BR-122021008131-B1 - Methods for controlling weeds and producing soybean products, as well as the use of plants, soybean seeds or their progeny, and soybean products.

BR122021008131B1BR 122021008131 B1BR122021008131 B1BR 122021008131B1BR-122021008131-B1

Abstract

METHODS FOR WEED CONTROL AND SOYBEAN PRODUCT PRODUCTION, AS WELL AS THE USE OF PLANTS, SOYBEAN SEEDS OR PROGENY THEREOF AND SOYBEAN PRODUCT. The invention relates to transgenic soybean plants, specific plant material and seeds, characterized in that these products encompass specific transformation events at specific locations in the soybean genome. Tools are also provided that allow for the rapid and unambiguous identification of these events in biological samples.

Inventors

JUSTIN THOMAS MASON
LESLIE JAMES LETTOW
MARK ALAN EBY
WILLIAM H. EBY
Günter Welz
Steven Verhaeghe
Marc De Beuckeleer
Veerle Habex
JEANMARC FERULLO

Assignees

BASF Agricultural Solutions Seed US LLC
MS TECHNOLOGIES, LLC

Dates

Publication Date: 20260310
Application Date: 20101123
Priority Date: 20091123

Claims (9)

1. A method for controlling weeds with no or reduced damage to a soybean crop, characterized in that it comprises the application of the herbicides glyphosate, glufosinate, or isoxaflutol to soybean plants, seeds, or progeny thereof, wherein said plants, seeds, or progeny comprise elite events EE-GM3 and EE-GM2 in their genome, wherein the reference seed comprising said elite event EE-GM3 is deposited under NCIMB deposit number 41659, and the reference seed comprising said elite event EE-GM2 is deposited under NCIMB deposit number 41660, and wherein said soybean plants, seeds, or progeny are tolerant to the herbicides isoxaflutol, glufosinate, and glyphosate.
2. Method according to claim 1, characterized in that said plants, seeds or progeny comprise elite EE-GM3 event and elite EE-GM2 event in their genome, wherein said elite EE-GM3 event comprises exogenous DNA comprising a chimeric gene encoding HPPD Pf W336 and a chimeric gene encoding 2mEPSPS, said elite EE-GM3 event comprising the nucleotide sequence 1 to 1451 of SEQ ID NO: 2 immediately upstream of, and contiguous to, said exogenous DNA and the nucleotide sequence 241 to 1408 of SEQ ID NO: 3 immediately downstream of, and contiguous to, said exogenous DNA and wherein said elite EE-GM2 event comprises exogenous DNA comprising a chimeric gene encoding phosphinothricin acetyltransferase, said elite EE-GM2 event comprising the sequence of nucleotides 1 to 311 of SEQ ID NO: 14 immediately upstream of, and contiguous to, said exogenous DNA and nucleotide sequences 508 to 1880 of SEQ ID NO: 15 immediately downstream of, and contiguous to, said exogenous DNA, wherein the reference seed comprising said elite event EE-GM3 is deposited under NCIMB deposit number 41659, and the reference seed comprising said elite event EE-GM2 is deposited under NCIMB deposit number 41660.
3. Method according to claim 1, characterized in that the exogenous DNA inserted into EE-GM3 comprises the nucleotide sequence of SEQ ID NO: 20, from nucleotide position 1452 to nucleotide position 16638, and in that the exogenous DNA inserted into EE-GM2 comprises the nucleotide sequence of SEQ ID NO: 14, from nucleotide 312 to nucleotide 810 and the nucleotide sequence of SEQ ID NO: 15, from nucleotide 1 to nucleotide 507.
4. Method according to claim 1, characterized in that said plants, seeds or progeny comprise in their genome, in order, the following nucleotide sequences: a) the nucleotide sequence of SEQ ID NO: 2, from nucleotide 1 to 1451; b) the nucleotide sequence of the complement of the nucleotide sequence of SEQ ID NO: 1, from nucleotide 6760 to nucleotide 6958; c) the nucleotide sequence of SEQ ID NO: 1, from nucleotide 6874 to nucleotide 7298; d) the nucleotide sequence of SEQ ID NO: 1, from nucleotide 7 to nucleotide 7291; e) the nucleotide sequence of SEQ ID NO: 1, from nucleotide 12 to nucleotide 7265; f) the nucleotide sequence with SEQ ID NO: 3, from nucleotide 217 to nucleotide 240; and g) the nucleotide sequence with SEQ ID NO: 3, from nucleotide 241 to nucleotide 1408; and further comprising, in order, the following nucleotide sequences: h) the nucleotide sequence with SEQ ID NO: 14, from nucleotide 1 to nucleotide 311; i) the nucleotide sequence with SEQ ID NO: 11, from nucleotide 3458 to nucleotide 3848; j) the nucleotide sequence with SEQ ID NO: 11, from nucleotide 413 to nucleotide 3457; (ek) the nucleotide sequence of SEQ ID NO: 15, from nucleotide 508 to nucleotide 1880.
5. Method according to claim 1, characterized in that said elite EE-GM3 event comprises a nucleic acid molecule with the nucleotide sequence SEQ ID NO: 20.
6. Use of plants, soybean seeds or progeny thereof, characterized in that said plants, soybean seeds or progeny thereof are used in a weed control method with no or reduced injury to a soybean crop, wherein said plants, seeds or progeny comprise in their genome the elite event EE-GM3, as defined in any of claims 1 to 5, and the elite event EE-GM2, as defined in any of claims 1 to 5, wherein said weed control method comprises the application of the herbicides isoxaflutol, glufosinate or glyphosate to said soybean plants, seeds or progeny thereof, and said soybean plants, seeds or progeny are tolerant to the herbicides isoxaflutol, glufosinate and glyphosate.
7. Method for weed control with no or reduced injury to a soybean crop in a soybean field comprising elite EE-GM3 and EE-GM2 events, as defined in any one of claims 1 to 5, characterized in that it comprises treating the field with an effective amount of a glufosinate-based and/or glyphosate-based and/or isoxaflutol-based herbicide, wherein such plants are tolerant to such herbicides.
8. Method for weed control with no or reduced damage to a soybean crop, characterized in that it comprises treating a field in which seeds containing elite events EE-GM3 and EE-GM2 were sown with an HPPD inhibitor herbicide, such as isoxaflutol, optionally followed by application of glyphosate and/or glufosinate as a post-emergence herbicide to the top of the plants, wherein said EE-GM2 and EE-GM3 are as defined in any of claims 1 to 5.
9. Method for weed control with no or reduced damage to a soybean crop, characterized in that it comprises treating soybean plants containing elite events EE-GM3 and EE-GM2 that were sown with an HPPD inhibitor herbicide, such as isoxaflutol, on top of the plants, applied together with, followed by, or preceded by treatment with glyphosate and/or glufosinate as a post-emergence herbicide on top of the plants, wherein said EE-GM2 and EE-GM3 are as defined in any of claims 1 to 5.

Description

Cross-referencing related requests [001] This application claims priority over provisional patent application US 61/367,251, filed July 23, 2010; provisional patent application US 61/263,707, filed November 23, 2009, and European patent application EP 09014565.7, filed November 23, 2009, the disclosures of each of which are incorporated herein by reference in their entirety. Field of Invention [002] This invention relates to transgenic soybean plants, plant material and seeds, characterized by harboring at least two specific transformation events, namely through the presence of at least two sets of genes encoding proteins that confer herbicide tolerance, each at a specific location in the soybean genome. The soybean plants of the invention combine the herbicide tolerance phenotype with agronomic performance, genetic stability and functionality in different genetic origins equivalent to the non-transformed soybean line in the absence of herbicide. This invention further provides methods and kits for identifying the presence of plant material specifically comprising EE-GM3 and EE-GM2 or EE-GM1 transformation events in biological samples. Background of the Invention [003] The phenotypic expression of a transgene in a plant is determined both by the structure of the gene or the genes themselves and by their locations in the plant genome. At the same time, the presence of transgenes or "foreign DNA" in different locations in the genome will influence the overall phenotype of the plant in different ways. The agronomically or industrially successful introduction of a commercially interesting trait into a plant through genetic manipulation can be a slow process dependent on different factors. The actual transformation and regeneration of genetically transformed plants is only the first of a series of selection steps that includes extensive genetic characterization, breeding and evaluation in field trials, eventually leading to the selection of an elite event. [004] The unequivocal identification of an elite event is becoming increasingly important in light of discussions on Novel Foods/Food, segregation of GMO and non-GMO products, and identification of proprietary material. Ideally, the identification method is quick and simple, without the need for an extensive laboratory setting. Furthermore, the method should provide results that allow for the unequivocal determination of the elite event without expert interpretation, but which can be performed under expert analysis if necessary. Specific tools for use in identifying elite events EE-GM3 and EE-GM1 or EE-GM2 in biological samples are described herein. [005] In this invention, EE-GM3 was identified as an elite event of a population of transgenic soybean plants in the development of herbicide-tolerant soybean (Glycine max) comprising a gene encoding for glyphosate tolerance combined with a gene conferring tolerance to 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitors, each under the control of an expressible plant promoter. [006] EE-GM1 and EE-GM2 were previously identified as elite events of a transgenic soybean plant population in the development of herbicide-tolerant soybean (Glycine max) comprising a gene encoding glufosinate tolerance under the control of a plant-expressible promoter and are described in WO2006/108674 and WO2006/108675 (both publications incorporated herein by reference). [007] Soybean plants comprising an herbicide tolerance gene have been disclosed in the art. However, none of the prior technical disclosures teaches or suggests the present invention. [008] It is known in the art that obtaining a commercial elite herbicide tolerance transformation event in soybean plants with acceptable agronomic performance, without yield drag, and providing sufficient herbicide tolerance, certainly to 3 different classes of herbicides, is not a simple matter. [009] In fact, it has been reported that the first soybean event (event 40-3-2) released on the market with herbicide tolerance had a significant yield drag compared to (near) isogenic lines (Elmore et al. (2001) Agron. J. 93:408-412). [0010] In addition, Optimum® GAT® soybeans (event 356043) were developed to combine glyphosate tolerance with ALS herbicide tolerance, but it has been reported that these soybeans did not meet the standards for glyphosate tolerance on their own (without combination with another glyphosate-tolerant soybean event (such as event 40-3-2) (see, for example, www.bloomberg.corri/apps/news?pid=newsarchive&sid=ad4L0hH9MKWE)). Summary of Preferred Embodiments of the Invention [0011] The present invention relates to a transgenic soybean plant, or seeds, cells or tissues thereof comprising stably integrated into its genome, an expression cassette comprising a herbicide tolerance gene comprising the coding sequence of the 2mEPSPS gene and another herbicide tolerance gene comprising the coding sequence of HPPD-PF W336 (both as described in Example 1.1 herein and as represented in