NOVEL SELECTABLE MARKER GENES
The subject invention relates to a novel gene referred to herein as DSM-2. This gene was identified in Streptomyces coelicolor A3. The DSM-2 protein is distantly related to PAT and BAR. The subject invention also provides plant-optimized genes encoding DSM-2 proteins. DSM-2 can be used as a transgenic trait to impart tolerance in plants and plant cells to the herbicides glufosinate and bialaphos. One preferred use of the subject genes are as selectable markers. The use of this gene as a selectable marker in a bacterial system can increase efficiency for plant transformations. Use of DSM-2 as the sole selection marker eliminates the need for an additional medicinal antibiotic marker (such as ampicillin resistance) during cloning. Various other uses are also possible according to the subject invention.
1 . A method of selecting or distinguishing an individual plant or a group of crop plants containing a DSM-2 gene from a population of plants of the same species not containing the gene.
2 . The method of claim 1 comprising applying glufosinate or bialaphos to the collection of plants.
3 . The method of claim 1 wherein said method is practiced in a greenhouse, a growth chamber, or a field.
4 . A method of controlling at least one weed in a field, said method comprising planting in said field seed of at least one transgenic plant comprising a plurality of cells comprising a polynucleotide that encodes a protein that has phosphinothricin acetyltransferase activity, said polynucleotide hybridizing under conditions of 6×SSC at 65° C. with the full complement of a nucleic acid probe that encodes SEQ ID NO:2, said plant producing said protein and comprising a heterologous polynucleotide that encodes an enzyme conferring resistance a glutamine synthetase inhibiting herbicide, and a second heterologous polynucleotide encodes an enzyme conferring resistance to at least one other herbicide; said method further comprising applying to at least a portion of said field a first herbicide selected from the group consisting of glufosinate, phosphinothricin and bialaphos; and applying said at least one other herbicide to said at least a portion of said field.
5 . The method of claim 4 wherein said herbicides are applied sequentially or concurrently.
6 . The method of claim 4 wherein said first herbicide is a glutamine synthetase inhibiting herbicide.
7 . The method of claim 4 wherein said at least one other herbicide is selected from the group consisting of 2,4-D, acetochlor, acifluorfen, alloxydim, amidosulfuron, aminopyralid, atrazine, beflubutamid, bispyribac, butafenacil, cafenstrole, carfentrazone, chlorimuron, chlorotoluron, cinidonethyl, clethodim, clodinafop, clomazone, cloproxydim, clopyralid, cloransulam, cyanazine, cyclosulfamuron, cycloxydim, cyhalofop, daimuron, dicamba, diclofop, dichlorprop, diclosulam, diflufenican, dimethenamid, diquat, dithiopyr, diuron, ethalfluralin, fenoxaprop, flazasulfuron, florasulam, fluazifop, flucarbazone, flufenacet, flufenican, flufenpyr, flumetsulam, flumiclorac, flumioxazin, fluroxypyr, fluthiacet, fomesafen, foramsulfuron, glufosinate, glyphosate, halosafen, halosulfuron, haloxyflop, imazamethabenz, imazamox, imazapic, imazapyr, imazaquin, imazethapyr. imazosulfuron, iodosulfuron, ioxynil, isoxaben, isoxaflutole, lactofen, linuron, MCPA, mecoprop, mefenacet, mefluidide, mesosulfuron, mesotrione, metamifop, metazachlor, metosulam, metribuzin, MSMA, napropamide, nicosulfuron, norflurazon, oryzalin, oxadiazon, oxyfluorfen, paraquat, pebulate, pendimethalin, penoxsulam, picloram, picolinafen, pinoxaden, primisulfuron, profoxydim, propanil, pyraflufen, pyrazosulfuron, pyribenzoxim, pyriminobac, pyrithiobac, pyroxasulfone, pyroxsulam, quinclorac, quinmerac, quizalofop, rimsulfuron, saflufenacil, sethoxydim, simazine, sulcotrione, sulfentrazone, sulfometuron, tefuryltrione, tembotrione, tepraloxydim, terbacil, thiazopyr, thidiazuron, thiencarbazone, thifensulfuron, thiobencarb, topramezone, tralkoxydim, triasulfuron, tribenuron, triclopyr, trifloxysulfuron, trifluralin, triflusulfuron, and tritosulfuron.
8 . A method of detecting whether a plant comprises a polynucleotide that encodes a protein that has phosphinothricin acetyltransferase activity, said polynucleotide hybridizing under conditions of 6×SSC at 65° C. with the full complement of a nucleic acid probe that encodes SEQ ID NO:2, wherein said method comprises collecting a sample from said plant and assaying said sample for the presence of said polynucleotide.
9 . The method of claim 8 wherein said method comprises assaying said sample for the presence of a protein encoded by said polynucleotide.
10 . The method of claim 8 wherein said method comprises using a PCR primer or probe to detect for the presence of said polynucleotide.
11 . The method of claim 8 wherein said method comprises using an antibody to detect for the presence of said protein.
12 . A method of controlling at least one weed in a field, said method comprising growing at least one plant in said field, said plant comprising a plurality of cells comprising a polynucleotide that encodes a protein that has phosphinothricin acetyltransferase activity, said polynucleotide hybridizing under conditions of 6×SSC at 65° C. with the full complement of a nucleic acid probe that encodes SEQ ID NO:2, and applying a glutamine synthetase inhibiting herbicide to at least a portion of said field.
13 . The method of claim 12 , further comprising herbicides glufosinate, bialaphos, and phosphinothricin.
14 . The method of claim 12 wherein said plant is resistant to a herbicide selected from the group consisting of glyphosate, glufosinate, 2,4-D, quizalofop, imazethapyr, chlorsulfuron, dicamba, mesotrione, isoxaflutole, and butafenacil.
15 . The method of claim 1 wherein said DSM-2 gene functions as a selectable marker and said method is practiced in a greenhouse, a growth chamber, or in a field.