Methods for plant transformation using spectinomycin selection

• US 8,466,345 B2
• Filed: 08/18/2011
• Issued: 06/18/2013
• Est. Priority Date: 03/09/2007
• Status: Active Grant

• Current Assignee: Monsanto Technology LLC (Bayer AG)
• Original Assignee: Monsanto Technology LLC (Bayer AG)
• Inventors: Martinell, Brian, Petersen, Michael, Somers, David, Wan, Yuechun, Williams, Edward, Ye, Xudong
• Primary Examiner(s): Fox, David T
• Application Number:
  US13/213,021
• Publication Number:
  US 20120077264A1
• Time in Patent Office: 670 Days
• Field of Search: None
• US Class Current: 800/300
• CPC Class Codes: A01H 4/003 Cutting apparatus specially adapted for tissue culture
• CPC Class Codes: A01H 4/008 Methods for regeneration to complete plants
• CPC Class Codes: A01H 6/202 Brassica napus [canola]
• CPC Class Codes: A01H 6/4684 Zea mays [maize]
• CPC Class Codes: A01H 6/542 Glycine max [soybean]
• CPC Class Codes: A01H 6/604 Gossypium [cotton]
• CPC Class Codes: C12N 1/205 Bacterial isolates
• CPC Class Codes: C12N 15/8201 Methods for introducing genetic material into plant cells, e.g. DNA, RNA, stable or transient incorporation, tissue culture methods adapted for transformation
• CPC Class Codes: C12N 15/8202 by biological means, e.g. cell mediated or natural vector
• CPC Class Codes: C12N 15/8205 Agrobacterium mediated transformation
• CPC Class Codes: C12N 15/8209 Selection, visualisation of transformants, reporter constructs, e.g. antibiotic resistance markers
• CPC Class Codes: C12N 15/8221 Transit peptides
• CPC Class Codes: C12N 15/8261 with agronomic (input) traits, e.g. crop yield
• CPC Class Codes: C12N 15/8265 Transgene containment, e.g. gene dispersal
• CPC Class Codes: C12N 15/8271 for stress resistance, e.g. heavy metal resistance
• CPC Class Codes: C12N 15/8274 for herbicide resistance
• CPC Class Codes: C12N 15/8275 Glyphosate
• CPC Class Codes: C12N 15/8277 Phosphinotricin
• CPC Class Codes: C12N 15/8281 for bacterial resistance
• CPC Class Codes: C12N 5/0025 Culture media for plant cell or plant tissue culture
• CPC Class Codes: C12N 5/04 : Plant cells or tissues culture media C12N5/0025
• CPC Class Codes: C12N 9/1029 : transferring groups other than amino-acyl groups (2.3.1)
• CPC Class Codes: C12R 2001/41 : Rhizobium
• CPC Class Codes: C12Y 203/01081 : Aminoglycoside N3'-acetyltransferase (2.3.1.81)
• CPC Class Codes: G01N 33/0098 : Plants or trees wood G01N33/46

Abstract

The present invention relates to methods and compositions for transforming soybean, corn, cotton, or canola explants using spectinomycin as a selective agent for transformation of the explants. The method may further comprise treatment of the explants with cytokinin during the transformation and regeneration process.

Claims

1. A transgenic soybean, cotton or canola plant cell transformed with a nucleic acid construct comprising two sequences conferring resistance to spectinomycin or streptomycin, wherein the first sequence comprises an aadA gene operably linked to a promoter active in a soybean, cotton or canola plant cell, and the second sequence is operably linked to a promoter active in a prokaryotic cell;

wherein the first sequence is operably linked to a polynucleotide sequence encoding a chloroplast transit peptide; and

wherein the plant cell exhibits resistance to said spectinomycin or streptomycin as a result of the first sequence.

2. The plant cell of claim 1, wherein both sequences conferring resistance to spectinomycin or streptomycin encode a polypeptide comprising aminoglycoside-3′

-adenyltransferase (aadA) activity.

3. The plant cell of claim 1, wherein at least one of the sequences comprises SEQ ID NO:

1 or SEQ ID NO;

2.

4. The plant cell of claim 1, wherein the construct further comprises a coding sequence that confers a trait of agronomic interest or improved end use.

5. The plant cell of claim 4, wherein the coding sequence confers resistance to a herbicide.

6. The plant cell of claim 5, wherein the coding sequence confers resistance to glyphosate, bialaphos, phosphinothricin, Basta, glufosinate, 2,4-D, an acetyl-coA carboxylase inhibitor, or dicamba.

7. A transgenic soybean, cotton or canola plant comprising the plant cell of claim 1.

8. A transgenic soybean, cotton or canola seed that comprises the plant cell of claim 1.