encoding at least one guide nucleic acid operably linked to a second heterologous promoter, wherein the at least one guide nucleic acid is capable of hybridizing to a target sequence within a genome of the plant;
• or
• (b) (i) a first nucleic acid sequence encoding a guided nuclease capable of generating a staggered cut in a double-stranded DNA molecule operably linked to a first heterologous promoter;
• and
• (ii) a second nucleic acid sequence encoding at least one guide nucleic acid operably linked to a heterologous floral tissue-preferred promoter or floral cell-preferred promoter,
• wherein the floral cell-preferred promoter, the floral tissue-preferred promoter, the first heterologous promoter, or the second heterologous promoter comprises a nucleic acid sequence at least 99% identical to the full-length of a nucleic acid sequence selected from the group consisting of SEQ ID NOs:
• 1 to 16, 18, 19, 21 to 30, and 49.

A method of editing a genome of a plant comprising:

• (a) introducing into a plant cell:
• (i) a first nucleic acid sequence encoding a guided nuclease capable of generating a staggered cut in a double-stranded DNA molecule operably linked to a heterologous floral cell-preferred promoter;
• and
• (ii) a second nucleic acid sequence encoding at least one guide nucleic acid operably linked to a second heterologous promoter, wherein the at least one guide nucleic acid is capable of hybridizing to a target sequence within the genome;
• and
• (b) regenerating at least one plant from the plant cell of step (a),
• wherein the guided nuclease and at least one guide nucleic acid form a ribonucleoprotein within at least one floral cell of the plant, and wherein the ribonucleoprotein generates at least one double-stranded break within the target sequence in the at least one floral cell, and
• wherein the heterologous floral cell-preferred promoter or the second heterologous promoter comprises a nucleic acid sequence at least 99% identical to the full-length of a nucleic acid sequence selected from the group consisting of SEQ ID NOs:
• 1 to 16, 18, 19, 21 to 30, and 49.

A method of generating a site-directed integration in a plant comprising:

• (a) introducing into a plant cell:
• (i) a first nucleic acid sequence encoding a guided nuclease capable of generating a staggered cut in a double-stranded DNA molecule operably linked to a heterologous floral cell-preferred promoter;
• (ii) a second nucleic acid sequence encoding one or more guide nucleic acids operably linked to a second heterologous promoter, wherein the one or more guide nucleic acids are
• (A) capable of hybridizing to a target sequence within a genome of the plant;
• and
• (B) capable of hybridizing to a first site and a second site flanking a nucleic acid sequence encoding a gene of interest;
• and
• (iii) a third nucleic acid sequence encoding the gene of interest;
• and
• (b) regenerating at least one plant from the plant cell of step (a);
• wherein the guided nuclease and at least one guide RNA form a ribonucleoprotein within at least one floral cell of the plant, wherein the ribonucleoprotein generates a double-stranded break within the target sequence molecule, the first site, and the second site, and wherein the gene of interest is integrated into the target sequence in the at least one floral cell, and
• wherein the heterologous floral cell-preferred promoter or the second heterologous promoter comprises a nucleic acid sequence at least 99% identical to the full-length of a nucleic acid sequence selected from the group consisting of SEQ ID NOs:
• 1 to 16, 18, 19, 21 to 30, and 49.