Compositions providing tolerance to multiple herbicides and methods of use thereof
Methods and compositions are provided related to improved plants that are tolerant to more than one herbicide. Particularly, the invention provides plants that are tolerant of glyphosate and are tolerant to at least one ALS inhibitor, and methods of use thereof. The glyphosate/ALS inhibitor-tolerant plants comprise a polynucleotide that encodes a polypeptide that confers tolerance to glyphosate and a polynucleotide that encodes an ALS inhibitor-tolerant polypeptide. In specific embodiments, a plant of the invention expresses a GAT polypeptide and an HRA polypeptide. Methods to control weeds, improve plant yield, and increase transformation efficiencies are provided.
1 . A method for controlling weeds in an area of cultivation comprising
a) planting the area with crop seeds or plants which comprise
i) a first polynucleotide encoding a polypeptide that can confer tolerance to glyphosate operably linked to a promoter active in said plant; and,
ii) a second polynucleotide encoding an ALS inhibitor-tolerant polypeptide operably linked to a promoter active in said plant;
b) applying to any crop, crop part, weed or area of cultivation thereof a combination of herbicides comprising at least an effective amount of a first herbicide and an effective amount of a second herbicide, wherein said combination of herbicides does not include glyphosate and wherein said effective amount of said first and said second herbicide is tolerated by the crop and controls weeds.
2 . The method of claim 1 , wherein said first herbicide comprises an ALS inhibitor.
3 . The method of claim 2 , wherein said second herbicide comprises a second ALS inhibitor.
4 . The method of claim 2 , wherein said effective amount of the first ALS inhibitor comprises about 0.1 to about 5000 g ai/hectare.
5 . The method of claim 4 , wherein said effective amount of the first ALS inhibitor comprises about 0.5 to about 350 g ai/hectare or about 1 to about 150 g ai/hectare.
6 . The method of claim 3 , wherein said effective amount of the second ALS inhibitor comprises about 0.1 to about 5000 g ai/hectare, about 0.5 to about 350 g ai/hectare or about 1 to about 150 g ai/hectare.
7 . The method of claim 1 , wherein said first polynucleotide encodes a glyphosate-N-acetyltransferase.
8 . The method of claim 1 , wherein said first polynucleotide encodes a glyphosate-tolerant 5-enolpyruvylshikimate-3-phosphate synthase or a glyphosate-tolerant glyphosate oxido-reductase.
9 . The method of claim 1 , wherein said ALS inhibitor-tolerant polypeptide comprises a mutated acetolactate synthase polypeptide.
10 . The method of claim 9 , wherein said mutated acetolactate synthase polypeptide comprises HRA.
11 . The method of claim 1 , wherein said combination of herbicides comprises an ALS inhibitor selected from the group consisting of a sulfonylurea, a triazolopyrimidine, a pyrimidinyloxy(thio)benzoate, an imidazolinone, and a sulfonylaminocarbonyltriazolinone.
12 . The method of claim 11 , wherein said ALS inhibitor is selected from the group consisting of:
a) Azimsulfuron;
b) Chlorimuron-ethyl;
c) Metsulfuron-methyl;
d) Nicosulfuron;
e) Rimsulfuron;
f) Sulfometuron-methyl;
g) Thifensulfuron-methyl;
h) Tribenuron-methyl;
i) Amidosulfuron;
j) Bensulfuron-methyl;
k) Chlorsulfuron;
l) Cinosulfuron;
m) Cyclosulfamuron;
n) Ethametsulfuron-methyl;
o) Ethoxysulfuron;
p) Flazasulfuron;
q) Flupyrsulfuron-methyl;
r) Foramsulfuron;
s) Imazosulfuron;
t) Iodosulfuron-methyl;
u) Mesosulfuron-methyl;
v) Oxasulfuron;
w) Primisulfuron-methyl;
x) Prosulfuron;
y) Pyrazosulfuron-ethyl;
z) Sulfosulfuron;
aa) Triasulfuron;
bb) Trifloxysulfuron;
cc) Triflusulfuron-methyl;
dd) Tritosulfuron;
ee) Halosulfuron-methyl;
ff) Flucarbazone;
gg) Procarbazone;
hh) Cloransulam-methyl;
ii) Flumetsulam;
jj) Diclosulam;
kk) Florasulam;
ll) Imazamox;
mm) Bispyribac;
nn) Pyriftalid;
oo) Pyribenzoxim;
pp) Pyrithiobac;
qq) Pyriminobac-methyl;
rr) Imazapyr;
ss) Imazethapyr;
tt) Imazaquin;
uu) Imazapic;
vv) Imazamethabenz-methyl;
ww) Imazamox;
xx) Flucetosulfuron;
yy) Metosulam;
zz) Penoxsulam; and,
aaa) Pyroxsulam.
13 . The method of claim 1 , wherein said combination of herbicides comprises at least a triazine or a phosphinic acid.
14 . The method of claim 1 , wherein said second herbicide and said first herbicide have different mechanisms of action.
15 . The method of claim 1 , wherein said second herbicide and said first herbicide have the same mechanism of action.
16 . The method of claim 1 , wherein said combination of herbicides comprises at least an effective amount of a third herbicide.
17 . The method of claim 1 , wherein
said first herbicide is applied pre-emergence, post-emergence or pre- and post-emergence to the weeds or the crop; and,
said second herbicide is applied pre-emergence, post-emergence or pre- or post-emergence to the weed or the crop.
18 . The method of claim 1 , wherein said first and said second herbicides are applied together or are applied separately.
19 . The method of claim 1 , wherein at least said first and said second herbicides comprise a homogeneous granule blend.
20 . The method of claim 1 , wherein said method further comprises applying an agricultural chemical selected from the group consisting of a fungicide, a nematicide, a growth regulator, a safener, an adjuvant, an insecticide, and any combination thereof.
21 . The method of claim 20 , wherein said agricultural chemical is applied pre-emergence, post-emergence or pre- and post-emergence to the weed or the crop.
22 . The method of claim 20 , wherein said agricultural chemical is applied together or separately with said first herbicide, said second herbicide, or said first and said second herbicide.
23 . The method of claim 1 , wherein the first herbicide comprises a sulfonylurea and the second herbicide comprises an ALS inhibitor.
24 . The method of claim 1 , wherein the first herbicide comprises a sulfonylurea and the second herbicide comprises an imidazolinone.
25 . The method of claim 23 , wherein the effective amount of said first or said second herbicide provides a safening effect.
26 . The method of claim 1 , wherein at least one of said first or said second polynucleotides is operably linked to at least one copy of the enhancer sequence set forth in SEQ ID NO: 1, 72, 85, 88 or 89 or an enhancer sequence having at least 90% sequence Identity to SEQ ID NO: 1, 72, 85, 88 or 89, wherein said enhancer sequence modulates the level of transcription.
27 . The method of claim 1 , wherein said crop is a dicot.
28 . The method of claim 27 , wherein said dicot is soybean, canola, sunflower, cotton, alfalfa, an ornamental, a fruit, a vegetable, a sugar beet, or an Arabidopsis.
29 . The method of claim 1 , wherein said plant is a monocot.
30 . The method of claim 29 , wherein said monocot is maize, wheat, rice, barley, sorghum, sugar cane, Switchgrass, or rye.
31 . The method of claim 1 , wherein said crop is maize and said first herbicide is applied at the V7 stage or later.
32 . The method of claim 1 , wherein step (b) is performed at least once prior to step (a).
33 . A method for controlling weeds in an area of cultivation comprising:
a) evaluating environmental conditions in an area of cultivation;
b) selecting an effective amount of a combination of herbicides comprising at least an effective amount of a first herbicide and an effective amount of a second herbicide, wherein said combination of herbicides does not include glyphosate and wherein said effective amount of said first and said second herbicide is tolerated by a crop and controls weeds; and,
c) applying the combination of herbicides to a crop, crop part, seed or an area of cultivation of said crop, wherein the crop comprises a plant having
i) a first polynucleotide encoding a polypeptide that can confer tolerance to glyphosate operably linked to a promoter active in said plant; and,
ii) a second polynucleotide encoding an ALS inhibitor-tolerant polypeptide operably linked to a promoter active in said plant.
34 . The method of claim 33 , wherein said first herbicide comprises an ALS inhibitor.
35 . The method of claim 34 , wherein said second herbicide comprises a second ALS inhibitor.
36 . The method of claim 35 , wherein said effective amount of the first or said second ALS inhibitor comprises about 0.1 to about 5000 g ai/hectare.
37 . The method of claim 35 , wherein said effective amount of the first or the second ALS inhibitor comprises about 1 to about 150 g ai/hectare.
38 . The method of claim 35 , wherein said effective amount of the first or the second ALS inhibitor comprises about 0.5 to about 350 g ai/hectare.
39 . The method of claim 33 , wherein said first polynucleotide encodes a glyphosate-N-acetyltransferase.
40 . The method of claim 33 , wherein said ALS inhibitor-tolerant polypeptide comprises a mutated acetolactate synthase polypeptide.
41 . The method of claim 40 , wherein said mutated acetolactate synthase polypeptide comprises HRA.
42 . The method of claim 33 , wherein said herbicide combination comprises an ALS inhibitor selected from the group consisting of a sulfonylurea, a triazolopyrimidine, a pyrimidinyloxy(thio)benzoate, an imidazolinone, and a sulfonylaminocarbonyltriazolinone.
43 . The method of claim 42 , wherein said ALS inhibitor is selected from the group consisting of:
a) Azimsulfuron;
b) Chlorimuron-ethyl;
c) Metsulfuron-methyl;
d) Nicosulfuron;
e) Rimsulfuron;
f) Sulfometuron-methyl;
g) Thifensulfuron-methyl;
h) Tribenuron-methyl;
i) Amidosulfuron;
j) Bensulfuron-methyl;
k) Chlorsulfuron;
l) Cinosulfuron;
m) Cyclosulfamuron;
n) Ethametsulfuron-methyl;
o) Ethoxysulfuron;
p) Flazasulfuron;
q) Flupyrsulfuron-methyl;
r) Foramsulfuron;
s) Imazosulfuron;
t) Iodosulfuron-methyl;
u) Mesosulfuron-methyl;
v) Oxasulfuron;
w) Primisulfuron-methyl;
x) Prosulfuron;
y) Pyrazosulfuron-ethyl;
z) Sulfosulfuron;
aa) Triasulfuron;
bb) Trifloxysulfuron;
cc) Triflusulfuron-methyl;
dd) Tritosulfuron;
ee) Halosulfuron-methyl;
ff) Flucarbazone;
gg) Procarbazone;
hh) Cloransulam-methyl;
ii) Flumetsulam;
jj) Diclosulam;
kk) Florasulam;
ll) Imazamox;
mm) Bispyribac;
nn) Pyriftalid;
oo) Pyribenzoxim;
pp) Pyrithiobac;
qq) Pyriminobac-methyl;
rr) Imazapyr;
ss) Imazethapyr;
tt) Imazaquin;
uu) Imazapic;
vv) Imazamethabenz-methyl;
ww) Imazamox.
xx) Flucetosulfuron;
yy) Metosulam;
zz) Penoxsulam; and,
aaa) Pyroxsulam.
44 . The method of claim 33 , wherein said second herbicide and said first herbicide have the same or a different mechanism of action.
45 . The method of claim 33 , wherein
said first herbicide is applied pre-emergence, post-emergence or pre- and post-emergence to the weeds or the crop; and,
said second herbicide is applied pre-emergence, post-emergence or pre- and post-emergence to the weed or the crop.
46 . The method of claim 33 , wherein said first and said second herbicides are applied together or are applied separately.
47 . The method of claim 33 , wherein at least said first and said second herbicides comprise a homogeneous granule blend.
48 . The method of claim 33 , wherein said method further comprises applying an agricultural chemical selected from the group consisting of a fungicide, a nematicide, a growth regulator, a safener, an adjuvant, an insecticide, or any combination thereof.
49 . The method of claim 33 , wherein at least one of said first or said second polynucleotides is operably linked to at least one copy of the enhancer sequence set forth in SEQ ID NO: 1, 72, 85, 88 or 89, or an enhancer sequence having at least 90% sequence identity to SEQ ID NO: 1, 72, 85, 88 or 89, wherein said enhancer sequence modulates the level of transcription.
50 . The method of claim 33 , wherein said crop is a dicot.
51 . The method of claim 50 , wherein said dicot is soybean, canola, sunflower, cotton, alfalfa, an ornamental, a fruit, a vegetable, a sugar beet, or an Arabidopsis.
52 . The method of claim 33 , wherein said plant is a monocot.
53 . The method of claim 52 , wherein said monocot is maize, wheat, rice, barley, sorghum, sugar cane, Switchgrass, or rye.
54 . A method for controlling weeds in an area of cultivation comprising
a) planting the area with crop seeds or plants which comprise
i) a first polynucleotide encoding a polypeptide that can confer tolerance to glyphosate operably linked to a promoter active in a plant; and,
ii) a second polynucleotide encoding an ALS inhibitor-tolerant polypeptide operably linked to a promoter active in a plant;
b) applying to any crop, crop part, weed or area of cultivation thereof an effective amount of a combination of herbicides selected from the group consisting of:
i) the combination of herbicides comprising glyphosate, imazapyr, chlorimuron-ethyl, quizalofop, and fomesafen, wherein said effective amount is tolerated by the crop and controls weeds; or,
ii) the combination of herbicides comprising glyphosate, chlorimuron-ethyl, and thifensulfuron-methyl, wherein said effective amount is tolerated by the crop and controls weeds.
55 . The method of claim 54 , wherein said crop is soybean, corn or cotton.
56 . The method of claim 54 , wherein the effective amount of the combination of herbicides of part (b)(i) comprises glyphosate comprising about 1110 to about 1130 g ai/hectare; the effective amount of imazapyr comprises about 7.5 to about 27.5 g ai/hectare; the effective amount of chlorimuron-ethyl comprises about 7.5 to about 27.5 g ai/hectare; the effective amount of quizalofop comprises about 50 to about 70 g ai/hectare; and, the effective amount of fomesafen comprises about 240 to about 260 g ai/hectare.
57 . A method of selecting cotton which has been transformed with a first polynucleotide encoding a first polypeptide which confers glyphosate tolerance and a second polynucleotide encoding an ALS inhibitor-tolerant polypeptide, said method comprising the step of growing transformed cotton callus while exposing it to glyphosate and/or an ALS inhibitor herbicide.
58 . The method of claim 57 , wherein said polypeptide which confers glyphosate tolerance comprise GAT and said ALS inhibitor-tolerant polypeptide comprises HRA.
59 . The method of claim 58 , wherein the callus is grown while exposing it to a medium containing chlorsulfuron at a concentration of 50-200 micrograms per liter.
60 . The method of claim 58 , wherein the callus is grown while exposing it to a medium containing glyphosate at a concentration of up to 450 μM.
61 . A method for controlling weeds in an area of cultivation comprising
a) planting the field with crop seeds or plants which comprise
i) a first polynucleotide encoding a polypeptide that can confer tolerance to glyphosate operably linked to a promoter active in a plant; and,
ii) a second polynucleotide encoding an ALS inhibitor resistance polypeptide operably linked to a promoter active in a plant;
b) applying to said crop, crop part, seed of said crop or the area of cultivation thereof an effective amount of an herbicide, wherein said effective amount is tolerated by the crop and controls weeds, wherein said herbicide is not glyphosate, chlorimuron-ethyl, rimsulfuron, tribenuron-methyl or thifensufuron-methyl.
62 . The method of claim 61 , wherein said herbicide is selected from the group consisting of
(i) a sulfonylaminocarbonyltriazolinone;
(ii) a triazolopyrimidine;
(iii) a pyrimidinyl(thio)benzoate;
(iv) an imidazolinone;
(v) a triazine; and
(vi) a phosphinic acid.
63 . A method for controlling weeds in a field containing a crop comprising
a) planting the field with crop seeds or plants which comprise
i) a first polynucleotide encoding a polypeptide that can confer tolerance to glyphosate; and,
ii) a second polynucleotide encoding an ALS inhibitor-tolerant polypeptide;
b) applying to any crop and weeds in the field an effective amount of a first herbicide comprising an ALS inhibitor and an effective amount of at least one additional agricultural agent selected from the group consisting of a fungicide, a nematicide, an insecticide, a safener, or an adjuvant,
wherein said effective amount of said ALS inhibitor herbicide is tolerated by the crop, and,
wherein said effective amount of said combination is not tolerated by a control plant which does not express both of said first and said second polypeptide.
64 . The method of claim 63 , wherein step (b) is performed at least once prior to step (a).