Methods and compositions for conferring and/or enhancing herbicide tolerance using protoporphyrinogen IX oxidase of various cyanobacteria or variant thereof

1 . A polypeptide comprising an amino acid sequence of modified SEQ ID NO: 1, comprising at least one amino acid mutation selected from the group consisting of:

(i) F360M, F360V, F360I, F360T or F360L,

(ii) A167C, A167L, or A167I,

(iii) V305M or V305L,

(iv) R89A,

(v) V165S or V165C,

(vi) L327T, and

(vii) 1408R, or 1408W,

in the amino acid sequence of SEQ ID NO: 1.

2 . A polynucleotide encoding the polypeptide of claim 1 .

3 . A recombinant vector comprising the polynucleotide of claim 2 .

4 . A recombinant cell comprising the recombinant vector of claim 3 .

5 . A composition for conferring or enhancing herbicide tolerance of a plant or algae, comprising one or more selected from the group consisting of:

(1) the polypeptide of claim 3 ;

(2) a polynucleotide encoding the polypeptide of (1);

(3) a recombinant vector comprising the polynucleotide of (2); and

(4) a recombinant cell comprising the recombinant vector of (3).

6 . The composition of claim 5 , wherein the herbicide is an herbicide inhibiting protoporphyrinogen IX oxidase.

7 . The composition of claim 5 , wherein the herbicide is at least one selected from the group consisting of pyrimidinediones, diphenyl-ethers, phenylpyrazoles, N-phenylphthalimides, phenylesters, thiadiazoles, oxadiazoles, triazinone, triazolinones, oxazolidinediones, pyraclonil, flufenpyr-ethyl, and profluazol.

8 . The composition of claim 5 , wherein the herbicide is at least one selected from the group consisting of butafenacil, saflufenacil, benzfendizone, tiafenacil, fomesafen, oxyfluorfen, aclonifen, acifluorfen, bifenox, ethoxyfen, lactofen, chlomethoxyfen, chlorintrofen, fluoroglycofen-ethyl, halosafen, pyraflufen-ethyl, fluazolate, flumioxazin, cinidon-ethyl, flumiclorac-pentyl, fluthiacet, thidiazimin, oxadiargyl, oxadiazon, carfentrazone, sulfentrazone, trifludimoxazin, azafenidin, pentoxazone, pyraclonil, flufenpyr-ethyl, profluazol, phenopylate, carbamate analogues of phenopylate, and agriculturally acceptable salt thereof.

9 . The composition of claim 5 , wherein the plant or algae further comprises a second herbicide-tolerant polypeptide or a gene encoding the same, and its tolerance to the second herbicide is conferred or enhanced.

10 . The composition of claim 9 , wherein the second herbicide is selected from the group consisting of glyphosate, glufosinate, dicamba, 2,4-D (2,4-Dichlorophenoxyacetic acid), isoxaflutole, ALS (acetolactate synthase)-inhibiting herbicide, photosystem II-inhibiting herbicide, phenylurea-based herbicide, bromoxynil-based herbicide, and combinations thereof.

11 . The composition of claim 9 , wherein the second herbicide-tolerant polypeptide is one or more selected from the group consisting of:

glyphosate herbicide-tolerant EPSPS (glyphosate resistant 5-enolpyruvylshikimate-3-phosphate synthase), GOX (glyphosate (glyphosate-N-acetyltransferase) or glyphosate decarboxylase; oxidase), GAT

glufosinate herbicide-tolerant PAT (phosphinothricin-N-acetyltransferase);

dicamba herbicide-tolerant DMO (dicamba monooxygenase);

2,4-D (2,4-dichlorophenoxyacetic acid) herbicide-tolerant 2,4-D monooxygenase or AAD (aryloxyalkanoate dioxygenase);

ALS (acetolactate synthase)-inhibiting sulfonylurea-based herbicide-tolerant ALS (acetolactate synthase), AHAS (acetohydroxyacid synthase) or AtAHASL ( Arabidopsis thaliana acetohydroxyacid synthase large subunit);

photosystem II-inhibiting herbicide-tolerant photosystem II protein D1;

phenylurea herbicide-tolerant Cytochrome P450;

plastid-inhibiting herbicide-tolerant HPPD (hydroxyphenylpyruvate dioxygenase);

bromoxynil herbicide-tolerant nitrilase; and

combinations thereof.

12 . The composition of claim 9 , wherein the gene encoding the second herbicide-tolerant polypeptide is one or more selected from the group consisting of:

glyphosate herbicide-tolerant cp4 epsps, mepsps, 2mepsps, goxv247, gat4601 or gat4621 gene;

glufosinate herbicide-tolerant BAR or PAT gene;

dicamba herbicide-tolerant dmo gene;

2,4-D (2,4-dichlorophenoxyacetic acid) herbicide-tolerant AAD-1 or AAD-12 gene;

isoxaflutole herbicide-tolerant HPPDPF W336 gene;

sulfonylurea herbicide-tolerant ALS, Csr1, Csr1-1, Csr1-2, GM-HRA, S4-HRA, Zm-HRA, SurA or SurB gene;

photosystem II-inhibiting herbicide-tolerant psbA gene;

phenylurea herbicide-tolerant CYP76B1 gene;

bromoxynil herbicide-tolerant bxn gene; and

combinations thereof.

13 . A transformant of a plant or algae having herbicide tolerance, or a clone or progeny thereof, comprising

(1) the polypeptide of claim 1 , or

(2) a polynucleotide encoding the polypeptide of (1).

14 . The transformant, clone, or progeny thereof of claim 13 , wherein the transformant is an alga, or a cell, protoplast, callus, hypocotyl, seed, cotyledon, shoot, or whole body of a plant.

15 . A method of preparing a transgenic plant or algae having herbicide tolerance, the method comprising introducing (1) the polypeptide of claim 3 , or (2) a polynucleotide encoding the polypeptide of (1), into an alga, or a cell, protoplast, callus, hypocotyl, seed, cotyledon, shoot, or whole body of a plant.

16 . A method of conferring or enhancing herbicide tolerance of a plant or algae, the method comprising introducing (1) the polypeptide of claim 3 , or (2) a polynucleotide encoding the polypeptide of (1), into an alga, or a cell, protoplast, callus, hypocotyl, seed, cotyledon, shoot, or whole body of a plant.

17 . A method of controlling weeds in a cropland, the method comprising:

providing the cropland with a plant comprising (1) the polypeptide of claim 1 , or (2) a polynucleotide encoding the polypeptide of (1), and

applying an effective dosage of protoporphyrinogen IX oxidase-inhibiting herbicide to the cropland or the plant.

18 . The method of claim 17 , wherein the step of applying an effective dosage of protoporphyrinogen IX oxidase-inhibiting herbicide to the cropland is performed by applying an effective dosage of two or more kinds of protoporphyrinogen IX oxidase-inhibiting herbicides sequentially or simultaneously.

19 . The method of claim 17 , wherein the plant further comprises a second herbicide-tolerant polypeptide or a gene encoding the same, and

the step of applying an effective dosage of protoporphyrinogen IX oxidase-inhibiting herbicide to the cropland is performed by applying effective dosages of the protoporphyrinogen IX oxidase-inhibiting herbicide and a second herbicide are applied sequentially or simultaneously.

20 . A method of removing an undesired aquatic organism from a culture media, the method comprising:

providing a culture media with algae comprising (1) the polypeptide of claim 1 , or (2) a polynucleotide encoding the polypeptide of (1), and

applying an effective dosage of protoporphyrinogen IX oxidase-inhibiting herbicide to the culture media.