Methods and compositions involving ALS variants with native substrate preference
Compositions and methods comprising polynucleotides and polypeptides having ALS activity and tolerance to at least one ALS inhibitor are provided. In specific embodiments, the sequence has an increased preference for 2-ketobutyrate, when compared to an appropriate control, such as for example, HRA, and/or a preference for 2-ketobutyrate similar to a native ALS. Further provided are nucleic acid constructs, plants, plant cells, explants, seeds and grain having the ALS inhibitor tolerant sequences. Various methods of employing the ALS inhibitor tolerant sequences are provided. Such methods include methods for producing an ALS inhibitor tolerant plant, plant cell, explant or seed and methods of controlling weeds in a field containing a crop employing the plants and/or seeds disclosed herein.
1. A method of producing an ALS inhibitor-tolerant soybean plant cell comprising:
a) transforming a soybean plant cell with a recombinant polynucleotide comprising a nucleotide sequence encoding a soybean acetolactate synthase (ALS) inhibitor-tolerant polypeptide comprising an amino acid sequence that is at least 97.5% identical to SEQ ID NO: 1, wherein the amino acid sequence contains a serine at the amino acid residue corresponding to position 178 of SEQ ID NO: 1, and wherein said polynucleotide is operably linked to an S-adenosylmethionine synthase promoter;
b) placing said cell in a medium that promotes growth;
c) regenerating a transgenic plant from said plant cell; and
d) selecting at least one plant produced from said cell that is tolerant to at least one ALS inhibitor in the imidazolinone class, wherein said plant produces at least one seed with less than 0.5% C17 fatty acids relative to the total amount of fatty acids present in the seed.
2. The method of claim 1 , wherein said transforming the plant cell results in the stable integration of the polynucleotide into the genome of the plant cell.
3. The method of claim 1 , wherein the ALS inhibitor-tolerant polypeptide comprises the amino acid sequence of SEQ ID NO: 15.
4. The method of claim 1 , wherein the at least one ALS inhibitor is an imazapyr.