Method for providing fertile plants via induction of BBM during transformation
A method for providing a transgenic plant, including transforming a plant cell or plant material with a nucleotide sequence coding for a BBM protein, wherein activity of the BBM protein is induced during transformation and/or regeneration of the transformed plant cell or plant material, and wherein the plant cell or plant material originates from a recalcitrant plant is provided. In addition, a transgenic plant or material thereof having a BBM protein, wherein the transgenic plant is a recalcitrant plant is also provided.
1. A method for providing a fertile transgenic plant comprising:
a) transforming a recalcitrant sweet pepper Capsicum annuum or Petunia hybrida plant cell with an expression vector encoding a babyboom protein (BBM), wherein said expression vector provides inducible nuclear transcriptional activity of said babyboom protein (BBM), and wherein babyboom protein BBM is SEQ ID NO: 1 under the condition that said babyboom protein (BBM) has transcriptional activity;
b) regenerating said recalcitrant transformed sweet pepper Capsicum annuum or Petunia hybrida plant cell into somatic embryos, shoot-like structures (SLS) and/or leaf-like structures (LLS) under inductive conditions resulting in nuclear transcriptional activity of said babyboom protein (BBM);
c) culturing said somatic embryos, shoot-like structures (SLS) and/or leaf-like structures (LLS) into a fertile transgenic sweet pepper Capsicum annuum or Petunia hybrida plant under non-inductive conditions resulting in the substantial absence of nuclear transcriptional activity of said babyboom protein (BBM).
2. The method according to claim 1 , wherein said inducible nuclear transcriptional activity is provided by a transcription regulation element, a nuclear targeting sequence, or a translation regulation element.
3. The method according to claim 2 , wherein said nuclear targeting sequence is encoded by a nucleotide sequence according to SEQ ID NO: 2.
4. The method according to claim 3 , wherein said inductive conditions are provided by dexamethasone.
5. The method according to claim 1 , wherein said expression vector further encodes one or more selectable markers, one or more proteins of interest and/or one or more transcription products of interest.
6. The method according to claim 1 , wherein said method further comprises:
d) obtaining a plant cell from said fertile transgenic sweet pepper Capsicum annuum or Petunia hybrida plant;
e) transforming said sweet pepper Capsicum annuum or Petunia hybrida plant cell with an expression vector encoding one or more proteins or transcription products of interest;
f) regenerating said transformed sweet pepper Capsicum annuum or Petunia hybrida plant cell into somatic embryos, shoot-like structures (SLS) and/or leaf-like structures (LLS) under inductive conditions resulting in nuclear transcriptional activity of said babyboom protein (BBM);
g) culturing said somatic embryos, shoot-like structures (SLS) and/or leaf-like structures into a fertile transgenic sweet pepper Capsicum annuum or Petunia hybrida plant under non-inductive conditions resulting in the absence of nuclear transcriptional activity of said babyboom protein (BBM);
h) optionally repeating steps (d) to (g) one or more times.
7. A fertile transgenic sweet pepper Capsicum annuum or Petunia hybrida plant comprising one or more expression vectors that encode a babyboom protein (BBM) having the sequence of SEQ ID NO: 1, wherein said expression vector provides inducible nuclear transcriptional activity of said babyboom protein (BBM).
8. A seed from the fertile plant according to claim 7 that comprises the expression vector of claim 7 .
9. The method according to claim 1 , wherein said plant cell is a Petunia hybrida plant cell.
10. The fertile transgenic plant according to claim 7 , wherein said plant is a Petunia hybrida.
11. The fertile transgenic plant according to claim 7 , wherein said plant is a sweet pepper Capsicum annum.
12. A fruit from the fertile plant according to claim 7 that comprises the expression vector of claim 7 .
13. The method according to claim 1 , wherein said plant cell is a Capsicum annum plant cell.
14. The fertile transgenic sweet pepper Capsicum annuum or Petunia hybrida plant according to claim 7 , wherein said plant is a sweet pepper Capsicum annuum deposited under number NCIMB 41732.
15. A plant part from the fertile plant according to claim 7 that comprises the expression vector of claim 7 .